王闯;王猛;聂影;文怡欣;刘野;鄂志野;鞠宝玲;张红军;

• 1:牡丹江医学院基础医学院免疫学教研室

摘要(Abstract)：

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。

关键词(KeyWords)： miR-9-3p;高通量测序;免疫性肝损伤;ConA

Abstract:

Keywords:

基金项目(Foundation): 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)

作者(Authors): 王闯;王猛;聂影;文怡欣;刘野;鄂志野;鞠宝玲;张红军;

DOI: 10.13799/j.cnki.mdjyxyxb.2021.02.001

参考文献(References)：

• [1] SUKUMARAN S,CHANDRASHEKARAN G.Role of miRNA and its potential as a novel diagnostic biomarker in drug-induced liver injury[J].European Journal of Clinical Pharmacology,2017,73(4):399-407.
• [2] ZHEN Z,CHEN Y L,KUN C.siRNA-and miRNA-based therapeutics for liver fibrosis[J].Translational Research,2019,21(4):17-29.
• [3] DAVID P B.MicroRNAs:Target Recognition and Regulatory Functions[J].Cell,2009,136(2):215-233.
• [4] RYAN M,O'CONNEL L.Physiological and pathological roles for microRNAs in the immune system[J].Nature Reviews Immunology,2010,10(2):111-122.
• [5] AMELIA C.miR-15 and miR-16 Induce Apoptosis by Targeting BCL2[J].Proceedings of the National Academy of Sciences of the United States of America,2005,102(39):13944-13949.
• [6] TANG J,LI Y,LIU K,et al.Exosomal miR-9-3p suppresses HBGF-5 expression and is a functional biomarker in hepatocellular carcinoma[J].International Journal of Cancer,2018,109(1):15-23.
• [7] BANDRES E,AGIRRE X,BITARTE N,et al.Epigenetic regulation of microRNA expression in colorectal cancer[J].International Journal of Cancer,2009,125(11):2737-2743.
• [8] LUJAMBIO A,CALIN G A,VILLANEVA A,et al.A microRNA DNA methylation signature for human cancer metastasis[J].Proceedings of the National Academy of Sciences USA,2008,105(36):13556-13561.
• [9] 苏静，薛凌飞，张晓彤，等.miR-9-5p靶向FGF9抑制胃癌细胞迁移侵袭的作用机制[J].现代肿瘤医学，2020,28(14):2401-2407.
• [10] ZHANG J,LIU Y,LIU Z,et al.Differential expression profiling and functional analysis of microRNAs through stage Ⅰ-Ⅲ papillary thyroid carcinoma[J].Int J Med Sci,2013,10(5):585-592.
• [11] LI D Y,CHEN W J,LOU L,et al.Prospective lncRNA-miRNA- mRNA regulatory network of long non-coding RNA LINC00968 in non-small cell lung cancer A549 cells:A miRNA microarray and bioinformatics investigation[J].Int J Mol Med,2017,40(6):1895-1906.
• [12] CAVIGLIA G P,FAGOONEE S.Role and function of exosomal miR-9-3p in hepatocellular carcinoma[J].Minerva Med,2018,109(1):4-6.
• [13] THIGASH I,HAYASHI H,TISHIMOT O,et al.miR-9-3p plays a tumour-suppressor role by targeting TAZ (WWTR1) in hepatocellular carcinoma cells[J].British Journal of Cancer,2015,113(18):252-258.
• [14] WANG Y,DU J,NIU X,et al.MiR-130a-3p attenuates activation and induces apoptosis of hepatic stellate cells in nonalcoholic fibrosing steatohepatitis by directly targeting TGFBR1 and TGFBR2[J].Cell Death Dis,2017,8(5):2792.