目的 基于单磷酸腺苷活化的蛋白激酶（adenosine monophosphate-activated protein kinase，AMPK）/沉默信息调节因子1（silent information regulator 1，SIRT1）/固醇调节元件结合蛋白1（sterol regulatory element-binding protein 1，SREBP1）信号通路观察膜肾颗粒对膜性肾病大鼠蛋白尿的影响及作用机制。方法 80只SD大鼠随机挑选15只为空白组，其余大鼠利用改良Border法建立膜性肾病大鼠模型，造模大鼠随机分为模型组、环孢素A（cyclosporin A，CsA）组和膜肾颗粒低、高剂量（1.85、7.38 g/kg）组。观察大鼠一般状态，采用双缩脲比色法检测大鼠造模前、造模后、给药结束后24 h尿蛋白定量（urinary protein quantity，UTP）；使用全自动生化仪检测大鼠尿素氮（blood urea nitrogen，BUN）、血肌酐（serum creatinine，Scr）、总胆固醇（total cholesterol，TC）、三酰甘油（triglyceride，TG）水平；采用苏木素-伊红（hematoxylin-eosin，HE）、基底膜六胺银染色（periodic acid-silver methenamine，PASM）染色观察肾脏病理学变化；采用Western blotting检测大鼠肾组织Nephrin、AMPK/SIRT1/SREBP1通路相关蛋白表达。结果 各组BUN、Scr水平无明显差异。与空白组比较，模型组大鼠24 h-UTP、TC、TG水平明显升高（P<0.01），肾组织病理改变明显，肾组织Nephrin、p-AMPKα/AMPKα和SIRT1表达显著降低（P<0.01），SREBP1蛋白表达水平显著升高（P<0.01）。与模型组比较，各给药组24 h-UTP、TC、TG水平明显降低（P<0.01），肾组织病理改变减轻，Nephrin、p-AMPKα/AMPKα、SIRT1蛋白表达水平明显升高（P<0.05、0.01），SREBP1蛋白表达水平显著降低（P<0.01）。结论 膜肾颗粒可以显著减少膜性肾病大鼠蛋白尿水平，可能与上调肾组织AMPKα/SIRT1信号通路表达，减少SREBP1合成，改善脂代谢重编程有关。

Objective To observe the effect and mechanism of Moshen Granules (膜肾颗粒) on proteinuria in rats with membranous nephropathy based on adenosine monophosphate-activated protein kinase (AMPK)/silent information regulator 1 (SIRT1)/sterol regulatory element-binding protein 1 (SREBP1) signal pathway. Methods Fifteen of 80 SD rats were randomly selected as the blank group, and the rest rats were used to establish the model with membranous nephropathy by the improved Border method. The rats were randomly divided into model group, cyclosporin A (CsA) group and low-, high-dose Moshen Granules (1.85, 7.38 g/kg) groups. The general state of rats was observed, and the urinary protein quantity (UTP) for 24 h was detected by biuret colorimetry before modeling, after modeling and after drug administration. The levels of blood urea nitrogen (BUN), serum creatinine (Scr), total cholesterol (TC) and triglyceride (TG) in rats were detected by automatic biochemical analyzer. Hematoxylin-eosin (HE) and periodic acid-silver methenamine (PASM) staining were used to observe the pathological changes of kidney. The expressions of Nephrin, AMPK/SIRT1/SREBP1 pathway-related proteins in rat kidney were detected by Western blotting. Results There was no significant difference in BUN and Scr levels among the groups. Compared with blank group, the levels of 24 h-UTP, TC and TG in model group were significantly increased (P < 0.01), the pathological changes of renal tissue were obvious, the expressions of Nephrin, p-AMPKα/AMPKα and SIRT1 in renal tissue were significantly decreased (P < 0.01), and the expression level of SREBP1 protein was significantly increased (P < 0.01). Compared with model group, the levels of 24 h-UTP, TC and TG in each treatment group were significantly decreased (P < 0.01), the pathological changes of renal tissue were alleviated, the expression levels of Nephrin, p-AMPKα/AMPKα and SIRT1 proteins were significantly increased (P < 0.05, 0.01), and the expression level of SREBP1 protein was significantly decreased (P < 0.01). Conclusion Moshen Granules can significantly reduce the proteinuria level in rats with membranous nephropathy, which may be related to up-regulating the expression of AMPKα/SIRT1 signal pathway in renal tissue, reducing the synthesis of SREBP1 and improving lipid metabolism reprogramming.

R285.5

国家重点研发计划项目（2019YFC1709401）；天津市卫生健康委员会天津市中医药管理局中医中西医结合科研课题（2021012）