降脂通络软胶囊调控AMPK/mTOR信号通路对膜性肾病大鼠自噬的影响  

Effect of Jiangzhi Tongluo Soft Capsules on Autophagy in Rats with Membranous Nephropathy via AMPK/mTOR Signaling Pathway

作　　者：郝文霞 高飞[1,2] 赵方 袁国栋 张倩[2] 檀金川 Hao Wenxia;Gao Fei;Zhao Fang;Yuan Guodong;Zhang Qian;Tan Jinchuan(Hebei University of Chinese Medicine,Shijiazhuang 050200;Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011)

机构地区：[1]河北中医学院,石家庄050200 [2]河北省中医院,石家庄050011

出　　处：《中药药理与临床》2021年第4期148-153,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：河北省中医药管理局资助项目(编号:2017027)。

摘　　要：目的:探讨降脂通络软胶囊对膜性肾病(MN)大鼠肾脏的干预作用及对足细胞自噬活性的影响。方法:60只SD大鼠随机抽取10只作为正常对照组,其余大鼠复制MN模型,造模成功后随机将大鼠分为模型对照组、盐酸贝那普利10 mg/kg组、降脂通络软胶囊25、50、100 mg/kg组。造模成功大鼠每日灌胃给药,连续给药4 w。末次药后检测各组大鼠24 h尿蛋白含量(UTP)、血清总胆固醇(TC)、三酰甘油(TG)、白蛋白(ALB)、尿素氮(BUN)、肌酐(Scr)水平变化,采用HE、PASM染色及电镜观察肾脏病理改变,免疫荧光观察肾组织IgG沉积,Western Blot法检测AMPK/mTOR信号通路相关蛋白表达,免疫组化法检测自噬蛋白LC3、Beclin-1、P62表达水平。结果:与正常对照组比较,模型对照组大鼠UTP、TC、TG含量显著升高,ALB含量显著下降(P<0.01);肾小球体积明显增大,PASM染色见基底膜不规则增厚,钉突形成,电镜示上皮细胞下电子致密物不规则沉积,足突广泛融合,免疫荧光染色示IgG沿系膜区及毛细血管壁呈颗粒样沉积。p-AMPK/AMPK比值显著降低(P<0.01),p-mTOR/mTOR、p-S6K/S6K比值显著升高(P<0.01);自噬相关蛋白LC3、Beclin-1表达显著下调,P62蛋白表达上调(P<0.01)。与模型对照组比较,各给药组UTP、TC、TG含量均显著下降,ALB含量显著升高(P<0.01);大鼠肾组织病理损伤均有缓解,以降脂通络软胶囊50、100 mg/kg组改善更加明显;p-AMPK/AMPK比值显著升高(P<0.01),p-mTOR/mTOR、p-S6K/S6K比值明显降低(P<0.05);自噬相关蛋白LC3、Beclin-1表达明显上调,P62蛋白表达明显下调(P<0.05)。结论:降脂通络软胶囊可能通过AMPK-mTOR通路增强足细胞自噬活性,减轻肾脏病理损伤,降低尿蛋白,延缓MN进展。Objective:To explore the effect of Jiangzhi Tongluo Soft Capsules(JZTL) on the kidney of membranous nephropathy(MN) rats and podocyte autophagic activity. Methods:Sixty SD rats were randomly divided into a normal control group(n = 10) and an experimental group(n = 50) for MN model induction. The model rats were then divided into a model group, a benazepril hydrochloride group(10 mg/kg), and low-(25 mg/kg/d), medium-(50 mg/kg/d), and high-dose(100 mg/kg/d) JZTL groups. Rats were administered intragastrically with corresponding drugs for four weeks. After the last administration, the levels of 24 h urine total protein(UTP), serum total cholesterol(TC), triacylglycerol(TG), albumin(ALB), urea nitrogen(BUN), and creatinine(SCr) were detected. HE and PASM staining and the electron microscopy were used to observe the pathological changes of the kidney, immunofluorescence assay to observe IgG deposition in renal tissues, Western blot to detect the protein expression related to the AMPK/mTOR signaling pathway, and immunohistochemistry to detect the expression levels of autophagy-related proteins LC3, Beclin-1, and P62. Results:Compared with the normal control group, the model group showed increased content of UTP, TC, and TG, decreased ALB content(P<0.01), and enlarged glomerular volume. The model group also displayed irregular thickening of the basement membrane and formation of spikes indicated by PASM staining, irregular deposition of electron compacts under epithelial cells and extensive fusion of foot processes revealed by the electron microscopy, and IgG deposition in a granular form along the mesangial area and capillary walls demonstrated by the immunofluorescence staining. Furthermore, p-AMPK/AMPK was down-regulated(P<0.01), while p-mTOR/mTOR and p-S6 K/S6 K were up-regulated(P<0.01);the expression of autophagy-related proteins LC3 and Beclin-1 was down regulated, while the expression of P62 was up regulated(P<0.01). Compared with the model group, the benazepril hydrochloride group and the JZTL groups showed dec

关 键 词：降脂通络软胶囊 膜性肾病 AMP依赖的蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路 自噬 

分 类 号：R285.5[医药卫生—中药学]