雷公藤多苷对糖尿病肾病大鼠肾脏细胞自噬的影响及相关机制

doi:10.16098/j.issn.0529-1356.2020.03.010

摘要/Abstract

摘要：

目的探讨雷公藤多苷对糖尿病肾病大鼠的肾脏细胞自噬的影响，并分析其相关分子机制。方法采用腹腔注射链脲佐菌素（STZ）的方法构建糖尿病肾病大鼠模型。将30只SD大鼠依据随机数表法分为5组，对照组、模型组、0.1 mg/kg药物组、0.5 mg/kg药物组和1.0 mg/kg药物组，每组各6只。糖尿病肾病模型建立成功后，0.1 mg/kg药物组、0.5 mg/kg药物组和1.0 mg/kg药物组分别灌胃0.1、0.5和1.0 mg/kg雷公藤多苷，对照组和模型组给予等量的生理盐水。比较各组间肾功能、氧化应激指标的水平。采用Western blotting法检测各组间磷酸化-哺乳动物雷帕霉素靶蛋白（p-mTOR）、哺乳动物雷帕霉素靶蛋白(mTOR)、微管相关蛋白1轻链3β-Ⅰ（LC3-Ⅰ）、微管相关蛋白1轻链3β-Ⅱ（LC3-Ⅱ）及Beclin1蛋白表达水平。使用Real-time PCR技术检测各组间LC3及Beclin1 mRNA表达水平。结果与对照组比较，模型组尿素氮（BUN）、血肌酐（Scr）、尿蛋白（Pro）及丙二醛（MDA）水平均升高，谷胱甘肽过氧化物酶（GSH-PX）及过氧化氢酶(CAT)水平均明显降低，差异具有统计学意义（P<0.05）；与模型组比较，药物组BUN、Scr、Pro及MDA水平均明显降低，谷胱甘肽过氧化物酶（GSH-PX）及CAT水平均明显升高（P<0.05），呈剂量依赖性。与对照组比较，模型组的肾组织p-mTOR蛋白表达水平升高，LC3-Ⅱ/LC3-Ⅰ及Beclin1蛋白表达水平降低，差异具有统计学意义（P<0.05）；与模型组比较，各剂量药物组p-mTOR蛋白表达水平降低，LC3-Ⅱ/LC3-Ⅰ蛋白表达水平显著升高（P<0.05），呈剂量依赖性，0.5 mg/kg及1.0 mg/kg药物组Beclin1蛋白表达水平显著高于对照组（P<0.05）。与对照组比较，模型组的肾组织，LC3-Ⅱ及Beclin1 mRNA表达水平降低，差异具有统计学意义（P<0.05）；与模型组比较，各剂量药物组LC3及Beclin1 mRNA表达水平显著升高，差异具有统计学意义（P<0.05）。结论雷公藤多苷对糖尿病性肾病大鼠肾功能具有一定的保护作用，其机制可能与抑制氧化应激和激活自噬功能有关。

关键词: 雷公藤多苷, 糖尿病肾病, 自噬, 免疫印迹法, 大鼠

Abstract:

Objective To investigate the effects of tripterygium glycosides on autophagy of renal cells in rats with diabetic nephropathy and to analyze its molecular mechanism. Methods A rat model of diabetic nephropathy was made by intraperitoneal injection of streptavidin. Thirty SD rats were randomly divided into 5 groups, control group, model group, 0.1 mg/kg drug group, 0.5 mg/kg drug group and 1.0 mg/kg drug group, and each group has 6 rat. After the successful establishment of the diabetic nephropathy model, the 0.1 mg/kg drug group, the 0.5 mg/kg drug group, and the 1.0 mg/kg drug group were intragastric administration with 0.1, 0.5, and 1.0 mg/kg tripterygium glycosides, respectively, and the control group and the model group were intraperitoneally injected with the same amount of normal saline. The levels of renal function and oxidative stress were compared among groups. The expression levels of p-mammalian target of rapamycin (p-mTOR),mammalian target of rapamycin(mTOR), microtubules associated protein 1 light chain 3β-Ⅱ/microtubules associated protein 1 light chain 3β-Ⅰ(LC3-Ⅱ/LC3-Ⅰ) and Beclin1 protein were detected by Western blotting. The expression levels of LC3, LC3-Ⅱ and Beclin1 mRNA in each group were detected by Real-time PCR. Results Compared with the control group, the serum creatinine(Scr), blood urea nitrogen(BUN), protein(Pro) and malondialdehyde(MDA) levels in the model group increased,and the the glutathione peroxidase(GSH-Px) and catalase(CAT) levels decreased significantly (P<0.05). Compared with the model group, the Scr, BUN, Pro and MDA levels of the drug group were significantly decreased, and GSH-Px and CAT levels were significantly increased in a dose-dependent manner (P<0.05). Compared with the control group, the expression level of p-mTOR protein in the renal tissue of the model group was increased, and the expression levels of LC3-Ⅱ/LC3-Ⅰ and Beclin1 protein were decreased (P<0.05). Compared with the model group, the expression level of p-mTOR protein was decreased in the dose group, and the expression levels of LC3-Ⅱ/LC3-Ⅰ protein in the dose group were significantly increased in a dose-dependent manner (P<0.05). The expression levels of Beclin1 protein in the 0.5 mg/kg drug group and 1.0 mg/kg drug group, were significantly higher than the model group (P<0.05). Compared with the control group, the expression levels of LC3-Ⅱ/LC3-Ⅰ and Beclin1 mRNA in the renal tissue of the model group were significantly lower (P<0.05). Compared with the model group, the expression levels of LC3 and Beclin1 mRNA in the drug groups of each dose group were significantly increased (P<0.05). Conclusion Tripterygium wilfordii glycosides can protect kidney function in rats with diabetic nephropathy, and its mechanism might be related to inhibition of oxidative stress and activation of autophagy.

Key words: Tripterygium glycoside, Diabetic nephropathy, Autophagy, Western blotting, Rat

中图分类号:

R587

张筠丁婷唐东兴王建平. 雷公藤多苷对糖尿病肾病大鼠肾脏细胞自噬的影响及相关机制[J]. 解剖学报, 2020, 51(3): 373-377.

ZHANG Jun DING Ting TANG Dong-xing WANG Jian-ping. Effect of tripterygium wilfordii polyglycoside on renal cell autophagy in rats with diabetic nephropathy and its related mechanism[J]. Acta Anatomica Sinica, 2020, 51(3): 373-377.

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