LncRNA NUTM2A-AS1调控miR-183-5p/TGFα影响骨关节炎软骨细胞增殖、凋亡的研究  

LncRNA NUTM2A-AS1 regulates the molecular mechanism of miR-183-5p/TGFαon the proliferation,apoptosis of osteoarthritis chondrocytes

作　　者：崔国峰 刘丹[3] 武军龙 魏戎 刘瑞宇[1] 王坤正[1] CUI Guofeng;LIU Dan;WU Junlong;WEI Rong;LIU Ruiyu;WANG Kunzheng(Department of Bone and Joint Surgery,The Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004;Department of Orthopedics,Luoyang Central Hospital Affiliated to Zhengzhou University,Luoyang 471009;Department of Rheumatology and Immunology,The First Affiliated Hospital of Xi’an Medical College,Xi’an 710077,China)

机构地区：[1]西安交通大学附属第二医院骨与关节外科,陕西西安710004 [2]郑州大学附属洛阳中心医院骨科,河南洛阳471009 [3]西安医学院第一附属医院风湿免疫科,陕西西安710077

出　　处：《中国骨质疏松杂志》2022年第6期794-801,共8页Chinese Journal of Osteoporosis

基　　金：洛阳市科技局医疗卫生重点项目(2101027A);河南省医学科技攻关计划联合共建项目(LHGJ20191226);陕西省自然科学基础研究计划资助项目(2021JZ-59);西安市卫生健康委员会项目(2020ms06)。

摘　　要：目的探讨LncRNA NUTM2A-AS1对白细胞介素-1β(IL-1β)诱导的软骨细胞损伤的影响及分子机制。方法将软骨细胞随机分为对照组、模型组(5μg/L的IL-1β)、miR-183-5p+模型组、miR-NC+模型组、si-LncRNA NUTM2A-AS1+模型组、si-TGFα+模型组、si-NC+模型组、pcDNA-TGFα+si-LncRNA NUTM2A-AS1+模型组、pcDNA+si-LncRNA NUTM2A-AS1+模型组;运用实时荧光定量PCR(RT-qPCR)检测LncRNA NUTM2A-AS1、miR-183-5p和TGFαmRNA的表达水平;运用细胞计数试剂盒8(CCK-8)检测细胞活性;运用流式细胞术检测软骨细胞凋亡情况;通过酶联免疫吸附法(ELISA)检测TNF-α、IL-6水平;通过双荧光素酶报告实验检测NUTM2A-AS1、miR-183-5p、TGFα之间的靶向关系。结果IL-1β诱导的软骨细胞中LncRNA NUTM2A-AS1和TGFα表达升高,miR-183-5p表达降低,软骨细胞活性降低,而凋亡率升高,TNF-α、IL-6水平升高。低表达LncRNA NUTM2A-AS1、低表达TGFα或过表达miR-183-5p后可促进细胞增殖及抑制细胞凋亡、炎症反应。结论低表达LncRNA NUTM2A-AS1通过调控miR-183-5p/TGFα抑制IL-1β诱导的软骨细胞凋亡及炎症反应,促进细胞存活。Objective To explore the effect of LncRNA NUTM2A-AS1 on interleukin-1β(IL-1β)-induced chondrocyte damage and its molecular mechanism.Methods Chondrocytes were randomly divided into control group,model group(5μg/L IL-1β),miR-183-5p+model group,miR-NC+model group,si-LncRNA NUTM2A-AS1+model group,si-TGFα+model group,si-NC+model group,pcDNA-TGFα+si-LncRNA NUTM2A-AS1+model group,pcDNA+si-LncRNA NUTM2A-AS1+model group;real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the expression levels of LncRNA NUTM2A-AS1,miR-183-5p and TGFαmRNA;cell counting kit 8(CCK-8)was used to detect cell viability;flow cytometry was used to detect chondrocyte apoptosis;enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of TNF-αand IL-6;dual luciferase reporter experiment was used to detect the targeting relationship between NUTM2A-AS1,miR-183-5p,and TGFα.Results In IL-1β-induced chondrocytes,the expressions of LncRNA NUTM2A-AS1 and TGFαwere increased,the expression of miR-183-5p was decreased,the activity of chondrocytes was decreased,the apoptosis rate was increased,and the levels of TNF-αand IL-6 were increased.Low expression of LncRNA NUTM2A-AS1,low expression of TGFαor overexpression of miR-183-5p could promote cell proliferation and inhibit apoptosis and inflammatory responses.Conclusion The low expression of LncRNA NUTM2A-AS1 can inhibit IL-1β-induced chondrocyte apoptosis and inflammation by regulating miR-183-5p/TGFα,and promote cell survival.

关 键 词：LncRNA NUTM2A-AS1 miR-183-5p TGFΑ 关节软骨细胞 凋亡 

分 类 号：R684.3[医药卫生—骨科学]