【作者】 黄娟；

【导师】 陈泽奇；

【作者基本信息】 中南大学 ， 中西医结合临床， 2009， 硕士

【摘要】 目的:观察加味补肝汤对糖尿病大鼠坐骨神经中p-JNK丝裂原活化蛋白激酶及c-jun mRNA表达的影响,探讨其对实验性糖尿病大鼠周围神经病变的防治作用。方法:1.65只清洁级雄性Wistar大鼠随机抽取20只为正常组,余下45只,采用链脲佐菌素(STZ)复制糖尿病神经病变大鼠模型,将造模成功后的40只大鼠,随机分为模型组(20只),加味补肝汤组(20只)。2.分别于造模前,造模后4周(4W)和8周(8W)三个时间点尾静脉采血测血糖值及体重。3.分别于造模后4W、8W两个时间点处死大鼠,用黄嘌呤氧化酶法测定血清SOD活力,用免疫组化技术测定坐骨神经p-JNK的表达,用逆转录-聚合酶链反应(RT-PCR)技术测定坐骨神经中c-jun mRNA的表达,并在处死之前检测各组大鼠坐骨神经传导速度。结果:1.加味补肝汤对大鼠体重与血糖的影响:与模型组相比加味补肝汤组对STZ诱导的糖尿病大鼠的体重和血糖无明显改善(P＞0.05)。2.加味补肝汤对坐骨神经传导速度的影响:与正常组相比,模型组大鼠坐骨神经传导速度在造模后4、8周明显减慢(P＜0.05);加味补肝汤干预均能减缓神经传导速度的减慢且与模型组间差异有显著意义(P＜0.05)。3.加味补肝汤对血清SOD活力的影响:模型组4W、8W时大鼠血清SOD含量低于同期正常组(P＜0.05),随病程的延长模型组大鼠血清SOD含量逐渐减少(P＜0.05)。加味补肝汤干预后血清SOD水平均明显高于模型组(P＜0.05)。4.加味补肝汤对大鼠坐骨神经中p-JNK及c-jun mRNA表达的影响:造模后4、8周模型组大鼠p-JNK及c-jun mRNA表达明显高于正常组(P＜0.05),加味补肝汤干预后p-JNK及c-jun mRNA表达明显低于模型组(P＜0.05)。结论:1.加味补肝汤可上调糖尿病大鼠机体SOD水平,具有抗氧化应激作用。2.加味补肝汤下调p-JNK的表达、抑制c-junmRNA的转录,减少细胞凋亡可能是通过调节机体氧化应激状态,从而减轻了周围神经细胞氧化应激损伤,这可能是该方防治DPN的作用机制之一。更多还原

【Abstract】 Objective:to observe the effect of jiaweibugan decoction on the expression of p-JNK mitogen-activated protein kinase and c-jun in sciatic nerve of experimental diabetic rats and explore the preventive and therapeutic effects of Jiaweibugan Decoction on peripheral neuropathy on experimental diabetic rats.Methods:1.Randomly selected 20 rats from 65 clean grade male Wistar rats as normal control group,and remnant 45 rats were established by streptozotocin（STZ） as diabetic model.The 40 rats which were successful induction were randomly divided into two groups including model-control group and JWBG decoction-treated group.2.Nonfasting plasma glucose and weight were measured on 4^th、8^th week and before treatment respectively.3.The rats were killed on 4^th or 8^th week from the beginning of treatment respectively:the blood serum and sciatic nerve were taken out, and then various parameters were measured as follows:serum SOD was determined by xanthine oxidasemethod;p-JNK expression of protein in sciatic nerve by immunohistochemistry;c-jun mRNA expression by reverse-transcriptase polymerase chain reaction（RT-PCR）,and detecting the speed of conduction in sciatic nerve before being killed respectively. Results:（1）The effects of JWBG decoction on blood glucose and weight: compared with the model group,JWBG decoction has no significantly ameliorating to weight and blood glucose of the rats with diabetes induced by streptozotocin（P＞0.05）.（2）The effects of JWBG decoction on the sciatic nerve conduction velocity:compared with the normal group,the sciatic nerve conduction velocity of the model group is significantly slow on the 4^th and 8^th week after successful induction by STZ（P＜0.01）.Intervention with JWBG decoction-treated group could slower the decreasing of nerve conduction velocity,and at the same time there also are a significant difference between the JWBG decoction-treated group and model group（p＜0.05）.（3） The effects of JWBG decoction on serum SOD activity:The results showed that SOD in the model group on 4^th or 8^th week was lower than that in the normal control group（P＜0.05）,and it became much lower with diabetic course prolonging gradually,while which in jiaweibugan decoction intervened group was markedly higher than that in model rats（P＜0.05）.（4） The effects of JWBG decoction on the expression of p-JNK mitogen-activated protein kinase and c-jun in sciatic nerve:The results showed that the expression of p-JNK and c-jun in the model group on 4^th or 8^th week was higher than that in the normal control group （P＜0.05）,while which in jiaweibugan decoction group was markedly lower than that in model rats（P＜0.05）.Conclusion:1.Jiaweibugan decoction could up-regulate the activity of SOD,and have the effect of anti-oxidative stress.2.JWBG decoction down-regulate the expression of p-JNK, inhibiting transcription of c-junmRNA and decreasing apoptosis may be through adjusting the oxidative stress state of organism,thus it can protect nerve cell from oxidative injury.This may be one of the function mechanisms of protective effects of Jiaweibugan Decoction on diabetic peripheral neuropathy.更多还原