lncRNA HOTAIR在白细胞介素1β介导骨关节炎中的作用机制  

The mechanism of IncRNA HOTAIR in interleukin-1beta-mediated osteoarthritis

作　　者：周亮 陈兴真 李振宇 张泽坤 段国庆[3] Zhou Liang;Chen Xingzhen;Li Zhenyu;Zhang Zekun;Duan Guoqing(School of Clinical Medicine,Jining Medical University,Jining 272067,Shandong Province,China;Department of Hand and Foot Surgery,Jining No.1 People’s Hospital,Jining 272067,Shandong Province,China;Deparment of Joint and Sports Medicine,Affiliated Hospital of Jining Medical University,Jining 272067,Shandong Province,China)

机构地区：[1]济宁医学院临床医学院,山东省济宁市272067 [2]济宁市第一人民医院手足外科,山东省济宁市272067 [3]济宁医学院附属医院关节与运动医学科,山东省济宁市272067

出　　处：《中国组织工程研究》2022年第35期5607-5613,共7页Chinese Journal of Tissue Engineering Research

基　　金：济宁医学院青年教师科研扶持基金(JY2017FS021),项目负责人:段国庆;贺林院士新医学科研基金项目(JYHL2019FMS13),项目负责人:段国庆。

摘　　要：背景:目前已有研究将骨关节炎患者关节软骨及正常人关节软骨进行研究后发现,lncRNA HOTAIR较对照组明显上调,可能与骨关节炎的发生有一定关系。目的:探索lncRNA HOTAIR在白细胞介素1β介导的骨关节炎中的作用机制。方法:(1)临床样本分组:选取20例关节炎行全膝关节表面置换患者的内侧髁软骨作为骨关节炎组,选取7例行下肢截肢或因股骨颈骨折行全髋关节置换患者的关节软骨作为正常软骨组;(2)实验动物分组:取48只SD大鼠随机分为骨关节炎组(36只)和正常对照组(12只),骨关节炎组建立膝关节骨关节炎模型;(3)细胞实验分组及处理:将体外培养的大鼠关节软骨细胞(RCCs-1)分为对照组、白细胞介素1β组(筛选出最佳质量浓度10μg/L模拟骨关节炎环境)、siRNA组、白细胞介素1β+siRNA组,对照组不做特殊处理,其他各组分别加入相应的溶液进行处理,培养48 h;(4)检测指标:分别于造模后4,6,8周取大鼠股骨行苏木精-伊红染色观察病理变化;采用RT-PCR分析患者关节软骨、大鼠关节软骨及白细胞介素1β处理的大鼠软骨细胞中lncRNA HOTAIR的mRNA表达水平;采用CCK-8法检测白细胞介素1β处理的软骨细胞活力;Western blot检测软骨细胞Ⅱ型胶原蛋白表达;流式细胞技术检测软骨细胞凋亡情况。结果与结论:(1)正常对照组大鼠软骨细胞排列整齐,软骨结构层次清晰,潮线完整;骨关节炎组随着造模时间的延长,软骨结构越紊乱,软骨细胞数量明显减少,软骨基质失染,潮线严重紊乱;(2)lncRNA HOTAIR在骨关节炎患者和骨关节炎组大鼠软骨细胞中表达增高;(3)白细胞介素1β降低大鼠膝关节软骨细胞活力,且促进HOTAIR的表达增加;(4)HOTAIR抑制大鼠膝关节软骨细胞活力及Ⅱ型胶原蛋白的表达;(5)HOTAIR的表达促进软骨细胞凋亡;(6)结果提示,抑制HOTAIR介导的软骨细胞凋亡可能是膝骨关节炎基因治疗的潜在机制。BACKGROUND:The a rticular ca rtilage of patients with osteoarthritis and normal human a rticular cartilage have been studied and found that long non-coding RNA(IncRNA)H OTAIR is significantly up-regulated compared with the control group,which may be related to the occurrence of osteoarthritis.OBJECTIVE:To explore the mechanism of IncRNA HOTAIR in interleukin-lβ-mediated osteoarthritis.METHODS:(1)Clinical grouping:The medial condyle ca rtilage samples of 20 patients with arthritis who underwent total knee replacement were selected as experimental group.The a rticular ca rtilage samples of seven patients with lower extremity amputation or undergoing total hip arthroplasty due to femoral neck fra cture were selected as control group.(2)Animal grouping:Fo rty-eight Sprague-Dawley rats were randomized into osteoarthritis group(n=36)and normal control group(n=12).Animal models of knee osteoarthritis were made in the osteoarthritis group.(3)Cell testing and treatment:The rat artic ular chondrocytes(RCCs-1)cultured in vitro were divided into control group,interleukin-1βgroup(the best mass concentration of 10μg/L was selected to simulate osteoarthritis environment),small interfering RNA(siRNA)group,interleukin-1β+siRNA group.Except for the control group with no special treatment,the other groups were treated with corresponding solutions and cultured for 48 hours.(4)Detection indicato rs:At 4,6,and 8 weeks after modeling,the rat femurs were collected for pathological observation using hematoxylin-eosin staining.RT-PCR was used to analyze the mRNA expression of IncRNA HOTAIR in interleukin-1β-mediated rat chondrocytes,rat articular ca rtilage,and the articular ca rtilage of patients.Chondrocyte viability was detected by cell counting kit-8 method.Expression of typeⅡcollagen in chondrocytes was detected by western blot.Apoptosis of chondrocytes was detected by flow cytometry.RESULTS AND CONCLUSION:(1)The chondrocytes of the rats in the normal control group were neatly arranged,the ca rtilage structure was clea r,and t

关 键 词：膝关节 骨关节炎 LncRNA HOTAIR 白细胞介素1Β 细胞凋亡 Ⅱ型胶原蛋白 细胞增殖 

分 类 号：R459.9[医药卫生—治疗学] R446[医药卫生—临床医学]