Rsk2沉默对大鼠骨髓基质干细胞的成骨分化的影响

Effect of Rsk2 silencing on osteogenic differentiation of rat bone marrow stromal cells

目的通过对核糖体S6激酶2(Rsk2)进行沉默,探讨Rsk2对大鼠骨髓基质干细胞(BMSCs)的成骨分化的影响。方法分离出BMSCs,空白对照组不做处理,模型组用Lipofectamine法将siRNA-NC转染到BMSCs细胞,实验组用Lipofectamine法将Rsk2 siRNA转染到BMSCs细胞用于沉默Rsk2,转染48 h后换用成骨诱导分化培养基,在诱导分化后第7 d检测碱性磷酸酶(ALP)相对活性,Rsk2、骨钙蛋白(OCN)、骨形成蛋白2(BMP2)、Ⅰ型胶原(COLL-Ⅰ)和成骨相关蛋白Runt相关因子2(Runx2)mRNA表达水平。结果在诱导分化第7 d,空白对照组、模型组和实验组的Rsk2 mRNA相对表达量分别为1.00±0.03,0.95±0.08和0.50±0.07;OCN mRNA相对表达量分别为1.00±0.09,1.07±0.11和0.43±0.06;BMP2 mRNA相对表达量分别为1.00±0.06,1.06±0.12和0.67±0.07;COLL-ⅠmRNA相对表达量分别为1.00±0.02,1.10±0.09和0.73±0.06;Runx2 mRNA相对表达量分别为1.00±0.08,1.04±0.04和0.52±0.08;实验组与空白对照组和模型组比较,上述指标的差异均有统计学意义(均P<0.05)。结论对大鼠Rsk2基因进行沉默后,BMSCs转化为成骨细胞的能力减弱。

Objective To investigate the effect of ribosomal S6 kinase 2(Rsk2)silencing on osteogenic differentiation of rat bone marrow stromal cells(BMSCs).Methods BMSCs were isolated and siRNA-NC was transfected into BMSCs cells by Lipofectamine method in model group and Rsk2 siRNA was transfected into BMSCs by Lipofectamine method to silence Rsk2.The blank group was not treated.After 48 hours of transfection,the BMSCs were transformed into osteoinductive differentiation medium.The relative activity of alkaline phosphatase(ALP),Rsk2,osteocalcin(OCN),bone morphogenetic protein 2(BMP2),collagen type I(COLL-I)and osteogenic protein Runt 2(Runx2)were detected on the 7 th day after induction of differentiation.Results On the 7 th day after differentiation,the relative expression levels of Rsk2 mRNA in the blank control group,the model group and the experimental group were 1.00±0.03,0.95±0.08 and 0.50±0.07,respectively.The relative expression levels of OCN mRNA in the blank control group,model group and experiment group were 1.00±0.09,1.07±0.11 and 0.43±0.06,respectively;the relative expression levels of BMP2 mRNA were 1.00±0.06,1.06±0.12 and 0.67±0.07,respectively;the relative expression levels of COLL-I mRNA were 1.00±0.02,1.10±0.09 and 0.73±0.06,respectively;the relative expression levels of Runx2 mRNA were 1.00±0.08,1.04±0.04 and 0.52±0.08,respectively.The difference of the above indicators between the experimental group and the blank control group,the model group were statistically significant(all P<0.05).Conclusion Rsk2 silencing can weak the osteogenic differentiation of rat BMSCs.