桂皮醛对miRNA-146a干扰的骨关节炎滑膜炎性反应影响的实验研究

摘要

Objective:To investigate the influence of miRNA-146a gene disturbances and cinnamic aldehyde (CA) on TLR4,NO,and MMP-13,which was released by synovioblast,and to explore their roles in the molecular mechanism of synovial inflammation in osteoarthritis (OA).Methods:Plasmids of miR-146a gene mimics and inhibitors were loaded into LPS-induced FLS using lipofection transfection.Then the interferential FLS were treated with CA,and the release amount of TRL4,NO and MMP-13 were detected.Results:After the miRNA-146a related plasmid transfection,the variation tendency of the concentrations of TLR4,NO and MMP-13 were almost the same.Compared with control group,the inhibitor group was elevated,and mimics group was declined (P ＜ 0.05).Compared with the control group,when the interferential FLS was treated with CA alone,TLR4,its release of TLR4,NO and MMP-13 were significantly decreased (P ＜ 0.05),whereas the mimics and CA coordinated intervention,the release was significantly decreased (P ＜ 0.05).Conclusion:Both of the miR-146a and CA have effects on synovial inflammation in OA,especially when mimics and CA were combined,the inhibition of inflammation was best.It could provide an experimental basis to clarify molecular mechanism,identify and develop novel drug targets of OA.%目的:研究miRNA-146a基因及桂皮醛对miRNA-146a干扰的滑膜成纤维细胞释放TLR4、NO、MMP-13的影响,探寻其在OA滑膜炎性反应分子学机制中的作用.方法:采用脂质体转染法将miR-146a基因模拟及抑制性质粒载体对经LPS诱导后的OA滑膜炎性反应效应细胞滑膜成纤维细胞进行基因导入,并用桂皮醛对干扰后的细胞进行干预,检测不同组间TLR4、NO及MMP-13表达的差异.结果:miRNA-146a相关质粒转染后的TLR4、NO、MMP-13浓度的变化趋势大致相同:与对照组比较,三者的inhibitor组均升高,mimics组则均呈下降趋势(P＜0.05).桂皮醛单独作用于miRNA-146a干扰后的细胞时,与对照组比较,TLR4、NO、MMP-13浓度均有明显下降(P＜0.05),而当mimics和CA联合干预时,三者释放量显著下降(P＜0.05).结论:miRNA-146a及桂皮醛均对骨关节滑膜炎性反应产生影响,当mimics和桂皮醛联合作用时,抑制炎性反应效果最好.为进一步阐明骨关节炎分子机制和药物靶点的识别与开发提供实验依据.