Zhang Wenwen, Wang Zexin, Sun Rong, Zeng Yi, Chen Yuan, Hu Qichao, Chen Lisheng, Ma Xiao, Guo Yaoguang, Zhao Yanling
State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Department of Pharmacy, Chinese PLA General Hospital, Beijing, China.
Inflammation. 2025 Jan 27. doi: 10.1007/s10753-025-02245-0.
Cholestasis is a multifactorial hepatobiliary disorder, characterized by obstruction of bile flow and accumulation of bile, which in turn causes damage to liver cells and other tissues. In severe cases, it can result in the development of life-threatening conditions, including cirrhosis and liver cancer. Paeoniflorin (PF) has been demonstrated to possess favourable therapeutic potential for the treatment of cholestasis. The objective of this research was to examine the molecular mechanism of PF in the treatment of ANIT-induced cholestasis and to propose novel avenues for further research on the pharmacological effects of PF. In vivo and in vitro models of cholestasis were developed. The histopathological changes in the bile ducts and liver were evaluated through the use of hematoxylin and eosin (HE) staining. The extent of apoptosis was evaluated through the use of immunofluorescence (IF), immunoblotting (WB), and electron microscopy. The JNK signalling pathway was identified as the direct mechanism of action of PF through the utilisation of HuProt™ 20 K chips and other technologies. The present study demonstrated that PF markedly alleviated liver injury in an ANIT-induced cholestasis model. Specifically, PF was observed to attenuate cholestasis-induced liver injury by reducing the abnormal elevation of liver function indices and suppressing the expression of inflammatory mediators. Furthermore, PF exhibited anti-apoptotic properties in both in vivo and in vitro experiments, thereby mitigating cholestasis-induced hepatocyte apoptosis. These protective effects are attributable to the fact that PF exerts its action through direct interaction with the JNK pathway. It has been demonstrated that PF is capable of binding directly to MAPK8 (JNK1) and MAPK9 (JNK2), thereby inhibiting JNK activation and reducing apoptosis. With regard to the protection of bile ducts, PF may indirectly inhibit hepatocyte apoptosis by maintaining the structural integrity and tight junctions of bile duct cells. PF improved cholestasis by inhibiting hepatocyte apoptosis directly by targeting the JNK signaling pathway and indirectly inhibited hepatocyte apoptosis by improving the tight junctions of bile duct cells to regulate the bile duct microenvironment.
胆汁淤积是一种多因素引起的肝胆疾病，其特征是胆汁流动受阻和胆汁积聚，进而导致肝细胞和其他组织受损。在严重情况下，它可导致危及生命的疾病，包括肝硬化和肝癌。芍药苷（PF）已被证明在治疗胆汁淤积方面具有良好的治疗潜力。本研究的目的是探讨PF治疗ANIT诱导的胆汁淤积的分子机制，并为进一步研究PF的药理作用提出新的途径。建立了胆汁淤积的体内和体外模型。通过苏木精和伊红（HE）染色评估胆管和肝脏的组织病理学变化。通过免疫荧光（IF）、免疫印迹（WB）和电子显微镜评估细胞凋亡程度。通过使用HuProt™ 20K芯片和其他技术，确定JNK信号通路为PF的直接作用机制。本研究表明，PF在ANIT诱导的胆汁淤积模型中显著减轻了肝损伤。具体而言，观察到PF通过降低肝功能指标的异常升高和抑制炎症介质的表达来减轻胆汁淤积诱导的肝损伤。此外，PF在体内和体外实验中均表现出抗凋亡特性，从而减轻胆汁淤积诱导的肝细胞凋亡。这些保护作用归因于PF通过与JNK途径直接相互作用发挥作用。已证明PF能够直接结合丝裂原活化蛋白激酶8（JNK1）和丝裂原活化蛋白激酶9（JNK2），从而抑制JNK活化并减少细胞凋亡。关于胆管的保护，PF可能通过维持胆管细胞的结构完整性和紧密连接间接抑制肝细胞凋亡。PF通过靶向JNK信号通路直接抑制肝细胞凋亡来改善胆汁淤积，并通过改善胆管细胞的紧密连接间接抑制肝细胞凋亡来调节胆管微环境。
