Lin Ziyang, Wang Zhaodi, Kim Thuong Van Pham, Zhang Xianlong, Liang Baien, Li Minyi, Li Mengqiu, Duan Tingting, Li Zhenghai, Li Ping, Wu Aihua, Yang Junzheng, Bao Kun, Liu Bo
The Second Clinical Medical College, and Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome, Guangzhou University of Chinese Medicine, Guangzhou, 510006, PR China; Guangdong Nephrotic Drug Engineering Technology Research Center, Institute of Consun Co. for Chinese Medicine in Kidney Diseases, Guangdong Consun Pharmaceutical Group, Guangzhou, PR China.
The Second Clinical Medical College, and Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome, Guangzhou University of Chinese Medicine, Guangzhou, 510006, PR China.
J Ethnopharmacol. 2025 May 28;348:119813. doi: 10.1016/j.jep.2025.119813. Epub 2025 Apr 14.
Sanqi Qushi Formula (SQQS), a clinically validated derivative of the Sanqi oral solution, integrates principles of traditional Chinese medicine (TCM) to treat membranous nephropathy (MN). Its efficacy in reducing proteinuria and preserving renal function has been observed in clinical practice.
This study aims to elucidate the therapeutic mechanisms, active components, and pathway-specific effects of SQQS in MN, providing a scientific foundation for its clinical use.
The components of SQQS were analyzed using UHPLC-MS/MS. A passive Heymann nephritis (PHN) rat model was induced by intravenous injection of anti-Fx1A serum. Rats received oral SQQS for 3 weeks, and urine/serum samples were collected to evaluate renal function and chemokine levels. Renal histopathology was assessed via immunofluorescence, PASM staining, and CD68 immunostaining. Network pharmacology integrated target prediction for SQQS compounds and differentially expressed genes from the Gene Expression Omnibus (GEO) database (MN patient glomeruli). Mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) pathway proteins and epithelial-mesenchymal transition (EMT) markers were analyzed by western blotting (WB). Molecular docking and molecular dynamics simulations evaluated compound-MEK interactions. Human glomerular podocytes were treated with SQQS-derived compounds; viability and migration were assessed using cell counting kit-8 assay and scratch assays.
UHPLC-MS/MS identified 129 compounds in SQQS. SQQS treatment significantly reduced renal injury markers, glomerular IgG deposition, and basement membrane thickening in PHN rats. GEO database analysis revealed 839 upregulated and 166 downregulated genes in MN glomeruli. Network pharmacology implicated the tumor necrosis factor (TNF) pathway, with 10 upregulated targets (e.g., MAP2K1, MMP3, CXCL10). WB confirmed SQQS suppressed MEK/ERK phosphorylation and decreased MMP3 and α-SMA levels. Renal CD68 macrophages and associated chemokines (CXCL10, CCL20) were reduced by SQQS. Methylnissolin-3-O-glucoside, a flavonoid from Astragalus mongholicus Bunge, dose-dependently inhibited TNF-α-induced MEK/ERK activation and migration. MEK agonists reversed methylnissolin-3-O-glucoside-mediated MEK/ERK suppression.
SQQS ameliorates MN progression by inhibiting the MEK/ERK pathway, suppressing EMT, and reducing macrophage recruitment, with methylnissolin-3-O-glucoside as a key bioactive component.
三七祛湿方（SQQS）是三七口服液经临床验证的衍生物，融合了中医理论用于治疗膜性肾病（MN）。其在降低蛋白尿和保护肾功能方面的疗效已在临床实践中得到观察。
本研究旨在阐明SQQS治疗MN的作用机制、活性成分及通路特异性效应，为其临床应用提供科学依据。
采用超高效液相色谱-串联质谱法（UHPLC-MS/MS）分析SQQS的成分。通过静脉注射抗Fx1A血清诱导建立被动型Heymann肾炎（PHN）大鼠模型。大鼠口服SQQS 3周，收集尿液/血清样本以评估肾功能和趋化因子水平。通过免疫荧光、PASM染色和CD68免疫染色评估肾脏组织病理学。利用网络药理学对SQQS化合物和来自基因表达综合数据库（GEO）（MN患者肾小球）的差异表达基因进行靶点预测。通过蛋白质印迹法（WB）分析丝裂原活化蛋白激酶（MEK）/细胞外信号调节激酶（ERK）通路蛋白和上皮-间质转化（EMT）标志物。分子对接和分子动力学模拟评估化合物与MEK的相互作用。用SQQS衍生化合物处理人肾小球足细胞；使用细胞计数试剂盒-8法和划痕试验评估细胞活力和迁移。
UHPLC-MS/MS鉴定出SQQS中的129种化合物。SQQS治疗显著降低了PHN大鼠的肾损伤标志物、肾小球IgG沉积和基底膜增厚。GEO数据库分析显示MN肾小球中有839个基因上调和166个基因下调。网络药理学表明肿瘤坏死因子（TNF）通路相关，有10个上调靶点（如MAP2K1、MMP3、CXCL10）。WB证实SQQS抑制MEK/ERK磷酸化并降低MMP3和α-SMA水平。SQQS减少了肾脏CD68巨噬细胞及相关趋化因子（CXCL10、CCL20）。黄芪甲苷-3-O-葡萄糖苷，一种来自蒙古黄芪的黄酮类化合物，剂量依赖性地抑制TNF-α诱导的MEK/ERK激活和迁移。MEK激动剂逆转了黄芪甲苷-3-O-葡萄糖苷介导的MEK/ERK抑制作用。
SQQS通过抑制MEK/ERK通路、抑制EMT和减少巨噬细胞募集来改善MN进展，黄芪甲苷-3-O-葡萄糖苷是关键生物活性成分。
