基于PTEN/Akt/mTOR通路探讨养肾方干预肾纤维化的自噬调控效应和机制  

Study on the Autophagy Regulation Effect and Mechanism of Yangshen Formula in the Intervention of Renal Fibrosis Based on PTEN/Akt/mTOR Pathway

作　　者：邓丽娥[1] 孙子锋 周玖瑶 郑绍琴 何新荣 李强[1] 卢汉祺 DENG Li-e;SUN Zi-feng;ZHOU Jiu-yao;ZHENG Shao-qin;HE Xin-rong;LI Qiang;LU Han-qi(Dongguan Hospital of Guangzhou University of Traditional Chinese Medicine,Dongguan 523000,China;Science and Technology Industrial Park,Guangzhou University of Traditional Chinese Medicine,Guangzhou 510080,China)

机构地区：[1]广州中医药大学东莞医院,广东东莞523000 [2]广州中医药大学科技产业园,广东广州510080

出　　处：《中药材》2023年第11期2825-2830,共6页Journal of Chinese Medicinal Materials

基　　金：广东省基础与应用基础研究基金项目之区域联合基金-青年基金项目(2020A1515110137);广东省中医药局项目(20221420);东莞市社会发展科技重点项目(20211800904412)。

摘　　要：目的:探讨养肾方干预肾纤维化的自噬调控效应和机制。方法:将40只C57BL/6小鼠随机分为假手术组、模型组及养肾方低、高剂量组。假手术组仅分离左侧输尿管,其余各组采用单侧输尿管结扎的方法制备肾纤维化模型。药物连续干预4 w后,检测小鼠24 h尿蛋白、Scr、UREA、Cys-C,计算肾脏指数,HE及Masson染色观察肾组织病理改变及肾纤维化程度,电镜观察小鼠细胞中自噬小体的数量,Western Blot检测肾组织P62、Beclin-1、LC3A/B、PTEN、p-Akt、Akt、p-mTOR、mTOR蛋白表达。结果:与假手术组比较,模型组肾脏指数、24 h尿蛋白、Scr、Cys-C水平显著升高(P<0.05或P<0.01),肾脏结构受到破坏,纤维化明显,肾组织LC3A/B、P62、Beclin-1、p-mTOR、mTOR、p-Akt蛋白表达显著降低(P<0.05或P<0.01)。与模型组比较,养肾方各剂量组24 h尿蛋白、Scr显著降低(P<0.05或P<0.01),肾组织结构的损伤程度减轻,自噬小体数量及LC3A/B蛋白表达显著升高,肾组织Akt蛋白表达显著降低,低剂量组肾组织mTOR、p-mTOR蛋白表达显著升高(P<0.05或P<0.01)。结论:养肾方可改善肾纤维化小鼠的肾功能、抗肾纤维化,其机制可能与养肾方调控PTEN/Akt/mTOR通路相关蛋白的表达,激活自噬有关。Objective:To explore the autophagy regulation effect and mechanism of Yangshen formula intervening renal fibrosis.Methods:40 C57BL/6 mice were randomly divided into sham operation group,model group,low-dose Yangshen formula group and highdose Yangshen formula group.In the sham operation group,only the left ureter was isolated,and in the other groups,renal fibrosis model was prepared by unilateral ureteral ligation.After 4 weeks of continuous drug intervention,the 24 hours urine protein,Scr,UREA,Cys-c of the mice were detected,the kidney index was calculated,HE staining and Masson staining were used to observe pathological changes of renal tissue and the degree of renal fibrosis were observed,the number of autophagosomes in mouse cells was observed by electron microscopy,Western Blot was used to detect the protein expressions of P62,Beclin-1,LC3A/B,PTEN,p-Akt,Akt,p-mTOR,mTOR in kidney tissue.Results:Compared with the sham operation group,the kidney index,24 hours urinary protein,Scr and Cys-C levels in the model group were significantly increased(P<0.05 or P<0.01),and the kidney structure was destroyed with obrious fibrosis.The protein expressions of LC3A/B,P62,Beclin-1,p-mTOR,mTOR,p-Akt were significantly decreased(P<0.05 or P<0.01).Compared with model group,the 24 hours urine protein,Scr of Yangshen formula groups were significantly decreased(P<0.05 or P<0.01),the damage degree of renal tissue structure was reduced,and the number of outophagosomes and LC3A/B protein expression were significantly increased.The protein expression of Akt in kidney was significantly decreased,the protein expressions of mTOP,p-mTOR in low-dose group were significantly increased(P<0.05 or P<0.01).Conclusion:Yangshen formula can improve renal function and inhibit renal fibrosis of renal fibrosis mice,the mechanism may be related to the regulation of PTEN/Akt/mTOR pathway related protein expression and activation of autophagy.

关 键 词：养肾方 慢性肾脏病 肾纤维化 AKT 

分 类 号：R285.5[医药卫生—中药学]