miR-21通过下调ADAMTS-1表达促进大鼠肺纤维化  

miR-21 promotes pulmonary fibrosis in rats via down-regulating the expression of ADAMTS-1

作　　者：刘理静 尹辉明 黄民江 贺兼斌[3] 易高众[3] 王在岩[4] 钱红 

机构地区：[1]湖南医药学院临床医学院,湖南怀化418000 [2]湖南医药学院第一附属医院呼吸内科,湖南怀化418000 [3]怀化市第一人民医院呼吸内科,湖南怀化418000 [4]南华大学附属第一医院呼吸内科,湖南衡阳421001

出　　处：《细胞与分子免疫学杂志》2016年第12期1636-1640,1645,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：湖南省教育厅资助优秀青年科研项目(14B142)

摘　　要：目的探讨微小RNA21(miR-21)对博莱霉素诱导的大鼠肺纤维化的影响及相关机制。方法收集特发性肺纤维化(IPF)患者及正常人的外周血各20份,采用荧光实时定量PCR检测miR-21表达。将45只SD大鼠随机分为对照组、miR-21激动剂(miR-21agomir)和miR-21拮抗剂(miR-21antagomir)组,每组15只。经气管内注入博莱霉素A5建立肺纤维化模型后,分别给予生理盐水、miR21 agomir、miR21 antagomir尾静脉注射,第28天处死所有大鼠。取出肺组织,行HE和Masson染色,通过荧光实时定量PCR和Western blot法分别检测含1型血小板结合蛋白基序的解聚蛋白样金属蛋白酶(ADAMTS-1)、1型胶原蛋白(Col1)、Col3 mRNA与蛋白表达,分离血清,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)的含量。结果 IPF患者外周血miR-21表达水平明显高于正常人。miR-21 agomir组大鼠肺泡炎和肺纤维化程度明显重于对照组,而miR-21 antagomir组大鼠肺泡炎和肺纤维化程度显著轻于对照组。与对照组相比,miR-21 agomir组ADAMTS-1mRNA与蛋白表达水平降低,Col1、Col3的mRNA与蛋白表达水平及血清P1CP、P3NP水平均增加。但miR-21 antagomir组ADAMTS-1 mRNA与蛋白表达水平较对照组升高,Col1、Col3的mRNA与蛋白表达水平及血清P1CP、P3NP含量较对照组减少。结论 miR-21促进博莱霉素诱导的大鼠肺纤维化进展,其机制可能与下调ADAMTS-1表达及随后增加肺组织Col1、Col3含量有关。Objective To observe the effect of miR-21 on bleomycin-induced pulmonary fibrosis in rats, and explore the related mechanism. Methods Peripheral blood was collected from idiopathic pulmonary fibrosis （IPF） patients （ n = 20） and healthy adults （n = 20）. Fluorescence quantitative real-time PCR was then used to measure miR-21 expression. Forty-five SD rats were randomly divided into control group, miR-21 agomir group and miR-21 antagomir group. Each group included 15 rats. After establishment of pulmonary fibrosis models by intratracheal administration with bleomycin A5, rats in control group, miR-21 agomir group and miR-21 antagomir group were injected at caudal vein with normal saline, miR21 agomir and miR21 antagomir, respectively. All rats were sacrificed on day 28 after modeling. Subsequently, the pulmonary tissues were removed for HE and Masson staining. The mRNA and protein expressions of a disintegrin-like and metallopreteinase with thrombospondin type 1 motif （ ADAMTS-1 ）, collagen type 1 （ Colt ） and collagen type 3 （ CoB ） were detected by fluorescence quantitative real-time PCR and Western blotting. Serum was separated to examine procollagen type 1 carboxyterminal propeptide （PICP） and procollagen type 3 aminoterminal propeptide （P3NP） concentrations by ELISA. Results The level of miR-21 in peripheral blood was higher in IPF patients than in healthy adults. The alveolitis and pulmonary fibrosis extent in miR-21 agomir group was heavier than that in the control group. However, the alveoUtis and pulmonary fibrosis extent in miR-2l antagomir group was improved when compared with the control group. In comparison with the control group, ADAMTS-t mRNA and protein expression was significantly downregulated, whereas the mRNA and protein expressions of Colt and Col3 were significantly upregulated and serum PICP and P3NP concentrations were elevated in miR-21 agomir group. On the contrary, the level of ADAMTS-t mRNA and protein expression in miR-21 antagomir group was higher than t

关 键 词：微小RNA21(miR-21) 肺纤维化 含1型血小板结合蛋白基序的解聚蛋白样金属蛋白酶(ADAMTS-1) 1型胶原蛋白(Col1) 3型胶原蛋白(Col3) 

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