四逆汤对内毒素休克大鼠脑损伤保护的分子机制研究
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摘要
感染性休克(Septic Shock)是现代危重病医学面临的普遍存在而又十分复杂的问题,成为直接影响患者预后,阻碍进一步提高救治成功率的突出难题。国外一组临床流行病学资料表明,2527例全身炎症反应患者中,感染性休克发生率为4%,其病死率为46%。我国的一项研究表明,烧伤面积大于30%者约40%并发感染性休克。对危重病患者的研究显示感染性休克发病率为15.7%,其中61.1%进一步发展为多器官功能障碍综合征(MODS),病死率为30.6%。由此可见,揭示感染性休克发病机理、探寻临床防治方法具有十分重要的理论价值和临床意义。在单纯西医治疗感染性休克难以取得进展性疗效时,积极地寻找中西医结合治疗感染性休克的方法就显得更为迫切,很多学者希望能从中医药方面对感染性休克治疗有所突破。根据其临床表现,感染性休克属厥证范围,多为脱证,故名之“厥脱”。中医数千年的发展在很大程度上是研究感染性休克的过程,在这方面积累了很多经验,但由于作用机制不明确,使得临床治疗缺乏依据。研究目的:
1.明确内毒素休克后脑损伤组织中炎症介质水平,中性粒细胞和脑血管内皮细胞等细胞表面黏附分子表达情况,脑血管平滑肌细胞、神经胶质细胞和神经细胞对细胞外炎症介质刺激产生的功能代谢变化以及结构改变等三者之间的规律性关系。
2.明确内毒素休克脑损伤中起作用的白细胞介素-1α(IL-1α)、肿瘤坏死因子-α(TNF-α)增加等信号异常是否可以上调中性粒细胞和脑血管内皮细胞等细胞表面黏附分子表达,脑血管平滑肌细胞、神经胶质细胞和神经细胞细胞内气体信使一氧化氮(NO)数量的异常等受体后信号转导通路的变化与脑部不同区域血管反应性及神经元损伤修复的关系。
3.明确四逆汤对以上指标有何作用,从而说明回阳救逆法改善感染性休克后脑微循环障碍和神经元损伤的分子细胞机制,发展中医急症理论。
研究方法:
1.以大鼠内毒素休克模型模拟感染性休克,首先运用基因芯片技术筛选出对感染性休克作用明显的受体前信号物质炎症细胞因子。
2.探讨四逆汤方对内毒素引起的受体前信号物质的作用,如组织中炎症介质IL-1α、TNF-α水平的变化。具体方法采用ELISA方法测定大鼠脑组织IL-1α、TNF-α的含量。
3.探讨四逆汤方对IL-1α、TNF-α引起的受体信号物质的作用,如中性粒细胞和脑血管内皮细胞等细胞表面黏附分子如细胞间黏附分子(ICAM-1,CD54)、神经细胞黏附分子(NCAM,CD56)、血管细胞黏附分子(VCAM-1,CD106)表达情况。具体方法采用免疫组化方法测定大鼠脑组织CD54、CD56、CD106分布与数量变化。
4.探讨四逆汤方对脑血管平滑肌细胞、神经胶质细胞和神经细胞接受细胞膜上受体刺激后细胞内信号物质的变化,如脑组织NO产量、超氧化物歧化酶(SOD)与丙二醛(MDA)含量等;用生化法测SOD与MDA含量,免疫组化法测诱导型一氧化氮合酶(iNOS)分布与数量变化。
5.最后探讨四逆汤方对内毒素休克脑损伤组织结构的改善作用。用TUNEL法测大鼠脑组织细胞凋亡,用电镜观察脑组织内各细胞的超微结构,用光镜观察脑组织结构变化。
研究结果:
1.假手术组大鼠平均动脉血压在实验过程于1h开始有轻度下降;在注射内毒素后,模型组从1h开始与假手术组相比有极显著差别(p＜0.01),至6h时血压降至初始值的40.75%。地塞米松组大鼠至6h时血压降至初始值的45.32%,与模型组相比血压有所升高(p＜0.05)。四逆汤组大鼠至6h时血压降至初始值的67.68%,与模型组相比血压升高极显著(p＜0.01)。
2.基因芯片筛查出Ccl2,4,7,20、Cxcl1,2,9,10,11、Ccr2,3,9;TNF-α、IL-1α、IL-1β等在内毒素休克大鼠脑组织出现非常显著的高表达;还有很少报道的IL-13、IL-15、I15ra也有差异表达。其中IL-13于1h无变化,于2h、3h、6h均上调,IL-15于1h、2h均下调,I15ra于1h、2h、3h、6h均下调。
3.内毒素休克大鼠脑组织中TNF-α与IL-1α从1h至6h的浓度变化规律与基因芯片技术检测出的mRNA水平变化规律基本相同。①模型组大鼠脑组织中IL-1α含量在1h较假手术组有所降低(p＜0.01),2h无显著差异(P＞0.05),3h(p＜0.05)、6h(p＜0.01)有所升高,并于6h达峰值。地塞米松组与四逆汤组大鼠脑组织中IL-1α含量在2h、3h、6h均较模型组有所下降(p＜0.01)。②模型组大鼠脑组织中TNF-α含量在四个时间段较假手术组均有所升高(p＜0.01),并于6h达峰值。地塞米松组大鼠脑组织中TNF-α含量在1h、2h、3h、6h均较模型组有所下降(p＜0.01)。四逆汤组大鼠脑组织中TNF-α含量在3h、6h时间段较模型组有所下降(p＜0.01)。
4.正常组、假手术组大鼠脑组织中CD54无表达,CD56、CD106有微弱表达。内毒素休克模型组大鼠脑组织中CD54、CD56、CD106于1h、2h、3h、6h等不同时间段均高于空白对照组与假手术组(p＜0.01),CD54、CD106于1h表达达峰值,CD56于6h表达达峰值。地塞米松组于1h、2h下调CD54的表达(p＜0.01),于1h上调CD56的表达(p＜0.01),于1h、2h、6h下调CD106的表达(p＜0.05)。四逆汤组于1h和2h(p＜0.01)、3h(p＜0.05)均下调CD54的表达,于3h(p＜0.05)、6h(p＜0.01)均上调CD56的表达,于1h、2h和3h(p＜0.01)、6h(p＜0.05)均下调CD106的表达。
5.正常组、假手术组大鼠脑组织iNOS无表达,模型组大鼠脑组织发现iNOS在1h表达量最高,2h、3h有所降低,6h时又有所回升。地塞米松组于1h(p＜0.01)、6h(p＜0.05)均下调iNOS的表达。四逆汤组仅于6h下调iNOS的表达(p＜0.01)。SOD在模型组大鼠脑组织中含量于四个时间段较假手术组均有所降低(p＜0.01),并于1h降至最低。地塞米松组与四逆汤组大鼠脑组织中SOD含量在1h、2h时间段较模型组有所升高(p＜0.01)。MDA在模型组大鼠脑组织中含量于四个时间段较假手术组均有所升高(p＜0.01),并于6h达峰值,其变化规律与SOD基本相同,地塞米松组大鼠脑组织中MDA含量在1h、2h、3h、6h时间段较模型组有所降低(p＜0.01),四逆汤组大鼠脑组织中MDA含量在2h、3h、6h时间段较模型组有所降低(p＜0.01)。
6.电镜与光镜观察结果显示:内毒素休克模型组大鼠于1h有反应性损伤,2h、3h损伤则有所恢复,6h损伤最严重;地塞米松组于1h较模型组损伤减轻;四逆汤组于6h较模型组损伤减轻。TUNEL法检测结果为:正常组与假手术组大鼠脑组织有极少量的凋亡细胞,内毒素休克模型组大鼠脑组织中的凋亡细胞较假手术组明显升高(p＜0.01),并于6h达峰值。地塞米松组于1h、2h较模型组凋亡减少(p＜0.01)。四逆汤组于2h、3h、6h较模型组凋亡减少(p＜0.01)。
研究结论:
1.内毒素休克后大鼠脑组织出现91个炎症细胞因子差异表达,其中36种细胞因子参与炎症应答,有较多报道的TNF-α、IL-1β、IL-10,还有很少报道的IL-1α、IL-13、IL-15、I15ra。
2.内毒素休克可以通过两条途径造成器官损伤:①LPS引起的受体前信号物质的作用,如组织中炎症介质TNF-α、IL-1α水平的升高→TNF-α、IL-1α引起的受体信号物质如中性粒细胞、脑血管内皮细胞等细胞表面黏附分子CD54表达→脑血管内皮细胞、神经胶质细胞和神经细胞接受细胞膜上受体刺激后细胞内信号物质的NO、MDA增加→脑组织结构损伤。②LPS直接引起受体信号物质CD54表达→CD54诱导TNF-α、IL-1α水平的变化→引发炎性介质的瀑布样效应,导致大量介质的激活→引起多种受体信号物质CD54、CD56、CD106表达→脑血管内皮细胞、神经胶质细胞和神经细胞接受细胞膜上受体刺激后细胞内信号物质的NO、MDA增加→脑组织结构损伤。
3.四逆注射液抢救休克是确有疗效的,而且其抗休克作用是多方面的,既可以升高血压、脑组织SOD含量、CD56的表达,又可以降低TNF-α、IL-1α、MDA的含量与CD54、CD106、iNOS、凋亡细胞的表达。但四逆汤的起效时间比较晚,作用效果稳定持久;地塞米松的起效时间比较早,但是作用效果不持久。
Septic shock is the widespread and complex problem concerned in modern Critical Care Medicine.It becomes the prominent issue to seriously impact on prognosis of patients and further improvement of successful treatment rate.One group of abroad clinical epidemiological research papers indicated that septic shock occurred at a rate of 4%among 2527 cases of patients with systemic inflammatory response,and 46%of septic shock patients were died.A study in China showed that 40%patients whose body surface areas are burned more than 30 percent had concurrently septic shock.Study on critically ill patients also showed that incidence of septic shock was 15.7%,61.1%of septic shock further developed to multiple organ dysfunction syndrome(MODS),fatality rate was 30.6%.Based on above studies,it has very important theoretical value and clinical significance to reveal septic shock pathogenesis and explore clinical prevention and treatment methods.When simplex western medicine treatment hardly makes progress in effect of septic shock,it appears urgently to look for Western & Chinese Traditional Medicine method to treat septic shock.Many medical scholars hope to find a breakthrough for septic shock with Chinese traditional medicine treatment.According to clinical manifestations, the septic shock is the scope of syncope.As most clinical manifestations are also collapse,it's also named as syncope and collapse syndrome.The development of traditional Chinese medicine in thousands of years was a research on septic shock and accumulated much experience,but the treatment is lack of evidence because of the unclear mechanism of septic shock.Much progress has been made in recent years through studying septic shock,however brain injury is rarely involved.This study takes the models of endotoxin-shock rats as research object to simulate septic shock.The study aims are to explore anomalous signal transduction in brain injury caused by septic shock and the effect of Sini Decoction in the process;define the molecular and cellular mechanisms of reviving yang for resuscitation method used to prevent and treat brain injury in septic shock and establish the scientific foundation for modernizing traditional Chinese medicine.
Objective
1.Clear level of inflammatory mediators,the expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,the changes of function metabolism and structural of brain vascular smooth muscle cells,glial cells,nerve cells stimulated by the extracellular inflammatory mediator of brain injury in septic shock.Then sum up regular relationship among them.
2.Clear if the role of increasing of IL-1α、TNF-αcan upregulate the expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,the relationship between the abnormal quantity of gas messenger-NO in brain vascular smooth muscle cells,glial cells and nerve cells and different brain regions vascular response and neuronal damage and repair.
3.Clear how Sini decoction act on above indicators in order to descript the method of reviving yang for resuscitation how improve molecular mechanisms of brain microcirculatory disturbances and neuronal damage.Develop Chinese medicine A&E theory.
Methods
1.This study takes the models of endotoxin-shock rats simulating septic shock for research object.First using technology of gene chip to screen prereceptor signal substances, inflammatory cytokines,which have an obvious role in septic shock.
2.To explore the role of Sini decoction on prereceptor signal substances induced by septic shock.Such as levels of tissue inflammatory mediators,IL-1α、TNF-α.The specific methods uses ELISA to measure IL-1α、TNF-αcontent in septic rat brain tissue.
3.To explore how Sini decoction act on receptor signal substances induced by increasing IL-1α、TNF-αcontent.Such as the expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,ICAM-1(CD54)、NCAM(CD56)、VCAM-1(CD106). The specific methods uses SABC to determine distribution and quantity of ICAM-1(CD54)、NCAM(CD56)、VCAM-1(CD106) in rat brain tissue.
4.To explore how Sini decoction act on the changes of signal substances after brain vascular smooth muscle cells,glial cells and nerve cells receiving membrane receptor stimulation. Such as NO production and SOD,MDA content in rat brain.The specific methods is to use biochemical methods to determine SOD,MDA content and SABC to determine distribution and quantity of iNOS rat brain tissue.
5.At last explore how Sini decoction improve tissue structure after septic brain injury. Using TUNEL to determine apoptosis in rat brain,ultrastructural changes of the cells with electron microscopy,structural changes of brain tissue with microscopy.
Results
1.Mean arterial blood pressure of sham-operated rats started to decline slightly at 1 hour. After i.v.injection of lipopolysaccharide,significant difference between model group and sham operated group at 1 hour(p<0.01),blood pressure fell to 40.75%of the initial values at 6 hour. Blood pressure of dexamethasone group fell to 45.32%of the initial values at 6 hour and had higher blood pressure compared with model group(p<0.05).Blood pressure of Sini decoction group fell to 67.68%of the initial values at 6 hour had higher blood pressure compared with model group(p<0.01).
2.Screening Ccl2,4,7,20、Cxcll,2,9,10,11、Ccr2,3,9;TNF-α、IL-1α、IL-1βwith very significant high expression in endotoxic rat brain.And some inflammatory cytokines such asIL-13、IL- 15、I15ra which were very few reports had differentially expressed. IL-13 has no changes at 1 hour,up-regulation at2,3,6 hour;IL-15 has down-regulation at 1,2 hour and I15ra has down-regulation at 1,2,3,6 hour.
3.Concentration changes of TNF-α、IL-1αin endotoxic rat brain were same as mRNA content changes detected by gene chip technology from 1 hour to 6hour.①Concentration of TNF-αin model group were higher than sham operated group on four time periods(p<0.01) and reached peak at 6 hour.Concentration of TNF-αof dexamethasone group were lower than model group from 1 hour to 6 hour(p<0.01).Concentration of TNF-αof Sini decoction group were lower than model group at 3,6 hour(p<0.01).②Concentration of IL-1αin model group were lower than sham operated group at 1 hour(p<0.01),no significant difference at 2 hour(P>0.05),higher at 3(p<0.05),6(p<0.01)hour and reached peak at 6 hour.Concentration of IL-1αof dexamethasone group were lower than model group from 1 hour to 6hour(p<0.01). Concentration of IL-1αof Sini decoction group were lower than model group at 2,3,6 hour(p<0.01).
4.No expression of CD54 and weak expression of CD56,CD106 in normal rat brain.The expression of CD54,CD56,CD106 in model group was higher than sham operated group and control group on four time periods(p<0.01).CD54,CD106 reached peak at 1 hour and CD56 reached peak at 6hour.The CD54 expressions of dexamethasone group were lower than model group at 1,2 hour(p<0.01).The CD56 expressions of dexamethasone group were higher than model group at 1 hour(p<0.01).The CD106 expressions of dexamethasone group were lower than model group at 1,2,6 hour(p<0.05).The CD54 expressions of Sini decoction group were lower than model group at 1,2(p<0.01),3(p<0.05) hour.The CD56 expressions of Sini decoction group were higher than model group at 3(p<0.05),6(p<0.01) hour.The CD106 expressions of Sini decoction group were lower than model group at 1,2,3(p<0.01),6(p<0.05) hour.
5.No expression of iNOS in normal rat brain.The expression of iNOS in model group was highest at 1 hour,decreased at 2,3 hour and rebounded at 6 hour.The iNOS expressions of dexamethasone group were lower than model group at 1(p<0.01),6(p<0.05) hour.The iNOS expression of Sini decoction group was lower than model group at 6 hour(p<0.01). Concentration of SOD in model group was lower than sham operated group on four time periods (p<0.01) and reached lowest valley at 1 hour.Concentration of SOD of dexamethasone group and Sini decoction group were higher than model group at 1,2 hour(p<0.01).Concentration of MDA in model group was higher than sham operated group on four time periods(p<0.01) and reached peak at 6 hour.It's changes regularities are same as SOD.Concentration of MDA of dexamethasone group was lower than model group at 1,2,3,6 hour(p<0.01).Concentration of MDA of Sini decoction group was lower than model group at 2,3,6 hour(p<0.01).
6.Electron microscopy and light microscopy showed:There was reactive injury at 1 hour, injury restoration at 2,3 hour,serious damage at 6 hour in endotoxic shock rat brain;rat brain damage of dexamethasone group reduced at 1 hour;rat brain damage of Sini decoction group reduced at 6 hour.TUNEL showed:There was a very small number of apoptotic cells in normal rat brain.Apoptotic cells of model group were less than sham operated group and reached peak at
6 hour.Apoptotic cells of dexamethasone group were less than model group atl,2 hour(p<0.01).Apoptotic cells of Sini decoction group were less than model group at2,3, 6hour(p<0.01).
Conclusion
1.There were 91 differentially expressed inflammatory cytokines in rat brain after endotoxic shock.There were 36 inflammatory cytokines involved with inflammatory response among them.TNF-α、IL-1β、IL-10 have been reported more and IL-1α、IL-13、IL-15、I15ra have been reported less.
2.Organ damage induced by two ways in endotoxic shock:①Increasing prereceptor signal substances such as TNF-α,IL-1αinduced by LPS.→The expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,CD54.→Increasing signal substances such as NO production and SOD,MDA content after brain vascular smooth muscle cells,glial cells and nerve cells receiving membrane receptor stimulation.→Rat brain tissue structure damage.②The expression of cell-surface adhesion molecules,CD54,induced by LPS.→CD54 increased content of TNF-α,IL-1α.→Lead to inflammatory mediator waterfall cascade reactions and a large number of mediator was activated.→The expression of many cell-surface adhesion molecules,CD54,CD56,CD106.→Increasing signal substances such as NO production and SOD,MDA content after brain vascular smooth muscle cells and nerve cells receiving membrane receptor stimulation.→Rat brain tissue structure damage.
3.Sini Tang Injection has efficacy at treating shock and multifaceted roles.It can up-regulate blood pressure,SOD content,CD56 expression and down-regulate content of TNF-α, IL-1α,MDA and CD54,CD106,iNOS,apoptotic cells expression.Sini decoction took effect relatively late but lasting stability and dexamethasone relatively early but unstability.
1.明确内毒素休克后脑损伤组织中炎症介质水平,中性粒细胞和脑血管内皮细胞等细胞表面黏附分子表达情况,脑血管平滑肌细胞、神经胶质细胞和神经细胞对细胞外炎症介质刺激产生的功能代谢变化以及结构改变等三者之间的规律性关系。
2.明确内毒素休克脑损伤中起作用的白细胞介素-1α(IL-1α)、肿瘤坏死因子-α(TNF-α)增加等信号异常是否可以上调中性粒细胞和脑血管内皮细胞等细胞表面黏附分子表达,脑血管平滑肌细胞、神经胶质细胞和神经细胞细胞内气体信使一氧化氮(NO)数量的异常等受体后信号转导通路的变化与脑部不同区域血管反应性及神经元损伤修复的关系。
3.明确四逆汤对以上指标有何作用,从而说明回阳救逆法改善感染性休克后脑微循环障碍和神经元损伤的分子细胞机制,发展中医急症理论。
研究方法:
1.以大鼠内毒素休克模型模拟感染性休克,首先运用基因芯片技术筛选出对感染性休克作用明显的受体前信号物质炎症细胞因子。
2.探讨四逆汤方对内毒素引起的受体前信号物质的作用,如组织中炎症介质IL-1α、TNF-α水平的变化。具体方法采用ELISA方法测定大鼠脑组织IL-1α、TNF-α的含量。
3.探讨四逆汤方对IL-1α、TNF-α引起的受体信号物质的作用,如中性粒细胞和脑血管内皮细胞等细胞表面黏附分子如细胞间黏附分子(ICAM-1,CD54)、神经细胞黏附分子(NCAM,CD56)、血管细胞黏附分子(VCAM-1,CD106)表达情况。具体方法采用免疫组化方法测定大鼠脑组织CD54、CD56、CD106分布与数量变化。
4.探讨四逆汤方对脑血管平滑肌细胞、神经胶质细胞和神经细胞接受细胞膜上受体刺激后细胞内信号物质的变化,如脑组织NO产量、超氧化物歧化酶(SOD)与丙二醛(MDA)含量等;用生化法测SOD与MDA含量,免疫组化法测诱导型一氧化氮合酶(iNOS)分布与数量变化。
5.最后探讨四逆汤方对内毒素休克脑损伤组织结构的改善作用。用TUNEL法测大鼠脑组织细胞凋亡,用电镜观察脑组织内各细胞的超微结构,用光镜观察脑组织结构变化。
研究结果:
1.假手术组大鼠平均动脉血压在实验过程于1h开始有轻度下降;在注射内毒素后,模型组从1h开始与假手术组相比有极显著差别(p＜0.01),至6h时血压降至初始值的40.75%。地塞米松组大鼠至6h时血压降至初始值的45.32%,与模型组相比血压有所升高(p＜0.05)。四逆汤组大鼠至6h时血压降至初始值的67.68%,与模型组相比血压升高极显著(p＜0.01)。
2.基因芯片筛查出Ccl2,4,7,20、Cxcl1,2,9,10,11、Ccr2,3,9;TNF-α、IL-1α、IL-1β等在内毒素休克大鼠脑组织出现非常显著的高表达;还有很少报道的IL-13、IL-15、I15ra也有差异表达。其中IL-13于1h无变化,于2h、3h、6h均上调,IL-15于1h、2h均下调,I15ra于1h、2h、3h、6h均下调。
3.内毒素休克大鼠脑组织中TNF-α与IL-1α从1h至6h的浓度变化规律与基因芯片技术检测出的mRNA水平变化规律基本相同。①模型组大鼠脑组织中IL-1α含量在1h较假手术组有所降低(p＜0.01),2h无显著差异(P＞0.05),3h(p＜0.05)、6h(p＜0.01)有所升高,并于6h达峰值。地塞米松组与四逆汤组大鼠脑组织中IL-1α含量在2h、3h、6h均较模型组有所下降(p＜0.01)。②模型组大鼠脑组织中TNF-α含量在四个时间段较假手术组均有所升高(p＜0.01),并于6h达峰值。地塞米松组大鼠脑组织中TNF-α含量在1h、2h、3h、6h均较模型组有所下降(p＜0.01)。四逆汤组大鼠脑组织中TNF-α含量在3h、6h时间段较模型组有所下降(p＜0.01)。
4.正常组、假手术组大鼠脑组织中CD54无表达,CD56、CD106有微弱表达。内毒素休克模型组大鼠脑组织中CD54、CD56、CD106于1h、2h、3h、6h等不同时间段均高于空白对照组与假手术组(p＜0.01),CD54、CD106于1h表达达峰值,CD56于6h表达达峰值。地塞米松组于1h、2h下调CD54的表达(p＜0.01),于1h上调CD56的表达(p＜0.01),于1h、2h、6h下调CD106的表达(p＜0.05)。四逆汤组于1h和2h(p＜0.01)、3h(p＜0.05)均下调CD54的表达,于3h(p＜0.05)、6h(p＜0.01)均上调CD56的表达,于1h、2h和3h(p＜0.01)、6h(p＜0.05)均下调CD106的表达。
5.正常组、假手术组大鼠脑组织iNOS无表达,模型组大鼠脑组织发现iNOS在1h表达量最高,2h、3h有所降低,6h时又有所回升。地塞米松组于1h(p＜0.01)、6h(p＜0.05)均下调iNOS的表达。四逆汤组仅于6h下调iNOS的表达(p＜0.01)。SOD在模型组大鼠脑组织中含量于四个时间段较假手术组均有所降低(p＜0.01),并于1h降至最低。地塞米松组与四逆汤组大鼠脑组织中SOD含量在1h、2h时间段较模型组有所升高(p＜0.01)。MDA在模型组大鼠脑组织中含量于四个时间段较假手术组均有所升高(p＜0.01),并于6h达峰值,其变化规律与SOD基本相同,地塞米松组大鼠脑组织中MDA含量在1h、2h、3h、6h时间段较模型组有所降低(p＜0.01),四逆汤组大鼠脑组织中MDA含量在2h、3h、6h时间段较模型组有所降低(p＜0.01)。
6.电镜与光镜观察结果显示:内毒素休克模型组大鼠于1h有反应性损伤,2h、3h损伤则有所恢复,6h损伤最严重;地塞米松组于1h较模型组损伤减轻;四逆汤组于6h较模型组损伤减轻。TUNEL法检测结果为:正常组与假手术组大鼠脑组织有极少量的凋亡细胞,内毒素休克模型组大鼠脑组织中的凋亡细胞较假手术组明显升高(p＜0.01),并于6h达峰值。地塞米松组于1h、2h较模型组凋亡减少(p＜0.01)。四逆汤组于2h、3h、6h较模型组凋亡减少(p＜0.01)。
研究结论:
1.内毒素休克后大鼠脑组织出现91个炎症细胞因子差异表达,其中36种细胞因子参与炎症应答,有较多报道的TNF-α、IL-1β、IL-10,还有很少报道的IL-1α、IL-13、IL-15、I15ra。
2.内毒素休克可以通过两条途径造成器官损伤:①LPS引起的受体前信号物质的作用,如组织中炎症介质TNF-α、IL-1α水平的升高→TNF-α、IL-1α引起的受体信号物质如中性粒细胞、脑血管内皮细胞等细胞表面黏附分子CD54表达→脑血管内皮细胞、神经胶质细胞和神经细胞接受细胞膜上受体刺激后细胞内信号物质的NO、MDA增加→脑组织结构损伤。②LPS直接引起受体信号物质CD54表达→CD54诱导TNF-α、IL-1α水平的变化→引发炎性介质的瀑布样效应,导致大量介质的激活→引起多种受体信号物质CD54、CD56、CD106表达→脑血管内皮细胞、神经胶质细胞和神经细胞接受细胞膜上受体刺激后细胞内信号物质的NO、MDA增加→脑组织结构损伤。
3.四逆注射液抢救休克是确有疗效的,而且其抗休克作用是多方面的,既可以升高血压、脑组织SOD含量、CD56的表达,又可以降低TNF-α、IL-1α、MDA的含量与CD54、CD106、iNOS、凋亡细胞的表达。但四逆汤的起效时间比较晚,作用效果稳定持久;地塞米松的起效时间比较早,但是作用效果不持久。
Septic shock is the widespread and complex problem concerned in modern Critical Care Medicine.It becomes the prominent issue to seriously impact on prognosis of patients and further improvement of successful treatment rate.One group of abroad clinical epidemiological research papers indicated that septic shock occurred at a rate of 4%among 2527 cases of patients with systemic inflammatory response,and 46%of septic shock patients were died.A study in China showed that 40%patients whose body surface areas are burned more than 30 percent had concurrently septic shock.Study on critically ill patients also showed that incidence of septic shock was 15.7%,61.1%of septic shock further developed to multiple organ dysfunction syndrome(MODS),fatality rate was 30.6%.Based on above studies,it has very important theoretical value and clinical significance to reveal septic shock pathogenesis and explore clinical prevention and treatment methods.When simplex western medicine treatment hardly makes progress in effect of septic shock,it appears urgently to look for Western & Chinese Traditional Medicine method to treat septic shock.Many medical scholars hope to find a breakthrough for septic shock with Chinese traditional medicine treatment.According to clinical manifestations, the septic shock is the scope of syncope.As most clinical manifestations are also collapse,it's also named as syncope and collapse syndrome.The development of traditional Chinese medicine in thousands of years was a research on septic shock and accumulated much experience,but the treatment is lack of evidence because of the unclear mechanism of septic shock.Much progress has been made in recent years through studying septic shock,however brain injury is rarely involved.This study takes the models of endotoxin-shock rats as research object to simulate septic shock.The study aims are to explore anomalous signal transduction in brain injury caused by septic shock and the effect of Sini Decoction in the process;define the molecular and cellular mechanisms of reviving yang for resuscitation method used to prevent and treat brain injury in septic shock and establish the scientific foundation for modernizing traditional Chinese medicine.
Objective
1.Clear level of inflammatory mediators,the expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,the changes of function metabolism and structural of brain vascular smooth muscle cells,glial cells,nerve cells stimulated by the extracellular inflammatory mediator of brain injury in septic shock.Then sum up regular relationship among them.
2.Clear if the role of increasing of IL-1α、TNF-αcan upregulate the expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,the relationship between the abnormal quantity of gas messenger-NO in brain vascular smooth muscle cells,glial cells and nerve cells and different brain regions vascular response and neuronal damage and repair.
3.Clear how Sini decoction act on above indicators in order to descript the method of reviving yang for resuscitation how improve molecular mechanisms of brain microcirculatory disturbances and neuronal damage.Develop Chinese medicine A&E theory.
Methods
1.This study takes the models of endotoxin-shock rats simulating septic shock for research object.First using technology of gene chip to screen prereceptor signal substances, inflammatory cytokines,which have an obvious role in septic shock.
2.To explore the role of Sini decoction on prereceptor signal substances induced by septic shock.Such as levels of tissue inflammatory mediators,IL-1α、TNF-α.The specific methods uses ELISA to measure IL-1α、TNF-αcontent in septic rat brain tissue.
3.To explore how Sini decoction act on receptor signal substances induced by increasing IL-1α、TNF-αcontent.Such as the expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,ICAM-1(CD54)、NCAM(CD56)、VCAM-1(CD106). The specific methods uses SABC to determine distribution and quantity of ICAM-1(CD54)、NCAM(CD56)、VCAM-1(CD106) in rat brain tissue.
4.To explore how Sini decoction act on the changes of signal substances after brain vascular smooth muscle cells,glial cells and nerve cells receiving membrane receptor stimulation. Such as NO production and SOD,MDA content in rat brain.The specific methods is to use biochemical methods to determine SOD,MDA content and SABC to determine distribution and quantity of iNOS rat brain tissue.
5.At last explore how Sini decoction improve tissue structure after septic brain injury. Using TUNEL to determine apoptosis in rat brain,ultrastructural changes of the cells with electron microscopy,structural changes of brain tissue with microscopy.
Results
1.Mean arterial blood pressure of sham-operated rats started to decline slightly at 1 hour. After i.v.injection of lipopolysaccharide,significant difference between model group and sham operated group at 1 hour(p<0.01),blood pressure fell to 40.75%of the initial values at 6 hour. Blood pressure of dexamethasone group fell to 45.32%of the initial values at 6 hour and had higher blood pressure compared with model group(p<0.05).Blood pressure of Sini decoction group fell to 67.68%of the initial values at 6 hour had higher blood pressure compared with model group(p<0.01).
2.Screening Ccl2,4,7,20、Cxcll,2,9,10,11、Ccr2,3,9;TNF-α、IL-1α、IL-1βwith very significant high expression in endotoxic rat brain.And some inflammatory cytokines such asIL-13、IL- 15、I15ra which were very few reports had differentially expressed. IL-13 has no changes at 1 hour,up-regulation at2,3,6 hour;IL-15 has down-regulation at 1,2 hour and I15ra has down-regulation at 1,2,3,6 hour.
3.Concentration changes of TNF-α、IL-1αin endotoxic rat brain were same as mRNA content changes detected by gene chip technology from 1 hour to 6hour.①Concentration of TNF-αin model group were higher than sham operated group on four time periods(p<0.01) and reached peak at 6 hour.Concentration of TNF-αof dexamethasone group were lower than model group from 1 hour to 6 hour(p<0.01).Concentration of TNF-αof Sini decoction group were lower than model group at 3,6 hour(p<0.01).②Concentration of IL-1αin model group were lower than sham operated group at 1 hour(p<0.01),no significant difference at 2 hour(P>0.05),higher at 3(p<0.05),6(p<0.01)hour and reached peak at 6 hour.Concentration of IL-1αof dexamethasone group were lower than model group from 1 hour to 6hour(p<0.01). Concentration of IL-1αof Sini decoction group were lower than model group at 2,3,6 hour(p<0.01).
4.No expression of CD54 and weak expression of CD56,CD106 in normal rat brain.The expression of CD54,CD56,CD106 in model group was higher than sham operated group and control group on four time periods(p<0.01).CD54,CD106 reached peak at 1 hour and CD56 reached peak at 6hour.The CD54 expressions of dexamethasone group were lower than model group at 1,2 hour(p<0.01).The CD56 expressions of dexamethasone group were higher than model group at 1 hour(p<0.01).The CD106 expressions of dexamethasone group were lower than model group at 1,2,6 hour(p<0.05).The CD54 expressions of Sini decoction group were lower than model group at 1,2(p<0.01),3(p<0.05) hour.The CD56 expressions of Sini decoction group were higher than model group at 3(p<0.05),6(p<0.01) hour.The CD106 expressions of Sini decoction group were lower than model group at 1,2,3(p<0.01),6(p<0.05) hour.
5.No expression of iNOS in normal rat brain.The expression of iNOS in model group was highest at 1 hour,decreased at 2,3 hour and rebounded at 6 hour.The iNOS expressions of dexamethasone group were lower than model group at 1(p<0.01),6(p<0.05) hour.The iNOS expression of Sini decoction group was lower than model group at 6 hour(p<0.01). Concentration of SOD in model group was lower than sham operated group on four time periods (p<0.01) and reached lowest valley at 1 hour.Concentration of SOD of dexamethasone group and Sini decoction group were higher than model group at 1,2 hour(p<0.01).Concentration of MDA in model group was higher than sham operated group on four time periods(p<0.01) and reached peak at 6 hour.It's changes regularities are same as SOD.Concentration of MDA of dexamethasone group was lower than model group at 1,2,3,6 hour(p<0.01).Concentration of MDA of Sini decoction group was lower than model group at 2,3,6 hour(p<0.01).
6.Electron microscopy and light microscopy showed:There was reactive injury at 1 hour, injury restoration at 2,3 hour,serious damage at 6 hour in endotoxic shock rat brain;rat brain damage of dexamethasone group reduced at 1 hour;rat brain damage of Sini decoction group reduced at 6 hour.TUNEL showed:There was a very small number of apoptotic cells in normal rat brain.Apoptotic cells of model group were less than sham operated group and reached peak at
6 hour.Apoptotic cells of dexamethasone group were less than model group atl,2 hour(p<0.01).Apoptotic cells of Sini decoction group were less than model group at2,3, 6hour(p<0.01).
Conclusion
1.There were 91 differentially expressed inflammatory cytokines in rat brain after endotoxic shock.There were 36 inflammatory cytokines involved with inflammatory response among them.TNF-α、IL-1β、IL-10 have been reported more and IL-1α、IL-13、IL-15、I15ra have been reported less.
2.Organ damage induced by two ways in endotoxic shock:①Increasing prereceptor signal substances such as TNF-α,IL-1αinduced by LPS.→The expression of cell-surface adhesion molecules of neutrophil and brain vascular endothelial cells,CD54.→Increasing signal substances such as NO production and SOD,MDA content after brain vascular smooth muscle cells,glial cells and nerve cells receiving membrane receptor stimulation.→Rat brain tissue structure damage.②The expression of cell-surface adhesion molecules,CD54,induced by LPS.→CD54 increased content of TNF-α,IL-1α.→Lead to inflammatory mediator waterfall cascade reactions and a large number of mediator was activated.→The expression of many cell-surface adhesion molecules,CD54,CD56,CD106.→Increasing signal substances such as NO production and SOD,MDA content after brain vascular smooth muscle cells and nerve cells receiving membrane receptor stimulation.→Rat brain tissue structure damage.
3.Sini Tang Injection has efficacy at treating shock and multifaceted roles.It can up-regulate blood pressure,SOD content,CD56 expression and down-regulate content of TNF-α, IL-1α,MDA and CD54,CD106,iNOS,apoptotic cells expression.Sini decoction took effect relatively late but lasting stability and dexamethasone relatively early but unstability.
引文
[1]黄帝内经.上海科学技术出版社,1959,337-342.
[2]难经.人民卫生出版社,1979,52.
[3]郭蔼春,张海玲.伤寒论校注语译.天津科学技术出版社,1996,216.
[4]李梃.医学入门.中国中医药出版社,1995,399-401.
[5]张介宾.景岳全书.山西科学技术出版社,2006,123-127.
[6]林珮琴.类证治裁.人民卫生出版社,1988,291-293.
[7]李中梓.医宗必读.上海科学技术出版社,1987,41-42.
[8]张锡纯.医学衷中参西录.河北科学技术出版社,1980,173-174.
[9]龚廷贤.万病回春.中国中医药出版社,1998,232.
[10]程杏轩.医述.安徽科学技术出版社,1959,672-676.
[11]马宏博,姜良铎.论攻补兼施治疗感染性休克.中华中医药杂志,2006,21(9):548.
[12]巢元方.诸病源候论.人民卫生出版社,1991,384-385.
[13]刘清泉.对脓毒症中医病机特点及治法的认识.北京中医,2007,26(4):198.
[14]周学平.厥脱证病机特征探析.中国中医急症,1994,3(3):129.
[15]皮持衡.中医内科急症手册.上海中医药大学出版社,1995,25.
[16]姜良铎.中医急诊学.中国中医药出版社,2003:18.
[17]郎继孝,李松林,唐由君.赵炳南益气养阴法治疗败血症探究.辽宁中医杂志,1997,24(2):541.
[18]马超英,耿耘,彭仁才,等.牛珀至宝丹拮抗内毒素的实验研究.中国危重病急救医学,1999,11(9):559-560.
[19]王左,方正龙,郑舜华.回阳救逆法治疗邪毒内陷所致厥脱证.中成药,1994,(1):28.
[20]万兰清,马超英,耿耘,等.开闭固脱法治疗流行性出血热休克100例临床研究.中西医结合实用临床急救学,1996,3(4):151.
[21]金妙文,周仲英,张世玮,等.理气活血开闭固脱法治疗感染性休克的研究.中国中西医结合杂志,1995,15(10):589-591.
[22]许锋,鲁焕章,吴咸中.脓毒性休克治疗研究进展.中国危重病急救医学,1996,8(3):187-189.
[23]胡德耀.抗休克药物研究状况及其展望.中国病理生理杂志,1998,14(3):329-330.
[24]冯栋年,娄彬,曹苏生.中西医结合治疗感染性休克42例临床观察.南京中医药大学学报,1995,11(6):17-18.
[25]付亚雷,谷明志.中西医结合治疗感染性休克30例临床观察.湖南中医药导报,1995,15(2):27.
[26]朱虹江.陈乔林谈中西医互补救治感染性休克.云南中医中药杂志,1998,19(2):6-8.
[27]杨光,李鸣真,叶望云,等.内毒素诱导肝损害家兔血浆和肝组织中肿瘤坏死因子-α白细胞介素-8水平变化及热毒清的保肝作用.中国危重病急救医学,1997,9(3):133-135.
[28]杨明炜,李鸣真,叶望云,等.热毒清的拆方研究体内及体外对内毒素的影响.同济医科大学学报,1996,25(5):392-394.
[29]邓文龙,徐嘉红,王文烈,等.热毒平的抗内毒素作用研究.中药药理与临床,1995,5(2):16-19.
[30]余林中,吴锐,黄泳,等.凉膈散对家兔内毒素温病模型的解毒作用研究.中药药理与临床,1996,(5):46.
[31]谢恬,凌一揆.清温败毒饮对内毒素诱发家兔温病气血两燔证的疗效和机理.中国中西医结合杂志,1993,13(2):94-97.
[32]杨明炜,李明真,叶望云,等.金叶败毒冲剂抗内毒素作用研究.中药药理与临床,1996,6(2):27-29.
[33]耿耘,马超英,肖诚,等.牛珀至宝丹对内毒素休克大鼠血液流变学的影响.中国危重病急救医学,1997,9(12):715-717.
[34]杜少辉,周大桥,张悦,等.牛珀至宝微丸对内毒素休克时蛋白激酶C调控一氧化氮生成的影响.中国中西医结合急救杂志,2002,9(4):189-193.
[35]王今达,雪琳.细菌、内毒素、炎性介质并治——治疗重症脓毒病的新对策.中国危重病急救医学,1998,10(6):323-325.
[36]曹书华,王今达.血必净对感染性多器官功能障碍综合征大鼠组织及内皮损伤保护作用的研究.中国危重病急救医学,2002,14(8):489-491.
[37]张爱民,黄启福.心脉灵注射液对内毒素休克狗红细胞流变学紊乱的影响.北京中医药大学学报,1994,17(2):5761.
[38]卫洪昌,张陈福.人参附子青皮对内毒素性休克的实验研究概况.上海中医药杂志,1989,(7):44.
[39]杨宁,赵诗云,蔡秉仁,等.逐瘀通腑合剂抗内毒素所致肠源性感染的实验研究.中国中西医结合杂志,1997,17(8):484.
[40]沈洪,周仲英,金妙文.凉血化瘀法对内毒素性DIC家兔血液流变学的影响.南京中医学院学报,1994,10(5):13-15.
[41]邓文龙.中医解毒实质的研究及内毒素性疾病的中医药防治.中药药理与临床,1992,5(4):40.
[42]曾祥发,陆辉,谢林,等.参麦注射液对大鼠伤寒内毒素脑水肿的作用观察.广西中医药,1989,13(1):44.
[43]方剑乔,高镇五,林咸明,等.不同腧穴针刺对内毒素致热家兔模型的影响.中国医药学报,1996,6(6):39.
[44]王宇华,杨长森.艾炷灸对大肠杆菌内毒素致热家兔体温及微循环的影响.中西医结合杂志,1988,16(9):541.
[45]黄为敏,黄坤厚,徐青燕等.电针对内毒素休克大鼠肝脏的影响(组织化学观察).针刺研究,1995,20(3):36.
[46]章韵.感染性休克的中医研究进展[J].陕西中医,2004,25(12):1150.
[1]Cavaillon M,Adib-Conquy M,Fitting C,et al.Cytokine cascade in sepsis[J].Scand J Infect Dis,2003,35(2):535-544.
[2]Ji XH,Sun KY,Feng YH,et al.Changes of inflammation-associated cytokine expressions during early phase of experimental endotoxic shock in macaques[J].World J Gastroentero,2004,10(6):3026-3301.
[3]Youn HS,Lee JY,Fitzgeiald KA,et al.Specific inhibition of MyD88-Independent Signaling pathways of TLR3 and TLR4 by Resveratrol Molecular Targets Are TBK1 and RIP1 in TRIF complex[J].J Immunol,2005,175(5):3339-3346.
[4]Rao N,Nguyen S,Ngo K,et al.A novel splice variant of interleukin-1 receptor(IL-1R)associated kinase 1 plays a negative regulatory role in toll/IL-1R-induced inflammatory signaling[J].Mal Cell Biol,2005,25(15):6521-6532.
[5]Shishcdin S,Koul D,Aggamal BB.Cycbooxygenase(COX)-2 inhibitor celecoxib sbrogates TNF-induced NF-kappa B activation through inhibitor of activation of 1 kappa B alpha kinase and Akt in human an non-small cell lung carcinoms.correlation with suppression of COX-2 synthesis[J].J Immunol 2004,173(3):2011-2022.
[6]Yoo NJ,Park WS,Kim SY,et al.Nod1,a CARD protein,enhances pro-interleukin-1beta processing through the interaction with pro-caspase-1[J].Biochem Biophys Res Commun,2002,299(4):652-658.
[7]Worrall NK,Chang K,LeJeune WS,et al.TNF-alpha causes reversible in vivo systemic vascular barrier dysfunction via NO-dependent and independent mechanisms[J].Am J Physiol,1997,273(6):22565-22574.
[8]Gornikiewicz A,Sautner T,Brostjan C,et al.Catecholamines up-regulate lipopolysaccharide induced IL-6 production in human microvascula endothelial cells[J].FASEB J,2000,14(9):10932-1100.
[9]李琦,钱桂生,张青,等.内毒素致伤大鼠白介素10表达及肺组织激活蛋白1活性的变化[J].中国危重病急救医学,2002,14(3):279-281.
[10]Blackwell TS,Christman JW.Sepsis and cytokines:current status[J].Br J Anaesth,1996,77(4):110-117.
[11]陈永华,蒋建新,谢国旗,等.内毒素血症时小鼠肺局部致炎及抗炎因子与急性肺损伤的关系[J].中国危重病急救医学,2000,12(6):331-333.
[12]Dinarello CA.Proinflammatory and anti-inflammatory cytokines as mediators in the pathgenesis of septic shock[J].Chest 1997,112(7):321-329.
[13]Worrall NK,Chang K,Lejeune WS,et al.TNF-alpha causes reversible in vivo systemic vascular barrier dysfunction via NO-dependent and independent mechanism[J].Am J Physial,1997,273(6):2565-2574.
[14]Yao YM,Redl H,Bahrami S,etal.The inflammatory basis of trauma/shock-associated multiple organ failure[J].Inflamm Res,1998,47(2):79.
[15]Amoit F,Fitting C,Tracey KJ,et al.Lipopolysaccharide-induced cytokine cascade and lethality in LT alpha/TNF alpha-deficient mice[J].Mol Med,1997,3(12):864-875.
[16]Murphey ED,Traber DL.Pretreatment with tumor necrosis factor-alpha attenuates arterial hypotension and mortality induced by endotox in pigs[J].Crit Care Med,2000,28(6):2015-2021.
[17]Wisnoski N,Chung CS,Chen Y,et al.The contribution of CD4+ CD25+ T-regulatory-cells to immune suppression in sepsis[J].Shock,2007,27(3):251-257.
[18]COHEN J.Recent developments in the identification of novel therapeutic targets for the treatment of patients with sepsis and septic shock[J].Scand J Infect Dis,2003,35(9):690-696.
[19]阎敬.多脏器功能障碍综合征研究进展.临床肺科杂志,2001,6(3):48-50.
[20]朱铁明,彭承宏,周君富,等1SIRS病人血浆自由基反应及其临床意义.中国实用外科杂志,2000,12(20):721-723.
[21]夏斌,邵福源.CD11/CD18在脑缺血/再灌注损伤中的作用[M].国外医学脑血管疾病分册,2000,8(3):134.
[22]Carrithers M,Tandon S,Canosa S,et al.Enhanced Susceptibility to Endotoxic Shock and Impaired STAT3 Signaling in CD31-Deficient Mice.Am J Pathol,2005,166(1):185-196.
[23]Yan WS,Kan WH,Hang QB,et al.Expression of intercellular adhesion molecule-1 in different organs of the mice with endotoxic shock induced by lipopolysaccharide.Sheng Li Xue Bao,2002,54(1):71-74.
[24]Weigand MA,Schmidt H,Pouimahmoud M,et al.Circulating intercellular adhesion molecule-1 as an carly predictor of hepatic failure in patients with septic shock.Crit Care Med,1999,27(12):2656-2661.
[25]章韵.感染性休克的中医研究进展[J].陕西中医,2004,25(12):1150.
[1]黄帝内经.上海科学技术出版社,1959.
[2]难经.人民卫生出版社,1979,52.
[3]郭蔼春,张海玲.伤寒论校注语译.天津科学技术出版社,1996,216.
[4]李梃.医学入门.中国中医药出版社,1995,399-401.
[5]张介宾.景岳全书.山西科学技术出版社,2006,123-127.
[6]林珮琴.类证治裁.人民卫生出版社,1988,291-293.
[7]李中梓.医宗必读.上海科学技术出版社,1987,41-42.
[8]张锡纯.医学衷中参西录.河北科学技术出版社,1980,173-174.
[9]龚廷贤.万病回春.中国中医药出版社,1998,232.
[10]程杏轩.医述.安徽科学技术出版社,1959,672-676.
[11]华佗.中藏经.学苑出版社,1995,453.
[12]张景岳.质疑录.江苏科学技术出版社,1981,13.
[13]王新华.中医基础理论.人民卫生出版社,2001,343.
[14]刘清泉,蓝海涛,赵红芳等.《重度脓毒症中医证候与APACHE Ⅱ评分相关性的临床研究》[J].北京中医,2007,26(4):208-210.
[15]刘清泉、陈志刚.麻黄汤类方治疗急性呼吸道感染并发SIRS的临床研究.中国中医急诊,2006,15(6):565-567.
[16]姜良铎.中医急诊学.中国中医药出版社,2003:18.
[17]马超英,耿耘,彭仁才,等.牛珀至宝丹拮抗内毒素的实验研究.中国危重病急救医学,1999,11(9):559-560.
[18]成无己.注解伤寒论.人民卫生出版社,2005,145.
[19]倪朱谟.本草汇言.上海科学技术出版社,2005,333.
[20]田建华.图解本草纲目.河北科学技术出版社,2007,127.
[21]汪昂.本草备要.天津科学技术出版社,1993,8.
[22]叶成炳,王明杰.伤寒明理论阐释.四川科学技术出版社,1988,248.
[23]黄竹齐.伤寒论集注.人民卫生出版社,1957,333.
[24]汪昂.医方集解.人民卫生出版社,2007,230-232.
[25]张璐.千金方衍义.中国中医药出版社,1995,440.
[26]吴谦.医宗金鉴.人民卫生出版社,2006:699.
[27]成无己.注解伤寒论.北京:人民卫生出版社,1978:168.
[28]陶节庵.伤寒六书.北京:人民卫生出版社,1990:36.
[29]柯琴.伤寒来苏集(卷四).上海:上海卫生出版社,1956:25-26.
[30]成都中医学院伤寒教研组.伤寒论讲义[M].北京:人民卫生出版社,1960:182-183.
[31]熊曼琪.伤寒学.北京:中国中医药出版社,2003:308.
[32]张志民,徐柏英.对张仲景用附子的研究[J].上海中医药杂志,1957(6):32.
[33]潘澄濂.附子在《伤寒杂病论》诸方配伍中的作用探讨[J].浙江中医学院学报,1990,14(6):4.
[34]陈明,张印生等.伤寒名医验案精选.学苑出版社,1998:405-406.
[35]黄宫绣.本草求真.中国中医药出版社,1997:109.
[36]段富津.方剂学[M].上海:上海科学技术出版社,1995:104.
[37]刘渡舟.新编伤寒论类方.山西人民出版社,1984:183.
[1]姚咏明,盛志勇.脓毒症研究的若干新动态.中国危重病急救医学[J],2000,12(6):323-325.
[2]Fink MP,Heard SO.Laboratory models of sepsis and septic shock[J].J Surg Res,1990,49(2):186-196.
[1]Cavaillon M,Adib-Conquy M,Fitting C,et al.Cytokine cascade in sepsis[J].Scand J Infect D is,2003,35(5):35-44.
[2]Ji XH,Sun KY,Feng YH,et al.Changes of inflammation-associated cytokine expressions during early phase of experimental endotoxic shock in macaques[J].World J Gastroentero,2004,10(4):3026-3301.
[3]Kumar A,Haery C,Parillo JE.Myocardial dysfunction in septic shock[J].Crit Care Clin,2000,16(2):251-287.
[4]Andrew D Luster.The role of chemokines in linking innate and adaptive immunity[J].Current Opinion in Immunology 2002,14(2):129-135.
[1]Blackwell TS,Christman JW.Sepsis and cytokines current status[J].Br J Anaesth,1996,77(2):110-117.
[2]陈永华,蒋建新,谢国旗,等.内毒素血症时小鼠肺局部敛炎及抗炎因子与急性肺损伤的关系[J].中国危重病急救医学,2000,12(4):331-333.
[3]Fink MP,Heard SO.Laboratory models of sepsis and septic shock[J].J Surg Res.1990,49(2):186-196.
[4]魏微,张微微,李远征.大鼠脑缺血冉灌注后肿瘤坏死因子及基质金属蛋白-9的表达[J].中华老年心脑血管病杂志,2004,6(6):406-408.
[5]Dinarello CA.Interleukin-1 and Interleukin-1 antagonism.Blood,1991,77(8):1627-1652.
[6]Heumann D,Roy DL,Zanetti G,et al.Contribution of TNF/TNFreceptor and of toxicity in murine models of endotoxemia and bacterial peritonitis.J Inflamm,1997,47(1):173-179.
[7]Amiot F,Fitting C,Tracey KJ,et al.Lipopolysaccharide-induced cyto-Kine cascade and lethality in LT alpha/TNF alpha-deficient mice[J].Mo Med,1997,3(12):864-875.
[8]Fujimura M,Cache Y.Early appearance of activated matrix metalloprotein ase-9 and blood brain barrier disruption in mice after focal cerebral ischemia and reperfusion [J].Brain Res,1999,842:92-100
[1]余惠平,张和平,田进军,等.ICAM-1在人脑胶质瘤的表达及临床意义[J].中国肿瘤临床与康复,2008,15(3):204-206.
[2]Zhang RL,Chopp M,Zaloga C,et al.The temporal profiles of ICAM-1 protein and mRNA expression after transientMCA occlusion in the rat[J].Brain Res,1995,682(1):182-186.
[3]郑绪鹏,刘班,骆平,等.大鼠脑缺血再灌注后NCAM基因表达的变化[J].青岛大学医学院学报,2006,42(2):140-144.
[4]Ning X Z,Liu Q,Zhao H.Nimodip ine for treatment of perifocal edema follwing asp iration and drainage in patients with cerebra lhemorrhage[J].Neur Regenerat Res,2007,2(5):310-313.
[5]WangW,Sun Y J,Xi T,et al.Effect of 1400W on neurogenesis after transient focal cerebral ischemia in the adult rat hippocampus[J].Chin Pharm acol Bull,2004,20(1):33-35.
[6]Memezawa H,Minamisawa H,SmithM L,et al.Ischemic penumbra in a model of reversible middle cerebral artery occlusion in the rat[J].Exp B rain Res,1992,89(1):67-78.
[7]Michael PB,Richard MN.Endothelial-leukocyte adhesion molecules in inflammation and etastasis[J].Thrombosis and Haemostasis,1993,70(1):152.
[8]Fukuda T.Role of interleukin-4 and vascular cell adhesion molecule-1 in selectiveeosinophil migration into the airways in allergic asthma[J].Am J Respir Cell Mol Biol,1996,14(1):84.
[1]Lehotsky J,Kap lan P,MatejovicovaM,et al.Iontransport systems as targets of free radicals during ischemia reperfusion injury[J].Gen Physiol B iophys,2002,21(1):31-37.
[2]Ravati A,Ahlemeyer B,Becker A,et al.Preconditioning-induced neurop rotection is mediated by reactive oxygen species and activation ofthe transcription factor nuclear factor-kappaB[J].Neurochem,2001,78(4):909-919.
[3]张敬伟.NO,SOD与缺血性脑血管病的关系[J].中国误诊学杂志,2005,5(5):9001.
[4]Tominaga T,Sato S,Ohnishi T et al.Potentiation of nitric oxide Formation following bilateral carotid occlusion and focal cerebral ischemia in the rat:in vivo detection of the nitric oxide radical by electron paramagnetic resonance spin trapping.Brain Res,1993,614(1-2):342-346.
[5]Nathan C,Xie Q W.Nitric oxide synthase:roles,tolls and controls.Cell,1994,7(8):915.
[6]Beray-Berthat V,Palmier B,Plotkine M et al.Neutrophils do not contribute to infarction,oxidative stress,and NO synthase activity in severe brain ischemia.Exp Neurol,2003,182(2):446-454.
[7]Franklin A,Parmentier-Batteur S,Walter L et al.Palmitoy-leth anolamide increases after focal cerebral ischemia and potentiates microglial cell motility.J Neuro Sci,2003,23(21):7767-7715.
[8]Takeyama Y.Significance of apop totic cell death in systemiccomp lications with severe acute pancreatitis[J].J Gastroen2 terol,2005,40(1):1210.
[1]Meyer R,Weissert R,Diem R,et al.Acute neuronal apoptosis in a rat model of multiple sclerosis[J].Neurosci,2001,21(16):6214-20.
[1]乔胃娟,冯全生.感染性休克动物模型评述[J].中外健康文摘,2008,4(2):23-24.
[2]张爱华,蔡毅.白细胞介素-13与肾脏疾病[J].国外医学:泌尿系统分册,1998,18(7):118-121.
[3]王大东,朱瑛梅,李秀智,等.白细胞介素-13功能研究进展[J].医学研究生报,2002,15(6):544-546.
[4]李鸿钧,苏婷.白细胞介素-13的研究进展[J].国外医学:临床生物化学与检验学分册,2001,22(4):187-188.
[5]Matsumto K.Interleukin-13 inhibits cytokine secretion by blood onocytes from patient s wit h IgA nephropat hy[J].Nephron,1997,75(3):295-302.
[6]Saura M,Martinezd R,Minty S,et al.Interleukin-13 inhibits inducible nit ric oxide synt hetase expression in human mesangial cells[J].Biochem J,1996,31(3):641-646.
[7]Gonz(?)lez Alvaro I,Domnguez Jim nez C,Ortiz AM,et al.Interleukin-15 and interfereon-γparticipate in the cross-talk between nat ural killer and monocytic cells required for turnout necrosis factor production[J].Art hritis Res Ther,2006,8(4):88.
[8]Dinarello CA.Proinflammatory and anti-inflammatory cytokines as mediators in the pathogenesis of septic shock[J].Chest,1997,11(2):321-329.
[9]Fukuda T.Role of interleukin-4 and vascular cell adhesion molecule-1 in selective eosinophil migration into the airways in allergic asthma[J].Am J Respir Cell Mol Biol,1996,14(1):84.
[10]El Azami M,Bauche J,Loucka R,Osicka,et al.Interaction of Bordetella adenylate cyclase with CD11b/CD18:role of toxin acylation and identification of the integrin interaction domain[J].J Biol Chem,2003,278(3):38514-38521.
[11]马宏博.论排毒解毒与扶正相结合治疗感染性休克.中国中医急症,2003,12(5):437-438.
[12]肖凤霞.四逆汤的药理研究进展.中医药研究,1999,15(4):60-62.
[13]陈信义,李峨,侯丽,等.乌头类生物碱研究进展与应用前景评述,中国中医药信息杂志,2004,11(10):922-923.
[14]邵春红,王晓良.四逆汤对失血性休克大鼠心功能和血压调节的肾上腺素受体机制研究.中国药学杂志,2003,38(11):847-850.
[2]难经.人民卫生出版社,1979,52.
[3]郭蔼春,张海玲.伤寒论校注语译.天津科学技术出版社,1996,216.
[4]李梃.医学入门.中国中医药出版社,1995,399-401.
[5]张介宾.景岳全书.山西科学技术出版社,2006,123-127.
[6]林珮琴.类证治裁.人民卫生出版社,1988,291-293.
[7]李中梓.医宗必读.上海科学技术出版社,1987,41-42.
[8]张锡纯.医学衷中参西录.河北科学技术出版社,1980,173-174.
[9]龚廷贤.万病回春.中国中医药出版社,1998,232.
[10]程杏轩.医述.安徽科学技术出版社,1959,672-676.
[11]马宏博,姜良铎.论攻补兼施治疗感染性休克.中华中医药杂志,2006,21(9):548.
[12]巢元方.诸病源候论.人民卫生出版社,1991,384-385.
[13]刘清泉.对脓毒症中医病机特点及治法的认识.北京中医,2007,26(4):198.
[14]周学平.厥脱证病机特征探析.中国中医急症,1994,3(3):129.
[15]皮持衡.中医内科急症手册.上海中医药大学出版社,1995,25.
[16]姜良铎.中医急诊学.中国中医药出版社,2003:18.
[17]郎继孝,李松林,唐由君.赵炳南益气养阴法治疗败血症探究.辽宁中医杂志,1997,24(2):541.
[18]马超英,耿耘,彭仁才,等.牛珀至宝丹拮抗内毒素的实验研究.中国危重病急救医学,1999,11(9):559-560.
[19]王左,方正龙,郑舜华.回阳救逆法治疗邪毒内陷所致厥脱证.中成药,1994,(1):28.
[20]万兰清,马超英,耿耘,等.开闭固脱法治疗流行性出血热休克100例临床研究.中西医结合实用临床急救学,1996,3(4):151.
[21]金妙文,周仲英,张世玮,等.理气活血开闭固脱法治疗感染性休克的研究.中国中西医结合杂志,1995,15(10):589-591.
[22]许锋,鲁焕章,吴咸中.脓毒性休克治疗研究进展.中国危重病急救医学,1996,8(3):187-189.
[23]胡德耀.抗休克药物研究状况及其展望.中国病理生理杂志,1998,14(3):329-330.
[24]冯栋年,娄彬,曹苏生.中西医结合治疗感染性休克42例临床观察.南京中医药大学学报,1995,11(6):17-18.
[25]付亚雷,谷明志.中西医结合治疗感染性休克30例临床观察.湖南中医药导报,1995,15(2):27.
[26]朱虹江.陈乔林谈中西医互补救治感染性休克.云南中医中药杂志,1998,19(2):6-8.
[27]杨光,李鸣真,叶望云,等.内毒素诱导肝损害家兔血浆和肝组织中肿瘤坏死因子-α白细胞介素-8水平变化及热毒清的保肝作用.中国危重病急救医学,1997,9(3):133-135.
[28]杨明炜,李鸣真,叶望云,等.热毒清的拆方研究体内及体外对内毒素的影响.同济医科大学学报,1996,25(5):392-394.
[29]邓文龙,徐嘉红,王文烈,等.热毒平的抗内毒素作用研究.中药药理与临床,1995,5(2):16-19.
[30]余林中,吴锐,黄泳,等.凉膈散对家兔内毒素温病模型的解毒作用研究.中药药理与临床,1996,(5):46.
[31]谢恬,凌一揆.清温败毒饮对内毒素诱发家兔温病气血两燔证的疗效和机理.中国中西医结合杂志,1993,13(2):94-97.
[32]杨明炜,李明真,叶望云,等.金叶败毒冲剂抗内毒素作用研究.中药药理与临床,1996,6(2):27-29.
[33]耿耘,马超英,肖诚,等.牛珀至宝丹对内毒素休克大鼠血液流变学的影响.中国危重病急救医学,1997,9(12):715-717.
[34]杜少辉,周大桥,张悦,等.牛珀至宝微丸对内毒素休克时蛋白激酶C调控一氧化氮生成的影响.中国中西医结合急救杂志,2002,9(4):189-193.
[35]王今达,雪琳.细菌、内毒素、炎性介质并治——治疗重症脓毒病的新对策.中国危重病急救医学,1998,10(6):323-325.
[36]曹书华,王今达.血必净对感染性多器官功能障碍综合征大鼠组织及内皮损伤保护作用的研究.中国危重病急救医学,2002,14(8):489-491.
[37]张爱民,黄启福.心脉灵注射液对内毒素休克狗红细胞流变学紊乱的影响.北京中医药大学学报,1994,17(2):5761.
[38]卫洪昌,张陈福.人参附子青皮对内毒素性休克的实验研究概况.上海中医药杂志,1989,(7):44.
[39]杨宁,赵诗云,蔡秉仁,等.逐瘀通腑合剂抗内毒素所致肠源性感染的实验研究.中国中西医结合杂志,1997,17(8):484.
[40]沈洪,周仲英,金妙文.凉血化瘀法对内毒素性DIC家兔血液流变学的影响.南京中医学院学报,1994,10(5):13-15.
[41]邓文龙.中医解毒实质的研究及内毒素性疾病的中医药防治.中药药理与临床,1992,5(4):40.
[42]曾祥发,陆辉,谢林,等.参麦注射液对大鼠伤寒内毒素脑水肿的作用观察.广西中医药,1989,13(1):44.
[43]方剑乔,高镇五,林咸明,等.不同腧穴针刺对内毒素致热家兔模型的影响.中国医药学报,1996,6(6):39.
[44]王宇华,杨长森.艾炷灸对大肠杆菌内毒素致热家兔体温及微循环的影响.中西医结合杂志,1988,16(9):541.
[45]黄为敏,黄坤厚,徐青燕等.电针对内毒素休克大鼠肝脏的影响(组织化学观察).针刺研究,1995,20(3):36.
[46]章韵.感染性休克的中医研究进展[J].陕西中医,2004,25(12):1150.
[1]Cavaillon M,Adib-Conquy M,Fitting C,et al.Cytokine cascade in sepsis[J].Scand J Infect Dis,2003,35(2):535-544.
[2]Ji XH,Sun KY,Feng YH,et al.Changes of inflammation-associated cytokine expressions during early phase of experimental endotoxic shock in macaques[J].World J Gastroentero,2004,10(6):3026-3301.
[3]Youn HS,Lee JY,Fitzgeiald KA,et al.Specific inhibition of MyD88-Independent Signaling pathways of TLR3 and TLR4 by Resveratrol Molecular Targets Are TBK1 and RIP1 in TRIF complex[J].J Immunol,2005,175(5):3339-3346.
[4]Rao N,Nguyen S,Ngo K,et al.A novel splice variant of interleukin-1 receptor(IL-1R)associated kinase 1 plays a negative regulatory role in toll/IL-1R-induced inflammatory signaling[J].Mal Cell Biol,2005,25(15):6521-6532.
[5]Shishcdin S,Koul D,Aggamal BB.Cycbooxygenase(COX)-2 inhibitor celecoxib sbrogates TNF-induced NF-kappa B activation through inhibitor of activation of 1 kappa B alpha kinase and Akt in human an non-small cell lung carcinoms.correlation with suppression of COX-2 synthesis[J].J Immunol 2004,173(3):2011-2022.
[6]Yoo NJ,Park WS,Kim SY,et al.Nod1,a CARD protein,enhances pro-interleukin-1beta processing through the interaction with pro-caspase-1[J].Biochem Biophys Res Commun,2002,299(4):652-658.
[7]Worrall NK,Chang K,LeJeune WS,et al.TNF-alpha causes reversible in vivo systemic vascular barrier dysfunction via NO-dependent and independent mechanisms[J].Am J Physiol,1997,273(6):22565-22574.
[8]Gornikiewicz A,Sautner T,Brostjan C,et al.Catecholamines up-regulate lipopolysaccharide induced IL-6 production in human microvascula endothelial cells[J].FASEB J,2000,14(9):10932-1100.
[9]李琦,钱桂生,张青,等.内毒素致伤大鼠白介素10表达及肺组织激活蛋白1活性的变化[J].中国危重病急救医学,2002,14(3):279-281.
[10]Blackwell TS,Christman JW.Sepsis and cytokines:current status[J].Br J Anaesth,1996,77(4):110-117.
[11]陈永华,蒋建新,谢国旗,等.内毒素血症时小鼠肺局部致炎及抗炎因子与急性肺损伤的关系[J].中国危重病急救医学,2000,12(6):331-333.
[12]Dinarello CA.Proinflammatory and anti-inflammatory cytokines as mediators in the pathgenesis of septic shock[J].Chest 1997,112(7):321-329.
[13]Worrall NK,Chang K,Lejeune WS,et al.TNF-alpha causes reversible in vivo systemic vascular barrier dysfunction via NO-dependent and independent mechanism[J].Am J Physial,1997,273(6):2565-2574.
[14]Yao YM,Redl H,Bahrami S,etal.The inflammatory basis of trauma/shock-associated multiple organ failure[J].Inflamm Res,1998,47(2):79.
[15]Amoit F,Fitting C,Tracey KJ,et al.Lipopolysaccharide-induced cytokine cascade and lethality in LT alpha/TNF alpha-deficient mice[J].Mol Med,1997,3(12):864-875.
[16]Murphey ED,Traber DL.Pretreatment with tumor necrosis factor-alpha attenuates arterial hypotension and mortality induced by endotox in pigs[J].Crit Care Med,2000,28(6):2015-2021.
[17]Wisnoski N,Chung CS,Chen Y,et al.The contribution of CD4+ CD25+ T-regulatory-cells to immune suppression in sepsis[J].Shock,2007,27(3):251-257.
[18]COHEN J.Recent developments in the identification of novel therapeutic targets for the treatment of patients with sepsis and septic shock[J].Scand J Infect Dis,2003,35(9):690-696.
[19]阎敬.多脏器功能障碍综合征研究进展.临床肺科杂志,2001,6(3):48-50.
[20]朱铁明,彭承宏,周君富,等1SIRS病人血浆自由基反应及其临床意义.中国实用外科杂志,2000,12(20):721-723.
[21]夏斌,邵福源.CD11/CD18在脑缺血/再灌注损伤中的作用[M].国外医学脑血管疾病分册,2000,8(3):134.
[22]Carrithers M,Tandon S,Canosa S,et al.Enhanced Susceptibility to Endotoxic Shock and Impaired STAT3 Signaling in CD31-Deficient Mice.Am J Pathol,2005,166(1):185-196.
[23]Yan WS,Kan WH,Hang QB,et al.Expression of intercellular adhesion molecule-1 in different organs of the mice with endotoxic shock induced by lipopolysaccharide.Sheng Li Xue Bao,2002,54(1):71-74.
[24]Weigand MA,Schmidt H,Pouimahmoud M,et al.Circulating intercellular adhesion molecule-1 as an carly predictor of hepatic failure in patients with septic shock.Crit Care Med,1999,27(12):2656-2661.
[25]章韵.感染性休克的中医研究进展[J].陕西中医,2004,25(12):1150.
[1]黄帝内经.上海科学技术出版社,1959.
[2]难经.人民卫生出版社,1979,52.
[3]郭蔼春,张海玲.伤寒论校注语译.天津科学技术出版社,1996,216.
[4]李梃.医学入门.中国中医药出版社,1995,399-401.
[5]张介宾.景岳全书.山西科学技术出版社,2006,123-127.
[6]林珮琴.类证治裁.人民卫生出版社,1988,291-293.
[7]李中梓.医宗必读.上海科学技术出版社,1987,41-42.
[8]张锡纯.医学衷中参西录.河北科学技术出版社,1980,173-174.
[9]龚廷贤.万病回春.中国中医药出版社,1998,232.
[10]程杏轩.医述.安徽科学技术出版社,1959,672-676.
[11]华佗.中藏经.学苑出版社,1995,453.
[12]张景岳.质疑录.江苏科学技术出版社,1981,13.
[13]王新华.中医基础理论.人民卫生出版社,2001,343.
[14]刘清泉,蓝海涛,赵红芳等.《重度脓毒症中医证候与APACHE Ⅱ评分相关性的临床研究》[J].北京中医,2007,26(4):208-210.
[15]刘清泉、陈志刚.麻黄汤类方治疗急性呼吸道感染并发SIRS的临床研究.中国中医急诊,2006,15(6):565-567.
[16]姜良铎.中医急诊学.中国中医药出版社,2003:18.
[17]马超英,耿耘,彭仁才,等.牛珀至宝丹拮抗内毒素的实验研究.中国危重病急救医学,1999,11(9):559-560.
[18]成无己.注解伤寒论.人民卫生出版社,2005,145.
[19]倪朱谟.本草汇言.上海科学技术出版社,2005,333.
[20]田建华.图解本草纲目.河北科学技术出版社,2007,127.
[21]汪昂.本草备要.天津科学技术出版社,1993,8.
[22]叶成炳,王明杰.伤寒明理论阐释.四川科学技术出版社,1988,248.
[23]黄竹齐.伤寒论集注.人民卫生出版社,1957,333.
[24]汪昂.医方集解.人民卫生出版社,2007,230-232.
[25]张璐.千金方衍义.中国中医药出版社,1995,440.
[26]吴谦.医宗金鉴.人民卫生出版社,2006:699.
[27]成无己.注解伤寒论.北京:人民卫生出版社,1978:168.
[28]陶节庵.伤寒六书.北京:人民卫生出版社,1990:36.
[29]柯琴.伤寒来苏集(卷四).上海:上海卫生出版社,1956:25-26.
[30]成都中医学院伤寒教研组.伤寒论讲义[M].北京:人民卫生出版社,1960:182-183.
[31]熊曼琪.伤寒学.北京:中国中医药出版社,2003:308.
[32]张志民,徐柏英.对张仲景用附子的研究[J].上海中医药杂志,1957(6):32.
[33]潘澄濂.附子在《伤寒杂病论》诸方配伍中的作用探讨[J].浙江中医学院学报,1990,14(6):4.
[34]陈明,张印生等.伤寒名医验案精选.学苑出版社,1998:405-406.
[35]黄宫绣.本草求真.中国中医药出版社,1997:109.
[36]段富津.方剂学[M].上海:上海科学技术出版社,1995:104.
[37]刘渡舟.新编伤寒论类方.山西人民出版社,1984:183.
[1]姚咏明,盛志勇.脓毒症研究的若干新动态.中国危重病急救医学[J],2000,12(6):323-325.
[2]Fink MP,Heard SO.Laboratory models of sepsis and septic shock[J].J Surg Res,1990,49(2):186-196.
[1]Cavaillon M,Adib-Conquy M,Fitting C,et al.Cytokine cascade in sepsis[J].Scand J Infect D is,2003,35(5):35-44.
[2]Ji XH,Sun KY,Feng YH,et al.Changes of inflammation-associated cytokine expressions during early phase of experimental endotoxic shock in macaques[J].World J Gastroentero,2004,10(4):3026-3301.
[3]Kumar A,Haery C,Parillo JE.Myocardial dysfunction in septic shock[J].Crit Care Clin,2000,16(2):251-287.
[4]Andrew D Luster.The role of chemokines in linking innate and adaptive immunity[J].Current Opinion in Immunology 2002,14(2):129-135.
[1]Blackwell TS,Christman JW.Sepsis and cytokines current status[J].Br J Anaesth,1996,77(2):110-117.
[2]陈永华,蒋建新,谢国旗,等.内毒素血症时小鼠肺局部敛炎及抗炎因子与急性肺损伤的关系[J].中国危重病急救医学,2000,12(4):331-333.
[3]Fink MP,Heard SO.Laboratory models of sepsis and septic shock[J].J Surg Res.1990,49(2):186-196.
[4]魏微,张微微,李远征.大鼠脑缺血冉灌注后肿瘤坏死因子及基质金属蛋白-9的表达[J].中华老年心脑血管病杂志,2004,6(6):406-408.
[5]Dinarello CA.Interleukin-1 and Interleukin-1 antagonism.Blood,1991,77(8):1627-1652.
[6]Heumann D,Roy DL,Zanetti G,et al.Contribution of TNF/TNFreceptor and of toxicity in murine models of endotoxemia and bacterial peritonitis.J Inflamm,1997,47(1):173-179.
[7]Amiot F,Fitting C,Tracey KJ,et al.Lipopolysaccharide-induced cyto-Kine cascade and lethality in LT alpha/TNF alpha-deficient mice[J].Mo Med,1997,3(12):864-875.
[8]Fujimura M,Cache Y.Early appearance of activated matrix metalloprotein ase-9 and blood brain barrier disruption in mice after focal cerebral ischemia and reperfusion [J].Brain Res,1999,842:92-100
[1]余惠平,张和平,田进军,等.ICAM-1在人脑胶质瘤的表达及临床意义[J].中国肿瘤临床与康复,2008,15(3):204-206.
[2]Zhang RL,Chopp M,Zaloga C,et al.The temporal profiles of ICAM-1 protein and mRNA expression after transientMCA occlusion in the rat[J].Brain Res,1995,682(1):182-186.
[3]郑绪鹏,刘班,骆平,等.大鼠脑缺血再灌注后NCAM基因表达的变化[J].青岛大学医学院学报,2006,42(2):140-144.
[4]Ning X Z,Liu Q,Zhao H.Nimodip ine for treatment of perifocal edema follwing asp iration and drainage in patients with cerebra lhemorrhage[J].Neur Regenerat Res,2007,2(5):310-313.
[5]WangW,Sun Y J,Xi T,et al.Effect of 1400W on neurogenesis after transient focal cerebral ischemia in the adult rat hippocampus[J].Chin Pharm acol Bull,2004,20(1):33-35.
[6]Memezawa H,Minamisawa H,SmithM L,et al.Ischemic penumbra in a model of reversible middle cerebral artery occlusion in the rat[J].Exp B rain Res,1992,89(1):67-78.
[7]Michael PB,Richard MN.Endothelial-leukocyte adhesion molecules in inflammation and etastasis[J].Thrombosis and Haemostasis,1993,70(1):152.
[8]Fukuda T.Role of interleukin-4 and vascular cell adhesion molecule-1 in selectiveeosinophil migration into the airways in allergic asthma[J].Am J Respir Cell Mol Biol,1996,14(1):84.
[1]Lehotsky J,Kap lan P,MatejovicovaM,et al.Iontransport systems as targets of free radicals during ischemia reperfusion injury[J].Gen Physiol B iophys,2002,21(1):31-37.
[2]Ravati A,Ahlemeyer B,Becker A,et al.Preconditioning-induced neurop rotection is mediated by reactive oxygen species and activation ofthe transcription factor nuclear factor-kappaB[J].Neurochem,2001,78(4):909-919.
[3]张敬伟.NO,SOD与缺血性脑血管病的关系[J].中国误诊学杂志,2005,5(5):9001.
[4]Tominaga T,Sato S,Ohnishi T et al.Potentiation of nitric oxide Formation following bilateral carotid occlusion and focal cerebral ischemia in the rat:in vivo detection of the nitric oxide radical by electron paramagnetic resonance spin trapping.Brain Res,1993,614(1-2):342-346.
[5]Nathan C,Xie Q W.Nitric oxide synthase:roles,tolls and controls.Cell,1994,7(8):915.
[6]Beray-Berthat V,Palmier B,Plotkine M et al.Neutrophils do not contribute to infarction,oxidative stress,and NO synthase activity in severe brain ischemia.Exp Neurol,2003,182(2):446-454.
[7]Franklin A,Parmentier-Batteur S,Walter L et al.Palmitoy-leth anolamide increases after focal cerebral ischemia and potentiates microglial cell motility.J Neuro Sci,2003,23(21):7767-7715.
[8]Takeyama Y.Significance of apop totic cell death in systemiccomp lications with severe acute pancreatitis[J].J Gastroen2 terol,2005,40(1):1210.
[1]Meyer R,Weissert R,Diem R,et al.Acute neuronal apoptosis in a rat model of multiple sclerosis[J].Neurosci,2001,21(16):6214-20.
[1]乔胃娟,冯全生.感染性休克动物模型评述[J].中外健康文摘,2008,4(2):23-24.
[2]张爱华,蔡毅.白细胞介素-13与肾脏疾病[J].国外医学:泌尿系统分册,1998,18(7):118-121.
[3]王大东,朱瑛梅,李秀智,等.白细胞介素-13功能研究进展[J].医学研究生报,2002,15(6):544-546.
[4]李鸿钧,苏婷.白细胞介素-13的研究进展[J].国外医学:临床生物化学与检验学分册,2001,22(4):187-188.
[5]Matsumto K.Interleukin-13 inhibits cytokine secretion by blood onocytes from patient s wit h IgA nephropat hy[J].Nephron,1997,75(3):295-302.
[6]Saura M,Martinezd R,Minty S,et al.Interleukin-13 inhibits inducible nit ric oxide synt hetase expression in human mesangial cells[J].Biochem J,1996,31(3):641-646.
[7]Gonz(?)lez Alvaro I,Domnguez Jim nez C,Ortiz AM,et al.Interleukin-15 and interfereon-γparticipate in the cross-talk between nat ural killer and monocytic cells required for turnout necrosis factor production[J].Art hritis Res Ther,2006,8(4):88.
[8]Dinarello CA.Proinflammatory and anti-inflammatory cytokines as mediators in the pathogenesis of septic shock[J].Chest,1997,11(2):321-329.
[9]Fukuda T.Role of interleukin-4 and vascular cell adhesion molecule-1 in selective eosinophil migration into the airways in allergic asthma[J].Am J Respir Cell Mol Biol,1996,14(1):84.
[10]El Azami M,Bauche J,Loucka R,Osicka,et al.Interaction of Bordetella adenylate cyclase with CD11b/CD18:role of toxin acylation and identification of the integrin interaction domain[J].J Biol Chem,2003,278(3):38514-38521.
[11]马宏博.论排毒解毒与扶正相结合治疗感染性休克.中国中医急症,2003,12(5):437-438.
[12]肖凤霞.四逆汤的药理研究进展.中医药研究,1999,15(4):60-62.
[13]陈信义,李峨,侯丽,等.乌头类生物碱研究进展与应用前景评述,中国中医药信息杂志,2004,11(10):922-923.
[14]邵春红,王晓良.四逆汤对失血性休克大鼠心功能和血压调节的肾上腺素受体机制研究.中国药学杂志,2003,38(11):847-850.