骨形成蛋白7对糖皮质激素诱导PC12细胞凋亡的作用研究

Effect of BMP-7 on glucocorticoid-induced apoptosis in PC12 cells

目的探究骨形成蛋白7(BMP-7)对糖皮质激素(GC)诱导PC12细胞凋亡的作用。方法采用不同浓度GC干预PC12细胞,明确不同浓度GC对PC12细胞凋亡的影响;在PC12细胞中感染BMP-7过表达/敲低慢病毒和对照病毒,然后采用DMSO或GC干预细胞,流式细胞仪检测细胞凋亡水平,实时荧光定量聚合酶链反应(qRT-PCR)检测Bax基因表达,Western blotting法检测Cleaved-Caspase-3蛋白表达。结果不同浓度GC处理后的细胞凋亡率、Bax基因和Cleaved-Caspase-3蛋白表达比较,差异有统计学意义(P<0.05);与0μmol/L GC比较,GC浓度为10μmol/L时,PC12细胞凋亡率明显升高(P<0.05);GC浓度为50μmol/L时,PC12细胞Bax基因和Cleaved-Caspase-3蛋白表达增加(P<0.05)。过表达或敲低BMP-7后4组细胞的细胞凋亡率、Bax基因和Cleaved-Caspase-3蛋白表达比较,差异有统计学意义(P<0.05);与GC-NC组比较,GC-oe BMP-7组细胞凋亡率、Bax基因和Cleaved-Caspase-3蛋白表达均降低(P<0.05);与DMSO-NC组比较,GC-oe BMP-7组细胞凋亡水平、Bax基因和Cleaved-Caspase-3蛋白表达差异无统计学意义(P>0.05)。与DMSO-NC组比较,DMSO-sh BMP-7组细胞凋水平、Bax基因和Cleaved-Caspase-3蛋白表达均增加(P<0.05);与GC-NC组比较,GC-sh BMP-7组细胞凋水平、Bax基因和Cleaved-Caspase-3蛋白表达显著增加(P<0.05)。结论高水平GC激素诱导PC12细胞凋亡增加,BMP-7可以拮抗GC诱导的细胞凋亡增加。

Objective To investigate the effect of Bone Morphogenetic Protein 7(BMP-7)on glucocorticoid(GC)-induced apoptosis in PC12 cells.Methods PC12 cells were treated with different concentrations of GC to determine the impact of GC on apoptosis.PC12 cells were then infected with BMP7 overexpression/knockdown lentivirus or control virus.Subsequently,cells were treated with DMSO or GC,and apoptosis levels were assessed using flow cytometry.Real-time quantitative polymerase chain reaction(qRT-PCR)was employed to measure Bax gene expression,and Western blotting was used to evaluate Cleaved-Caspase-3 protein expression.Results There were statistically significant differences in apoptosis rate,Bax gene expression,and Cleaved-Caspase-3 protein expression among cells treated with different concentrations of GC(P<0.05).Compared to 0μmol/L GC,PC12 cell apoptosis significantly increased at a GC concentration of 10μmol/L(P<0.05),and at a concentration of 50μmol/L,Bax gene expression and Cleaved-Caspase-3 protein expression increased(P<0.05).Apoptosis rate,Bax gene expression,and Cleaved-Caspase-3 protein expression differed significantly among the four groups with BMP-7 overexpression or knockdown(P<0.05).Compared to the GC-negative control(GC-NC)group,cells in the GC-overexpressing BMP-7 group showed decreased apoptosis,Bax gene expression,and Cleaved-Caspase-3 protein expression(P<0.05).No significant differences were observed in apoptosis level,Bax gene expression,and Cleaved-Caspase-3 protein expression between the GC-overexpressing BMP-7 group and the DMSO-negative control(DMSO-NC)group(P>0.05).In comparison to the DMSO-NC group,cells in the DMSO-knockdown BMP-7 group exhibited increased apoptosis,Bax gene expression,and Cleaved-Caspase-3 protein expression(P<0.05).The GC-knockdown BMP-7 group showed a significant increase in apoptosis level,Bax gene expression,and Cleaved-Caspase-3 protein expression compared to the GC-NC group(P<0.05).Conclusion High levels of GC induce increased apoptosis in PC12 cells,and BMP-7 can counteract GC-induced apoptosis.