miR-27a-3p通过阻断Wnt3a/β-catenin途径抑制大鼠肺纤维化  

miR-27a-3p inhibits pulmonary fibrosis by blocking Wnt3a/β-catenin pathway in rats

作　　者：刘理静 钱红 胡柯 王乐 张自珍[2] 尹辉明 贺兼斌[4] LIU Lijing;QIAN Hong;HU Ke;WANG Le;ZHANG Zizhen;YIN Huiming;HE Jianbm(Sehool of Medicine,Hunan University of Medicine,Huaihua 418000;School of Medicine,Hunan Polytechnic of Environment and Biology,Hengyang 421001;Department of Respiration,First Affiliated Hospital,Hunan University of Medicine,Huaihua 418000;Department of Respiration,Huaihua First People's Hospital,Huaihua 418000,China)

机构地区：[1]湖南医药学院医学院,湖南怀化418000 [2]湖南环境生物职业技术学院医学院,湖南衡阳421001 [3]湖南医药学院第一附属医院呼吸内科,湖南怀化418000 [4]怀化市第一人民医院呼吸内科,湖南怀化418000

出　　处：《细胞与分子免疫学杂志》2018年第11期1015-1020,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：湖南省自然科学基金(2018JJ2279);怀化市科技计划资助项目(2017G3305)

摘　　要：目的探讨微小RNA-27a-3p(miR-27a-3p)对博莱霉素A5所致大鼠肺纤维化(PF)的影响及分子机制。方法雄性SD大鼠45只,随机分为对照组、miR-27a-3p激动剂(miR-27a-3p agomir)组和miR-27a-3p拮抗剂(miR-27a-3p antagomir)组,每组15只。经气管内注入博莱霉素A5建立PF模型,给药后次日分别予生理盐水、miR-27a-3p agomir、miR-27a-3p antagomir尾静脉注射,每3 d注射1次,共9次。第28天收集血液,ELISA检测血清1型前胶原蛋白羧基端前肽(P1CP)和3型前胶原蛋白氨基端前肽(P3NP)水平;处死大鼠,取肺组织,HE染色和Masson染色评价PF病变程度,实时荧光定量PCR检测miR-27a-3p、1型胶原蛋白(Col1)、Col3的mRNA水平,Western blot法检测Col1、Col3、Wnt3a、β联蛋白(β-catenin)的蛋白水平。结果miR-27a-3p agomir处理明显增加肺组织miR-27a-3p表达,miR-27a-3p antagomir则降低miR-27a-3p表达,提示miR-27a-3pagomir/antagomir转染效率较高。与对照组相比,miR-27a-3p agomir显著减轻大鼠肺泡炎症和PF程度,而miR-27a-3p antagomir则明显加重大鼠肺泡炎症和PF程度。miR-27a-3p agomir组血清P1CP、P3NP水平降低、下调肺组织Col1、Col3、Wnt3a、β-catenin水平,miR-27a-3p antagomir组则作用相反。结论 miR-27a-3p通过抑制Wnt3a/β-catenin信号通路,下调Col1、Col3表达,发挥抗PF作用。Objective To observe the effect of miR-27 a-3 p on bleomycin A5-induced pulmonary fibrosis( PF) in rats and explore the underlying mechanism. Methods Forty-five male SD rats were randomly divided into control group,miR-27 a-3 pagomir group and miR-27 a-3 p antagomir group. Each group contained 15 animals. All rats were injected intratracheally withbleomycin A5 to establish PF models. On the first day after bleomycin A5 administration,the rats in the control group,miR-27 a-3 p agomir group and miR-27 a-3 p antagomir group were injected at the caudal vein with physiological saline,agomir andantagomir,respectively. Injection was given one time each three days,totally nine times. On day 28,blood samples werecollected and then underwent enzyme linked immunosorbent assay for procollagen type 1 carboxyterminal propeptide( P1 CP)and procollagen type 3 aminoterminal propeptide( P3 NP) concentrations. Subsequently,all rats were sacrificed to removepulmonary tissue. Both HE and Masson staining were performed to evaluate the pathological changes of PF. The expressionof miR-27 a-3 p,collagen type 1( Col1),and collagen type 3( Col3) were detected using fluorescence real time quantitativePCR. Western blotting was used to examine Col1,Col3,Wnt3 a and β-catenin levels. Results The miR-27 a-3 p agomirmarkedly increased miR-27 a-3 p expression in the pulmonary tissue,whereas its antagomir decreased it,showing highertransfection efficacy. The pulmonary inflammation and fibrosis degree was alleviated in the miR-27 a-3 p agomir group whileaggravated in the miR-27 a-3 p antagomir group. In comparison with control group,serum P1 CP and P3 NP levels decreasedin the miR-27 a-3 p agomir group but increased in the miR-27 a-3 p antagomir group. Treatment with miR-27 a-3 p agomirdown-regulated the expression of Col1,Col3,Wnt3 a and β-catenin in the pulmonary tissue,while miR-27 a-3 p antagomirup-regulated their expression. Conclusion The miR-27 a-3 p inhibits the Wnt3 a/β-catenin signaling pathway,leading to thedown-regulation of Col1 and

关 键 词：miR-27a-3p 肺纤维化 1型胶原蛋白(Col1) 3型胶原蛋白 WNT3A β联蛋白(β-catenin) 

分 类 号：R563[医药卫生—呼吸系统] R392-33[医药卫生—内科学]