miR-200c-3p通过靶向Zeb1调控骨关节炎小鼠模型中软骨细胞凋亡和炎症反应  

MiR-200c-3p regulates chondrocyte apoptosis and inflammation in a mouse model of osteoarthritis by targeting Zeb1

作　　者：韩楠 乔晓峰[1] 朱晓文[2] 倪健[2] HAN Nan;QIAO Xiaofeng;ZHU Xiaowen;NI Jian(Department of Osteology,The First Affiliated Hospital of Jiamusi University,Jiamusi 154000,Heilongjiang,China;Department of General Surgery,The First Affiliated Hospital of Jiamusi University,Jiamusi 154000,Heilongjiang,China)

机构地区：[1]佳木斯大学附属第一医院骨一科,黑龙江佳木斯154000 [2]佳木斯大学附属第一医院普外科,黑龙江佳木斯154000

出　　处：《西部医学》2022年第12期1760-1765,共6页Medical Journal of West China

摘　　要：目的研究miR-200c-3p在骨关节炎(OA)中的作用及其下游的分子机制。方法通过破坏内侧半月板的稳定性构建骨关节炎小鼠模型,采用脂多糖(LPS)处理小鼠原代软骨细胞构建骨关节炎细胞模型。采用RT-qPCR检测骨关节炎小鼠模型样本以及对照组小鼠样本中miR-200c-3p的表达水平。采用RT-qPCR检测LPS处理前后小鼠原代软骨细胞中miR-200c-3p和Zeb1的表达水平变化。CCK-8实验检测miR-200c-3p过表达对骨关节炎细胞模型增殖能力的影响;流式细胞术检测miR-200c-3p过表达对骨关节炎细胞模型凋亡水平的影响;酶联免疫吸附试验检测miR-200c-3p过表达对骨关节炎细胞模型中炎症因子(IL-1β、IL-6、TNF-α)含量的影响。生物信息学分析RNA pull down实验和荧光素酶报告实验验证miR-200c-3p和下游靶基因相互作用。功能拯救实验验证Zeb1对miR-200c-3p作用的影响。结果miR-200c-3p在OA小鼠关节软骨组织和LPS处理的软骨细胞中显著下调。Zeb1在LPS处理的软骨细胞中显著上调。miR-200c-3p上调显著抑制了脂多糖诱导的软骨细胞凋亡和炎症损伤。Zeb1是miR-200c-3p的下游靶基因。Zeb1过表达逆转miR-200c-3p过表达对骨关节炎小鼠模型中软骨细胞凋亡和炎症反应的影响。结论miR-200c-3p通过靶向Zeb1抑制骨关节炎小鼠模型中软骨细胞凋亡并且缓解炎症反应,有助于发现骨关节炎治疗的有效靶点。Objective To investigate the role of miR-200c-3p in osteoarthritis(OA)and explore its downstream molecular mechanism.Methods OA animal model was established through destabilizing the medial meniscus.OA cell model was established through treating primary mouse chondrocytes with LPS.RT-qPCR was used to measure the expression level of miR-200c-3p in OA animal model and control animal model.The expression levels of miR-200c-3p and Zeb1 in primary mouse chondrocytes treated with or without LPS were detected by RT-qPCR.CCK-8 assay was applied to measure the proliferation of OA cells with miR-200c-3p overexpression.Flow cytometry was performed to analyze the apoptotic rate of OA cells after miR-200c-3p was overexpressed.ELISA assay was conducted to examine the concentration of inflammatory factors(IL-1β,IL-6,TNF-α)in OA cells with miR-200c-3p overexpression.Bioinformatics analysis,RNA pull down and luciferase reporter assay were conducted to demonstrate the interaction between miR-200c-3p and its downstream target.Functional rescue assays were used to verify the effect of Zeb1 on the functional role of miR-200c-3p.Results MiR-200c-3p was significantly down-regulated in OA mice articular cartilage tissues and lipopolysaccharide(LPS)-treated primary mouse chondrocytes.Zeb1 was up-regulated in LPS-treated primary mouse chondrocytes.Upregulation of miR-200c-3p significantly inhibited lipopolysaccharide induced chondrocyte apoptosis and inflammatory injury.Zeb1 is the downstream target of miR-200c-3p.Zeb1 overexpression reversed the effect of miR-200c-3p overexpression on chondrocyte apoptosis and inflammation in an OA mouse model.Conclusion MiR-200c-3p suppresses chondrocyte apoptosis and attenuates inflammation in OA mouse model by targeting Zeb1,which contributes to finds effective therapeutic target for OA.

关 键 词：骨关节炎 miR-200c-3p Zeb1 炎症反应 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学]