退黄合剂通过调控法尼醇X受体抑制NLRP3炎症小体改善α-萘异硫氰酸酯诱导的大鼠胆汁淤积  

Tuihuang Mixture improvesα-naphthylisothiocyanate-induced cholestasis in rats by inhibiting NLRP3 inflammasomes via regulating farnesoid X receptor

作　　者：朱正望 王琳琳 赵静涵 马瑞雪 余雨春 蔡庆春[3] 王兵 朱平生 苗明三 ZHU Zhengwang;WANG Linlin;ZHAO Jinghan;MA Ruixue;YU Yuchun;CAI Qingchun;WANG Bing;ZHU Pingsheng;MIAO Mingsan(First Clinical Medical College of Henan University of Chinese Medicine,Zhengzhou 450046,China;Collaborative Innovation Center of Research and Development on the Whole Industry Chain of Yu-Yao,Zhengzhou 450046,China;Department of Pathology,Third Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450008,China;Department of Traditional Chinese Medicine,Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200233,China)

机构地区：[1]河南中医药大学第一临床医学院,河南郑州450046 [2]豫药全产业链研发河南省协同创新中心,河南郑州450046 [3]河南中医药大学第三附属医院病理科,河南郑州450008 [4]上海交通大学医学院附属第六人民医院中医科,上海200233

出　　处：《南方医科大学学报》2025年第4期718-724,共7页Journal of Southern Medical University

基　　金：国家自然科学基金(82074340);河南省“双一流”创建学科中医学科学研究专项(HSRP-DFCTCM-2023-1-19,HSRP-DFCTCM-2023-8-32);河南省科技创新人才计划-杰出青年项目(154100510020)。

摘　　要：目的研究退黄合剂通过调控法尼醇X受体(FXR)抑制NLRP3炎症小体活化改善胆汁淤积的作用机制。方法将48只Wistar大鼠随机分为空白组、模型组、熊去氧胆酸组、退黄合剂组(12只/组),除空白组外,其余大鼠采用α-萘异硫氰酸酯(ANIT)灌胃复制胆汁淤积模型。造模后,各组大鼠分别给予相应药物灌胃。检测血清中ALT、AST、ALP、γ-GT、TBA、TBIL水平;肝组织中IL-1β、IL-18水平及mRNA表达;Western blotting和q-PCR检测肝组织中FXR、NLRP3、ASC、Caspase-1、GSDMD蛋白及mRNA表达;HE染色观察肝组织病理学变化。结果与空白组相比,模型组大鼠血清中ALT、AST、ALP、γ-GT、TBA、TBIL水平明显升高,肝组织中IL-1β、IL-18水平及mRNA表达明显升高,FXR蛋白及mRNA表达明显降低,NLRP3、Caspase-1蛋白及NLRP3、ASC、Caspase-1、GSDMD mRNA表达明显升高(P<0.05,P<0.01);肝细胞排列紊乱,胆管上皮细胞增生,炎症细胞浸润。与模型组相比,退黄合剂组大鼠血清中ALT、AST、ALP、γ-GT、TBA、TBIL水平明显降低,肝组织中IL-1β、IL-18水平及mRNA表达明显降低,FXR蛋白及mRNA表达明显升高,NLRP3、ASC、Caspase-1、GSDMD蛋白及NLRP3、ASC、Caspase-1 mRNA表达明显降低(P<0.05,P<0.01);肝细胞损伤、胆管上皮细胞增生、炎症细胞浸润减轻。结论退黄合剂可有效改善胆汁淤积,其机制可能与调控FXR抑制NLRP3炎症小体活化介导的细胞焦亡有关。Objective To study the therapeutic mechanism of Tuihuang Mixture against cholestasis.Methods Forty-eight Wistar rats were randomized equally into blank group,model group,ursodeoxycholic acid group and Tuihuang Mixture group.Except for those in the blank group,all the rats were givenα‑naphthylisothiocyanate(ANIT)to establish rat models of cholestasis,followed by treatments with indicated drugs or distilled water.Serum levels of ALT,AST,ALP,γ-GT,TBA and TBIL of the rats were determined,and hepatic expressions IL-1β,IL-18,FXR,NLRP3,ASC,Caspase-1 and GSDMD were detected using q-PCR,ELISA or Western blotting.Histopathological changes of the liver tissues were observed using HE staining.Results The rat models of cholestasis had significantly increased serum levels of ALT,AST,ALP,γ-GT,TBA and TBIL with increased mRNA and protein expressions of IL-1βand IL-18,decreased protein and mRNA expressions of FXR,and increased protein expressions of NLRP3 and Caspase-1 and mRNA expressions of NLRP3,ASC,Caspase-1 and GSDMD in the liver tissue,showing also irregular arrangement of liver cells,proliferation of bile duct epithelial cells and inflammatory cells infiltration.Treatment of the rat models with Tuihuang Mixture significantly decreased serum levels of ALT,AST,ALP,γ-GT,TBA and TBIL,lowered IL-1βand IL-18 and increased FXR protein and mRNA expressions,and reduced NLRP3,ASC,Caspase-1 and GSDMD proteins and NLRP3,ASC and Caspase-1 mRNA expressions in the liver tissue.Tuihuang Mixture also significantly alleviated hepatocyte injury,bile duct epithelial cell proliferation and inflammatory cell infiltration in the liver of the rat models.Conclusion Tuihuang Mixture can effectively improve cholestasis in rats possibly by inhibiting NLRP3 inflammatosome-mediated pyroptosis via regulating FXR.

关 键 词：退黄合剂 胆汁淤积 FXR NLRP3炎症小体 细胞焦亡 

分 类 号：R285.5[医药卫生—中药学]