microRNA-144影响成纤维样滑膜细胞增殖的机制研究  

Mechanism of microRNA-144 regulating the proliferation of fibroblast-like synoviocytes

作　　者：钱进 龚子臣 张义娜 郑伟 康夏 QIAN Jin;GONG Zi-chen;ZHANG Yi-na;ZHENG Wei;KANG Xia(Department of Orthopedics,General Hospital of Western Theater Command,Chengdu Sichuan 610000,China)

机构地区：[1]西部战区总医院骨科,四川成都610000

出　　处：《局解手术学杂志》2020年第11期855-861,共7页Journal of Regional Anatomy and Operative Surgery

基　　金：国家自然科学基金(81601940)。

摘　　要：目的探讨microRNA-144(mir-144)通过调控MDM4对炎症环境中成纤维样滑膜细胞(FLS)增殖和表达谱的影响。方法从C57BL/6小鼠中提取原代FLS和软骨细胞,免疫荧光染色和蛋白印迹检测MDM4在FLS中的蛋白表达情况,BrdU法和CCK-8法检测MDM4对FLS增殖的调控作用,双荧光素酶报告质粒检测mir-144与MDM4的靶向关系,实时荧光定量PCR检测mir-144对FLS表达谱的影响,Tunel法检测mir-144对软骨细胞凋亡的影响。结果MDM4在IL-1β刺激的FLS中高表达,mir-144表达下调;mir-144能够结合MDM4的3’UTR端。MDM4调控FLS的增殖,mir-144可显著抑制FLS的增殖,过表达MDM4可拮抗此抑制作用。mir-144过表达可降低FLS中MMP-9、TNF-α和IL-6的表达水平。mir-144过表达的FLS分泌物可抑制软骨细胞的凋亡。结论mir-144可通过调控MDM4抑制FLS在炎症环境中的增殖。Objective To explore the effect of microRNA-144(mir-144)on the proliferation and expression profile of fibroblast-like synoviocytes(FLS)in an inflammatory environment through regulating MDM4.Methods Primary FLS and chondrocytes were isolated from C57BL/6 mice.Immunofluorescence staining and Western blot were performed to test the protein expression of MDM4 in FLS.BrdU and CCK-8 methods were used to detect the regulation of MDM4 on FLS proliferation.Dual-luciferase assay was used to detect the targeting relationship between mir-144 and MDM4.Real-time fluorescent quantitative PCR was used to detect the effect of mir-144 on FLS expression profile.Tunel method was used to detect the effect of mir-144 on the apoptosis of chondrocytes.Results The level of MDM4 was upregulated and mir-144 was downregulated in FLS stimulated by IL-1β.mir-144 could bind to the 3’UTR region of MDM4.MDM4 could regulate the proliferation of FLS,and mir-144 could significantly inhabit the proliferation of FLS,but the overexpression of MDM4 could antagonize this inhibitory effect.Overexpression of mir-144 could reduce the expression levels of MMP-9,TNF-αand IL-6 in FLS.FLS secretion of mir-144 overexpression could inhibit the apoptosis of chondrocytes.Conclusion mir-144 can inhibit the proliferation of FLS in inflammatory environment through regulating MDM4.

关 键 词：关节炎 MICRORNA MDM4 成纤维样滑膜细胞 增殖 

分 类 号：R684.3[医药卫生—骨科学]