长链非编码LncHECW1-IT1对滑膜间充质干细胞成软骨分化的调控机制  

The regulatory mechanism of long chain non coding LncHECW1-IT1 on cartilage differentiation of synovial mesenchymal stem cells

作　　者：王彬 范世珍 郭艳玲 赵硕 林松青 WANG Bin;FAN Shizhen;GUO Yanling;ZHAO Shuo;LIN Songqing(Shenzhen Hospital of Guangzhou University of Chinese Medicine(Futian),Shenzhen 518034,China)

机构地区：[1]广州中医药大学深圳医院(福田),广东深圳518034 [2]深圳市福田区妇幼保健院

出　　处：《中国实验诊断学》2024年第2期191-200,共10页Chinese Journal of Laboratory Diagnosis

基　　金：广东省深圳市科创委课题(No.201803013000229);深圳市福田区科技局课题(No.FTWS2021049)。

摘　　要：目的 本文旨在探讨长链非编码RNAHECW1-IT1对滑膜间充质干细胞(SMSCs)成软骨分化的调控机制。方法 选取广州中医药大学深圳医院骨伤科关节镜手术或是关节置换手术患者的膝关节内滑膜,分离培养及诱导软骨分化,利用各种实验室方法,检测滑膜间充质干细胞成软骨分化的情况及分化过程中信号通路的表达。结果 从供体的滑膜组织中分离出SMSCs,并通过流式细胞术、RT-qPCR和WB鉴定出SMSCs;通过阿利新蓝染色分析了SMSCs的高软骨分化能力;采用高通量RNA测序技术分析了软骨分化诱导后lncRNA和mRNA的不同表达水平,最终共发现121个lncRNA和2539个mRNA,对所有DEmRNAs进行了GO富集和KEGG通路分析,我们发现MAPK和Wnt信号通路在软骨分化过程中显著富集。通过构建竞争的内源性RNA网络,鉴定发现LncHECW1-IT1和ATF2在SMSCs软骨分化中的重要作用,是治疗及干预骨关节炎的调节因子和重要靶点。结论 LncHECW1-IT1通过上调ATF2的表达来促进人SMSCs的软骨生成分化,LncHECW1-IT1-ATF2轴是调控SMSCs成软骨分化的新机制。Objective The purpose of the present study was to investigate the regulatory mechanism of long-chain non-coding RNAHECW1-IT1 on chondrogenic differentiation of synovial mesenchymal stem cells(SMSCs).Methods The synovium in the knee joint of patients undergoing arthroscopic surgery or joint replacement surgery in the Department of orthopedics and traumatology of Shenzhen Hospital of Guangzhou University of traditional Chinese medicine was selected to isolate, culture and induce cartilage differentiation.Various laboratory methods were used to detect the chondrogenic differentiation of synovial mesenchymal stem cells and the expression of signal pathways in the differentiation process.Results SMSCs were isolated from synovial tissue of donors and identified by flow cytometry, RT qPCR and WB;The high cartilage differentiation ability of SMSCs was analyzed by alcian blue staining;High throughput RNA sequencing technology was used to analyze the different expression levels of lncrna and mRNA after cartilage differentiation induction.A total of 121 lncrna and 2539 mRNA were finally found.Go enrichment and KEGG pathway analysis were carried out for all demrnas.We found that MAPK and Wnt signaling pathways were significantly enriched in the process of cartilage differentiation.By constructing a competitive endogenous RNA network, it was identified that LncHECW1-IT1 and ATF2 play an important role in cartilage differentiation in SMSCs, serving as regulatory factors and important targets for the treatment and intervention of osteoarthritis.Conclusion LncHECW1-IT1 promotes chondrogenic differentiation of human SMSCs by upregulating the expression of ATF2.The LncHECW1-IT1-ATF2 axis is a novel mechanism for regulating chondrogenic differentiation of SMSCs.

关 键 词：骨关节炎 长链非编码RNAHECW1-IT1 软骨细胞分化 滑膜间充质干细胞 ATF2 

分 类 号：R681.3[医药卫生—骨科学]