LncRNA GAS5通过miR-21/ADAMTS-1途径抑制大鼠肺纤维化  

LncRNA GAS5 inhibits pulmonary fibrosis through miR-21/ADAMTS-1 pathway in rats

作　　者：刘理静 钱红 王乐 胡柯 贺兼斌[5] 田玉梅 LIU Li-jing;QIAN Hong;WANG Le;HU Ke;HE Jian-bin;TIAN Yu-mei(School of Nursing,Hunan University of Medicine,Huaihua Hunan 418000,China;School of Medicine,Changsha Social Work College,Changsha 410004,China;School of Medicine,Hunan University of Medicine,First People’s Hospital of Huaihua,Huaihua Hunan 418000,China;Hunan Provincial Key Lab of Dong Medicine,Hunan University of Medicine,First People’s Hospital of Huaihua,Huaihua Hunan 418000,China;Dept of Respiratory and Critical Care Medicine,First People’s Hospital of Huaihua,Huaihua Hunan 418000,China)

机构地区：[1]湖南医药学院护理学院,湖南怀化418000 [2]长沙民政职业技术学院医学院,湖南长沙410004 [3]湖南医药学院医学院,湖南怀化418000 [4]湖南医药学院侗医药研究湖南省重点实验室,湖南怀化418000 [5]怀化市第一人民医院呼吸与危重症医学科,湖南怀化418000

出　　处：《中国药理学通报》2020年第12期1737-1743,共7页Chinese Pharmacological Bulletin

基　　金：湖南省自然科学基金资助项目(No 2018JJ2279)。

摘　　要：目的探讨生长阻滞特异性转录本5(growth arrest-specific transcript 5,GAS5)对肺纤维化的影响及分子机制。方法采用生物信息学、荧光素酶报告基因分析GAS5、miR-21、ADAMTS-1间的竞争性内源RNA(ceRNA)关系。SD大鼠分为对照组、LV-GAS5组和LV-GAS5+miR-21 agomir组,经气管内注入博莱霉素A5建立肺纤维化模型后,分别给予尾静脉注射0.2 mL的PBS、LV-GAS5、LV-GAS5+miR-21 agomir。d 28,处死大鼠,收集血液,采用ELISA分析血清PICP和PⅢNP水平,提取肺组织,HE染色和Masson染色观察病理改变,并用qRT-PCR检测GAS5、miR-21、ADAMTS-1表达,Western blot检测ADAMTS-1、ColⅠ、ColⅢ表达。结果GAS5、miR-21、ADAMTS-1之间存在ceRNA调控模式。与对照组比较,GAS5过表达减轻肺组织病理改变,降低血清PICP、PⅢNP水平,上调肺组织GAS5、ADAMTS-1表达,降低肺组织miR-21、ColⅠ、ColⅢ水平(P <0.01)。此外,miR-21agomir处理逆转了GAS5过表达对肺纤维化大鼠肺组织病理改变、胶原沉积和ADAMTS-1表达的影响(P <0.01)。结论GAS5通过内源性吸附miR-21,上调ADAMTS-1表达,促进ColⅠ、ColⅢ降解,减轻肺纤维化病变。Aim To explore the effects of GAS5 on pulmonary fibrosis and the underlying mechanisms.Methods Both bioinformatics and dual luciferase reporter gene were used to analyze the competitive endogenous RNA(ceRNA)relationship among GAS5,miR-21 and ADAMTS-1.SD rats were randomly divided into control group,LV-GAS5 group and LV-GAS5+miR-21 agomir group.All rats were injected intratracheally with bleomycin A5 to establish pulmonary fibrosis model.Rats in control group,LV-GAS5 group and LV-GAS5+miR-21 agomir group were injected at caudal vein with 0.2 mL of PBS,LV-GAS5,and LV-GAS5+miR-21 agomir,respectively.On day 28,all rats were sacrificed to collect blood and extract pulmonary tissue.ELISA was used to analyze serum levels of PICP and PⅢNP.Both HE and Masson staining were performed to observe the pathological changes in pulmonary tissues.Pulmonary GAS5,miR-21 and ADAMTS-1 expressions were detected using qRT-PCR.In addition,ADAMTS-1,ColⅠand ColⅢwere detected by Western blot.Results There was a ceRNA among GAS5,miR-21 and ADAMTS-1.In comparison with control group,GAS5 overexpression relieved the pathological changes in pulmonary tissues,reduced serum PICP and PⅢNP levels,up-regulated GAS5 and ADAMTS-1 expression,and decreased pulmonary miR-21,ColⅠand ColⅢlevels(P<0.01).Importantly,treatment with miR-21 agomir reversed the effects of GAS5 overexpression on pulmonary pathological changes,collagen deposition and ADAMTS-1 expression(P<0.01).Conclusions GAS5 sponges miR-21,up-regulates ADAMTS-1 expression and then promotes ColⅠand ColⅢdegradation,leading to the alleviation of pulmonary fibrosis.

关 键 词：肺纤维化 GAS5 MIR-21 ADAMTS-1 Ⅰ型胶原 Ⅲ型胶原 

分 类 号：R-332[医药卫生] R322.35