荜茇酰胺对四氯化碳诱导肝纤维化大鼠模型肠道屏障功能障碍的影响

doi:10.19803/j.1672-8629.20260012

摘要/Abstract

摘要： 目的探讨荜茇酰胺（Piperlongumine,PPLGM）对四氯化碳（CCl₄）诱导的大鼠肝纤维化的作用机制及肠道屏障保护作用。方法将50只SD大鼠随机分为对照组,模型组及PPLGM低、中、高剂量组（2.5、5、10 mg·kg^-1）。除对照组外,其余各组均采用皮下注射50% CCl₄-橄榄油溶液构建肝纤维化模型。造模成功后,各给药组予以相应剂量的PPLGM灌胃治疗4周。干预结束后,腹主动脉取血并采集肝、肠组织。通过苏木精-伊红（HE）和Masson染色评估肝、肠组织病理变化及肝脏胶原沉积;生化法检测血清丙氨酸氨基转氨酶（ALT）、天冬氨酸氨基转移酶（AST）肝功能指标;免疫组化、蛋白免疫印迹（Westen Blot,WB）和RT-qPCR技术检测肝肠组织中TLR4-NF-κB信号通路关键分子、纤维化指标（α-SMA, Collagen Ⅰ）及肠道紧密连接蛋白（Claudin-1）的表达水平;ELISA法检测血清中内毒素（LPS）含量。分子对接技术模拟分析PPLGM与TLR4、NF-κB、TNF-α的结合能力。结果与正常组相比,模型组大鼠肝组织α-SMA与CollagenⅠ表达上调,血清ALT、AST含量升高,同时伴有结肠组织Claudin-1蛋白表达下调及血清LPS水平升高（P<0.05）。免疫组化和WB实验结果显示肝肠组织中TLR4、NF-κB和TNF-α表达均显著上升（P<0.05）。PPLGM干预后,上述异常得到显著逆转：肝纤维化病理特征明显改善,血清ALT、AST显著下降,α-SMA、Collagen Ⅰ及通路（TLR4/NF-κB/TNF-α）表达下调,Claudin-1表达回升,血清LPS水平降低（P<0.05）。分子对接结果显示PPLGM与TLR4、NF-κB、TNF-α靶点的结合能均< -5 kcal·mol^-1。结论 PPLGM可显著改善CCl₄诱导的肝纤维化及肠道屏障功能障碍,其抗纤维化作用的核心机制在于调控“肠-肝轴”。该化合物可能通过抑制肠道屏障损伤所触发的TLR4/NF-κB信号通路,阻断肠道源性炎性信号向肝脏的传递,从而减轻肝脏炎性微环境与纤维化进程。

关键词: 荜茇酰胺, 肝纤维化, TLR4, 肠肝轴, 血清中内毒素（LPS）, 肠道屏障, 大鼠

Abstract: Objective To investigate the mechanism of piperlongumine (PPLGM) against carbon tetrachloride (CCl₄)-induced liver fibrosis in rats and its protective effect on the intestinal barrier. Methods Fifty SD rats were randomly divided into the control group, model group, and PPLGM low, medium and high-dose groups (2.5, 5, 10 mg·kg^-1). Except the control group, all these groups were subcutaneously injected with 50% CCl₄-olive oil solution to construct a liver fibrosis model. After modeling, each administration group was given the corresponding dose of PPLGM by gavage for 4 weeks. After the intervention, blood was taken from the abdominal aorta before liver and intestinal tissues were collected. Hematoxylin and Eosin (HE) and Masson staining were used to assess the pathological changes of liver and intestinal tissues and liver collagen deposition. Such liver function indexes as serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected using biochemical methods. Immunohistochemistry, Western blot, and RT-qPCR techniques were adopted to detect expression levels of key molecules in the TLR4-NF-κB signaling pathway, fibrosis markers (α-SMA, CollagenⅠ), and intestinal tight junction protein (Claudin-1) in liver and intestinal tissues. The content of endotoxin (LPS) in serum was detected by ELISA. The binding ability of PPLGM to TLR4, NF-κB and TNF-α was analyzed via molecular docking technology. Results Compared with the control group, α-SMA and collagenⅠ expression levels were upregulated in liver tissues and serum ALT and AST levels were elevated in the model group, accompanied by downregulation of claudin-1 protein expressions in colon tissues and increases in serum endotoxin levels (P<0.05). Immunohistochemistry and Western blot analysis suggested significantly increased expression levels of TLR4, NF-κB, and TNF-α in liver and intestinal tissues (P<0.05). Following treatment with PPLGM, the above abnormalities were significantly reversed in that liver fibrosis was markedly improved pathologically, serum ALT and AST levels were decreased significantly, expression levels of α-SMA, collagen Ⅰ, and the TLR4/NF-κB/TNF-α pathway were downregulated, claudin-1 expression was recovered, and serum LPS levels were decreased (P<0.05). Molecular docking results indicated that the binding affinity of PPLGM to TLR4, NF-κB, and TNF-α was less than -5 kcal·mol^-1. Conclusion PPLGM can significantly improve CCl₄-induced liver fibrosis and intestinal barrier dysfunction by regulating the gut-liver axis. This compound may block the transmission of intestinal-derived inflammatory signals to the liver by inhibiting the TLR4/NF-κB signaling pathway triggered by intestinal barrier damage, thereby reducing the liver inflammatory microenvironment and fibrosis process.

Key words: Piperlongumine, Liver Fibrosis, TLR4, Enterohepatic Axis, Endotoxin（LPS）, Intestinal Barrier, Rat

中图分类号:

张宁芳, 赵丽, 杨勇, 杨宏新. 荜茇酰胺对四氯化碳诱导肝纤维化大鼠模型肠道屏障功能障碍的影响[J]. 中国药物警戒, 2026, 23(10): 1123-1130.

Zhang Ningfang, Zhao Li, Yang Yong, Yang Hongxin. Effects of piperlongumine on gut barrier dysfunction in a rat model of carbon tetrachloride-induced liver fibrosis[J]. Chinese Journal of Pharmacovigilance, 2026, 23(10): 1123-1130.

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