Lu Huan, Luo Yuezhong, Su Baolin, Tang Shuifu, Chen Gangyi, Zhang Licai, Song Chao, Wang Chao, Tu Haitao, Wu Xinbo
Department of Nephrology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
Graduate College, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
Evid Based Complement Alternat Med. 2020 May 12;2020:6813760. doi: 10.1155/2020/6813760. eCollection 2020.
Wenyang Lishui decoction (WYD) has been frequently used to treat patients with membranous nephropathy (MN) in China. Our previous study showed that WYD aqueous extract could alleviate F-actin reorganization of podocytes induced by serum from idiopathic membranous nephropathy (IMN) patients. This study aims to investigate the effects and molecular mechanisms of WYD on MN. MN rat models were induced by cationic bovine serum albumin. Experimental rats were divided into four groups: normal, model, WYD, and benazepril. The normal group consisted of normal rats receiving distilled water for four weeks, while the model, WYD, and benazepril groups consisted of MN rats receiving distilled water, 16.5 g/kg/day WYD aqueous extract, and 10 mg/kg/day benazepril, respectively. Alanine aminotransferase, kidney function, albumin, and 24 h urine total protein (UTP) were measured. Hematoxylin-eosin and electron microscopy analyses were performed. Mouse podocytes were induced to develop cell models by serum from IMN patients with antibody to the M-type phospholipase A2 receptor and spleen and kidney Yang deficiency syndrome. They were divided into five groups: control, model, 2 mg/ml WYD, 4 mg/ml WYD, and 8 mg/ml WYD. CCK-8 assays, flow cytometry, qRT-PCR, and Western blot analyses were performed. In the animal experiment, side effects of WYD were not found. Also, there was no significant difference in kidney function among the groups. In addition, UTP level was significantly reduced, and kidney histological damage was restored in both WYD and benazepril groups but difference in UTP level between them was not found. In the cell experiment, apoptosis rate was increased in the model group while it was decreased by coincubation with WYD. Besides, mRNA and protein levels of p53 were decreased, and those of Bcl-2 were increased by treatment using WYD. In conclusion, WYD could reduce proteinuria and ameliorate podocyte injury by regulating the expression of p53 and Bcl-2. The study is registered in the Chinese Clinical Trial Registry (ChiCTR-OCH-14005137).
温阳利水汤（WYD）在中国常用于治疗膜性肾病（MN）患者。我们之前的研究表明，WYD水提取物可减轻特发性膜性肾病（IMN）患者血清诱导的足细胞F-肌动蛋白重排。本研究旨在探讨WYD对MN的作用及分子机制。采用阳离子牛血清白蛋白诱导MN大鼠模型。将实验大鼠分为四组：正常组、模型组、WYD组和苯那普利组。正常组由接受蒸馏水四周的正常大鼠组成，而模型组、WYD组和苯那普利组分别由接受蒸馏水、16.5 g/kg/天WYD水提取物和10 mg/kg/天苯那普利的MN大鼠组成。检测丙氨酸氨基转移酶、肾功能、白蛋白和24小时尿总蛋白（UTP）。进行苏木精-伊红染色和电子显微镜分析。用M型磷脂酶A2受体抗体和脾肾阳虚证IMN患者血清诱导小鼠足细胞建立细胞模型。将其分为五组：对照组、模型组、2 mg/ml WYD组、4 mg/ml WYD组和8 mg/ml WYD组。进行CCK-8检测、流式细胞术、qRT-PCR和蛋白质印迹分析。在动物实验中，未发现WYD有副作用。此外，各组间肾功能无显著差异。此外，WYD组和苯那普利组的UTP水平均显著降低，肾脏组织学损伤均得到恢复，但两组UTP水平差异未发现。在细胞实验中，模型组凋亡率升高，而与WYD共同孵育可降低凋亡率。此外，WYD处理可降低p53的mRNA和蛋白水平，增加Bcl-2的mRNA和蛋白水平。总之，WYD可通过调节p53和Bcl-2的表达来降低蛋白尿并改善足细胞损伤。该研究已在中国临床试验注册中心注册（ChiCTR-OCH-14005137）。
