糖肾煎对糖尿病肾病大鼠足细胞自噬及mTOR/P70S6K通路的影响研究  

Study on the regulatory effect of Tangshenjian decoction on podocyte autophagy and mTOR/P70S6K pathway in rats with diabetic nephropathy

作　　者：牛晓静[1] 张利芳[1] 赵静[1] 叶寒露[1] 晏玲[1] Niu Xiaojing;Zhang Lifang;Zhao Jing;Ye Hanlu;Yan Ling(Department of Endocrinology,Wuhan Hospital of Traditional Chinese Medicine,WuHan 430014,China)

机构地区：[1]武汉市中医医院内分泌科,武汉430014

出　　处：《广西医科大学学报》2021年第11期2064-2070,共7页Journal of Guangxi Medical University

基　　金：湖北省知识创新专项(自然科学基金)项目(No.2018CFB707)。

摘　　要：目的:研究糖肾煎对糖尿病肾病大鼠足细胞自噬及mTOR/P70S6K通路的影响。方法:大鼠按照随机数字表法分为空白对照组(6只)及造模组(35只),采用高糖高脂饲料喂养联合腹腔注射链脲佐菌素(STZ)法建立糖尿病肾病大鼠模型,将造模成功大鼠随机分为模型组,糖肾煎低、中、高剂量组及二甲双胍组。糖肾煎低、中、高剂量组分别按照5 g/kg、10 g/kg、20 g/kg的剂量灌胃给予大鼠糖肾煎,二甲双胍组按照100 mg/kg灌胃给予大鼠二甲双胍,空白对照组及模型组灌胃给予等剂量生理盐水,所有组别给药1次/d,连续给药16周。使用尿蛋白检测试剂盒测定各组大鼠24 h尿蛋白量;使用全自动生化分析仪测定大鼠空腹血糖及糖化血红蛋白水平;使用酶联免疫吸附(ELISA)试剂盒测定血清肌酐、血清尿素氮水平及肾组织超氧化物歧化酶(SOD)、丙二醛(MDA)、总抗氧化能力(T-AOC)水平;苏木精—伊红(HE)染色法检查肾组织病理学变化;免疫荧光法测定足细胞Synaptopodin及LC3水平;免疫印记法(western blotting)检测肾组织p-mTOR、mTOR、p-P70S6K、P70S6K蛋白水平。结果:与空白对照组比较,模型组大鼠空腹血糖、糖化血红蛋白、24 h尿蛋白量、血清肌酐、血清尿素氮、肾组织MDA水平、肾组织pmTOR/mTOR及p-P70S6K/P70S6K水平显著升高(P<0.05),肾组织SOD及T-AOC水平、足细胞Synaptopodin及LC3荧光强度显著降低(P<0.05);与模型组比较,糖肾煎各剂量组大鼠空腹血糖、糖化血红蛋白、24 h尿蛋白量、血清肌酐、血清尿素氮、肾组织MDA水平、肾组织p-mTOR/mTOR及p-P70S6K/P70S6K水平显著降低(P<0.05),肾组织SOD及T-AOC水平、足细胞Synaptopodin及LC3荧光强度显著升高,并呈剂量依赖性(P<0.05)。结论:糖肾煎能够显著改善糖尿病肾病大鼠肾功能,修复足细胞损伤,调节足细胞自噬水平,其机制可能与调节mTOR/P70S6K通路有关。Objective:To study the regulative effect of Tangshenjian decoction(TSJD) on podocyte autophagy and mTOR/P70 S6 K pathway in rats with diabetic nephropathy.Methods:Rats were randomly divided into blank control group(6 rats) and model group(35 rats),A diabetic nephropathy rat model was established by high-fat and high-sugar feeding combined with intraperitoneal injection of streptozotocin(STZ).Successfully modeled rats were randomly divided into,model group,metformin(100 mg/kg) group and low-dose(5 g/kg),mediumdose(10 g/kg),and high-dose(20 g/kg) TSJD groups.The drugs were given by gavage,once a day for 16 weeks.The 24 h urine protein,were measured by urine protrin test kit;fasting blood glucose and glycosylated hemoglobin contents were measured by automatic biochemical analyzer.The serum creatinine,serum urea nitrogen,and superoxide dismutase(SOD),malondialdehyde(MDA)and total antioxidant capacity(T-AOC) levels in renal tissues were measured by ELISA.The histopathological changes of kidney were observed by hematoxylineosin(HE) staining.The levels of Synaptopodin and LC3 in podocytes were detected by immunofluorescence.The protein expressions of p-mTOR,mTOR,p-P70 S6 K,and P70 S6 K in renal tissues were determined by western blotting.Results:Compared with the blank control group,the levels of fasting blood glucose,glycosylated hemoglobin,24 h urine protein,serum creatinine,serum urea nitrogen,and MDA as well as the ratios of p-mTOR/mTOR and p-P70 S6 K/P70 S6 K in renal tissues in the model group were significantly increased,while the levels of SOD and T-AOC in renal tissues and the fluorescence intensity of Synaptopodin and LC3 in podocytes were significantly decreased(P<0.05).Compared with the model group,each dose of TSJD treatment led to significant decreases in fasting blood glucose,glycosylated hemoglobin,24 h urine protein,serum creatinine,serum urea nitrogen and MDA,and ratios of p-mTOR/mTOR and p-P70 S6 K/P70 S6 K,whereas increases in the levels of SOD and T-AOC,and the fluorescence intensity of Synaptopodin

关 键 词：糖肾煎 mTOR/P70S6K 足细胞自噬 

分 类 号：R73[医药卫生—肿瘤]