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

摘要:

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0 μmol/L GC比较，GC浓度为10 μmol/L时，PC12细胞凋亡率明显升高（P <0.05）；GC浓度为50 μmol/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。

Abstract:

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 0 μmol/L GC, PC12 cell apoptosis significantly increased at a GC concentration of 10 μmol/L (P < 0.05), and at a concentration of 50 μmol/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.