College of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, 450046, China.
The Engineering and Technology Center for Chinese Medicine Development of Henan Province, 156 Jinshui East Road, Zhengzhou, 450046, China.
BMC Complement Med Ther. 2023 Jun 23;23(1):207. doi: 10.1186/s12906-023-04012-w.
Renal fibrosis with Renin-angiotensin-aldosterone system (RAAS) activation and oxidative stress are one of the major complications in hypertension. 2-phenylacetamide (PA), a major active component of Lepidium apetalum Willd. (L.A), has numerous pharmacological effects. Its analogues have the effect of anti-renal fibrosis and alleviating renal injury. This study aims to explore the underlying mechanism of PA for regulating the renal fibrosis in SHR based on the MAPK pathway mediated RAAS and oxidative stress.
The SHR rats were used as the hypertension model, and the WKY rats were used as the control group. The blood pressure (BP), urine volume were detected every week. After PA treatment for 4 weeks, the levels of RAAS, inflammation and cytokines were measured by Enzyme-Linked Immunosorbnent Assay (ELISA). Hematoxylin-Eosin staining (HE), Masson and Immunohistochemistry (IHC) were used to observe the renal pathology, collagen deposition and fibrosis. Western blot was used to examine the MAPK pathway in renal. Finally, the SB203580 (p38 MAPK inhibitor) antagonism assay in the high NaCl-induced NRK52e cells was used, together with In-Cell Western (ICW), Flow Cytometry (FCM), High Content Screening (HCS) and ELISA to confirm the potential pharmacological mechanism.
PA reduced the BP, RAAS, inflammation and cytokines, promoted the urine, and relieved renal pathological injury and collagen deposition, repaired renal fibrosis, decreased the expression of NADPH Oxidase 4 (NOX4), transforming growth factor-β (TGF-β), SMAD3 and MAPK signaling pathway in SHR rats. Meanwhile,,the role of PA could be blocked by p38 antagonist SB203580 effectively in the high NaCl-induced NRK52e cells. Moreover, molecular docking indicated that PA occupied the ligand binding sites of p38 MAPK.
PA inhibited renal fibrosis via MAPK signalling pathway mediated RAAS and oxidative stress in SHR Rats.
肾纤维化伴肾素-血管紧张素-醛固酮系统（RAAS）激活和氧化应激是高血压的主要并发症之一。2-苯乙酰胺（PA）是 Lepidium apetalum Willd.（L.A）的主要活性成分之一，具有多种药理学作用。其类似物具有抗肾纤维化和减轻肾损伤的作用。本研究旨在探讨 PA 通过 MAPK 通路介导的 RAAS 和氧化应激调节 SHR 肾纤维化的潜在机制。
用 SHR 大鼠作为高血压模型，WKY 大鼠作为对照组。每周检测血压（BP）、尿量。PA 治疗 4 周后，用酶联免疫吸附试验（ELISA）检测 RAAS、炎症和细胞因子水平。用苏木精-伊红染色（HE）、Masson 和免疫组织化学（IHC）观察肾脏病理、胶原沉积和纤维化。用 Western blot 检测肾内 MAPK 通路。最后，用 SB203580（p38 MAPK 抑制剂）拮抗高盐诱导的 NRK52e 细胞，结合 In-Cell Western（ICW）、流式细胞术（FCM）、高内涵筛选（HCS）和 ELISA 验证潜在的药理学机制。
PA 降低 BP、RAAS、炎症和细胞因子，促进尿，缓解 SHR 大鼠肾病理损伤和胶原沉积，修复肾纤维化，降低 NADPH 氧化酶 4（NOX4）、转化生长因子-β（TGF-β）、SMAD3 和 MAPK 信号通路表达。同时，p38 拮抗剂 SB203580 可有效阻断 PA 在高盐诱导的 NRK52e 细胞中的作用。此外，分子对接表明 PA 占据了 p38 MAPK 的配体结合位点。
PA 通过 MAPK 信号通路介导的 RAAS 和氧化应激抑制 SHR 大鼠肾纤维化。
