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目的:观察中药黄芪菟箭合剂对STZ诱导的糖尿病大鼠肾脏PI3K-Akt-m TOR信号通路的影响,并与雷帕霉素的作用进行比较。方法:糖尿病大鼠分为糖尿病对照组(DM组)、黄芪菟箭合剂治疗组(CM组)、雷帕霉素治疗组(RAPA组),并设正常大鼠对照组(CON组),分别给予不同治疗,12周后用ELISA法检测大鼠的尿白蛋白排泄率,免疫组化法观察肾小球ZO-1、nephrin蛋白表达;Western-blot法检测肾脏磷酸化Akt(Thr308)、磷酸化PTEN、磷酸化S6K的蛋白表达;应用qRT-PCR检测肾脏Akt、PTEN、S6K的mRNA表达。结果:CM组与RAPA组大鼠尿微量白蛋白排泄率均显著低于DM组,ZO-1、nephrin阳性表达显著高于DM组,且CM组与RAPA组之间无显著差别;CM组与RAPA组肾脏磷酸化S6K蛋白表达、S6KmRNA表达均显著低于DM组;CM组磷酸化PTEN蛋白及PTEN mRNA表达显著高于DM组,磷酸化Akt(Thr308)蛋白、Akt mRNA则低于DM组;但RAPA组磷酸化PTEN蛋白及PTEN mRNA表达以及磷酸化Akt(Thr308)蛋白、Akt mRNA表达与DM差异无统计学意义,RAPA组体重显著低于DM组及CM组、死亡率显著高于DM组及CM组。结论:雷帕霉素和黄芪菟箭合剂都可以有效地保护STZ诱导的糖尿病大鼠足细胞的完整性、减少尿微量白蛋白排泄率,但雷帕霉素存在显著毒副作用。
OBJECTIVE: To observe the effect of Huangqizhijian mixture on PI3K-Akt-m TOR signaling pathway in STZ-induced diabetic rats and to compare its effect with rapamycin. Methods: Diabetic rats were divided into DM group, CM group, RAPA group and control group (CON group), respectively After 12 weeks, the urinary albumin excretion rate was measured by ELISA. The expression of ZO-1 and nephrin in glomerulus was observed by immunohistochemistry. The expressions of phosphorylated Akt (Thr308), phosphorylated PTEN and phospho-S6K were detected by immunohistochemical method. The mRNA expressions of Akt, PTEN and S6K in kidney were detected by qRT-PCR. Results: The urinary albumin excretion rate of rats in CM group and RAPA group was significantly lower than that in DM group. The positive expression of ZO-1 and nephrin in CM group and RAPA group was significantly higher than that in DM group. There was no significant difference between CM group and RAPA group. The expression of phosphorylated S6K protein and S6K mRNA in kidney of RAPA group were significantly lower than those in DM group. The phosphorylation of PTEN protein and PTEN mRNA in CM group were significantly higher than that in DM group. The Akt mRNA expression was lower in DM group than in DM group. However, the phosphorylation of PTEN protein and PTEN mRNA, phosphorylated Akt (Thr308) protein and Akt mRNA in RAPA group were not significantly different from those in DM group. The body weight in RAPA group was significantly lower than that in DM group and CM group, and the mortality was significantly higher than that in DM group And CM group. Conclusion: Both rapamycin and Huangqijiejian mixture can effectively protect the integrity of podocytes in STZ-induced diabetic rats and decrease the urinary albumin excretion rate, but rapamycin has significant toxic and side effects.