周期性机械应力对大鼠软骨细胞增殖的影响

Effect of cyclic mechanical stress on the proliferation of chondrocyte in rats

目的:探讨周期性机械应力对大鼠软骨细胞增殖的影响。方法:通过Ⅱ型胶原酶消化分离大鼠原代软骨细胞,并将软骨细胞分为对照组和实验组。MTT法检测细胞活力,实时荧光定量PCR(Realtime PCR)法检测蛋白聚糖(Aggrecan)及Ⅱ型胶原酶(ColⅡ)mRNA的表达,western blot检测B细胞淋巴瘤/白血病-2(Bcl-2)、增殖细胞抗原(PCNA)及细胞周期蛋白D1(CyclinD1)蛋白表达水平与细胞外调节蛋白激酶(ERK)磷酸化水平。结果:与对照组比较,实验组1 h、2 h的软骨细胞活力均明显提高(P<0.05),Aggrecan及ColⅡmRNA表达量均明显提高(P<0.05),Bcl-2、PCNA及CyclinD1蛋白表达量均明显上调(P<0.05),ERK1/2磷酸化水平明显上调(P<0.05)。结论:周期性机械应力能显著促进软骨细胞增殖。

Objective:To explore the effect of cyclic mechanical stress on the proliferation of chondrocyte in rats.Methods:The primary chondrocytes of rats were isolated by digestion with type II collagenase.The chondrocytes were divided into control group and experimental group.The cell viability was detected by MTT assay.The expression of Aggrecan and Col II mRNA was detected by Realtime PCR.The expression of B cell lymphoma/lewkmia-2(Bcl-2),Proliferating Cell Nuclear Antigen(PCNA),CyclinD1 and the phosphorylation level of extracellular regulated protein kinases(ERK)was detected by Western blot.Results:Compared with control group,the activity of chondrocytes in the experimental group increased significantly at 1 h and 2 h(P<0.05),the expression of Aggrecan and Col II mRNA was increased(P<0.05),the expression of Bcl-2,PCNA,CyclinD1 and p-ERK1/2 was up-regulated in experimental group(P<0.05).Conclusion:Cyclic mechanical stress can significantly promote chondrocyte proliferation.