低剂量白细胞介素-2对自身免疫性肝炎小鼠的肝保护作用及其机制  

Protective effect and its mechanism of low-dose interleukin-2 against hepatocyte injury in mice with autoimmune hepatitis

作　　者：陈晨 李竹蓉 吕思学 郭地 武嘉文 杨娜 刘杨 Chen Chen;Li Zhu-Rong;Lyu Si-Xue;Guo Di;Wu Jia-Wen;Yang Na;Liu Yang(School of Basic Medical Sciences,Shanxi University of Traditional Chinese Medicine,Jinzhong,Shanxi 030619,China)

机构地区：[1]山西中医药大学基础医学院,山西晋中030619

出　　处：《解放军医学杂志》2025年第2期214-220,共7页Medical Journal of Chinese People's Liberation Army

基　　金：山西省科学技术厅山西省重点国别科技合作项目(202104041101013);中西医结合防治风湿免疫病山西省科技创新人才重点团队(202204051002033);山西中医药大学科技创新团队项目(2022TD2003);山西中医药大学学科建设项目(2023XKJS-03)。

摘　　要：目的探讨低剂量白细胞介素-2(IL-2)对刀豆蛋白A(Con A)诱导的自身免疫性肝炎(AIH)小鼠的肝保护作用及其机制。方法SPF级雌性C57BL/6小鼠18只随机分为正常组、模型组与治疗组,每组6只。治疗组小鼠皮下注射300μl 10000 U IL-2,给药频率为1次/d,连续12 d。末次给药2 h后,模型组和治疗组均尾静脉注射Con A(15 mg/kg)。造模8 h后,采用HE染色观察小鼠肝脏病理形态变化,ELISA法检测小鼠血清中谷草转氨酶(AST)、谷丙转氨酶(ALT)、肿瘤坏死因子-α(TNF-α)和γ干扰素(IFN-γ)水平,Western blotting检测凋亡蛋白胱天蛋白酶(caspase 8/9/3)的表达情况,流式细胞术检测调节性T细胞(Treg)、Th1细胞百分比。结果与正常组比较,模型组小鼠肝脏指数,脾脏指数,肝组织坏死面积百分比,血清ALT、AST、TNF-α、IFN-γ水平,凋亡蛋白caspase 8/9/3的表达均明显升高(P<0.05或P<0.01);与模型组比较,治疗组小鼠肝脏指数,脾脏指数,肝组织坏死面积百分比,血清ALT、TNF-α、IFN-γ水平,凋亡蛋白caspase 8/9/3的表达均明显降低(P<0.05或P<0.01)。流式细胞术检测结果显示,与正常组比较,模型组小鼠Treg、Th1细胞百分比以及Th1/Treg比例均明显升高(P<0.05或P<0.01);与模型组比较,治疗组小鼠Treg细胞百分比进一步升高(P<0.01),Th1细胞百分比差异无统计学意义(P>0.05),Th1/Treg比例明显降低(P<0.05)。结论低剂量IL-2可有效缓解AIH小鼠的肝损伤,其作用机制可能与诱导Tregs活化有关。Objective To investigate the protective effect and its mechanism of low-dose interleukin-2(IL-2)against hepatocyte injury in Concanavalin A(Con A)-induced autoimmune hepatitis(AIH)mice.Methods Eighteen SPF female C57BL/6 mice were randomly divided into normal group,model group and treatment group,each group with 6 mice.Mice in the treatment group were subcutaneously injected with 300μl 10,000 U IL-2 for 12 d,once a day.2 h after the last dose,Con A(15 mg/kg)was injected through the tail vein in the model group and treatment group.After 8 h of modeling,the histopathological changes in the mouse liver were observed using HE staining,and the serum levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),tumor necrosis factor-α(TNF-α),and interferon-γ(IFN-γ)were detected using ELISA method;the expression of apoptotic protein caspase 8/9/3 was detected by Western blotting;and the percentages of Treg and Th1 cells were observed by flow cytometry.Results Compared with normal group,the liver index,spleen index,the percentage of necrotic area of liver tissue,the serum levels of ALT,AST,TNF-α and IFN-γ,and the expression of apoptosis protein caspase 8/9/3 significantly increased in the model group(P<0.05 or P<0.01);Compared with model group,the liver index,spleen index,the percentage of necrotic area of liver tissue,the serum levels of ALT,TNF-α and IFN-γ,and the expression of apoptosis protein caspase 8/9/3 significantly decreased in the treatment group(P<0.05 or P<0.01).The flow cytometry results showed that compared with normal group,the percentages of Treg and Th1 cells and Th1/Treg ratio increased in the model group(P<0.05 or P<0.01);Compared with the model group,the percentage of Treg cells further increased(P<0.01),Th1/Treg ratio decreased significantly in the treatment group(P<0.05),but there was no significant difference in the percentage of Th1 cells between two groups(P>0.05).Conclusion Low-dose of IL-2 can effectively improve liver injury in AIH mice,and the mechanism of action may be re

关 键 词：自身免疫性肝炎 白细胞介素-2 调节性T细胞 TH1细胞 

分 类 号：R575[医药卫生—消化系统]