摘要
目的探讨内毒素血症下调肝细胞核因子4α(HNF4α)表达介导肝损伤进展的机制。方法144只BALB/c小鼠采用随机数字表法分组进行以下实验:(1)四氯化碳(CCl_(4))诱导小鼠急性肝损伤。对照组和0.5 mL/kg、1.0 mL/kg、2.0 mL/kg CCl_(4)组。(2)筛选脂多糖(LPS)干预小鼠的剂量。对照组和0.1 mg/kg、0.5 mg/kg、2.5 mg/kg LPS组。(3)LPS干预CCl_(4)(1.0 mL/kg)诱导急性肝损伤模型小鼠。对照组、CCl_(4)组、0.1 mg/kg LPS+CCl_(4)组、0.5 mg/kg LPS+CCl_(4)组。每组12只。诱导24 h后处死小鼠,采用酶速率法检测血清丙氨酸转氨酶(ALT),重氮法检测总胆红素(TBil)水平;Western blot法检测肝组织HNF4α、胱天蛋白酶3剪切体(Cleaved caspase-3)蛋白表达;原位末端标记法检测肝细胞凋亡情况。结果0.5、1.0、2.0 mL/kg CCl_(4)组的血清ALT、TBil及肝组织HNF4α、Cleaved caspase-3蛋白表达水平高于对照组,且呈剂量依赖性增高。2.5 mg/kg LPS组血清ALT、TBil及肝组织Cleaved caspase-3蛋白表达水平高于对照组和0.1、0.5 mg/kg LPS组,肝组织HNF4α蛋白表达水平低于对照组和0.1、0.5 mg/kg LPS组(P<0.05)。CCl_(4)组和0.1、0.5 mg/kg LPS+CCl_(4)组的血清ALT、TBil,肝组织HNF4α、Cleaved caspase-3蛋白表达水平及肝细胞凋亡指数均高于对照组(P<0.05);0.1、0.5 mg/kg LPS+CCl_(4)组的血清ALT、TBil,肝组织Cleaved caspase-3蛋白表达水平及肝细胞凋亡指数高于CCl_(4)组,HNF4α蛋白表达水平低于CCl_(4)组(P<0.05)。结论内毒素血症通过下调HNF4α表达增加肝细胞凋亡,可能是其介导肝损伤进展的机制之一。
Objective To investigate the mechanism of endotoxemia down-regulating the expression of hepatocyte nuclear factor 4α(HNF4α)and mediating the progression of liver injury.Methods A total of 144 BALB/c mice were divided into the following experiments using the digital table method.(1)carbon tetrachloride(CCl_(4))induced acute liver injury in mice:the control group,the 0.5 mL/kg,the 1.0 mL/kg and the 2.0 mL/kg groups;(2)screening doses of lipopolysaccharide(LPS)intervention in mice:the control group,the 0.1 mg/kg,the 0.5 mg/kg and the 2.5 mg/kg LPS groups;(3)LPS intervention in CCl_(4)(1.0 mL/kg)-induced acute liver injury model mice:the control group,the CCl_(4) group,the 0.1 mg/kg LPS+CCl_(4) group and the 0.5 mg/kg LPS+CCl_(4) group.There were 12 mice in each dose group.Mice were sacrificed after 24 hours induction,and blood samples of mice were collected to detect serum alanine aminotransferase(ALT)by enzyme rate method,and total bilirubin(TBil)level by diazo method.The protein contents of HNF4αand Cleaved caspase-3 in liver tissue were detected by Western blot assay.TUNEL assay was used to detect the apoptosis of hepatocytes.Results The serum levels of ALT,TBil,and liver tissue HNF4αand Cleaved caspase-3 protein expression levels were higher in the 0.5,1.0 and 2.0 mL/kg CCl_(4) groups than those in the control group in a dose-dependent manner.The serum levels of ALT and TBil,and liver tissue Cleaved caspase-3 protein were higher in the 2.5 mg/kg LPS group than that in the control group and the 0.1 and 0.5 mg/kg LPS groups.The expression level of HNF4αprotein in liver tissue was lower in the 2.5 mg/kg LPS group than that in the control group and the 0.1,0.5 mg/kg LPS group(P<0.05).Serum levels of ALT and TBil, liver tissue HNF4α and Cleaved caspase-3 protein expression levels and hepatocyte apoptosis index were higher in the CCl_(4) group and the 0.1 and 0.5 mg/kg LPS + CCl_(4) groups than those in the control group (P<0.05). The serum ALT, TBil, liver tissue Cleaved caspase-3 protein expression level and hepatocyte apoptosis index were higher in the 0.1, 0.5 mg/kg LPS+CCl_(4) groups than those of CCl_(4) group. The protein expression level of HNF4α was lower than that of the CCl_(4) group (P< 0.05). Conclusion Endotoxemia increases hepatocyte apoptosis by downregulating the expression of HNF4α, which may be one of the machanisms mediating the progression of liver injury.
作者
唐永静
陈云芬
廖月
程其娇
张桂娟
王云
何毅怀
TANG Yongjing;CHEN Yunfen;LIAO Yue;CHENG Qijiao;ZHANG Guijuan;WANG Yun;HE Yihuai(Department of Infectious Diseases,the Affiliated Hospital of Zunyi Medical University,Zunyi 563000,China;Department of Gastroenterology,Dafang County People's Hospital)
出处
《天津医药》
CAS
北大核心
2022年第10期1026-1030,共5页
Tianjin Medical Journal
基金
国家自然科学基金资助项目(81560110)
贵州省科技厅科技支撑计划项目[黔科合支撑(2019)2803号]
贵州省科学技术基金项目(黔科合平台人才[2017]5733-013)
贵州省卫生健康委科学技术基金项目(gzwjkj2020-1-041)。