茵陈蒿汤调控Fas/Caspase-8/Caspase-3信号通路改善胆汁淤积性肝损伤的机制OA
Mechanism Study of Yinchenhao Tang Regulating Fas/Caspase-8/Caspase-3 Signaling Pathway to Improve Cholestatic Liver Injury
目的:探究茵陈蒿汤调控肿瘤坏死因子受体超家族成员6(Fas)/胱天蛋白酶(Caspase)-8/Caspase-3信号通路抑制肝细胞凋亡改善胆汁淤积性肝损伤(CLI)的作用机制.方法:48只Wistar大鼠,随机选取12只为空白组,其余大鼠一次性给予α-萘异硫氰酸酯(ANIT)灌胃复制胆汁淤积性肝损伤模型.造模大鼠随机分为模型组、熊去氧胆酸组(0.1 g·kg-1)、茵陈蒿汤组(9.23 g·kg-1),每组12只,各组大鼠给予相应药物灌胃,连续3 d.检测血清中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、γ-谷氨酰转肽酶(γ-GT)、总胆红素(TBil)、总胆汁酸(TBA)水平;肝组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平;苏木素-伊红(HE)染色观察肝组织病理学变化;蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应(Real-time PCR)检测肝组织中Fas、Caspase-8、Caspase-3、B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)、Bcl-2蛋白及mRNA表达.结果:与空白组比较,模型组ALT、AST、ALP、γ-GT、TBA、TBil水平显著升高(P<0.01);TNF-α、IL-1β含量及mRNA表达显著升高(P<0.01);肝细胞排列紊乱,可见炎性细胞浸润,胆管上皮细胞增生;Fas、Caspase-8、Caspase-3、Bax蛋白及mRNA表达明显升高,Bcl-2蛋白及mRNA表达明显降低(P<0.05,P<0.01).与模型组比较,熊去氧胆酸组ALP、γ-GT、TBA、TBil水平明显降低,TNF-α、IL-1β水平及mRNA表达明显降低(P<0.05,P<0.01),Fas、Caspase-8、Caspase-3、Bax蛋白及mRNA表达明显降低(P<0.05,P<0.01),Bcl-2 mRNA表达明显升高(P<0.05);茵陈蒿汤组ALT、AST、γ-GT、TBA、TBil水平显著降低(P<0.01),TNF-α、IL-1β水平及mRNA表达明显降低(P<0.05,P<0.01),Fas、Bax蛋白及Fas、Caspase-8、Caspase-3、Bax mRNA表达明显降低(P<0.05,P<0.01),Bcl-2蛋白及mRNA表达明显升高(P<0.05,P<0.01);肝细胞损伤、炎性细胞浸润减轻,胆管上皮细胞增生减少.结论:茵陈蒿汤可改善胆汁淤积性肝损伤,其作用机制可能与抑制Fas/Caspase-8/Caspase-3通路介导的肝细胞凋亡有关.
Objective:To explore the mechanism of Yinchenhao Tang regulating the tumor necrosis factor receptor superfamily member 6(Fas)/cysteine protease-8(Caspase-8)/cysteine protease-3(Caspase-3)signaling pathway to inhibit hepatocyte apoptosis and improve cholestatic liver injury(CLI).Methods:Among 48 Wistar rats,12 rats were randomly selected as the blank group,and the other rats were administered alpha-naphthalene isothiocyanate(ANIT)by gavage to induce a CLI model.The modeling rats were randomly divided into the model group,the ursodeoxycholic acid group(0.1 g·kg-1)and the Yinchenhao Tang group(9.23 g·kg-1),with 12 rats in each group.The rats in each group were given corresponding drugs by gavage for three consecutive days.The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),gamma-glutamyl transpeptidase(γ-GT),total bilirubin(TBil)and total bile acid(TBA)in serum were detected.The levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in liver tissue were detected.The histopathological changes of the liver were observed by hematoxylin-eosin(HE)staining.The protein and mRNA expressions of Fas,Caspase-8,Caspase-3,B-cell lymphoma-2(Bcl-2)associated X protein(Bax)and Bcl-2 in liver tissue were detected by Western blot and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Results:Compared with those in the blank group,the levels of ALT,AST,ALP,γ-GT,TBA and TBil in serum of the model group were significantly increased(P<0.01).The levels and mRNA expressions of TNF-α and IL-1β in liver tissue were significantly increased(P<0.01).The arrangement of hepatocytes was disordered,and inflammatory cell infiltration and bile duct epithelial cell proliferation were observed.The protein and mRNA expressions of Fas,Caspase-8,Caspase-3 and Bax in liver tissue were significantly increased(P<0.05,P<0.01),while the protein and mRNA expressions of Bcl-2 were significantly decreased(P<0.05,P<0.01).Compared with those in the model group,the levels of ALP,γ-GT,TBA and TBil in the serum of rats in the ursodeoxycholic acid group were significantly decreased.The levels and mRNA expressions of TNF-α and IL-1β in liver tissue were significantly decreased(P<0.05,P<0.01).The protein and mRNA expressions of Fas,Caspase-8,Caspase-3 and Bax in liver tissue were significantly decreased(P<0.05,P<0.01),while the mRNA expression of Bcl-2 was significantly increased(P<0.05,).The levels of ALT,AST,γ-GT,TBA and TBil in the serum of rats in the Yinchenhao Tang group were significantly decreased(P<0.01).The levels and mRNA expressions of TNF-α and IL-1β in liver tissue were significantly decreased(P<0.05,P<0.01).The protein expression of Fas and Bax and the mRNA expression of Fas,Caspase-8,Caspase-3 and Bax in liver tissue were significantly decreased(P<0.05,P<0.01),while the protein and mRNA expression of Bcl-2 were significantly increased(P<0.05,P<0.01).Hepatocyte injury,inflammatory cell infiltration and proliferation of bile duct epithelial cells were reduced.Conclusion:Yinchenhao Tang can ameliorate CLI,and its mechanism may be related to inhibiting hepatocyte apoptosis mediated by the Fas/Caspase-8/Caspase-3 signaling pathway.
ZHU Zhengwang;WANG Linlin;ZHAO Jinghan;SHE Linjing;TANG Yinpei;CAI Qingchun;WANG Bing;ZHU Pingsheng;MIAO Mingsan
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,ChinaHenan University of Chinese Medicine,Zhengzhou 450046,China||Collaborative Innovation Center of Research and Development on the Whole Industry Chain of Yu-Yao,Zhengzhou 450046,ChinaHenan University of Chinese Medicine,Zhengzhou 450046,China||Collaborative Innovation Center of Research and Development on the Whole Industry Chain of Yu-Yao,Zhengzhou 450046,ChinaHenan University of Chinese Medicine,Zhengzhou 450046,China||Collaborative Innovation Center of Research and Development on the Whole Industry Chain of Yu-Yao,Zhengzhou 450046,ChinaHenan University of Chinese Medicine,Zhengzhou 450046,ChinaThird Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450008,ChinaShanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200233,ChinaHenan University of Chinese Medicine,Zhengzhou 450046,China||Collaborative Innovation Center of Research and Development on the Whole Industry Chain of Yu-Yao,Zhengzhou 450046,ChinaHenan 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
医药卫生
茵陈蒿汤胆汁淤积肝损伤肿瘤坏死因子受体超家族成员6(Fas)/胱天蛋白酶(Caspase)-8/Caspase-3通路
Yinchenhao Tangcholestasisliver injurytumor necrosis factor receptor superfamily member 6(Fas)/cysteinyl aspartate specific proteinase(Caspase)-8/Caspase-3 signaling pathway
《中国实验方剂学杂志》 2026 (1)
39-46,8
国家自然科学基金项目(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)
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