荜茇酰胺抑制肝星状细胞焦亡发挥肝保护作用及关键分子对接研究  

Study on the Hepatoprotective Effect and Key Molecule Docking of Piperlongumine in Inhibiting Hepatic Stellate Cell Pyroptosis

作　　者：赵丽 郭梓琪 杨勇 杨宏新 ZHAO Li;GUO Ziqi;YANG Yong(Department of Basic Medical,The Inner Mongolia Medical University,Inner Mongolia 010110,China)

机构地区：[1]内蒙古医科大学基础医学院,呼和浩特010110

出　　处：《医学研究杂志》2024年第10期39-45,共7页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81960772);内蒙古自治区卫生健康委员会科技计划项目(202202010);内蒙古医科大学重点项目(YKD2022ZD016);内蒙古医科大学肿瘤类器官基因编辑转化医学科技创新团队项目。

摘　　要：目的 探究荜茇提取物中有效成分荜茇酰胺(piperlongumine, PL)在体外对肝星状细胞损伤的保护作用及机制。方法 通过加入脂多糖(lipopolysaccharide, LPS)对大鼠肝星状细胞(HSC-T6)造模激活,用不同浓度的荜茇酰胺处理肝星状细胞,采用Western blot法和qPCR法检测细胞中IL-18、GSDMD、NLRP3、caspase-1的蛋白及mRNA表达水平;免疫荧光染色技术显示IL-18、GSDMD、NLRP3、caspase-1在细胞中定位及表达情况;采用分子对接技术分析荜茇酰胺作用的靶点分子。结果体外实验显示,荜茇酰胺能抑制IL-18、GSDMD、NLRP3、caspase-1的蛋白及mRNA表达,呈现浓度依赖性;分子对接的结果验证了其相关性,荜茇酰胺可与IL-18、GSDMD、caspase-1结合发挥作用,与NLRP3结合效果较差。结论 荜茇酰胺可通过阻遏HSC-T6细胞的caspase-1信号通路阻止细胞焦亡发生,从而发挥肝脏损伤的保护作用。Objective Exploring the protective effect and mechanism of piperlongumine(PL),an active ingredient in the extract of Piper longum,on hepatic stellate cell damage in vitro.Methods The liver stellate cells(HSC-T6)were activated by adding lipopolysaccharide(LPS),and the liver stellate cells were treated with different concentrations of piperlongumine,and the protein and mRNA expression levels of IL-18,GSDMD,NLRP3 and caspase-1 in the cells were detected by Western blot and qPCR.Immunofluorescence staining showed the localization and expression of IL-18,GSDMD,NLRP3 and caspase-1 in cells.Molecular docking technology was used to analyze the target molecules of the action of piperlongumine.Results In vitro experiments showed that piperlongumine could inhibit the protein and mRNA expression of IL-18,GSDMD,NLRP3 and caspase-1 in a concentration-dependent manner.The results of molecular docking verified the correlation,and piperlongumine could bind to IL-18,GSDMD and caspase-1,but had a poor binding effect with NLRP3.Conclusion Piperlongumine can prevent pyroptosis by inhibiting the caspase-1 signaling pathway of HSC-T6 cells,thereby exerting a protective effect on liver injury.

关 键 词：荜茇酰胺 细胞焦亡 肝星状细胞 分子对接 GSDMD 

分 类 号：R331[医药卫生—人体生理学]