盲肌样蛋白1(MBNL1)对血小板衍生生长因子诱导大鼠血管平滑肌细胞表型转换的作用研究

Effect of Muscleblind-like 1(MBNL1)on Phenotypic Switching of Rat Vascular Smooth Muscle Cells Induced by Platelet Derived Growth Factor

目的对盲肌样蛋白1(MBNL1)在血管平滑肌细胞(VSMCs)表型转换中的作用进行研究。方法分离并鉴定大鼠血管平滑肌细胞,分组为对照组、10 ng/ml组、20 ng/ml组、40 ng/ml组、80 ng/ml组,给予不同浓度血小板衍生生长因子(PDGF-bb),通过Western blot检测各组MBNL1表达水平改变情况。使用慢病毒在细胞水平过表达MBNL1,分为Lv-nc组、Lv-nc+PDGF-bb组、Lv-mbnl1组、Lv-mbnl1+PDGF-bb组,并给予PDGF-bb刺激。使用siRNA在细胞水平敲低MBNL1,分为siRNA-nc组、siRNA-nc+PDGF-bb组、siRNA-mbnl1组、siRNA-mbnl1+PDGF-bb组,并给予PDGF-bb刺激,通过Transwell实验检测细胞迁移能力。结果给予PDGF-bb后10 ng/ml组、20 ng/ml组、40 ng/ml组、80 ng/ml组MBNL1表达量较对照组增高(P<0.05);Lv-nc组细胞迁移数与Lv-mbnl1组比较,差异无统计学意义(P>0.05);Lv-nc+PDGF-bb组细胞迁移数较Lv-mbnl1组高(P<0.05);siRNA-nc组细胞迁移数与siRNA-mbnl1组比较,差异有统计学意义(P<0.05);siRNA-nc+PDGF-bb组细胞迁移数低于siRNA-mbnl1组(P<0.05)。结论MBNL1抑制VSMCs迁移能力进而影响细胞表型转换。

Objective To investigate the role of muscleblind-like 1(MBNL1)in phenotypic switching of vascular smooth muscle cells(VSMCs).Methods Rat vascular smooth muscle cells were isolated and identified,and divided into control group,10 ng/ml group,20 ng/ml group,40 ng/ml group,and 80 ng/ml group.Different concentrations of platelet-derived growth factor(PDGF-bb)were given,and the change of MBNL1 expression level in each group was detected by Western blot.The lentivirus was used to overexpress MBNL1 at the cell level,which was divided into Lv-nc group,Lv-nc+PDGF-bb group,Lv-mbnl1 group and Lv-mbnl1+PDGF-bb group,and PDGF-bb stimulation was given.The MBNL1 was knocked down by siRNA at the cell level,which was divided into siRNA-nc group,siRNA-nc+PDGF-bb group,siRNA-mbnl1 group,and siRNA-mbnl1+PDGF-bb group.PDGF-bb stimulation was given,and the cell migration ability was detected by Transwell experiment.Results The expression levels of MBNL1 in the 10 ng/ml group,20 ng/ml group,40 ng/ml group and 80 ng/ml group after PDGF-bb administration were higher than those in the control group(P<0.05).There was no significant difference in cell migration between Lv-nc group and Lv-mbnl1 group(P>0.05).The number of cell migration in Lv-nc+PDGF-bb group was higher than that in Lv-mbnl1 group(P<0.05).The number of cell migration in siRNA-nc group was significantly different from that in siRNA-mbnl1 group(P<0.05).The number of cell migration in siRNA-nc+PDGF-bb group was lower than that in siRNA-mbnl1 group(P<0.05).Conclusion MBNL1 inhibits the migration ability of VSMCs and further affects cell phenotype conversion.