微小RNA-543对大鼠骨关节炎软骨细胞的作用机制研究

Study on the Mechanism of Effect of MicroRNA-543 on Osteoarthritis Chondrocytes in Rats

目的:研究微小RNA-543对大鼠骨关节炎软骨细胞存在的保护作用机制。方法:此次研究选取20只健康SD(Sprague-Dawley)大鼠进行研究,采取随机数字表法将入组的大鼠分成研究组和对照组,每组10只,对照组大鼠不进行特殊处理,研究组大鼠接受关节内侧副韧带切断术,构建建立大鼠骨关节炎(OA)模型,采用PCR检测方法分别检测两组大鼠软骨组织的microRNA-543表达水平。对大鼠正常膝关节软骨细胞进行体外分离,10ng/L的IL-1β培养24h形成体外OA模拟状态,使用miR-543mimics进行转染,观察分析miR-543过表达对IL-1β诱导条件下软骨细胞增殖及凋亡相关基因表达水平的影响。结果:研究组大鼠的软骨组织中miR-543的表达水平显著低于对照组(P<0.05)。miR-543mimics转染后软骨细胞的增殖率明显高于对照组(P<0.05),经过IL-1β诱导处理的软骨细胞增殖能力显著低于对照组(P<0.05),经过miR-543mimics转染后IL-1β诱导处理过的软骨细胞增殖能力显著上升(P<0.05)。miR-543mimics转染细胞MDM4mRNA表达水平明显低于对照组(P<0.05),Akt1和Bcl-2mRNA的表达显著高于对照组(P<0.05),经过IL-1β诱导处理后的软骨细胞MDM4mRNA表达显著高于对照组(P<0.05),Akt1和Bcl-2mRNA表达则显著低于对照组(P<0.05),经过miR-543mimics转染后,IL-1β诱导处理过的软骨细胞MDM4、Akt1和Bcl-2mRNA表达水平的变化能够得到显著缓解(P<0.05)。结论:OA大鼠的软骨组织miR-543表达会显著下降,通过miR-543mimics转染促进miR-543表达上调,能够使软骨细胞的增殖得到提升,加之对MDM4、Akt1、Bcl-2表达的影响,能够有效对抗IL-1β诱导引发的软骨细胞损伤。

Objective:To study the protective mechanism of microRNA-543 on the presence of chondrocytes in rat osteoarthritis.Method:In this study, 20 healthy SD(Sprague-Dawley) rats were randomly divided into study group(n=10) and control group(n=10).The rats in the study group were treated without special treatment.The rats in the study group underwent medial collateral ligament cutting,and the model of osteoarthritis(OA) was established.The expression of microRNA-543 in cartilage tissue of the two groups was detected by PCR.Rat normal knee cartilage cells were isolated in vitro and cultured with 10 ng/L IL- 1β for 24 h to form OA simulation in vitro.The effect of miR-543 overexpression on the proliferation of chondrocytes and the expression level of apoptosisrelated genes under the induction of IL-1 under the induction of IL-1 was observed.Result:The expression level of miR-543 in cartilage tissue of the study group was significantly lower than that of the control group(P<0.05).The proliferation rate of cartilage cells induced by miR-543 mimics was significantly higher than that of the control group(P<0.05).The proliferation ability of cartilage cells induced by IL-1β was significantly lower than that of the control group(P<0.05). The proliferation ability of treated cartilage cells induced by IL-1β was significantly increased after miR-543 mimics transfection(P<0.05).The expression of MDM4 mRNA in miR-543 mimics transfected cells was significantly lower than that in the control group(P<0.05).The expression of Akt1 and Bcl-2 mRNA was significantly higher than that of the control group(P<0.05).The expression of MDM4 mRNA in chondrocytes induced by IL-1β was significantly higher than that in the control group(P<0.05),and the expression of Akt1 and Bcl-2 mRNA.The expression of MDM4,Akt1 and Bcl-2 mRNA in chondrocytes induced by IL-1β was significantly relieved after transfection with miR-543 mimics(P<0.05).Conclusion:The expression of miR-543 in cartilage tissue of OA rats is significantly decreased.The expression of miR-543 is up-regulated by miR-543 mimics transfection,which can enhance the proliferation of chondrocytes and the expression of MDM4,Aktl and Bcl-2.The effect is effective against the chondrocyte damage induced by IL-1β.