Xue Wenjing, Liu Haiqing, Su Ziheng, Wang Siqi, Cheng Junping, Pan Yunzhi, Zhang Lurong
Key Laboratory for Evaluation and Transformation of Wu Men Medical School's Empirical Prescriptions, Suzhou Traditional Chinese Medicine Hospital, Affiliated to Nanjing University of Chinese Medicine, Suzhou, China.
Department of Pharmacy, Affiliated Hospital of Hebei University, Baoding, China.
Front Pharmacol. 2025 Jan 24;16:1525914. doi: 10.3389/fphar.2025.1525914. eCollection 2025.
Qinggan Yipi Capsule (QgYp) is a hospital preparation that has been used for many years in the treatment of chronic liver diseases. However, the mechanism of QgYp in ameliorating hepatic fibrosis (HF) remains unclear. This study aims to clarify the anti-liver fibrosis effect of QgYp and its mechanism of action.
This study uses a carbon tetrachloride (CCl) induced HF rat model and TGF-β1 stimulated HSC-T6 cell line (rat HSCs) as experimental models. The therapeutic effects were evaluated through pathology, biochemical tests, and ELISA. The therapeutic mechanism of QgYp for HF was predicted through network pharmacology. The expression of TGF-β1/Smad2/3 related proteins was detected by qPCR analysis and Western blot analysis. The composition of the gut microbiota was analyzed using 16S rRNA gene sequencing.
Histopathological analysis, serum biochemical tests, and ELISA measurements showed that QgYp effectively decreased the levels of ALT, AST, HA, LN, PCIII, and IV-C while improving collagen deposition and hepatocyte necrosis. Protein-protein interaction (PPI) network analysis screened HF-related genes, including peroxisome proliferator-activated receptor gamma (PPARG), tumor necrosis factor (TNF), and TGF-β1. GO and KEGG analyses indicated that QgYp significantly affects TGF-β signaling pathway. In addition, the results of qPCR and Western blot analysis from both and experiments indicated that QgYp significantly downregulated the expression of proteins and mRNA associated with the TGF-β1/Smad2/3 pathway. The 16S rDNA gene sequencing results showed that QgYp can increase the diversity and richness of the gut microbiota in HF rats and alter the composition of the gut microbiota.
QgYp could effectively ameliorate HF, and this effect might be connected to the downregulation of the TGF-β1/Smad2/3 pathway, the suppression of HSCs activation, and regulation of gut microbiota dysbiosis.
清肝抑脾胶囊（QgYp）是一种在治疗慢性肝病方面已使用多年的医院制剂。然而，QgYp改善肝纤维化（HF）的机制尚不清楚。本研究旨在阐明QgYp的抗肝纤维化作用及其作用机制。
本研究采用四氯化碳（CCl）诱导的HF大鼠模型和转化生长因子-β1（TGF-β1）刺激的HSC-T6细胞系（大鼠肝星状细胞）作为实验模型。通过病理学、生化检测和酶联免疫吸附测定（ELISA）评估治疗效果。通过网络药理学预测QgYp治疗HF的机制。采用实时定量聚合酶链反应（qPCR）分析和蛋白质免疫印迹法检测TGF-β1/Smad2/3相关蛋白的表达。利用16S核糖体RNA（rRNA）基因测序分析肠道微生物群的组成。
组织病理学分析、血清生化检测和ELISA测量结果表明，QgYp能有效降低丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、透明质酸（HA）、层粘连蛋白（LN）、Ⅲ型前胶原（PCIII）和Ⅳ型胶原（IV-C）水平，同时改善胶原沉积和肝细胞坏死。蛋白质-蛋白质相互作用（PPI）网络分析筛选出与HF相关的基因，包括过氧化物酶体增殖物激活受体γ（PPARG）、肿瘤坏死因子（TNF）和TGF-β1。基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析表明，QgYp显著影响TGF-β信号通路。此外，体内和体外实验的qPCR和蛋白质免疫印迹分析结果表明，QgYp显著下调与TGF-β1/Smad2/3通路相关的蛋白质和信使核糖核酸（mRNA）的表达。16S核糖体DNA（rDNA）基因测序结果表明，QgYp可增加HF大鼠肠道微生物群的多样性和丰富度，并改变肠道微生物群的组成。
QgYp可有效改善HF，这种作用可能与下调TGF-β1/Smad2/3通路、抑制肝星状细胞激活以及调节肠道微生物群失调有关。
