Feng Qunying, Zhao Qinghua, Qu Shuaiyong, Zhao Yiju, Li Kunlun, Yuan Bo, Chang Qinzheng, Xu Jingjiang, Wang Hongxia, Zhu Yongqin, Fu Kai, Liu Jingsheng
Kaifeng Hospital of Traditional Chinese Medicine, Kaifeng, Henan, 475000, China.
Kaifeng Hospital of Traditional Chinese Medicine, Kaifeng, Henan, 475000, China.
J Ethnopharmacol. 2025 Jan 30;337(Pt 2):118893. doi: 10.1016/j.jep.2024.118893. Epub 2024 Oct 2.
Liver fibrosis is a critical pathological process in the progression of chronic liver injury, ultimately resulting in cirrhosis, for which currently available therapeutic interventions remain inadequate. Among these, the Qianggan Ruanjian Pill (QGRJP) has emerged as a clinically experienced formula with notable therapeutic efficacy against liver fibrosis. However, the precise underlying mechanisms require further investigation.
In this study, we investigated the key pathways and target genes of QGRJP that attenuate liver fibrosis and elucidated the underlying mechanisms.
High-performance liquid chromatography-mass spectrometry (HPLC-MS) was used to identify the major components of the QGRJP. Mouse models of liver fibrosis were established by injecting olive oil containing 25% carbon tetrachloride (CCl4), which was administered at different doses of QGRJP by gavage. Liver damage and function were assessed using serum biochemical detection, ultrasound imaging, and histopathological examination. The anti-fibrosis effect was assessed using immunohistochemistry, western blotting, and quantitative real-time PCR (qRT-PCR). The in vivo safety of the QGRJP was evaluated using weight monitoring and biopsy. Potential anti-liver fibrosis signalling pathways and key targets of QGRJP were identified using RNA-seq analysis and network pharmacology. The predicted targets and pathways were validated using in vitro and in vivo experiments.
QGRJP significantly ameliorated CCl-induced liver fibrosis, and its mechanism was correlated with the inhibition of hepatic stellate cell (HSC) activation and the inflammatory response via inhibition of the TGF-β1/Smad and PI3K/AKT pathways, leading to a significant reduction in the expression of collagen and other fibrosis-related proteins. Additionally, no obvious toxic side effects were observed in the major organs of the mice or in activated HSCs (aHSCs).
This study demonstrated that QGRJP mitigated liver injury, inflammation, and fibrosis by inhibiting the TGF-β1/Smad and PI3K/AKT signalling pathways.
肝纤维化是慢性肝损伤进展中的一个关键病理过程，最终会导致肝硬化，目前可用的治疗干预措施仍然不足。其中，强肝软坚丸（QGRJP）已成为一种临床经验丰富的方剂，对肝纤维化具有显著的治疗效果。然而，其确切的潜在机制仍需进一步研究。
在本研究中，我们研究了QGRJP减轻肝纤维化的关键途径和靶基因，并阐明了其潜在机制。
采用高效液相色谱-质谱联用（HPLC-MS）法鉴定QGRJP的主要成分。通过注射含25%四氯化碳（CCl4）的橄榄油建立肝纤维化小鼠模型，并通过灌胃给予不同剂量的QGRJP。使用血清生化检测、超声成像和组织病理学检查评估肝损伤和肝功能。使用免疫组织化学、蛋白质印迹和定量实时聚合酶链反应（qRT-PCR）评估抗纤维化作用。通过体重监测和活检评估QGRJP的体内安全性。使用RNA测序分析和网络药理学确定QGRJP潜在的抗肝纤维化信号通路和关键靶点。使用体外和体内实验验证预测的靶点和途径。
QGRJP显著改善了CCl诱导的肝纤维化，其机制与通过抑制TGF-β1/Smad和PI3K/AKT途径抑制肝星状细胞（HSC）活化和炎症反应有关，导致胶原蛋白和其他纤维化相关蛋白的表达显著降低。此外，在小鼠的主要器官或活化的肝星状细胞（aHSCs）中未观察到明显的毒副作用。
本研究表明，QGRJP通过抑制TGF-β1/Smad和PI3K/AKT信号通路减轻肝损伤、炎症和纤维化。
