中药提取物烯甙对高糖条件下雪旺细胞功能变化的影响
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摘要
目的
1、建立具有糖尿病性损害特点的雪旺细胞模型;此细胞模型可作为治疗糖尿病神经病有效药物筛选及其机理研究的实验工具和检测体系。
2、进一步观察高糖条件下雪旺细胞功能的变化特点,为阐明糖尿病神经病的发病机理提供直接的实验依据。
3、观察中药提取物烯甙是否具有改善雪旺细胞实验性糖尿病性损伤的作用。
4、探讨烯甙发挥作用的可能机制,为中医药的现代化以及中医理论的创新积累实验资料。
方法
1、雪旺细胞的分离、培养与鉴定:
采用brockes改良法从新生wistar乳鼠坐骨神经组织分离纯化雪旺细胞,并以10%胎牛血清和含0.1%GS(5.0mmol/L)的DMEM培养液培养。鉴定方法:将细胞贴壁于载玻片上,经固定(10%福尔马林5分钟)后分别采用抗S100和抗MBP的免疫细胞化学染色(ABC法)和苏木素复染,计算免疫染色着色细胞的阳性率。
2、高糖培养对雪旺细胞活性与增殖能力的影响
雪旺细胞贴壁培养后弃培养液,分为正常组、高糖组和高糖高渗组,分别采用含正糖(5.0 mmol/L GS)、高糖(30.0 mmol/L Gs)和高糖高渗(含30.0 mmol/LGS+100 mmol/L NaCl)的培养液培养48小时。采用XTT法测定三组不同培养条件的雪旺细胞活性:各组细胞每孔加50ulXTT/PMS应用液,避光振摇培养4h后,在酶标仪以450-500nm波长测定各孔OD值。采用~3H-TdR掺入法检测三组不同条件培养雪旺细胞的增殖能力;各组细胞每孔分别加入0.5uci/10ul的~3H-TdR,继续培养16小时。细胞裂解处理后取裂解液100ul加于含有4ml闪烁液的检测瓶中,在闪烁计数仪上检测CPM读数。
3、烯甙对高糖培养雪旺细胞活性与增殖能力的影响
雪旺细胞贴壁培养后弃培养液,分为正常组(LG)、高糖组(HG)、烯甙组和参麦组;分别加入含正糖(5.0 mmol/L GS)、高糖(30.0 mmol/L GS)、高糖加烯甙(30.0 mmol/L GS+8mg/ml稀甙)、高糖加参麦(30.0 mmol/L GS+10mg/ml参麦)的培养液培养,48小时后四组分别采用XTT法和~3H-TdR掺入法测定雪旺细胞的活性和增殖能力,并记录检测结果。
4、烯甙对高糖培养雪旺细胞增殖能力、NGF的表达与合成影响的观察
雪旺细胞贴壁培养12小时后弃培养液,分为正常组(LG)、高糖组(HG)、烯甙组和参麦组;各组分别加入含正糖(5.0 mmol/L GS)、高糖(30.0 mmol/lGS)、高糖加烯甙(30.0 mmol/L GS+8mg/ml烯甙)、高糖加参麦(30.0 mmol/L GS+10mg/ml参麦)的培养液培养,48小时后四组分别收集上清液采用Elisa法测定NGF的合成水平。收集雪旺细胞分别采用~3H-TdR掺入法测定雪旺细胞的增殖能力,采用实时萤光定量PCR法检测各组雪旺细胞NGFmRNA的表达水平。
结果
1、雪旺细胞的分离、培养与鉴定
从Wistar乳鼠坐骨神经分离雪旺细胞,每只大鼠原代约可取得1×10~6个细胞;高倍镜下观察,细胞胞体较小,大多数成梭形。取样经ABC法染色,抗S100和抗MBP染色着色细胞阳性率分别为92±2%和93±3%(N=10),两种方法阳性率比较无显著差异。
2、高糖培养对雪旺细胞活性和增殖能力的影响
采用XTT法检测雪旺细胞活性;高糖组同正常组比较,细胞活性明显降低,差别具有显著性(P＜0.05);高糖高渗组同正常组比较,细胞活性更低(P＜0.01);高糖组与高糖高渗组比较,差别也具有极显著意义。表明高糖对雪旺细胞活性具有显著抑制作用,但较高渗存在时为轻。
采用~3H-TdR掺入法检测雪旺细胞增殖能力;高糖组同正常组比较,细胞增殖能力明显抑制,差别具有极显著性(P＜0.01);高糖高渗组与正常组比较,细胞增殖能力也明显抑制,差别具有极显著性(P＜0.01);高糖组与高糖高渗组比较,差别也具有极显著意义。表明高糖培养对于雪旺细胞功能具有抑制性作用,但程度较高渗存在时为轻。
3、烯甙对高糖培养雪旺细胞活性与增殖能力变化的影响
采用XTT法检测雪旺细胞活性;高糖组同正常组比较,细胞活性明显降低,差别具有极显著性(P＜0.01);烯甙组同高糖组比较,细胞活性无明显改善,差别无显著性(P＜0.05);参麦组同高糖组比较,细胞活性也无明显改善,差别无显著性。表明烯甙和参麦组治疗对高糖导致的细胞活性下降无显著改善作用。
采用~3H-TdR掺入法检测雪旺细胞增殖能力;高糖组同正常组比较,细胞增殖能力明显下降,差别具有显著性(P＜0.01);烯甙组与高糖组比较,细胞增殖能力明显改善,差别具有显著性(P＜0.01);而参麦组与高糖组比较差别无显著性(P＞0.05)。表明烯甙对高糖条件下雪旺细胞增殖能力的抑制性变化具有显著的改善作用。
4、烯甙对高糖条件下雪旺细胞增殖能力,NGF表达与合成变化的影响
如前所述,烯甙对高糖条件下雪旺细胞增殖能力的抑制性改变具有显著改善作用,而参麦则无明显影响。高糖组与正常组比较、NGF合成显著减少(P＜0.01),表明高糖对雪旺细胞的增殖能力以及NGF的合成均具抑制作用;烯甙组与高糖组比较,NGF的合成功能明显改善(P＜0.01)。同时NGFmRNA的表达也显著上调(P＜0.01),表明烯甙对高糖时雪旺细胞的增殖能力、NGF表达及合成损害均具有改善作用。参麦组同高糖组比较则无显著差异(P＞0.05),表明参麦治疗对雪旺细胞的增殖能力、NGF合成与表达无明显影响。
结论
1、采用brockes改良法从新生wistar乳鼠双侧坐骨神经分离雪旺细胞,每只乳鼠分离细胞数可达1×10~6个细胞(原代),分别应用抗S100和抗MBP免疫染色(ABC法)鉴定,阳性率均大于90%,表明此种分离雪旺细胞的方法无论是效率还是纯度,均能满足实验研究的需要。
2、高糖培养对雪旺细胞的活性和增殖能力均具有显著的抑制作用,表明高糖培养的雪旺细胞作为实验性糖尿病性损伤的细胞模型已达到成模要求;进一步研究其病理改变特征,可帮助我们从一个侧面来了解糖尿病神经病的发病机制,并可用作有效药物筛选的实验工具和检测体系。
3、烯甙对高糖雪旺细胞的活性变化虽未产生作用,但对其增殖能力的抑制性变化却具有显著的改善作用,而对照治疗药物参麦则无此作用。
4、烯甙对高糖条件下雪旺细胞的增殖能力、NGF表达与合成影响的同步观察显示:(1)高糖条件下雪旺细胞在增殖能力发生显著变化的同时,其NGF的合成也发生显著的变化,表现为NGF合成下降,其病理意义十分明显。(2)烯甙可使高糖条件下雪旺细胞的增殖能力、NGF的合成的抑制变化同时得到明显改善以及NGFmRNA表达上调。由于NGF介导的信号转导通路与雪旺细胞的增殖活动密切相关,所以,有理由推测烯甙有可能通过影响NGF合成与作用的某个环节,并经由其介导的胞内信号级联机制,逆转高糖培养下雪旺细胞增殖能力的下降,这对于雪旺细胞在病理环境中发挥救他与自救潜能将有重要帮助。
Objective
1. To establish Schwann cell model having the features of diabetic lesion ; this cell model can act as an experimental tool and a test system of screening effective drugs and its mechanism research.
2. To make further observations on the functional change features of Schwann cells to provide a direct experimental basis for clarifying the mechanism of diabetic neuropathy.
3. To test and verify if allyl glycoside, which is extracted from traditional Chinese medicine, has the effect of improving experimental diabetic lesion of Schwann cells.
4. To research the possible mechanism of action of allyl glycoside to accumulate experimental data for modernization of traditional Chinese medicine and bring forth new ideas of theory of traditional Chinese medicine.
Methods
1. Isolation, purification, culture, and identification of Schwann cells:
Adopting Brokes modification, Schwann cells were isolated from the sciatic nerves of newborn Wistar sucking rats and purified. They were cultured in DMEM culture solution, containing 10% fetus bovine serum and 0. 1% GS. Identification method: the cells were adhered to the slide and fixed; the immune cytochemical staining of S100 antibody and MBP antibody (ABC method) and hematoxylin counter staining were made in order to calculate the positive rate of immunostaining positive cells.
2. Effect of culture with high glucose on activity and proliferation of Schwann cells:
After strapping-wall culture of Schwann cells, the culture solution was thrown aside. They were divided into the normal group, the high glucose group, and the high glucose and hyperosmotic group. The culture solution, containing normal glucose (5.0 mmol/L GS), high glucose (30. 0 mmol/L GS), high and hyperosmotic glucose (30. 0 mmol/L GS +100 mmol/L NaCl), was cultured for 48 hours. XTT method to measure the activity of Schwann cells under three different cultural circumstances was adopted. 50μl XTT/PMS applied solution was added to each pore of the cells of each group; after the cells were protected from light, shaken, and cultured for 4 hours, 450-500 mm wavelength on the enzyme labelling apparatus was adopted to measure 0D value of each pore. By the use of ~3H-TdR incorporative method, the proliferative ability of Schwann's cells which were cultured under different cultural circumstances in the three groups was measured. ~3H-TdR of 0. 5 uci/10μl was added to each pore of cells of the three groups; these cells were continually cultured for 16 hours. After cell-cleavage disposal, 100μl of lytic solution of cell-cleavage were added to a test bottle which contains 4 ml of scintillation liquid to measure CPM reading on the scintillation counter.
3. Effect of allyl glycoside on the activity and change of proliferative ability of Schwann cells cultured by high glucose:
After strapping-wall culture of Schwann cells, the culture solution was thrown aside. They were divided into the normal group (LG), the high glucose group (HG), the allyl glycoside group, and the radix ophiopogonis group, normal glucose (5. 0 mmol/L GS), high glucose (30. 0 mmol/L GS), high glucose plus allyl glycoside (30. 0 mmol/L GS+8mg/ml AG) , and high glucose plus radix ophiopogonis(30.0 mmol/L GS+10mg/ml RO) were added to the culture solution to be cultured. After 48 hours in the four groups the activity and proliferative ability of Schwann cells were measured using XTT method and ~3H-TdR incorporative method, and the test results were recorded.
4. Synchronous observations on the effect of allyl glycoside on the proliferative ability NGF synthesis, and expressive change of Schwann cells cultured by high glucose:
After strapping-wall culture of Schwann cells for 12 hours, the culture solution was thrown aside. They were divided into the normal group, the high glucose group, the allyl glycoside group, and the radix ophiopogonis group. normal glucose (5. 0 mmol/L GS), high glucose (30.0 mmol/L GS), high glucose plus allyl glycoside (30. 0 mmol/L GS +8mg/ml mgAG), and high glucose plus radix ophiopogonis (30. 0 mmol/L GS +10mg/ml R0) were respectively added to the culture solution for culture. After 48 hours, the supernatant fluid was collected. OD values of reading on the enzyme labelling in all group were measured with Elisa method. Schwann's cells were collected and their proliferative ability was measured by ~3H-TdR in- corporative method. The general RNA was prepared; the expressive levels of NGFmRNA in all groups were measured by the real-time fluorescent quantitative PCR method.
Results
1. Isolation, purification, culture, and identification of Schwann cells:
Schwann's cells were isolated from the sciatic nerves of Wistar sucking rats; about 1×10~6 cells were obtained from each rat; under high power lens observations were made, the cellular body was less; the cell was spindle in shape. The specimen was taken and stained by ABC method; in the staining cells of anti-S100 and anti-MBP, the positive rates were respectively 92±2% and 93±3% (N=10). When the positive rates of two methods were compared, there was no significant difference.
2. Effect of high glucose culture on activity and proliferative ability of Schwann cells: The activity of Schwann cells measured with XTT method. As compared with the normal group, the cellular activity in the high glucose group was markedly attenuated and there was a very marked difference (P<0.01). As compared with the normal group, the cellular activity in the high glucose and hyperosmotic group was markedly attenuated, and there was a very obvious difference (P<0. 01). When the high glucose group was compared with the high glucose and hyperosmotic group, there was also a very obvious difference. It showed that high glucose had a marked inhibitory effect on the activity of Schwann cells, but was less than in the presence of hyperosmotic state.
The proliferative ability of Schwann cells was measured with ~3H-TdR incorporative method. As compared with the normal group, the cellular proliferative ability in the high glucose group was obviously inhibited, and there was a very marked difference (P<0.01), as compared with the normal group, the cellular proliferative ability in the high glucose and hyperosmotic group was also markedly inhibited, and there was a very obvious differenct (P<0. 01). When the high glucose group was compared with the high glucose and hyperosmotic group, there was a significant meaning. It showed that the culture with high glucose had an inhibitory effect on the function of Schwann cells, but the degree was less in the presence of hyperosmotic state.
3. Effect of allyl glycoside on the change of the activity and proliferative ability of Schwann cells cultured by high glucose: The activity of Schwann cells was measured with XTT method. As compared with the normal group, the cellular activity in the high glucose group was markedly attenuated, and there was a very marked difference (P<0. 01). As compared with the high glucose group, the cellular activity in the allyl glycoside group was not obviously improved, and there was no obvious difference (P>0.05). As compared with the high glucose group, the cellular activity in the radix ophiopogonis group was also not markedly improved, and there was no significant difference. It showed that there was no improving effect when the lowering of the cellular activity caused by high glucose was treated by allyl glycoside and radix ophiopogonis.
The proliferative ability of Schwann cells was measured with ~3H-TdR incorporative method. As compared with the normal group, the cellular proliferative ability in the high glucose group was markedly lowered, and there was a significant difference (P<0. 01). As compared with the high glucose group, the cellular proliferative ability in the allyl glycoside group was obviously improved, and there was an obvious difference (P<0. 01). When the radix ophiopogonis group was compared with the high glucose group, there was no significant difference (P<0. 05). It showed that the inhibitory change of the proliferative ability of Schwann cells caused by high glucose was improved by allyl glycoside.
4. Effect of allyl glycoside on the proliferative ability, NGF expression and synthesis change of Schwann cells in the circumstances of high glucose:
As stated above, allyl glycoside had an obvious improving effect on the inhibitory change of the proliferative ability of Schwann cells under the circumstances of high glucose, but radix ophiopogonis had no marked influence on the inhibitory action. As compared with the normal group, NGF synthesis in the high glucose group was obviously reduced (P<0. 01). It showed that high glucose had an inhibitory effect on the proliferative ability and NGF synthesis of Schwann cells. As compared with the high glucose group, the function of NGF synthesis in the allyl glycoside group was markedly improved (P<0.01).At the same time the expression of NGFmRNA was obviously raised (P<0.01). It showed that allyl glycoside had an improving effect on the proliferation, NGF espression , and synthesis lesion of Schwann cells. As compared with the high glucose group, in the radix ophiopogonis group there was no significant difference (P>0. 05). It showed that there was no marked influence on the proliferation, NGF expression, and synthesis of Schwann cells treated with radix ophiopogonis.
Conclusion
1. Adopting Brockes modification, Schwann cells were isolated from the bilateral sciatic nerves of newborn Wistar sucking rats. The cell number isolated from sucking rats amounts to 1×10~6 cells. They are immunized, stained, and identified with ABC method of S100 antibody and MBP antiblody. The positive rate accounts for over 90%. It shows that this method isolating Schwann cells can meet the needs of experimental research either in efficiency or in purity.
2. The culture of high glucose has marked inhibitory effect on both the activity or proliferative ability of Schwann cells. It shows that the modelling condition that Schwann cells cultured by high glucose serve as the lesion model of experimental diabetic neuropathy has been possessed. Further study of the charactiristics of pathologic changes can serve as a experimental tool and test system for helping us to know the explanation of diabetic neuropathic pathologic mechanism from one aspectand screening effective drugs.
3. Although allyl glycoside does not play a role in activity change of Schwann cells cultured by high glucose, its inhibitory change of the proliferative ability has a marked improving effect. The control-treatment drug radix ophiopogonis has not such effect.
4. The synchronous observative results of the proliferation, NGF synthesis, and influence of expressive change of Schwann cells cultured by high glucose show that (1) As the proliferative ability of expression Schwann cells cultured by high glucose develops marked changes, their NGF and synthesis also develop obvious changes. It shows that NGFmRNA expression goes down and NGF synthesis drops. The pathologic significance is apparent. (2) Allyl glycoside can markedly improve the inhibitory change of proliferative ability. NGFmRNA expression, NGF synthesis of Schwann cells cultured by high glucose. Because the relay way of the signals mediated by NGF is closely related to proliferation of Schwann cells, there is a reason for guessing that allyl glycoside may influence a given link of NGF synthesis andthrough of intracellular cascade mechanism reverse the lowering of proliferative ability of Schwann cells cultured by high glucose. It will greatly help to develop the latent energy of saving others and themselves of Schwann's in the pathologic conditions.
1、建立具有糖尿病性损害特点的雪旺细胞模型;此细胞模型可作为治疗糖尿病神经病有效药物筛选及其机理研究的实验工具和检测体系。
2、进一步观察高糖条件下雪旺细胞功能的变化特点,为阐明糖尿病神经病的发病机理提供直接的实验依据。
3、观察中药提取物烯甙是否具有改善雪旺细胞实验性糖尿病性损伤的作用。
4、探讨烯甙发挥作用的可能机制,为中医药的现代化以及中医理论的创新积累实验资料。
方法
1、雪旺细胞的分离、培养与鉴定:
采用brockes改良法从新生wistar乳鼠坐骨神经组织分离纯化雪旺细胞,并以10%胎牛血清和含0.1%GS(5.0mmol/L)的DMEM培养液培养。鉴定方法:将细胞贴壁于载玻片上,经固定(10%福尔马林5分钟)后分别采用抗S100和抗MBP的免疫细胞化学染色(ABC法)和苏木素复染,计算免疫染色着色细胞的阳性率。
2、高糖培养对雪旺细胞活性与增殖能力的影响
雪旺细胞贴壁培养后弃培养液,分为正常组、高糖组和高糖高渗组,分别采用含正糖(5.0 mmol/L GS)、高糖(30.0 mmol/L Gs)和高糖高渗(含30.0 mmol/LGS+100 mmol/L NaCl)的培养液培养48小时。采用XTT法测定三组不同培养条件的雪旺细胞活性:各组细胞每孔加50ulXTT/PMS应用液,避光振摇培养4h后,在酶标仪以450-500nm波长测定各孔OD值。采用~3H-TdR掺入法检测三组不同条件培养雪旺细胞的增殖能力;各组细胞每孔分别加入0.5uci/10ul的~3H-TdR,继续培养16小时。细胞裂解处理后取裂解液100ul加于含有4ml闪烁液的检测瓶中,在闪烁计数仪上检测CPM读数。
3、烯甙对高糖培养雪旺细胞活性与增殖能力的影响
雪旺细胞贴壁培养后弃培养液,分为正常组(LG)、高糖组(HG)、烯甙组和参麦组;分别加入含正糖(5.0 mmol/L GS)、高糖(30.0 mmol/L GS)、高糖加烯甙(30.0 mmol/L GS+8mg/ml稀甙)、高糖加参麦(30.0 mmol/L GS+10mg/ml参麦)的培养液培养,48小时后四组分别采用XTT法和~3H-TdR掺入法测定雪旺细胞的活性和增殖能力,并记录检测结果。
4、烯甙对高糖培养雪旺细胞增殖能力、NGF的表达与合成影响的观察
雪旺细胞贴壁培养12小时后弃培养液,分为正常组(LG)、高糖组(HG)、烯甙组和参麦组;各组分别加入含正糖(5.0 mmol/L GS)、高糖(30.0 mmol/lGS)、高糖加烯甙(30.0 mmol/L GS+8mg/ml烯甙)、高糖加参麦(30.0 mmol/L GS+10mg/ml参麦)的培养液培养,48小时后四组分别收集上清液采用Elisa法测定NGF的合成水平。收集雪旺细胞分别采用~3H-TdR掺入法测定雪旺细胞的增殖能力,采用实时萤光定量PCR法检测各组雪旺细胞NGFmRNA的表达水平。
结果
1、雪旺细胞的分离、培养与鉴定
从Wistar乳鼠坐骨神经分离雪旺细胞,每只大鼠原代约可取得1×10~6个细胞;高倍镜下观察,细胞胞体较小,大多数成梭形。取样经ABC法染色,抗S100和抗MBP染色着色细胞阳性率分别为92±2%和93±3%(N=10),两种方法阳性率比较无显著差异。
2、高糖培养对雪旺细胞活性和增殖能力的影响
采用XTT法检测雪旺细胞活性;高糖组同正常组比较,细胞活性明显降低,差别具有显著性(P＜0.05);高糖高渗组同正常组比较,细胞活性更低(P＜0.01);高糖组与高糖高渗组比较,差别也具有极显著意义。表明高糖对雪旺细胞活性具有显著抑制作用,但较高渗存在时为轻。
采用~3H-TdR掺入法检测雪旺细胞增殖能力;高糖组同正常组比较,细胞增殖能力明显抑制,差别具有极显著性(P＜0.01);高糖高渗组与正常组比较,细胞增殖能力也明显抑制,差别具有极显著性(P＜0.01);高糖组与高糖高渗组比较,差别也具有极显著意义。表明高糖培养对于雪旺细胞功能具有抑制性作用,但程度较高渗存在时为轻。
3、烯甙对高糖培养雪旺细胞活性与增殖能力变化的影响
采用XTT法检测雪旺细胞活性;高糖组同正常组比较,细胞活性明显降低,差别具有极显著性(P＜0.01);烯甙组同高糖组比较,细胞活性无明显改善,差别无显著性(P＜0.05);参麦组同高糖组比较,细胞活性也无明显改善,差别无显著性。表明烯甙和参麦组治疗对高糖导致的细胞活性下降无显著改善作用。
采用~3H-TdR掺入法检测雪旺细胞增殖能力;高糖组同正常组比较,细胞增殖能力明显下降,差别具有显著性(P＜0.01);烯甙组与高糖组比较,细胞增殖能力明显改善,差别具有显著性(P＜0.01);而参麦组与高糖组比较差别无显著性(P＞0.05)。表明烯甙对高糖条件下雪旺细胞增殖能力的抑制性变化具有显著的改善作用。
4、烯甙对高糖条件下雪旺细胞增殖能力,NGF表达与合成变化的影响
如前所述,烯甙对高糖条件下雪旺细胞增殖能力的抑制性改变具有显著改善作用,而参麦则无明显影响。高糖组与正常组比较、NGF合成显著减少(P＜0.01),表明高糖对雪旺细胞的增殖能力以及NGF的合成均具抑制作用;烯甙组与高糖组比较,NGF的合成功能明显改善(P＜0.01)。同时NGFmRNA的表达也显著上调(P＜0.01),表明烯甙对高糖时雪旺细胞的增殖能力、NGF表达及合成损害均具有改善作用。参麦组同高糖组比较则无显著差异(P＞0.05),表明参麦治疗对雪旺细胞的增殖能力、NGF合成与表达无明显影响。
结论
1、采用brockes改良法从新生wistar乳鼠双侧坐骨神经分离雪旺细胞,每只乳鼠分离细胞数可达1×10~6个细胞(原代),分别应用抗S100和抗MBP免疫染色(ABC法)鉴定,阳性率均大于90%,表明此种分离雪旺细胞的方法无论是效率还是纯度,均能满足实验研究的需要。
2、高糖培养对雪旺细胞的活性和增殖能力均具有显著的抑制作用,表明高糖培养的雪旺细胞作为实验性糖尿病性损伤的细胞模型已达到成模要求;进一步研究其病理改变特征,可帮助我们从一个侧面来了解糖尿病神经病的发病机制,并可用作有效药物筛选的实验工具和检测体系。
3、烯甙对高糖雪旺细胞的活性变化虽未产生作用,但对其增殖能力的抑制性变化却具有显著的改善作用,而对照治疗药物参麦则无此作用。
4、烯甙对高糖条件下雪旺细胞的增殖能力、NGF表达与合成影响的同步观察显示:(1)高糖条件下雪旺细胞在增殖能力发生显著变化的同时,其NGF的合成也发生显著的变化,表现为NGF合成下降,其病理意义十分明显。(2)烯甙可使高糖条件下雪旺细胞的增殖能力、NGF的合成的抑制变化同时得到明显改善以及NGFmRNA表达上调。由于NGF介导的信号转导通路与雪旺细胞的增殖活动密切相关,所以,有理由推测烯甙有可能通过影响NGF合成与作用的某个环节,并经由其介导的胞内信号级联机制,逆转高糖培养下雪旺细胞增殖能力的下降,这对于雪旺细胞在病理环境中发挥救他与自救潜能将有重要帮助。
Objective
1. To establish Schwann cell model having the features of diabetic lesion ; this cell model can act as an experimental tool and a test system of screening effective drugs and its mechanism research.
2. To make further observations on the functional change features of Schwann cells to provide a direct experimental basis for clarifying the mechanism of diabetic neuropathy.
3. To test and verify if allyl glycoside, which is extracted from traditional Chinese medicine, has the effect of improving experimental diabetic lesion of Schwann cells.
4. To research the possible mechanism of action of allyl glycoside to accumulate experimental data for modernization of traditional Chinese medicine and bring forth new ideas of theory of traditional Chinese medicine.
Methods
1. Isolation, purification, culture, and identification of Schwann cells:
Adopting Brokes modification, Schwann cells were isolated from the sciatic nerves of newborn Wistar sucking rats and purified. They were cultured in DMEM culture solution, containing 10% fetus bovine serum and 0. 1% GS. Identification method: the cells were adhered to the slide and fixed; the immune cytochemical staining of S100 antibody and MBP antibody (ABC method) and hematoxylin counter staining were made in order to calculate the positive rate of immunostaining positive cells.
2. Effect of culture with high glucose on activity and proliferation of Schwann cells:
After strapping-wall culture of Schwann cells, the culture solution was thrown aside. They were divided into the normal group, the high glucose group, and the high glucose and hyperosmotic group. The culture solution, containing normal glucose (5.0 mmol/L GS), high glucose (30. 0 mmol/L GS), high and hyperosmotic glucose (30. 0 mmol/L GS +100 mmol/L NaCl), was cultured for 48 hours. XTT method to measure the activity of Schwann cells under three different cultural circumstances was adopted. 50μl XTT/PMS applied solution was added to each pore of the cells of each group; after the cells were protected from light, shaken, and cultured for 4 hours, 450-500 mm wavelength on the enzyme labelling apparatus was adopted to measure 0D value of each pore. By the use of ~3H-TdR incorporative method, the proliferative ability of Schwann's cells which were cultured under different cultural circumstances in the three groups was measured. ~3H-TdR of 0. 5 uci/10μl was added to each pore of cells of the three groups; these cells were continually cultured for 16 hours. After cell-cleavage disposal, 100μl of lytic solution of cell-cleavage were added to a test bottle which contains 4 ml of scintillation liquid to measure CPM reading on the scintillation counter.
3. Effect of allyl glycoside on the activity and change of proliferative ability of Schwann cells cultured by high glucose:
After strapping-wall culture of Schwann cells, the culture solution was thrown aside. They were divided into the normal group (LG), the high glucose group (HG), the allyl glycoside group, and the radix ophiopogonis group, normal glucose (5. 0 mmol/L GS), high glucose (30. 0 mmol/L GS), high glucose plus allyl glycoside (30. 0 mmol/L GS+8mg/ml AG) , and high glucose plus radix ophiopogonis(30.0 mmol/L GS+10mg/ml RO) were added to the culture solution to be cultured. After 48 hours in the four groups the activity and proliferative ability of Schwann cells were measured using XTT method and ~3H-TdR incorporative method, and the test results were recorded.
4. Synchronous observations on the effect of allyl glycoside on the proliferative ability NGF synthesis, and expressive change of Schwann cells cultured by high glucose:
After strapping-wall culture of Schwann cells for 12 hours, the culture solution was thrown aside. They were divided into the normal group, the high glucose group, the allyl glycoside group, and the radix ophiopogonis group. normal glucose (5. 0 mmol/L GS), high glucose (30.0 mmol/L GS), high glucose plus allyl glycoside (30. 0 mmol/L GS +8mg/ml mgAG), and high glucose plus radix ophiopogonis (30. 0 mmol/L GS +10mg/ml R0) were respectively added to the culture solution for culture. After 48 hours, the supernatant fluid was collected. OD values of reading on the enzyme labelling in all group were measured with Elisa method. Schwann's cells were collected and their proliferative ability was measured by ~3H-TdR in- corporative method. The general RNA was prepared; the expressive levels of NGFmRNA in all groups were measured by the real-time fluorescent quantitative PCR method.
Results
1. Isolation, purification, culture, and identification of Schwann cells:
Schwann's cells were isolated from the sciatic nerves of Wistar sucking rats; about 1×10~6 cells were obtained from each rat; under high power lens observations were made, the cellular body was less; the cell was spindle in shape. The specimen was taken and stained by ABC method; in the staining cells of anti-S100 and anti-MBP, the positive rates were respectively 92±2% and 93±3% (N=10). When the positive rates of two methods were compared, there was no significant difference.
2. Effect of high glucose culture on activity and proliferative ability of Schwann cells: The activity of Schwann cells measured with XTT method. As compared with the normal group, the cellular activity in the high glucose group was markedly attenuated and there was a very marked difference (P<0.01). As compared with the normal group, the cellular activity in the high glucose and hyperosmotic group was markedly attenuated, and there was a very obvious difference (P<0. 01). When the high glucose group was compared with the high glucose and hyperosmotic group, there was also a very obvious difference. It showed that high glucose had a marked inhibitory effect on the activity of Schwann cells, but was less than in the presence of hyperosmotic state.
The proliferative ability of Schwann cells was measured with ~3H-TdR incorporative method. As compared with the normal group, the cellular proliferative ability in the high glucose group was obviously inhibited, and there was a very marked difference (P<0.01), as compared with the normal group, the cellular proliferative ability in the high glucose and hyperosmotic group was also markedly inhibited, and there was a very obvious differenct (P<0. 01). When the high glucose group was compared with the high glucose and hyperosmotic group, there was a significant meaning. It showed that the culture with high glucose had an inhibitory effect on the function of Schwann cells, but the degree was less in the presence of hyperosmotic state.
3. Effect of allyl glycoside on the change of the activity and proliferative ability of Schwann cells cultured by high glucose: The activity of Schwann cells was measured with XTT method. As compared with the normal group, the cellular activity in the high glucose group was markedly attenuated, and there was a very marked difference (P<0. 01). As compared with the high glucose group, the cellular activity in the allyl glycoside group was not obviously improved, and there was no obvious difference (P>0.05). As compared with the high glucose group, the cellular activity in the radix ophiopogonis group was also not markedly improved, and there was no significant difference. It showed that there was no improving effect when the lowering of the cellular activity caused by high glucose was treated by allyl glycoside and radix ophiopogonis.
The proliferative ability of Schwann cells was measured with ~3H-TdR incorporative method. As compared with the normal group, the cellular proliferative ability in the high glucose group was markedly lowered, and there was a significant difference (P<0. 01). As compared with the high glucose group, the cellular proliferative ability in the allyl glycoside group was obviously improved, and there was an obvious difference (P<0. 01). When the radix ophiopogonis group was compared with the high glucose group, there was no significant difference (P<0. 05). It showed that the inhibitory change of the proliferative ability of Schwann cells caused by high glucose was improved by allyl glycoside.
4. Effect of allyl glycoside on the proliferative ability, NGF expression and synthesis change of Schwann cells in the circumstances of high glucose:
As stated above, allyl glycoside had an obvious improving effect on the inhibitory change of the proliferative ability of Schwann cells under the circumstances of high glucose, but radix ophiopogonis had no marked influence on the inhibitory action. As compared with the normal group, NGF synthesis in the high glucose group was obviously reduced (P<0. 01). It showed that high glucose had an inhibitory effect on the proliferative ability and NGF synthesis of Schwann cells. As compared with the high glucose group, the function of NGF synthesis in the allyl glycoside group was markedly improved (P<0.01).At the same time the expression of NGFmRNA was obviously raised (P<0.01). It showed that allyl glycoside had an improving effect on the proliferation, NGF espression , and synthesis lesion of Schwann cells. As compared with the high glucose group, in the radix ophiopogonis group there was no significant difference (P>0. 05). It showed that there was no marked influence on the proliferation, NGF expression, and synthesis of Schwann cells treated with radix ophiopogonis.
Conclusion
1. Adopting Brockes modification, Schwann cells were isolated from the bilateral sciatic nerves of newborn Wistar sucking rats. The cell number isolated from sucking rats amounts to 1×10~6 cells. They are immunized, stained, and identified with ABC method of S100 antibody and MBP antiblody. The positive rate accounts for over 90%. It shows that this method isolating Schwann cells can meet the needs of experimental research either in efficiency or in purity.
2. The culture of high glucose has marked inhibitory effect on both the activity or proliferative ability of Schwann cells. It shows that the modelling condition that Schwann cells cultured by high glucose serve as the lesion model of experimental diabetic neuropathy has been possessed. Further study of the charactiristics of pathologic changes can serve as a experimental tool and test system for helping us to know the explanation of diabetic neuropathic pathologic mechanism from one aspectand screening effective drugs.
3. Although allyl glycoside does not play a role in activity change of Schwann cells cultured by high glucose, its inhibitory change of the proliferative ability has a marked improving effect. The control-treatment drug radix ophiopogonis has not such effect.
4. The synchronous observative results of the proliferation, NGF synthesis, and influence of expressive change of Schwann cells cultured by high glucose show that (1) As the proliferative ability of expression Schwann cells cultured by high glucose develops marked changes, their NGF and synthesis also develop obvious changes. It shows that NGFmRNA expression goes down and NGF synthesis drops. The pathologic significance is apparent. (2) Allyl glycoside can markedly improve the inhibitory change of proliferative ability. NGFmRNA expression, NGF synthesis of Schwann cells cultured by high glucose. Because the relay way of the signals mediated by NGF is closely related to proliferation of Schwann cells, there is a reason for guessing that allyl glycoside may influence a given link of NGF synthesis andthrough of intracellular cascade mechanism reverse the lowering of proliferative ability of Schwann cells cultured by high glucose. It will greatly help to develop the latent energy of saving others and themselves of Schwann's in the pathologic conditions.
引文
[1]陈灏珠主编.实用内科学[M].第12版,北京:人民卫生出版社,2005.1015-1030.
[2]Vinik AL,Park TS,Stansberryt KB,et al.Diabetic neuropathies[J].Diabetologia.2000,43:957-973.
[3]Nishikawa T,Edelstein D,Du XL,et al.Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage[J].Nature,2000.404:787-790.
[4]Kang D,Hamasaki N,Mitochondrial oxidative stress and mitochondrial DNA[J].Clin Chem Lab Med,2003.41(10):1281-1288.
[5]Eckersley L.Role of the Schwann cell in diabetic neuropathy[J].International review of neurobiology.2002.50:293-312.
[6]Feldman.et al.Diabetic neuropathy.In:Current review of diabetes[J].Current medicine,Philadelphia,1999.71-83.
[7]James M.Kennedy,et al.Impaired peripheral nerve regeneration in diabetes mellitus[J].Journal of the peripheral nervous system,2005.10:144-157
[8]徐科.神经生物学纲要(M).北京:科学出版社,2000.45-47。
[9]Luke Eckersley.Role of the Schwann cell in diabetic neuropathy.International review of neurobiology,2002,50:293-312.
[10]Grinspan,JB.Axonal interactions regulate Schwann cell apoptosis in developing peripheral nerve:Neuregulation receptors and the role of neuregulin[J].Neurosci,1996.16:6107-6118.
[11]Gulati A K.Peripheral nerve regeneration through short and longterm degenerated nerve transplants[J].Brain Res,1996.742:265-270.
[12]Ansselin,A D.Peripheral nerve regeneration through nerve guides seeded with adult Schwann cells[J].Neuropathol Appl,Neurobiol,1997.23:387-398.
[13]Korching.The neurotrophic factor concept:a reexamination[J].Neurosci,1993,13:2793.
[14]Jiang X,Zhang JS,Jakobsen J.Differential effect of p75~(NTR) on Expression of pro-apoptotic proteins C-Jun,and caspase-3 in DRG cells After axotomy in experimental diabetes[J].Neuroscience,2005. 132:1083-1092.
[15] Meyer M, Matsuoka I, Wetmore C, et al. Enhanced synthesis of brain neurotrophic factor in lesioned peripheral nerve: different mechanism are responsible for the regulation of BDNFand NGFmRNA[J].J Cell Biol, 1992.199:45-50.
[16] Hammcerberg H, Piehl F, Cullheims, et al. GDNFmRNA in Schwann cell and DRG satellite cell after chronic sciatic nerve injure[J].Neuroreport, 1996.22:857-860.
[17] Grothe C. meisinger C. Hertenstein A, et al. Expression of FGF-2 and FGFR-1 mRNA in spinal gonglial an sciatic nerve regeneration after peripheral lesion, [J] Neurosci, 1996,76:123-125.
[18] Hiroshi F. Frigen J. Difterential expression of mRNA for neurotrophins and their receptors after axotomy of the sciatic nerve[J]. J Cell Biol, 1992,123:455-457.
[19] Kuffler DD. Promoting and directing axon autogrowth [J]. Mol Neurobiol, 1994.9:233-235.
[20] Cheng C,. in vivo proliferation, migration and phenotypic changes of Schwann cells in the presence of myelinated fibers[J].2002.115:321-329.
[21] Eckersley L. Effects of experimental diabetes on axonal and Schwann cell changes in sciatic nerve isografts(J). Brain Res,2001.92:128-137.
[22] Ferri C C. Improved survival of injuried sciatic nerve Schwann cells in mice lacking the P75 receptor[J]. Neurosci. Lett, 1999.272:191-194.
[23] Leinninger GM, Vincent AM, Feldman LF. The role of growth factors in diabetic peripheral neuropathy [J]. J Peripheral Nervous System.2004.9:26-53.
[24] Kaplan DR, HempsteadBL, Martin-Zanca D, etal. The trk proto-oncogene product:a signal transducing receptor for nerve growth factor [J].Science, 1991. 252:554-558.
[25] Patapoution A, Reichardt LF. Trk receptors: mediators of neurotrophin action [J]. Curr Opin Neurobiol, 2001.11:272-280.
[26] Vinik AI. Diabetic neuropathy: pathogenesis and therapy[J]. The American Journal of Medicine,1999.107:17-26.
[27]王学华,汪少颖主编.消渴病古今名家验案全析(M).北京:科学技术文献出版社,2006.480
[28]曾莉,陈卫平.疑病病中西诊疗指南(M).江苏:东南大学出版社,2004.206.
[29]姜喆.内分泌代谢病临床诊治(M).北京:科学技术文献出版社,2005.71.
[30]季钟朴.中西医结合思路与方法(M).上海:上海科学技术出版社,2006.199.
[31]胡熙明.中国中医秘方大全(M).上海:文汇出版社出版,1989.573.
[32]张家庆.中西医结合防治糖尿病研究的思路与方法[J].中国中西医结合杂志,1995.15(5):315-316.
[33]张家庆.中医治糖尿病不仅仅是降糖[J].中医杂志,1995,36(7):428.
[34]高从容,张家庆,黄庆玲.黄连素增加胰岛素抵抗大鼠横型胰岛素敏感性的实验研究[J].中国中西医结合杂志,1997,17(3):163-164.
[35]潘明政,郭富珊,梁晓春等.中药仙贞片对气阴两虚肾虚血瘀型糖尿病患者和细胞膜Na、K-ATP酶等的影响[J].中国中西医结合杂志,1997.17(1):13-16.
[36]熊曼琪,林安钟,朱章志等.加味挑仁承气汤对2型糖尿病大鼠胰岛素抵抗的影响[J],中国中西医结合杂志,1997.17(3):165-138.
[37]姜兆顺,张胜兰,蔡新告等.复方丹参注射液对2型糖尿病患者抗脂质过养化损伤的影响[J].中国中西医结合杂志,1997.7(1):32-34.
[1]薛庆善,主编.体外培养的原理与技术[M].北京:科学出版社,2001.151.
[2]汪谦,主编.临床医学实验方法学[M].北京:北京科学出版社,2002.19-22.
[3]Vinik AI.Diabetic Neuropathy:Pathogenesis and therapy[J].The American Journal of Medicaine,1999.107:17-23.
[4]James MK and Diygkas WZ.Impaired peripheral nerve regeneration in diabetes mellitus[J].Journal of the Peripheral Nervous System,2005.10:144-157.
[5]Brockes JP,Fields KL,Raff MC.A surfare marker for rat Schwann cells[J].Nature,1977.266:364-366.
[6]Brockes JP,Fields KL,Raft MC.Studies on cultured rat Schwann cells[J].Brain Res,1979.165(1):105-118.
[7]Askanas V,Engel WK,Dalakas MC,et al.Human Schwann cells in culture:Histochemical and ultrastructratl studies[J].Arch Neurol,1980.37(6):329-337.
[8]Oda Y,Okada Y,Katsuda S,et al.A simple method for the Schwann cell preparation from newborn rat sciatic nerves[J].J Neurosci meth.1989.28(3):163-169.
[9]Kaff ME,Abney E,Brockes JP,et al.Schwann cell growth factors(J).Cell 1978.5(3):813-822.
[10]Wood PM.Bunge CP.Evidence that sensory axons are mitogenic for Schwan cells[J].Nature,1975.256:662-664.
[11]Salzer JL,Williams AK,Glaser L,et al.Studies of Schwann cell proliferation.Ⅱ.Characterization of the stimulation and specificity of the response to neurite membrane fraction[J].J Cell Biol,1980.84(3):753-766.
[12]Raft MC,Abney ER,Cohen J,et al.Two types of astrocytes in cultures of developing rat white matter:Difference in morphology,surface ganglioside,and growth characteristics(J).J Nenrosci,1983.3(6):1289-1300
[13]Mirsky R.Cell-type-specific markers in the nervous system cultures(J).TINS.1980.3(8):190-192.
[14]严恒林,沈馨亚,刘才栋等.人胎儿雪旺氏细胞的体外培养及其纯化的研究(J).神经解剖学杂志,1994.10(4):289-293
[15]徐科,主编.神经生物学纲要[M].北京:科学出版社,2001.47.
[1]Keiko Maekawa,Tsuyoshi Tanimoto,Satoshi Okada,et al.Expression of aldose reductase and sorbitol dehydrogenase genes in Schwann cells isolated from rat:effects of high glucose and osmotic stress[J].Molecular Brain Research,2001.251-256.
[2]Takeshi Suzuki,Hiromi sekido,Noriaki kato,et al.Neurotrophin-3-induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose:involvement of the polyol pathway[J].Journal of Neurochemistry,2004.91:1430-1438.
[3]Christopher R,Pierson,Zhang Weixian,et al.Early gene responses of trophic factor in nerve regeneration differ in experimental type 1 and type 2 diabetic polyneuropathies[J].Journal of Neuropathology and Experimental Neurology,2002.10:857-871.
[4]郑志宏,林玲.神经细胞培养理论与实践[M].北京:科学出版社,2002.196-197.
[5]薛庆善,主编.体外培养的原理与技术[M].北京:科学出版社,2001.151.
[6]蔡文琴,李海标,主编.发育神经生物学[M].北京:科学出版社,2001.171-174
[7]LB.莱维坦,LK.卡茨玛克著,舒斯云、包新民主译.神经元:细胞和分子生物学[M].北京:科学出版社,2001.247-267.
[8]许绍芬,主编.神经生物学[M].第2版,上海:上海医科大学出版社,1999.96-111。
[9]罗学港,主编.神经科学基础[M].湖南:中南大学出版社,2002.82-111.
[10]吴克夏,主编.细胞通讯与疾病[M].北京:科学出版社,2006.1-45.
[11]Takeshi Suzuki,Hiromi sekido,Noriaki kato,et al.Neurotrophin-3-induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose:involvement of the polyol pathway[J].Journal of Neurochemistry,2004.91:1430-1438.
[12]Meier C,Parmantier E,Brennan A,et al.Developing Schwann cells acquire the ability to survive without axons by establishing an autocrine circuit involving insulin-like growth factor,neurotrophin-3,and platelet-derived growth factor-BB[J].Neurosci,1999.19:3847-3859.
[1]季钟朴,主编.中西医结合思路与方法[M].上海:上海科学技术出版社,2006.199-209.
[2]曾莉,陈卫平.主编.疑难病中西诊疗指南[M].江苏:东南大学出版社,2004.206-225.
[3]张家庆.中西医结合防治糖尿病研究的思路与方法[J].中国中西医结合杂志,1995.15(5):315-316.
[4]姜喆,主编.内分泌代谢病临床诊治[M].北京:科学技术文献出版社,2005.71-117.
[5]王学华,汪少颖,主编..消渴病古今名家验案全析[M].北京:科学技术文献出版社,2006.480-527.
[6]胡熙明,主编.中国中医秘方大全[M].上海:文汇出版社出版,1989.573-587.
[7]王林丽.红景天及复方制剂药理和临床研究进展[J].中医药学报,2003.31(1):51-54.
[8]王强,阮晓,方兰等.资源植物红景天研究综述[J].新疆农业大学学报,2002.25(4):57-62.
[9]许绍芬,主编.神经生物学[M].上海:上海医科大学出版社,1999.96-111.
[10]罗学港,主编.神经科学基础[M].湖南:中南大学出版社,2002.82-111.
[11]LB.莱维坦,LK.卡茨玛克著,舒斯云、包新民主译.神经元:细胞和分子生物学[M].北京:科学出版社,2001.247-267
[12]郑志宏,林玲编著.神经细胞培养理论与实践[M].北京:科学出版社,2002.196-197.
[13]薛庆善.主编.体外培养的原理与技术[M].北京:科学出版社,2001.151-157.
[14]汪谦.主编.临床医学实验方法学[M].北京:北京科学出版社,2002.19-227.
[15]吴克夏,主编.细胞通讯与疾病[M].北京:科学出版社,2006.1-45
[16]黄文林,朱孝峰,主编.信号转导[M].北京:人民卫生出版社,2005.274-306.
[17]方秀斌,主编.神经肽与神经营养因子[M].北京:人民卫生出版社,2002.274-306.
[1]薛庆善,主编.体外培养的原理与技术[M].北京:科学出版社,2001:151.
[2]黄文林,朱孝峰,主编.信号转导(M).北京:人民卫生出版社,2005:274-306.
[3]方秀斌,主编.神经肽与神经营养因子(M).北京:人民卫生出版社,2002:274-306.
[4]Taniuchi M.clark HB.Johnson EM.Induction of nerve growth factor receptor in Schwann cells after axotomy.Proc Nati Acad Sci USA.1986,83:4094-4098
[5]raniuchi M.Clark HB.Schweitzer JB.Expression of nerve growth factor receptors by Schwann cell of axotomized peripheral nerves:Ultrastructural Location.Suppression by axonal contact and binding properties[J].J Neurosci.1988,8:664-681.
[6]Rozakis-Adcock M,Mcglade J,Mbamalu G.Association of the Shc and Brb2/sem5 SH2-containing proteins is implicated in activation of the Ras pathway by tyrosine kinases[J].Nature 1992,360:689-692.
[7]Xing J,Ginty DD,Greenbery ME.Coupling of the RAS-MAPK path way to gene activation by RSK2,a growth factor-regulated CREB kinase[J].Science 1996,273:959-963.
[8]Atwal JK,Massie B,Miller FD.The TrkB-Shc site signals neuronal survival and local axon growth via mEk and P13-kinase[J].Neuron,2000,27:265-277.
[9]Sharma AK,Thomas PK.Peripheral nerve regeneration in experimental diabetes[J].Neurol sci,1975,24:417-424.
[10]Ekstron AR,Kanje M,Skottner A.Nerve regeneration and serum levels of insulin-like growth factor-1 in rats with streptoxotocin-induced insulin deficiency[J].Brain Res,1989,496:141-147.
[11]James Mk Diygkas WZ.Impaired peripheral nerve regeneration in diabetes mellitus[J].Journal of the Peripheral Nervous System,2005,10:144-157.
[12]Luke Eckersley.Role of the Schwann cell in diabetic neuropathy.International Review of Neurobiology[J].2002,50:293-312.
[13]Korching.The neurotrophic factor concept:a reexamination[J].Neurosci,1993,13:2793.
[14]罗学港,主编,神经科学基础[M].湖南:中南大学出版社,2002,100-101.
[15] Terada M, Yasuda H, Kikkawa R, et al. Tolrestat improves nerve regeneration after crush injury in streptozocin-induced diabetic rats[J]. Metabolism, 1996,45:1189-1195.
[16] Perry VH , Brown MC. Macrophages and nerve regeneration[J]. Curr Opin Neurobiol, 1992, 2:679-682.
[17] Reichert F, saada A. Rotshenker S. Peripheral nerve injury induces Schwann cells to express two macrophage phenotypes: phagocytosis and the galactose-specitic lectin MAC-2[J].Neurosci, 1994,14:3231-3245.
[18] Cheng C, Zochodne DW. In vivo proliferation, migration and phenotypic changes of Schwann cells in the presence of myelinated fibers[J].Neurosci, 2002,115:321-329.
[19] Hiroshi F, Frigen J. Differential expression of mRNA for neurotrophins and their receptors after axotomy of the sciatic nerve[J].Cell Biol, 1992,123:455.
[20] Vinik Al, Park Ts, Stansberryt KB, et al. Diabetic neuropathies [J].Diabetologia, 2000,43:957-973.
[21] Nishikawa T, Edelstein D, Du XL, et al. Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage[J]. Nature, 2000,404:787-790.
[22] Kang D, Hamasaki N. Mitochondrialoxidative stress and mitochondrial DNA[J]. Clin Chem Lab Med, 2003, 41(10):1281-1288.
[23] Taniuchi M, clark HB, Johnson EM. Induction of nerve growth factor receptor in Schwann cells after axotomy[J]. Proc Nati Acad Sci USA,1986,83:4094-4098.
[24] Taniuchi M, Clark HB, Schweitzer JB. Expression of nerve growth factor receptors by Schwann cell of axotomized peripheral nerves:Ultrastructural Location. Suppression by axonal contact and binding properties[J]. Neurosci, 1988, 8:664-681.
[25] Hellweg R, Hartung HD. Endogenous levels of nerve gronth factor (NGF) are altered in experimental diabetes mellitus: a possible role for NGF in the pathogenesis of diabetic neuropathy[J]. Neurosci Res,1990,26:258-267.
[26] Takeshi Suzuki, Hiromi sekido, Noriaki kato, et al. Neurotrophin-3- induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose:involvement of the polyol pathway[J].Journal of Neurochemistry,2004,91:1430-1438.
[27]吴克夏,主编.细胞通讯与疾病[M].北京:科学出版社,2006,1-45.
[28]vinik AI.Diabetic Neuropathy:Pathogenesis and Therapy[J].The American Journal of Medicine,1999,107:17-25
[29]王学华,汪少颖,主编.消渴病古今名家验案全析[M].北京:科学技术文献出版社,2006,480.
[30]曾莉、陈卫平,主编.疑难病中西诊疗指南[M].江苏:东南大学出版社,2004,206.
[31]姜喆,主编.内分泌代谢病临床诊治[M].北京:科学技术文献出版社,2005,71.
[32]季钟朴,主编.中西医结合思路与方法[M].上海:上海科学技术出版社,2006,199.
[33]胡熙明,主编.中国中医秘方大全[M].上海:文汇出版社,1989,573.
[34]张家庆.中西医结合防治糖尿病研究的思路与方法[J].中国中西医结合杂志,1995,15(5):315-316.
[35]张家庆.中医治糖尿病不仅仅是降糖[J].中医杂志,1995,36(7):428.
[36]高从容,张家庆,黄庆玲.黄连素增加胰岛素抵抗大鼠横型胰岛素敏感性的实验研究[J].中国中西医结合杂志,1997,17(3):162-164
[37]潘明政,郭富珊,梁晓春,等.中药仙贞片对气阴两虚肾虚血瘀型糖尿病患者和细胞膜Na、K-ATP酶等的影响[J].中国中西医结合杂志,1997,17(1):13-16.
[38]熊曼琪,林安钟,朱章志,等.加味挑仁承气汤对2型糖尿病大鼠胰岛素抵抗的影响[J].中国中西医结合杂志,1997,17(3):165-138
[39]姜兆顺,张胜兰,蔡新告,等.复方丹参注射液对2型糖尿病患者抗脂质过养化损伤的影响[J].中国中西医结合杂志,1997,7(1):32-34
[40]王林丽,红景天及复方制剂药理和临床研究进展[J].中医药学报,2003,31(1):51-54
[41]王强,阮晓,方兰,等.资源植物红景天研究综述[J].新疆农业大学学报,2002,25(4):57-62
[42]陈秋丽,邓伟国,王毅.红景天皂苷对雌性大鼠蛋白质和脂肪代谢的影响[J].吉林大学学报医学版,2003,29(4):411-413.
[43]赵文,王庭欣,蒋东升.蔷薇红景天提取液对糖尿病模型小鼠胰腺功能的修 复作用[J].中国比较医学杂志,2003,13(5):309.
[44]赵文,王庭欣,蒋东升,等.蔷薇红景天对四氧嘧啶糖尿病小鼠血糖的调节作用[J].卫生研究,1999,28(6):362-363.
[45]胡敏,于金德.红景天苷对心血管系统作用的研究[J].国外医学.心血管疾病分册,2003,30(5):398-300.
[1]Feldman EL,Steven MJ,Russell JW,et al.Diabetic neuropathy.In:Current Review of Diabetes[J].Current medicine.Philadelphia,1999,71-83.
[2]James Mk,Diygkas WZ.Impaired peripheral nerve regeneration in diabetes mellitus[J].Journal of the Peripheral Nervous System,2005,10:144-157.
[3]徐科,主编.神经生物学纲要(M).北京:科学出版社,2000,45-47.
[4]罗学港,唐建华,熊燕编,主编.神经科学基础(M).湖南:中南大学出版社,2002,96-98.
[5]Luke Eckersley.Role of the Schwann cell in diabetic neuropathy[J].International Review of Neurobiology,2002,50:293-312。
[6]Grinspan JB,Marchionni MA,Reeves M,et al.Axonal interactions regulate Schwann cell apoptosis in developing periphesal nerve:Neuregulation receptors and the role of neuregulin[J].Neurosci,1996,16:6107-6118.
[7]Gulati AK.Peripheral nerve regeneration through short-and longterm degenerated nerve transplants[J].Brain Res,1996,742:265-270.
[8]Ansselin AD.Peripheral nerve regeneration through nerve guides seeded with adult schwann cells[J].Neuropathol.Appl.Neurobiol,1997,23:387-398.
[9]Ferri CC,Bisby MA.Improved survival of injuried Sciatic nerve schwann cells in mice lacking the P75 receptor[J].Neurosci,Lett.1999,272:191-194.
[10]Sharma AK,Thomas PK.Peripheral nerve regeneration in experimental diabetes(J).Neurol sci,1975,24:417-424.
[11]Ekstrom AR,Kanje M,Skottner A.Nerve regeneration and serum levels of insulin-like growth factor-1 in rats with streptoxotocin-induced insulin deficiency(J).Brain Res,1989,496:141-147.
[12]Cheng C,Zochodne DW.in vivo proliferation,migration and phenotypic changes of Schwann cells in the presence of myelinated fibers[J].Neurosci,2002,115:321-329.
[13] Eckersley L, Ansselin AD, Tomlinson DR. Effects of experimental diabetes on axonal and Schwann cell changes in sciatic nerve isografts[J]. Brain Res, 2001,92:128-137.
[14] Hellweg R, and Hartung HD. Endogenous levels of nerve growth factor (NGF) are altered in experimental diabetes mellitus: a possible role for NGF in the pathogenesis of diabetic neuropathy[J]. J. Neurosci.Res, 1990,26:258-267.
[15] Vo PA and Tomlinson DR. The regeneration of peripheral noradrenergic nerves after chemical sympathectomy in diabetic rats: effects of nerve growth factor[J]. Exp. Neurol, 1999, 157(:)127- 134
[16] Arthur JC, Yiannoutsos C, Simpson DM, et al. A phase 11 trial of nerve growth factor for sensory neuropathy associated with HIV infection[J]. Neurology, 2000,54:1080-1088.
[17] Goss JR, Goins WF, Lacomis D, et al. Herpes simplex-mediated gene transfer of nerve growth factor protects against peripheral neuropathy in streptozotocin-induced diabetes in the mouse[J].Diabetes, 2002, 51:2227-2232.
[18] Takeshi Suzuki, Hiromi sekido, Noriaki Kato, et al. Neurotrophin-3-induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose: involvement of the polyol pathway[J]. Journal of Neurochemistry. 2004,91:1430-1438.
[19] Kaplan DR. and Miller FD. Neurotrophin signal transduction in the nervous system[J]. Curr. Opin. Neurobiol, 2002,10:381-391.
[20] Segal RA. Selectivity in neurotrophin signaling: theme and variations[J]. Annu. Rev. Neurosci, 2003,26:299-330.
[21] Anton ES. Weskamp G, Reichardt LF, et al. Nerve growth factor and ist low-affinity receptor promote Schwann cell migration(J). proc.Nati Acad. Sci, 1994,91:2795-2799.
[22] Bentley CA. and Lee KF. p75 is important for axon growth and Schwann cell migration during development[J]. Neurosci. 2000,20:7706-7715.
[23] Meier C, Parmantier E, Brennan A, et al. Developing Schwann cells acquire the ability to survive with out axons by establishing an autocrine circurt involving insulin-like growth factor, neurotrophin-3. and platelet-derived growth factor-BB[J]. Neurosci,1999, 19:3847-3859.
[24] Obrosuva IG, Van Huysen C, Fathallan L, et al. Aldose reductase inhibitor fidarestat prevents retina oxidative stress and vascular endothelial growth factor overexpression in streptozotocin-diabetic rats[J]. Diabetes, 2003, 52:864-871.
[25] Christopher RP, Weixian Z, Yuichi M, et al. Early gene responses of trophic factor in nerve regeneration differ in experimental type 1 and type 2 diabetic polyneuropathies[J]. Journal of Neuropathology and Experimental Neurology, 2002, 10:857-871.
[1] Luke Eckersley. Role of the Schwann cell in diabetic neuropathy[J].International Review of Neurobiology,2002, 50:293-312
[2] Grinspan JB, Marchionni MA , Reeves M, et al. Axonal interactions regulate Schwann cell apoptosis in developing peripheral nerve:Neuregulation receptors and the role of neuregulins[J]. Neurosci,1996,16:6107-6118.
[3] Kunjorr, Hirohito FT, Akira T, et al. Basic behavior of migratory Schwann cells in peripheral nerve regeneration[J]. Exp Neuro,1996,137:301-304.
[4] Korching. The neurotrophic factor concept: a reexamination[J].Neurosci, 1993.13:2793-2795.
[5] Meyer M, Matsuoka I, Wetmore C, et al. Enhanced synthesis of brain neurotrophic factor in lesioned peripheral nerve: different mechanism are responsible for the regulation of BDNFand NGFmRNA[J].J Cell Biol, 1992,199:45-47.
[6] Hammcerberg H, Piehl FC. GDNFmRNA in Schwann cell and DRG satellite cell after chronic sciatic nerve injure[J]. Neuroreport, 1996,22:857-859.
[7] Grothe C, meisinger C, Hertenstein A, et al. Expression of FGF-2 and FGFR-1 mRNA in spinal ganglial an sciatic nerve regeneration after peripheral lesion[J]. Neurosci, 1996,76:123-125.
[8] Hiroshi F, Frigen J. Difterential expression of mRNA for neurotrophins and their receptors after axotomy of the sciatic nerve[J]. J Cell Biol, 1992,123:455-456.
[9] Kuff ler DD. Promoting and directing axon autogrowth[J]. Mol Neurobiol,1994,9:233-235.
[10] Oi zhi Gong, Michael T. Expression of excellular matrix and cell surface molecules in the olfactory nerve pathway during early development[J]. J Comp Neuro, 1996,366:1-5.
[11] Jiang X, Zhang JS, Jakobsen J. Differential effect of p75~(NTR) on expression of pro-apoptotic proteins C-Jun, p38 and caspase-3 in DRG cells after axotomy in experimental diabetes[J]. Neuroscience, 2005,132:1083-1093.
[12] Taniuchi M, Clark HB, Johnson Em. Induction of nerve growth factor receptor in Schwann cells after axotomy. Proc Nati Acad Sci USA,1986,83:4094-4098.
[13] Taniuchi M, Clark HB, Schweitzer JB, Expression of nerve growth factor receptors by Schwann cell of axotomized peripheral nerves:Ultrastructural Location. Suppression by axonal contact and binding properties[J]. J Neurosci, 1988, 8:664-681.
[14] Leinninger Gm, Vincent AM, Feldman LF. The role of growth factors in diabetic peripheral neuropathy[J]. J Peripheral Nervous System,2004,9:26-53.
[15] Kaplan DR. Hempstead BL. Martin-Zanca D et al. The trk proto-oncogene product:a signal transducing receptor for nerve growth factor [J]. Science, 1991, 252:554-558.
[16] Patapoution A. Reichardt LF. Trk receptors: mediators of neurotrophin action[J]. Curr Opin Neurobiol, 2001,11:272-280.
[17] Phee SG. Regulation of phosphoinositide-specific phospholipase C[J].Anny Rev Biochem. 2001,70:281-312.
[18] Dikic L, Batzer AG, Blaikie P, et al. Shc binding to nerve growth factor receptor is mediated by the phosphotyrosine interaction domain[J].J Biol Chem., 1995,270:15125-15129.
[19] Rozakis-Adcock M, McGlade J, Mbamalu G. Association of the Shc and Brb2/sem5 SH2-containing proteins is implicated in activation of the Ras pathway by tyrosine kinases[J]. Nature, 1992,360:689-692.
[20] Atwal JK, Massie B, Miller FD. The TrkB-Shc site signals neuronal survival and local axon growth via mEk and P13-kinase[J]. Neuron,2000,27:265-277.
[21] Xing J, Ginty DD, Greenbery ME. Coupling of the RAS-MAPK path way to gene activation by RSK2, a growth factor-regulated CREB kinase[J].Science, 1996,273:959-963.
[22] Zheng NH, Kar S, Quirion R. FKHRL 1 and its homoloys are new targets of nerve growth factor Trk receptor signaling. J Neurochem,2002,80:1049-1061.
[23] Holgado-madruga M, Moscatello DK, Emlet DR. et al. Grb2-associated binding-1 mediates phosphatidy linositol 3 kinase activation and the promotion of cell survival by nerve growth factor[J]. proc Acad Sci USA, 1997,94:12419-12424.
[24] Datta SR. Brunnet A. Greenbery ME. Cellular survival: a play in three Akts[J]. Gene Dev, 1999,13:2905-2927.
[25] Datta SR, Dudek H, Jao X. et al. Akt phoshorylation of BAD couples survival signals to the cell-intrinsic death machinery [J]. Cell,1997,91:231-41.
[26] Ozdinler PH, ErzurumluRS. Regulation of neurotrophin-induced axonal responses via Pho GTPases[J]. J Comp Neurol, 2001,438:377-387.
[27] Takahushi M. the GDNF/RET signaling pathway and human diseases[J].Cytokine Growth Factor Rev 2001,12:361-373.
[28] O' connor R, Kauftmann-Zeh A, Liu YM, et al. Identification of domains of the insulin-like growth factor I receptor that are required for protection from apoptosis[J]. Mol Cell Biol.2001,17:427-435.
[29] Sharma AK, Thomas PK. Peripheral nerve regeneration in experimental diabetes[J].Neurol sci, 1975, 24:417-424.
[30] Ekstron AR, Kanje M, Skottner A. Nerve regeneration and serum levels of insulin-like growth factor-1 in rats with streptoxotocin-induced insulin deficiency[J]. Brain Res, 1989,496:141-147.
[31] James Mk, Diygkas WZ. Impaired peripheral nerve regeneration in diabetes mellitus[J]. Journal of the Peripheral Nervous System,2005,10:144-157.
[32] Terada M, Yasuda H, Kikkawa R, et al. Tolrestat improves nerve regeneration after crush injury in streptozocin-induced diabetic rats. Metabolism. 1996,45:1189-1195.
[33] Perry VH. , Brown MC. Macrophages and nerve regeneration[J]. Curr Opin Neurobiol, 1992,2:679-682.
[34] Reichert F, saada A. Rotshenker S. Peripheral nerve injury induces Schwann cells to express two macrophage phenotypes: phagocytosis and the galactose-specfitic lectin MAC-2[J]. J Neurosci ,1994,14:3231-3245.
[35] Cheng C. Zochodne DW. In vivo proliferation, migration and phenotypic changes of Schwann cells in the presence of myelinated fibers[J]. Neurosci,2002,115:321-329.
[36] Eckersley L, Ansselin AD, Tomlinson DR. Effects of experimental diabetes on axonal and Schwann cell changes in sciatic nerve isografts[J]. Brain Res, 2001,92:128-137.
[37] Takeshi Suzuki, Hiromi sekido, Noriaki kato, et al.Neurotrophin-3-induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose: involvement of the polyol pathway[J]. Journal of Neurochemistry. 2004,91:1430-1438.
[38] Kaplan DR. Miller FD. Neurotrophin signal transduction in the nervous system[J]. Curr. Opin. Neurobiol, 2002,10:381-391.
[39] Segal RA. Selectivity in neurotrophin signaling: theme and variations[J]. Annu. Rev. Neurosci, 2003,26:299-330.
[40] Anton ES, Weskamp G, Reichard LF, et al. Nerve growth factor and its low-affinity receptor promote Schwann cell migration[J]. Proc. Nati Acad. Sci, 1994,91:2795-2799.
[41] Bentley CA, and Lee KF. p75 is important for axon growth and Schwann cell migration during development(J). Neurosci. 2000,20:7706-7715.
[42] Meier C, Parmantier E, Brennan A,et al. Developing Schwann cells acquire the ability to survive without axons by establishing an autocrine circuit involving insulin-like growth factor,neurotrophin-3. and platelet-derived growth factor-BB[J]. Neurosci.1999, 19:3847-3859.
[43] Christopher R, Pierson MD, Weixian MD, et al. Early gene responses of trophic factor in nerve regeneration differ in experimental type 1 and type 2 diabetic polyneuropathies[J]. Journal of Neuropathology and Experimental Neurology. 2002, 10:857-871.
[44] Vinik Al, Park TS, Stansberryt KB, et al. Diabetic neuropathies [J] Diabetologia, 2000,43:957-973.
[45] Nishikawa T, Edelstein D, Du XL, et al. Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage Nature, 2000,404:787-790.
[46] Kang D, Hamasaki N. Mitochondrial oxidative stress and mitochondrial DNA[J].. Clin Chem Lab Med, 2003,41(10):1281-1288.
[2]Vinik AL,Park TS,Stansberryt KB,et al.Diabetic neuropathies[J].Diabetologia.2000,43:957-973.
[3]Nishikawa T,Edelstein D,Du XL,et al.Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage[J].Nature,2000.404:787-790.
[4]Kang D,Hamasaki N,Mitochondrial oxidative stress and mitochondrial DNA[J].Clin Chem Lab Med,2003.41(10):1281-1288.
[5]Eckersley L.Role of the Schwann cell in diabetic neuropathy[J].International review of neurobiology.2002.50:293-312.
[6]Feldman.et al.Diabetic neuropathy.In:Current review of diabetes[J].Current medicine,Philadelphia,1999.71-83.
[7]James M.Kennedy,et al.Impaired peripheral nerve regeneration in diabetes mellitus[J].Journal of the peripheral nervous system,2005.10:144-157
[8]徐科.神经生物学纲要(M).北京:科学出版社,2000.45-47。
[9]Luke Eckersley.Role of the Schwann cell in diabetic neuropathy.International review of neurobiology,2002,50:293-312.
[10]Grinspan,JB.Axonal interactions regulate Schwann cell apoptosis in developing peripheral nerve:Neuregulation receptors and the role of neuregulin[J].Neurosci,1996.16:6107-6118.
[11]Gulati A K.Peripheral nerve regeneration through short and longterm degenerated nerve transplants[J].Brain Res,1996.742:265-270.
[12]Ansselin,A D.Peripheral nerve regeneration through nerve guides seeded with adult Schwann cells[J].Neuropathol Appl,Neurobiol,1997.23:387-398.
[13]Korching.The neurotrophic factor concept:a reexamination[J].Neurosci,1993,13:2793.
[14]Jiang X,Zhang JS,Jakobsen J.Differential effect of p75~(NTR) on Expression of pro-apoptotic proteins C-Jun,and caspase-3 in DRG cells After axotomy in experimental diabetes[J].Neuroscience,2005. 132:1083-1092.
[15] Meyer M, Matsuoka I, Wetmore C, et al. Enhanced synthesis of brain neurotrophic factor in lesioned peripheral nerve: different mechanism are responsible for the regulation of BDNFand NGFmRNA[J].J Cell Biol, 1992.199:45-50.
[16] Hammcerberg H, Piehl F, Cullheims, et al. GDNFmRNA in Schwann cell and DRG satellite cell after chronic sciatic nerve injure[J].Neuroreport, 1996.22:857-860.
[17] Grothe C. meisinger C. Hertenstein A, et al. Expression of FGF-2 and FGFR-1 mRNA in spinal gonglial an sciatic nerve regeneration after peripheral lesion, [J] Neurosci, 1996,76:123-125.
[18] Hiroshi F. Frigen J. Difterential expression of mRNA for neurotrophins and their receptors after axotomy of the sciatic nerve[J]. J Cell Biol, 1992,123:455-457.
[19] Kuffler DD. Promoting and directing axon autogrowth [J]. Mol Neurobiol, 1994.9:233-235.
[20] Cheng C,. in vivo proliferation, migration and phenotypic changes of Schwann cells in the presence of myelinated fibers[J].2002.115:321-329.
[21] Eckersley L. Effects of experimental diabetes on axonal and Schwann cell changes in sciatic nerve isografts(J). Brain Res,2001.92:128-137.
[22] Ferri C C. Improved survival of injuried sciatic nerve Schwann cells in mice lacking the P75 receptor[J]. Neurosci. Lett, 1999.272:191-194.
[23] Leinninger GM, Vincent AM, Feldman LF. The role of growth factors in diabetic peripheral neuropathy [J]. J Peripheral Nervous System.2004.9:26-53.
[24] Kaplan DR, HempsteadBL, Martin-Zanca D, etal. The trk proto-oncogene product:a signal transducing receptor for nerve growth factor [J].Science, 1991. 252:554-558.
[25] Patapoution A, Reichardt LF. Trk receptors: mediators of neurotrophin action [J]. Curr Opin Neurobiol, 2001.11:272-280.
[26] Vinik AI. Diabetic neuropathy: pathogenesis and therapy[J]. The American Journal of Medicine,1999.107:17-26.
[27]王学华,汪少颖主编.消渴病古今名家验案全析(M).北京:科学技术文献出版社,2006.480
[28]曾莉,陈卫平.疑病病中西诊疗指南(M).江苏:东南大学出版社,2004.206.
[29]姜喆.内分泌代谢病临床诊治(M).北京:科学技术文献出版社,2005.71.
[30]季钟朴.中西医结合思路与方法(M).上海:上海科学技术出版社,2006.199.
[31]胡熙明.中国中医秘方大全(M).上海:文汇出版社出版,1989.573.
[32]张家庆.中西医结合防治糖尿病研究的思路与方法[J].中国中西医结合杂志,1995.15(5):315-316.
[33]张家庆.中医治糖尿病不仅仅是降糖[J].中医杂志,1995,36(7):428.
[34]高从容,张家庆,黄庆玲.黄连素增加胰岛素抵抗大鼠横型胰岛素敏感性的实验研究[J].中国中西医结合杂志,1997,17(3):163-164.
[35]潘明政,郭富珊,梁晓春等.中药仙贞片对气阴两虚肾虚血瘀型糖尿病患者和细胞膜Na、K-ATP酶等的影响[J].中国中西医结合杂志,1997.17(1):13-16.
[36]熊曼琪,林安钟,朱章志等.加味挑仁承气汤对2型糖尿病大鼠胰岛素抵抗的影响[J],中国中西医结合杂志,1997.17(3):165-138.
[37]姜兆顺,张胜兰,蔡新告等.复方丹参注射液对2型糖尿病患者抗脂质过养化损伤的影响[J].中国中西医结合杂志,1997.7(1):32-34.
[1]薛庆善,主编.体外培养的原理与技术[M].北京:科学出版社,2001.151.
[2]汪谦,主编.临床医学实验方法学[M].北京:北京科学出版社,2002.19-22.
[3]Vinik AI.Diabetic Neuropathy:Pathogenesis and therapy[J].The American Journal of Medicaine,1999.107:17-23.
[4]James MK and Diygkas WZ.Impaired peripheral nerve regeneration in diabetes mellitus[J].Journal of the Peripheral Nervous System,2005.10:144-157.
[5]Brockes JP,Fields KL,Raff MC.A surfare marker for rat Schwann cells[J].Nature,1977.266:364-366.
[6]Brockes JP,Fields KL,Raft MC.Studies on cultured rat Schwann cells[J].Brain Res,1979.165(1):105-118.
[7]Askanas V,Engel WK,Dalakas MC,et al.Human Schwann cells in culture:Histochemical and ultrastructratl studies[J].Arch Neurol,1980.37(6):329-337.
[8]Oda Y,Okada Y,Katsuda S,et al.A simple method for the Schwann cell preparation from newborn rat sciatic nerves[J].J Neurosci meth.1989.28(3):163-169.
[9]Kaff ME,Abney E,Brockes JP,et al.Schwann cell growth factors(J).Cell 1978.5(3):813-822.
[10]Wood PM.Bunge CP.Evidence that sensory axons are mitogenic for Schwan cells[J].Nature,1975.256:662-664.
[11]Salzer JL,Williams AK,Glaser L,et al.Studies of Schwann cell proliferation.Ⅱ.Characterization of the stimulation and specificity of the response to neurite membrane fraction[J].J Cell Biol,1980.84(3):753-766.
[12]Raft MC,Abney ER,Cohen J,et al.Two types of astrocytes in cultures of developing rat white matter:Difference in morphology,surface ganglioside,and growth characteristics(J).J Nenrosci,1983.3(6):1289-1300
[13]Mirsky R.Cell-type-specific markers in the nervous system cultures(J).TINS.1980.3(8):190-192.
[14]严恒林,沈馨亚,刘才栋等.人胎儿雪旺氏细胞的体外培养及其纯化的研究(J).神经解剖学杂志,1994.10(4):289-293
[15]徐科,主编.神经生物学纲要[M].北京:科学出版社,2001.47.
[1]Keiko Maekawa,Tsuyoshi Tanimoto,Satoshi Okada,et al.Expression of aldose reductase and sorbitol dehydrogenase genes in Schwann cells isolated from rat:effects of high glucose and osmotic stress[J].Molecular Brain Research,2001.251-256.
[2]Takeshi Suzuki,Hiromi sekido,Noriaki kato,et al.Neurotrophin-3-induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose:involvement of the polyol pathway[J].Journal of Neurochemistry,2004.91:1430-1438.
[3]Christopher R,Pierson,Zhang Weixian,et al.Early gene responses of trophic factor in nerve regeneration differ in experimental type 1 and type 2 diabetic polyneuropathies[J].Journal of Neuropathology and Experimental Neurology,2002.10:857-871.
[4]郑志宏,林玲.神经细胞培养理论与实践[M].北京:科学出版社,2002.196-197.
[5]薛庆善,主编.体外培养的原理与技术[M].北京:科学出版社,2001.151.
[6]蔡文琴,李海标,主编.发育神经生物学[M].北京:科学出版社,2001.171-174
[7]LB.莱维坦,LK.卡茨玛克著,舒斯云、包新民主译.神经元:细胞和分子生物学[M].北京:科学出版社,2001.247-267.
[8]许绍芬,主编.神经生物学[M].第2版,上海:上海医科大学出版社,1999.96-111。
[9]罗学港,主编.神经科学基础[M].湖南:中南大学出版社,2002.82-111.
[10]吴克夏,主编.细胞通讯与疾病[M].北京:科学出版社,2006.1-45.
[11]Takeshi Suzuki,Hiromi sekido,Noriaki kato,et al.Neurotrophin-3-induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose:involvement of the polyol pathway[J].Journal of Neurochemistry,2004.91:1430-1438.
[12]Meier C,Parmantier E,Brennan A,et al.Developing Schwann cells acquire the ability to survive without axons by establishing an autocrine circuit involving insulin-like growth factor,neurotrophin-3,and platelet-derived growth factor-BB[J].Neurosci,1999.19:3847-3859.
[1]季钟朴,主编.中西医结合思路与方法[M].上海:上海科学技术出版社,2006.199-209.
[2]曾莉,陈卫平.主编.疑难病中西诊疗指南[M].江苏:东南大学出版社,2004.206-225.
[3]张家庆.中西医结合防治糖尿病研究的思路与方法[J].中国中西医结合杂志,1995.15(5):315-316.
[4]姜喆,主编.内分泌代谢病临床诊治[M].北京:科学技术文献出版社,2005.71-117.
[5]王学华,汪少颖,主编..消渴病古今名家验案全析[M].北京:科学技术文献出版社,2006.480-527.
[6]胡熙明,主编.中国中医秘方大全[M].上海:文汇出版社出版,1989.573-587.
[7]王林丽.红景天及复方制剂药理和临床研究进展[J].中医药学报,2003.31(1):51-54.
[8]王强,阮晓,方兰等.资源植物红景天研究综述[J].新疆农业大学学报,2002.25(4):57-62.
[9]许绍芬,主编.神经生物学[M].上海:上海医科大学出版社,1999.96-111.
[10]罗学港,主编.神经科学基础[M].湖南:中南大学出版社,2002.82-111.
[11]LB.莱维坦,LK.卡茨玛克著,舒斯云、包新民主译.神经元:细胞和分子生物学[M].北京:科学出版社,2001.247-267
[12]郑志宏,林玲编著.神经细胞培养理论与实践[M].北京:科学出版社,2002.196-197.
[13]薛庆善.主编.体外培养的原理与技术[M].北京:科学出版社,2001.151-157.
[14]汪谦.主编.临床医学实验方法学[M].北京:北京科学出版社,2002.19-227.
[15]吴克夏,主编.细胞通讯与疾病[M].北京:科学出版社,2006.1-45
[16]黄文林,朱孝峰,主编.信号转导[M].北京:人民卫生出版社,2005.274-306.
[17]方秀斌,主编.神经肽与神经营养因子[M].北京:人民卫生出版社,2002.274-306.
[1]薛庆善,主编.体外培养的原理与技术[M].北京:科学出版社,2001:151.
[2]黄文林,朱孝峰,主编.信号转导(M).北京:人民卫生出版社,2005:274-306.
[3]方秀斌,主编.神经肽与神经营养因子(M).北京:人民卫生出版社,2002:274-306.
[4]Taniuchi M.clark HB.Johnson EM.Induction of nerve growth factor receptor in Schwann cells after axotomy.Proc Nati Acad Sci USA.1986,83:4094-4098
[5]raniuchi M.Clark HB.Schweitzer JB.Expression of nerve growth factor receptors by Schwann cell of axotomized peripheral nerves:Ultrastructural Location.Suppression by axonal contact and binding properties[J].J Neurosci.1988,8:664-681.
[6]Rozakis-Adcock M,Mcglade J,Mbamalu G.Association of the Shc and Brb2/sem5 SH2-containing proteins is implicated in activation of the Ras pathway by tyrosine kinases[J].Nature 1992,360:689-692.
[7]Xing J,Ginty DD,Greenbery ME.Coupling of the RAS-MAPK path way to gene activation by RSK2,a growth factor-regulated CREB kinase[J].Science 1996,273:959-963.
[8]Atwal JK,Massie B,Miller FD.The TrkB-Shc site signals neuronal survival and local axon growth via mEk and P13-kinase[J].Neuron,2000,27:265-277.
[9]Sharma AK,Thomas PK.Peripheral nerve regeneration in experimental diabetes[J].Neurol sci,1975,24:417-424.
[10]Ekstron AR,Kanje M,Skottner A.Nerve regeneration and serum levels of insulin-like growth factor-1 in rats with streptoxotocin-induced insulin deficiency[J].Brain Res,1989,496:141-147.
[11]James Mk Diygkas WZ.Impaired peripheral nerve regeneration in diabetes mellitus[J].Journal of the Peripheral Nervous System,2005,10:144-157.
[12]Luke Eckersley.Role of the Schwann cell in diabetic neuropathy.International Review of Neurobiology[J].2002,50:293-312.
[13]Korching.The neurotrophic factor concept:a reexamination[J].Neurosci,1993,13:2793.
[14]罗学港,主编,神经科学基础[M].湖南:中南大学出版社,2002,100-101.
[15] Terada M, Yasuda H, Kikkawa R, et al. Tolrestat improves nerve regeneration after crush injury in streptozocin-induced diabetic rats[J]. Metabolism, 1996,45:1189-1195.
[16] Perry VH , Brown MC. Macrophages and nerve regeneration[J]. Curr Opin Neurobiol, 1992, 2:679-682.
[17] Reichert F, saada A. Rotshenker S. Peripheral nerve injury induces Schwann cells to express two macrophage phenotypes: phagocytosis and the galactose-specitic lectin MAC-2[J].Neurosci, 1994,14:3231-3245.
[18] Cheng C, Zochodne DW. In vivo proliferation, migration and phenotypic changes of Schwann cells in the presence of myelinated fibers[J].Neurosci, 2002,115:321-329.
[19] Hiroshi F, Frigen J. Differential expression of mRNA for neurotrophins and their receptors after axotomy of the sciatic nerve[J].Cell Biol, 1992,123:455.
[20] Vinik Al, Park Ts, Stansberryt KB, et al. Diabetic neuropathies [J].Diabetologia, 2000,43:957-973.
[21] Nishikawa T, Edelstein D, Du XL, et al. Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage[J]. Nature, 2000,404:787-790.
[22] Kang D, Hamasaki N. Mitochondrialoxidative stress and mitochondrial DNA[J]. Clin Chem Lab Med, 2003, 41(10):1281-1288.
[23] Taniuchi M, clark HB, Johnson EM. Induction of nerve growth factor receptor in Schwann cells after axotomy[J]. Proc Nati Acad Sci USA,1986,83:4094-4098.
[24] Taniuchi M, Clark HB, Schweitzer JB. Expression of nerve growth factor receptors by Schwann cell of axotomized peripheral nerves:Ultrastructural Location. Suppression by axonal contact and binding properties[J]. Neurosci, 1988, 8:664-681.
[25] Hellweg R, Hartung HD. Endogenous levels of nerve gronth factor (NGF) are altered in experimental diabetes mellitus: a possible role for NGF in the pathogenesis of diabetic neuropathy[J]. Neurosci Res,1990,26:258-267.
[26] Takeshi Suzuki, Hiromi sekido, Noriaki kato, et al. Neurotrophin-3- induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose:involvement of the polyol pathway[J].Journal of Neurochemistry,2004,91:1430-1438.
[27]吴克夏,主编.细胞通讯与疾病[M].北京:科学出版社,2006,1-45.
[28]vinik AI.Diabetic Neuropathy:Pathogenesis and Therapy[J].The American Journal of Medicine,1999,107:17-25
[29]王学华,汪少颖,主编.消渴病古今名家验案全析[M].北京:科学技术文献出版社,2006,480.
[30]曾莉、陈卫平,主编.疑难病中西诊疗指南[M].江苏:东南大学出版社,2004,206.
[31]姜喆,主编.内分泌代谢病临床诊治[M].北京:科学技术文献出版社,2005,71.
[32]季钟朴,主编.中西医结合思路与方法[M].上海:上海科学技术出版社,2006,199.
[33]胡熙明,主编.中国中医秘方大全[M].上海:文汇出版社,1989,573.
[34]张家庆.中西医结合防治糖尿病研究的思路与方法[J].中国中西医结合杂志,1995,15(5):315-316.
[35]张家庆.中医治糖尿病不仅仅是降糖[J].中医杂志,1995,36(7):428.
[36]高从容,张家庆,黄庆玲.黄连素增加胰岛素抵抗大鼠横型胰岛素敏感性的实验研究[J].中国中西医结合杂志,1997,17(3):162-164
[37]潘明政,郭富珊,梁晓春,等.中药仙贞片对气阴两虚肾虚血瘀型糖尿病患者和细胞膜Na、K-ATP酶等的影响[J].中国中西医结合杂志,1997,17(1):13-16.
[38]熊曼琪,林安钟,朱章志,等.加味挑仁承气汤对2型糖尿病大鼠胰岛素抵抗的影响[J].中国中西医结合杂志,1997,17(3):165-138
[39]姜兆顺,张胜兰,蔡新告,等.复方丹参注射液对2型糖尿病患者抗脂质过养化损伤的影响[J].中国中西医结合杂志,1997,7(1):32-34
[40]王林丽,红景天及复方制剂药理和临床研究进展[J].中医药学报,2003,31(1):51-54
[41]王强,阮晓,方兰,等.资源植物红景天研究综述[J].新疆农业大学学报,2002,25(4):57-62
[42]陈秋丽,邓伟国,王毅.红景天皂苷对雌性大鼠蛋白质和脂肪代谢的影响[J].吉林大学学报医学版,2003,29(4):411-413.
[43]赵文,王庭欣,蒋东升.蔷薇红景天提取液对糖尿病模型小鼠胰腺功能的修 复作用[J].中国比较医学杂志,2003,13(5):309.
[44]赵文,王庭欣,蒋东升,等.蔷薇红景天对四氧嘧啶糖尿病小鼠血糖的调节作用[J].卫生研究,1999,28(6):362-363.
[45]胡敏,于金德.红景天苷对心血管系统作用的研究[J].国外医学.心血管疾病分册,2003,30(5):398-300.
[1]Feldman EL,Steven MJ,Russell JW,et al.Diabetic neuropathy.In:Current Review of Diabetes[J].Current medicine.Philadelphia,1999,71-83.
[2]James Mk,Diygkas WZ.Impaired peripheral nerve regeneration in diabetes mellitus[J].Journal of the Peripheral Nervous System,2005,10:144-157.
[3]徐科,主编.神经生物学纲要(M).北京:科学出版社,2000,45-47.
[4]罗学港,唐建华,熊燕编,主编.神经科学基础(M).湖南:中南大学出版社,2002,96-98.
[5]Luke Eckersley.Role of the Schwann cell in diabetic neuropathy[J].International Review of Neurobiology,2002,50:293-312。
[6]Grinspan JB,Marchionni MA,Reeves M,et al.Axonal interactions regulate Schwann cell apoptosis in developing periphesal nerve:Neuregulation receptors and the role of neuregulin[J].Neurosci,1996,16:6107-6118.
[7]Gulati AK.Peripheral nerve regeneration through short-and longterm degenerated nerve transplants[J].Brain Res,1996,742:265-270.
[8]Ansselin AD.Peripheral nerve regeneration through nerve guides seeded with adult schwann cells[J].Neuropathol.Appl.Neurobiol,1997,23:387-398.
[9]Ferri CC,Bisby MA.Improved survival of injuried Sciatic nerve schwann cells in mice lacking the P75 receptor[J].Neurosci,Lett.1999,272:191-194.
[10]Sharma AK,Thomas PK.Peripheral nerve regeneration in experimental diabetes(J).Neurol sci,1975,24:417-424.
[11]Ekstrom AR,Kanje M,Skottner A.Nerve regeneration and serum levels of insulin-like growth factor-1 in rats with streptoxotocin-induced insulin deficiency(J).Brain Res,1989,496:141-147.
[12]Cheng C,Zochodne DW.in vivo proliferation,migration and phenotypic changes of Schwann cells in the presence of myelinated fibers[J].Neurosci,2002,115:321-329.
[13] Eckersley L, Ansselin AD, Tomlinson DR. Effects of experimental diabetes on axonal and Schwann cell changes in sciatic nerve isografts[J]. Brain Res, 2001,92:128-137.
[14] Hellweg R, and Hartung HD. Endogenous levels of nerve growth factor (NGF) are altered in experimental diabetes mellitus: a possible role for NGF in the pathogenesis of diabetic neuropathy[J]. J. Neurosci.Res, 1990,26:258-267.
[15] Vo PA and Tomlinson DR. The regeneration of peripheral noradrenergic nerves after chemical sympathectomy in diabetic rats: effects of nerve growth factor[J]. Exp. Neurol, 1999, 157(:)127- 134
[16] Arthur JC, Yiannoutsos C, Simpson DM, et al. A phase 11 trial of nerve growth factor for sensory neuropathy associated with HIV infection[J]. Neurology, 2000,54:1080-1088.
[17] Goss JR, Goins WF, Lacomis D, et al. Herpes simplex-mediated gene transfer of nerve growth factor protects against peripheral neuropathy in streptozotocin-induced diabetes in the mouse[J].Diabetes, 2002, 51:2227-2232.
[18] Takeshi Suzuki, Hiromi sekido, Noriaki Kato, et al. Neurotrophin-3-induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose: involvement of the polyol pathway[J]. Journal of Neurochemistry. 2004,91:1430-1438.
[19] Kaplan DR. and Miller FD. Neurotrophin signal transduction in the nervous system[J]. Curr. Opin. Neurobiol, 2002,10:381-391.
[20] Segal RA. Selectivity in neurotrophin signaling: theme and variations[J]. Annu. Rev. Neurosci, 2003,26:299-330.
[21] Anton ES. Weskamp G, Reichardt LF, et al. Nerve growth factor and ist low-affinity receptor promote Schwann cell migration(J). proc.Nati Acad. Sci, 1994,91:2795-2799.
[22] Bentley CA. and Lee KF. p75 is important for axon growth and Schwann cell migration during development[J]. Neurosci. 2000,20:7706-7715.
[23] Meier C, Parmantier E, Brennan A, et al. Developing Schwann cells acquire the ability to survive with out axons by establishing an autocrine circurt involving insulin-like growth factor, neurotrophin-3. and platelet-derived growth factor-BB[J]. Neurosci,1999, 19:3847-3859.
[24] Obrosuva IG, Van Huysen C, Fathallan L, et al. Aldose reductase inhibitor fidarestat prevents retina oxidative stress and vascular endothelial growth factor overexpression in streptozotocin-diabetic rats[J]. Diabetes, 2003, 52:864-871.
[25] Christopher RP, Weixian Z, Yuichi M, et al. Early gene responses of trophic factor in nerve regeneration differ in experimental type 1 and type 2 diabetic polyneuropathies[J]. Journal of Neuropathology and Experimental Neurology, 2002, 10:857-871.
[1] Luke Eckersley. Role of the Schwann cell in diabetic neuropathy[J].International Review of Neurobiology,2002, 50:293-312
[2] Grinspan JB, Marchionni MA , Reeves M, et al. Axonal interactions regulate Schwann cell apoptosis in developing peripheral nerve:Neuregulation receptors and the role of neuregulins[J]. Neurosci,1996,16:6107-6118.
[3] Kunjorr, Hirohito FT, Akira T, et al. Basic behavior of migratory Schwann cells in peripheral nerve regeneration[J]. Exp Neuro,1996,137:301-304.
[4] Korching. The neurotrophic factor concept: a reexamination[J].Neurosci, 1993.13:2793-2795.
[5] Meyer M, Matsuoka I, Wetmore C, et al. Enhanced synthesis of brain neurotrophic factor in lesioned peripheral nerve: different mechanism are responsible for the regulation of BDNFand NGFmRNA[J].J Cell Biol, 1992,199:45-47.
[6] Hammcerberg H, Piehl FC. GDNFmRNA in Schwann cell and DRG satellite cell after chronic sciatic nerve injure[J]. Neuroreport, 1996,22:857-859.
[7] Grothe C, meisinger C, Hertenstein A, et al. Expression of FGF-2 and FGFR-1 mRNA in spinal ganglial an sciatic nerve regeneration after peripheral lesion[J]. Neurosci, 1996,76:123-125.
[8] Hiroshi F, Frigen J. Difterential expression of mRNA for neurotrophins and their receptors after axotomy of the sciatic nerve[J]. J Cell Biol, 1992,123:455-456.
[9] Kuff ler DD. Promoting and directing axon autogrowth[J]. Mol Neurobiol,1994,9:233-235.
[10] Oi zhi Gong, Michael T. Expression of excellular matrix and cell surface molecules in the olfactory nerve pathway during early development[J]. J Comp Neuro, 1996,366:1-5.
[11] Jiang X, Zhang JS, Jakobsen J. Differential effect of p75~(NTR) on expression of pro-apoptotic proteins C-Jun, p38 and caspase-3 in DRG cells after axotomy in experimental diabetes[J]. Neuroscience, 2005,132:1083-1093.
[12] Taniuchi M, Clark HB, Johnson Em. Induction of nerve growth factor receptor in Schwann cells after axotomy. Proc Nati Acad Sci USA,1986,83:4094-4098.
[13] Taniuchi M, Clark HB, Schweitzer JB, Expression of nerve growth factor receptors by Schwann cell of axotomized peripheral nerves:Ultrastructural Location. Suppression by axonal contact and binding properties[J]. J Neurosci, 1988, 8:664-681.
[14] Leinninger Gm, Vincent AM, Feldman LF. The role of growth factors in diabetic peripheral neuropathy[J]. J Peripheral Nervous System,2004,9:26-53.
[15] Kaplan DR. Hempstead BL. Martin-Zanca D et al. The trk proto-oncogene product:a signal transducing receptor for nerve growth factor [J]. Science, 1991, 252:554-558.
[16] Patapoution A. Reichardt LF. Trk receptors: mediators of neurotrophin action[J]. Curr Opin Neurobiol, 2001,11:272-280.
[17] Phee SG. Regulation of phosphoinositide-specific phospholipase C[J].Anny Rev Biochem. 2001,70:281-312.
[18] Dikic L, Batzer AG, Blaikie P, et al. Shc binding to nerve growth factor receptor is mediated by the phosphotyrosine interaction domain[J].J Biol Chem., 1995,270:15125-15129.
[19] Rozakis-Adcock M, McGlade J, Mbamalu G. Association of the Shc and Brb2/sem5 SH2-containing proteins is implicated in activation of the Ras pathway by tyrosine kinases[J]. Nature, 1992,360:689-692.
[20] Atwal JK, Massie B, Miller FD. The TrkB-Shc site signals neuronal survival and local axon growth via mEk and P13-kinase[J]. Neuron,2000,27:265-277.
[21] Xing J, Ginty DD, Greenbery ME. Coupling of the RAS-MAPK path way to gene activation by RSK2, a growth factor-regulated CREB kinase[J].Science, 1996,273:959-963.
[22] Zheng NH, Kar S, Quirion R. FKHRL 1 and its homoloys are new targets of nerve growth factor Trk receptor signaling. J Neurochem,2002,80:1049-1061.
[23] Holgado-madruga M, Moscatello DK, Emlet DR. et al. Grb2-associated binding-1 mediates phosphatidy linositol 3 kinase activation and the promotion of cell survival by nerve growth factor[J]. proc Acad Sci USA, 1997,94:12419-12424.
[24] Datta SR. Brunnet A. Greenbery ME. Cellular survival: a play in three Akts[J]. Gene Dev, 1999,13:2905-2927.
[25] Datta SR, Dudek H, Jao X. et al. Akt phoshorylation of BAD couples survival signals to the cell-intrinsic death machinery [J]. Cell,1997,91:231-41.
[26] Ozdinler PH, ErzurumluRS. Regulation of neurotrophin-induced axonal responses via Pho GTPases[J]. J Comp Neurol, 2001,438:377-387.
[27] Takahushi M. the GDNF/RET signaling pathway and human diseases[J].Cytokine Growth Factor Rev 2001,12:361-373.
[28] O' connor R, Kauftmann-Zeh A, Liu YM, et al. Identification of domains of the insulin-like growth factor I receptor that are required for protection from apoptosis[J]. Mol Cell Biol.2001,17:427-435.
[29] Sharma AK, Thomas PK. Peripheral nerve regeneration in experimental diabetes[J].Neurol sci, 1975, 24:417-424.
[30] Ekstron AR, Kanje M, Skottner A. Nerve regeneration and serum levels of insulin-like growth factor-1 in rats with streptoxotocin-induced insulin deficiency[J]. Brain Res, 1989,496:141-147.
[31] James Mk, Diygkas WZ. Impaired peripheral nerve regeneration in diabetes mellitus[J]. Journal of the Peripheral Nervous System,2005,10:144-157.
[32] Terada M, Yasuda H, Kikkawa R, et al. Tolrestat improves nerve regeneration after crush injury in streptozocin-induced diabetic rats. Metabolism. 1996,45:1189-1195.
[33] Perry VH. , Brown MC. Macrophages and nerve regeneration[J]. Curr Opin Neurobiol, 1992,2:679-682.
[34] Reichert F, saada A. Rotshenker S. Peripheral nerve injury induces Schwann cells to express two macrophage phenotypes: phagocytosis and the galactose-specfitic lectin MAC-2[J]. J Neurosci ,1994,14:3231-3245.
[35] Cheng C. Zochodne DW. In vivo proliferation, migration and phenotypic changes of Schwann cells in the presence of myelinated fibers[J]. Neurosci,2002,115:321-329.
[36] Eckersley L, Ansselin AD, Tomlinson DR. Effects of experimental diabetes on axonal and Schwann cell changes in sciatic nerve isografts[J]. Brain Res, 2001,92:128-137.
[37] Takeshi Suzuki, Hiromi sekido, Noriaki kato, et al.Neurotrophin-3-induced production of nerve growth factor is suppressed in Schwann cells exposed to high glucose: involvement of the polyol pathway[J]. Journal of Neurochemistry. 2004,91:1430-1438.
[38] Kaplan DR. Miller FD. Neurotrophin signal transduction in the nervous system[J]. Curr. Opin. Neurobiol, 2002,10:381-391.
[39] Segal RA. Selectivity in neurotrophin signaling: theme and variations[J]. Annu. Rev. Neurosci, 2003,26:299-330.
[40] Anton ES, Weskamp G, Reichard LF, et al. Nerve growth factor and its low-affinity receptor promote Schwann cell migration[J]. Proc. Nati Acad. Sci, 1994,91:2795-2799.
[41] Bentley CA, and Lee KF. p75 is important for axon growth and Schwann cell migration during development(J). Neurosci. 2000,20:7706-7715.
[42] Meier C, Parmantier E, Brennan A,et al. Developing Schwann cells acquire the ability to survive without axons by establishing an autocrine circuit involving insulin-like growth factor,neurotrophin-3. and platelet-derived growth factor-BB[J]. Neurosci.1999, 19:3847-3859.
[43] Christopher R, Pierson MD, Weixian MD, et al. Early gene responses of trophic factor in nerve regeneration differ in experimental type 1 and type 2 diabetic polyneuropathies[J]. Journal of Neuropathology and Experimental Neurology. 2002, 10:857-871.
[44] Vinik Al, Park TS, Stansberryt KB, et al. Diabetic neuropathies [J] Diabetologia, 2000,43:957-973.
[45] Nishikawa T, Edelstein D, Du XL, et al. Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage Nature, 2000,404:787-790.
[46] Kang D, Hamasaki N. Mitochondrial oxidative stress and mitochondrial DNA[J].. Clin Chem Lab Med, 2003,41(10):1281-1288.
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