miR-9-3p对ConA诱导的小鼠慢性实验性肝损伤的影响  

Effect of Mir-9-3p on ConA induced chronic experimental liver injury in mice

作　　者：王闯 王猛 聂影 文怡欣 刘野 鄂志野 鞠宝玲 张红军 WANG Chuang(Mudanjiang Medical University,Mudanjiang 157011,China)

机构地区：[1]牡丹江医学院基础医学院免疫学教研室,黑龙江牡丹江157011

出　　处：《牡丹江医学院学报》2021年第2期1-5,共5页Journal of Mudanjiang Medical University

基　　金：黑龙江省省属高等学校基本科研业务费支持项目(2020-KYYWF-0781;2018-KYYWFMY-0024)。

摘　　要：目的探讨miR-9-3p对刀豆蛋白A(ConA)诱导所致小鼠慢性实验性肝损伤的影响。方法ConA构建小鼠慢性实验性肝损伤模型,高通量测序筛查模型动物肝组织miRNA的差异表达。BALB/c小鼠40只,随机分成对照组、ConA损伤组、过表达组(ConA+慢病毒包装miR-9-3p组)、过表达对照组(ConA+慢病毒空载体组)每组10只;各模型组小鼠经尾静脉按8 mg/kg体重注射ConA每周1次(对照组注射等体积生理盐水),连续8周;过表达组于第7周经尾静脉注射,将慢病毒包装的miR-9-3p过表达质粒(2×107 TU/mL)注射到小鼠体内,每周1次,连续2周;过表达对照组注射等量慢病毒空载体;于ConA末次给药7 d后处死小鼠收集样本,电子天平检测肝脏指数变化,血清酶学检测AST,ALT水平,HE染色检测模型小鼠肝脏组织病理改变;蛋白印迹(Western Blot)检测模型小鼠肝脏组织中COL1A1及α-SMA蛋白的表达变化。结果高通量测序显示慢性肝损伤模型中miR-9-3p表达明显下调。在肝损伤鼠体内过表达miR-9-3p后,血清AST和ALT水平明显下降;HE染色检测观察到过表达的miR-9-3p明显减轻肝ConA诱导的肝脏组织病理损伤,与ConA损伤组比较,细胞肿胀坏死减弱,小叶结构恢复;Western Blot检测过表达miR-9-3p后COL1A1及α-SMA蛋白的表达明显减低。结论miR-9-3p可减弱ConA诱导所致小鼠慢性实验性肝损伤。Objective To investigate the effect of miR-9-3p on chronic experimental liver injury induced by ConA in mice.Methods The mice model of chronic experimental liver injury was established by ConA,and the differential expression of miRNAs in liver tissue of model animals was screened by high-throughput sequencing.40 BALB/c mice were randomly divided into control group,ConA injury group,overexpression group(ConA+lentivirus-packaged miR-9-3p group)and overexpression control group(ConA+lentivirus empty vector group)with 10 mice in each group.Mice in each model group were injected with ConA via tail vein at 8 mg/kg body weight once a week(the control group was injected with the same volume of normal saline)for consecutive 8 weeks,and the overexpression group was injected via tail vein at the 7th week.The overexpression plasmid of miR-9-3p packaged in chronic disease was injected into mice,once a week for consecutive 2 weeks.The control group was given with empty lentivirus vector.The mice were sacrificed at day 7 after the last administration of ConA,electronic balance was used to detected the liver index changes,AST and ALT levels were detected by serum enzymology,and pathological changes of liver tissue were detected by HE staining.Western Blot was used to measure the expression of liver fibrosis effector protein COL1A1 andα-SMA in the middle liver of chronic liver injury.Results High-throughput sequencing showed that miR-9-3p expression was significantly down-regulated in the chronic liver injury model.After the overexpression of miR-9-3p in mice with liver injury,the serum AST and ALT levels decreased significantly.HE staining showed that the overexpression of miR-9-3p significantly eased the pathological damage of liver tissue induced by ConA.Compared with the ConA injury group,the cell swelling and necrosis were reduced and it restored the lobular structure.The expression changes of COL1A1 andα-SMA proteins were significantly decreased after the overexpression of miR-9-3p detected by Western Blot.Conclusion miR-9

关 键 词：miR-9-3p 高通量测序 免疫性肝损伤 CONA 

分 类 号：R392[医药卫生—免疫学]