滋阴益气熄风活血通络法对DPN的临床观察及对糖尿病大鼠坐骨神经雪旺细胞凋亡的实验研究
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摘要
目的:观察滋阴益气活血熄风通络法对糖尿病周围神经病变的临床疗效,探讨滋阴益气活血熄风通络法治疗糖尿病周围神经病变的分子机制,为中医药治疗糖尿病周围神经病变提供理论依据。
方法:临床部分:将入选病人随机分为治疗组和对照组各30例,两组中医辨证均属于气阴两虚兼有血瘀型DPN患者。治疗组服用降糖舒络方,对照组服用弥可保片。治疗前后观察一般情况、临床疗效、证候疗效、症状积分、血糖、糖化血红蛋白和神经传导速度情况。两组均在西药降糖的基础上进行治疗。一个月为一疗程,共观察一个疗程。实验部分:将48只SD雌性大鼠随机分为正常组、模型组、西药组、降糖舒络方高剂量组、降糖舒络方中剂量组、降糖舒络方低剂量组6组,采用STZ腹腔注射,建立SD大鼠糖尿病模型。以实验大鼠一般情况、血糖、糖化血红蛋白、血清胰岛素、坐骨神经雪旺细胞凋亡及介导细胞凋亡的caspase-3和调控细胞凋亡的基因蛋白bcl-2、bax的表达为观察指标,以达美康+弥可保为对照进行观察。
结果:临床部分:治疗组病人临床疗效、证候疗效和临床主要症状改善与对照组疗效相当(P＞0.05),但是治疗组的临床疗效显效率、证候痊愈率以及对次要症状改善均优于对照组(P＜0.05)。两组治疗后与治疗前对FBG、PBG、HbA_1C和神经传导速度的影响均有统计学意义(P＜0.01),两组治疗后比较无统计学(P＞0.05)。实验部分:各组大鼠造模前饮水量、体重、食量、尿量、血糖均无明显改变(P＞0.05);造模后各组大鼠的饮水量、体重、食量、尿量、血糖跟模型组相比均有显著差异(P＜0.01);随着用药的进行,西药组和中药组DM大鼠的饮水量、尿量及食量均较模型组明显减少,血糖逐渐下降,而体重较模型组则明显增加(P＜0.01)。实验结束后西药组和中药组一般情况、血糖较模型组均有明显改善,而对于饮水量、食量、尿量的改善中药组优于西药组,而且呈剂量依赖性。西药组对DM大鼠的体重及血糖影响优于中药组(P＜0.01),中药各组对大鼠体重影响以中药大剂量组疗效占优(P＜0.05),但是对血糖影响无组间差异(P＞0.05)。模型组SINS、HbA1c跟正常组比较有显著差异(P＜0.01),治疗组和对照组SINS、HbA1c与模型组相比均有显著意义(P＜0.01),中药高剂量组和中剂量组对SINS、HbA1c的影响明显优于对照组(P＜0.05或0.01),对照组对SINS、HbA1c的疗效则优于中药小剂量组(P＜0.01),中药组间比较以中药大剂量组和中剂量组疗效占优(P＜0.01)。模型组大鼠坐骨神经雪旺细胞凋亡、caspase-3、bax基因蛋白表达较正常组明显增强,bcl-2明显下降(p＜0.01)。中药组和西药组对雪旺细胞凋亡、caspase-3、bax和bcl-2基因蛋白的影响与模型组相比均有统计学意义(p＜0.01);中药中剂量组和大剂量组组对于雪旺细胞凋亡、caspase-3、bax和bcl-2的影响优于西药组(P＜0.05)。中药组间比较以中药大剂量组疗效占优(P＜0.05或0.01)。
结论:滋阴益气熄风活血通络法可以稳定血糖,改善糖尿病周围神经病变患者临床症状和神经传导速度,估计其治疗作用与改善糖代谢、抑制醛糖还原酶活性和多元醇通路代谢增强有关。滋阴益气熄风活血通络法可以改善糖尿病大鼠血清胰岛素水平,降低糖化血红蛋白水平,抑制雪旺细胞凋亡,认为其作用机制与抑制caspase-3活性和bax蛋白表达及促进bcl-2蛋白表达有关,并且对于各观察指标影响基本呈剂量依赖性。
Objective:To observe the clinical effect of nourishing Yin,invigorating Qi,extinguishing wind,promoting blood circulation and draining collateral method to DPN,to discuss the mechanism that nourishing Yin, invigorating Qi,extinguishing wind,promoting blood circulation,Extinguishing wind and draining collateral method to treat DPN,to provide the theory basis for TCM to treat DPN.
Method:Clinical part:The selected patients were randomly devided into treatment group and control group,each group has 30 patients.The two groups patients were belong to symptoms of accumulation of blood, deficiency of both qi and yin.The treatment group patients take jiangtangshuluo formula,the control group patients take methycobal tablets,observe the ordinary circumstances,clinical effect,syndrome effect,Symptom integral,BG,HbA_1c and NCV between before treatment and after treatment.The two groups carry on the treatment based on controlling BG by the western medicine.One month is a treatment course,altogether observe a treatment course.Experiment part:48 SD femail rats were randomly devided normal group,model group,western medicine group,jiangtangshuluo fomula high dosage group,jiangtangshuluo formula middle dosage group,jiangtangshuluo formula low dosage group into 6 groups,had abdominal cavity injection by STZ to make SD rats DM model.Compared with methycobal and gliclazide,took ordinary circumstances,BG,HbA_1c,SINS,sciatic nerve schwann cells apotosis,the expression of caspase-3(caused apotosis)-and bcl-2,bax(adjusted and controlled apotosis) as observational targets.
Results:Clinical part:The clinical and syndrome effect and the mayor symptoms of the treatment group patients were equalled with the control group(P＞0.05),but the effect of the treatment group to the clinical apparent rate,syndrome heal rate and the secondary symptoms were superior to the control group(P＜0.05).There was statistics significance of the FBG,PBG,HbA_1C and NCV,the two groups compared after treatment with before treatment(P＜0.01),and there is no statistics significance after treatment of the two groups compared(P＞0.05).Experiment part:There were no obviously changement of water intake,body weight,food quantity, urine volume,BG of each group DM rats before making models(P＞0.05).There were obviously significance of water intake,body weight,food quantity,urine volume,BG of each group DM rats after making models(P＜0.01),with the process of using medicines,the water intake,food quantity,urine volume of the western medicine group and the TCM group DM rats obviously reduced compared with the model group DM rats,the levels of BG dropped gradually,but the body weight obviously increased compared with the model group(P＜0.01).After the experiment ended,the ordinary circumstances and BG of the western medicine group and the TCM group had obviously changement compared with the model group,the effect of TCM group to warer intake,food quantity,urine volume was better than the western medicine group,and assumed the dosage dependence.The effect of body weight and BG of western medicine group DM rats restored better than the TCM group(P＜0.01),the curative effect of each TCM group compared,the high dosage group was better than the other two groups(P＜0.05),but there was no statistical significance among the TCM groups(P＞0.05).The levels of serum insulin,HbA_1c of the model group compared with the normal group had remarkable difference(P＜0.01),the serum insulin,HbA_1c of the treatment group and the control group all had statistics significance than the model group after treatment(P＜0.01),the effect of the high dosage TCM group and the middle dosage TCM group to the surem insulin,HbA1c was superior than the western group obviously(P＜0.05or 0.01),the curative effect of the control group to the serum insulin,HbA_1c was better than the low dosage TCM group(P＜0.01),the curative effect of each TCM group compared,the high dosage group and the middle dosage group was better than the low dosage groups(P＜0.01).The DM rats sciatic nerve schwann cells apotosis,caspase-3,bax gene protine expressed of the model group rats sciatic nerve higher and the bcl-2 gene protine expressed lower than the normal group obviously(P＜0.01).There were statistics significance both TCM group and western medicine group compared with the model group to the sciatic nerve schwann cells apotosis,caspase-3,bax and bcl-2(P＜0.01).The effect of each TCM high and middle dosage group to the schwann cells apotosis,caspase-3,bax and bcl-2 was superior to the western medicine group(P＜0.05),the curative effect of each TCM group compared,the high dosage group was better than the other two groups(P＜0.05or0.01).
Conclusion:Nourishing Yin,invigorating Qi,extinguishing wind, promoting blood circulation and draining collateral method can stabilizate dlood glucoge,improve the clinical symptoms and nerve conductive velocity of the DPN patients,estimate it's curative effect has something to do with improving sugar metabolism,inhibiting the activity of aldose reductase.Nourishing Yin,invigorating Qi,extinguishing wind,promoting blood circulation and draining collateral method can improve the levels of Insulin and reduce the level of HbA1c on DM rats,inhibit schwann cells apoptosis,it was thought that the function mechanism was concerned on inhibiting the activeness of caspase-3,the expressing of bax and promoting the expressing of bcl-2,and the effect of the method to each observation target show the dosage dependence.
方法:临床部分:将入选病人随机分为治疗组和对照组各30例,两组中医辨证均属于气阴两虚兼有血瘀型DPN患者。治疗组服用降糖舒络方,对照组服用弥可保片。治疗前后观察一般情况、临床疗效、证候疗效、症状积分、血糖、糖化血红蛋白和神经传导速度情况。两组均在西药降糖的基础上进行治疗。一个月为一疗程,共观察一个疗程。实验部分:将48只SD雌性大鼠随机分为正常组、模型组、西药组、降糖舒络方高剂量组、降糖舒络方中剂量组、降糖舒络方低剂量组6组,采用STZ腹腔注射,建立SD大鼠糖尿病模型。以实验大鼠一般情况、血糖、糖化血红蛋白、血清胰岛素、坐骨神经雪旺细胞凋亡及介导细胞凋亡的caspase-3和调控细胞凋亡的基因蛋白bcl-2、bax的表达为观察指标,以达美康+弥可保为对照进行观察。
结果:临床部分:治疗组病人临床疗效、证候疗效和临床主要症状改善与对照组疗效相当(P＞0.05),但是治疗组的临床疗效显效率、证候痊愈率以及对次要症状改善均优于对照组(P＜0.05)。两组治疗后与治疗前对FBG、PBG、HbA_1C和神经传导速度的影响均有统计学意义(P＜0.01),两组治疗后比较无统计学(P＞0.05)。实验部分:各组大鼠造模前饮水量、体重、食量、尿量、血糖均无明显改变(P＞0.05);造模后各组大鼠的饮水量、体重、食量、尿量、血糖跟模型组相比均有显著差异(P＜0.01);随着用药的进行,西药组和中药组DM大鼠的饮水量、尿量及食量均较模型组明显减少,血糖逐渐下降,而体重较模型组则明显增加(P＜0.01)。实验结束后西药组和中药组一般情况、血糖较模型组均有明显改善,而对于饮水量、食量、尿量的改善中药组优于西药组,而且呈剂量依赖性。西药组对DM大鼠的体重及血糖影响优于中药组(P＜0.01),中药各组对大鼠体重影响以中药大剂量组疗效占优(P＜0.05),但是对血糖影响无组间差异(P＞0.05)。模型组SINS、HbA1c跟正常组比较有显著差异(P＜0.01),治疗组和对照组SINS、HbA1c与模型组相比均有显著意义(P＜0.01),中药高剂量组和中剂量组对SINS、HbA1c的影响明显优于对照组(P＜0.05或0.01),对照组对SINS、HbA1c的疗效则优于中药小剂量组(P＜0.01),中药组间比较以中药大剂量组和中剂量组疗效占优(P＜0.01)。模型组大鼠坐骨神经雪旺细胞凋亡、caspase-3、bax基因蛋白表达较正常组明显增强,bcl-2明显下降(p＜0.01)。中药组和西药组对雪旺细胞凋亡、caspase-3、bax和bcl-2基因蛋白的影响与模型组相比均有统计学意义(p＜0.01);中药中剂量组和大剂量组组对于雪旺细胞凋亡、caspase-3、bax和bcl-2的影响优于西药组(P＜0.05)。中药组间比较以中药大剂量组疗效占优(P＜0.05或0.01)。
结论:滋阴益气熄风活血通络法可以稳定血糖,改善糖尿病周围神经病变患者临床症状和神经传导速度,估计其治疗作用与改善糖代谢、抑制醛糖还原酶活性和多元醇通路代谢增强有关。滋阴益气熄风活血通络法可以改善糖尿病大鼠血清胰岛素水平,降低糖化血红蛋白水平,抑制雪旺细胞凋亡,认为其作用机制与抑制caspase-3活性和bax蛋白表达及促进bcl-2蛋白表达有关,并且对于各观察指标影响基本呈剂量依赖性。
Objective:To observe the clinical effect of nourishing Yin,invigorating Qi,extinguishing wind,promoting blood circulation and draining collateral method to DPN,to discuss the mechanism that nourishing Yin, invigorating Qi,extinguishing wind,promoting blood circulation,Extinguishing wind and draining collateral method to treat DPN,to provide the theory basis for TCM to treat DPN.
Method:Clinical part:The selected patients were randomly devided into treatment group and control group,each group has 30 patients.The two groups patients were belong to symptoms of accumulation of blood, deficiency of both qi and yin.The treatment group patients take jiangtangshuluo formula,the control group patients take methycobal tablets,observe the ordinary circumstances,clinical effect,syndrome effect,Symptom integral,BG,HbA_1c and NCV between before treatment and after treatment.The two groups carry on the treatment based on controlling BG by the western medicine.One month is a treatment course,altogether observe a treatment course.Experiment part:48 SD femail rats were randomly devided normal group,model group,western medicine group,jiangtangshuluo fomula high dosage group,jiangtangshuluo formula middle dosage group,jiangtangshuluo formula low dosage group into 6 groups,had abdominal cavity injection by STZ to make SD rats DM model.Compared with methycobal and gliclazide,took ordinary circumstances,BG,HbA_1c,SINS,sciatic nerve schwann cells apotosis,the expression of caspase-3(caused apotosis)-and bcl-2,bax(adjusted and controlled apotosis) as observational targets.
Results:Clinical part:The clinical and syndrome effect and the mayor symptoms of the treatment group patients were equalled with the control group(P＞0.05),but the effect of the treatment group to the clinical apparent rate,syndrome heal rate and the secondary symptoms were superior to the control group(P＜0.05).There was statistics significance of the FBG,PBG,HbA_1C and NCV,the two groups compared after treatment with before treatment(P＜0.01),and there is no statistics significance after treatment of the two groups compared(P＞0.05).Experiment part:There were no obviously changement of water intake,body weight,food quantity, urine volume,BG of each group DM rats before making models(P＞0.05).There were obviously significance of water intake,body weight,food quantity,urine volume,BG of each group DM rats after making models(P＜0.01),with the process of using medicines,the water intake,food quantity,urine volume of the western medicine group and the TCM group DM rats obviously reduced compared with the model group DM rats,the levels of BG dropped gradually,but the body weight obviously increased compared with the model group(P＜0.01).After the experiment ended,the ordinary circumstances and BG of the western medicine group and the TCM group had obviously changement compared with the model group,the effect of TCM group to warer intake,food quantity,urine volume was better than the western medicine group,and assumed the dosage dependence.The effect of body weight and BG of western medicine group DM rats restored better than the TCM group(P＜0.01),the curative effect of each TCM group compared,the high dosage group was better than the other two groups(P＜0.05),but there was no statistical significance among the TCM groups(P＞0.05).The levels of serum insulin,HbA_1c of the model group compared with the normal group had remarkable difference(P＜0.01),the serum insulin,HbA_1c of the treatment group and the control group all had statistics significance than the model group after treatment(P＜0.01),the effect of the high dosage TCM group and the middle dosage TCM group to the surem insulin,HbA1c was superior than the western group obviously(P＜0.05or 0.01),the curative effect of the control group to the serum insulin,HbA_1c was better than the low dosage TCM group(P＜0.01),the curative effect of each TCM group compared,the high dosage group and the middle dosage group was better than the low dosage groups(P＜0.01).The DM rats sciatic nerve schwann cells apotosis,caspase-3,bax gene protine expressed of the model group rats sciatic nerve higher and the bcl-2 gene protine expressed lower than the normal group obviously(P＜0.01).There were statistics significance both TCM group and western medicine group compared with the model group to the sciatic nerve schwann cells apotosis,caspase-3,bax and bcl-2(P＜0.01).The effect of each TCM high and middle dosage group to the schwann cells apotosis,caspase-3,bax and bcl-2 was superior to the western medicine group(P＜0.05),the curative effect of each TCM group compared,the high dosage group was better than the other two groups(P＜0.05or0.01).
Conclusion:Nourishing Yin,invigorating Qi,extinguishing wind, promoting blood circulation and draining collateral method can stabilizate dlood glucoge,improve the clinical symptoms and nerve conductive velocity of the DPN patients,estimate it's curative effect has something to do with improving sugar metabolism,inhibiting the activity of aldose reductase.Nourishing Yin,invigorating Qi,extinguishing wind,promoting blood circulation and draining collateral method can improve the levels of Insulin and reduce the level of HbA1c on DM rats,inhibit schwann cells apoptosis,it was thought that the function mechanism was concerned on inhibiting the activeness of caspase-3,the expressing of bax and promoting the expressing of bcl-2,and the effect of the method to each observation target show the dosage dependence.
引文
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