Glt25d1基因敲低对刀豆蛋白A诱导的自身免疫性肝炎小鼠巨噬细胞和中性粒细胞的影响  

Effects of Glt25d1 gene knockdown on macrophages and neutrophils of mice with autoimmune hepatitis induced by concanavalin A

作　　者：高美欣 何玲玲 叶小慧 杨君茹 李佳 张曼卡 张健[3] 肖凡 魏红山[1,2] Gao Meixin;He Lingling;Ye Xiaohui;Yang Junru;Li Jia;Zhang Manka;Zhang Jian;Xiao Fan;Wei Hongshan(Department of Gastroenterology,Peking University Ditan Teaching Hospital,Beijing 100015,China;Department of Gastroenterology,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China;Department of Center of Integrated Traditional Chinese and Western Medicine,Peking University Ditan Teaching Hospital,Beijing 100015,China;Department of Institute of Infectious Diseases,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China.)

机构地区：[1]北京大学地坛医院教学医院消化科,北京100015 [2]首都医科大学附属北京地坛医院消化科,北京100015 [3]北京大学地坛医院教学医院中西医结合中心,北京100015 [4]首都医科大学附属北京地坛医院传染病学研究所,北京100015

出　　处：《中国肝脏病杂志（电子版）》2020年第1期36-43,共8页Chinese Journal of Liver Diseases：Electronic Version

基　　金：首都临床特色应用研究(Z181100001718084);北京市医院管理中心消化内科学科协同发展中心专项(XXZ0404);北京市自然科学基金(7152073);国家自然科学基金(81271901)。

摘　　要：目的研究Glt25d1基因在刀豆蛋白A(concanavalin A,Con A)诱导的自身免疫性肝炎中的作用及其对巨噬细胞和中性粒细胞的影响。方法随机抽取6~8周无特定病原体雌性Glt25d1+/-小鼠及与其同窝出生的野生型(wild type,WT)小鼠,体质量(20±2)g,分为WT对照组、Glt25d1+/-对照组、WT造模组和Glt25d1+/-造模组,每组8只。造模组通过内眦静脉注射Con A,剂量为10 mg/kg,对照组给予相同剂量生理盐水注射。造模12 h后处死小鼠。检测小鼠血浆中丙氨酸氨基转移酶(alanine aminotransferase,ALT)和天门冬氨酸氨基转移酶(aspartateaminotransferase,AST)水平,观察肝组织病理。采用流式细胞术检测小鼠外周血、脾脏和肝脏中巨噬细胞和中性粒细胞的比例。采用定量逆转录PCR(quantitative reverse transcriptase-mediated PCR,qRT-PCR)检测肝脏中单核-巨噬细胞趋化蛋白-1(monocyte/macrophage chemotaxis proteinⅠ,MCP-Ⅰ)、巨噬细胞炎性蛋白1α(macrophage inflammatory proteinⅠα,MIP-Ⅰα)、巨噬细胞炎性蛋白2(macrophage inflammatoryproteinⅡ,MIP-Ⅱ)及淋巴细胞抗原6G(lymphocyteantigen6complexlocusG,Ly6G)等mRNA的相对表达量。结果 WT对照组和Glt25d1+/-对照组小鼠ALT [(52.00±18.19)U/L vs (45.00±6.85)U/L]和AST [(168.17±21.33)U/L vs(276.17±83.37)U/L]水平差异无统计学意义(t值分别为0.360、1.255,P值分别为0.7263、0.2589)。Con A造模12 h后,WT造模组小鼠ALT和AST水平分别为(3089.67±663.92)U/L、(3099.50±519.15)U/L,Glt25d1+/-造模组小鼠ALT和AST水平分别(11565.17±1381.38)U/L、(10875.17±1558.68)U/L,均显著高于未造模组(P均<0.05),Glt25d1+/-造模组小鼠ALT和AST水平均显著高于WT造模组(t值分别为5.530、4.733,P值分别为0.0003、0.0032)。肝组织HE染色示:WT对照组和Glt25d1+/-对照组小鼠肝组织未见明显损伤,Con A诱导12 h后,与WT Con A造模组小鼠相比,Glt25d1+/-造模组小鼠肝脏坏死面积更广、汇管区炎性细胞浸润更显著。Glt25d1+/-对照组�Objective To investigate the role of Glt25d1 in concanavalin A(Con A) induced autoimmune hepatitis, and its effects on macrophages and neutrophils in this progression. Methods The specific pathogen free Glt25d1+/-mice and wild type(WT) mice born in the same litter were randomly selected, the mice were all female and 6~8 weeks, the body mass were(20 ± 2) g. The mice were divided into WT control group, Glt25d1+/-control group, WT Con A administration group and Glt25d1+/-Con A administration group, 8 mice in each group. Mice in administered group were given Con A via the internal iliac venous plexus with a dose of 10 mg/kg body mass, while mice in control group were injected with the same dose of saline. Mice were sacrificed after Con A challenged for 12 h. The levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were detected and the liver histopathology were observed. Flow cytometry were used to detect the percentage of macrophages and neutrophils in blood, spleen and liver single-cell suspension of mice. Quantitative reverse transcriptase-mediated PCR(qRT-PCR) were used to detect the relative expression of monocyte/macrophage chemotaxis protein Ⅰ(MCP-Ⅰ)、macrophage inflammatory protein Ⅰα(MIP-Ⅰα)、macrophage inflammatory protein Ⅱ(MIP-Ⅱ) and lymphocyte antigen 6 complex locus G(Ly6G) mRNA. Results There were no statistically significant differences of ALT [(52.00 ± 18.19) U/L vs(45.00 ± 6.85) U/L ] and AST [(52.00 ± 18.19) U/L vs(45.00 ± 6.85) U/L ] in mice between WT control group and Glt25d1+/-control group(t = 0.360, 1.255, P = 0.7263, 0.2589). After Con A administrated for 12 h, the ALT and AST levels of mice in WT Con A administration group were(3089.67 ± 663.92) U/L and(3099.50 ± 519.15) U/L, respectively, and in Glt25d1+/-Con A administration group were(11565.17 ± 1381.38) U/L and(10875.17 ± 1558.68) U/L, respectively, the levels of ALT and AST were significantly higher than those in control group(all P < 0.05). In addition, the ALT and AST levels of Glt25d1+

关 键 词：Glt25d1 自身免疫性肝炎 巨噬细胞 中性粒细胞 

分 类 号：R575.1[医药卫生—消化系统]