摘要
目的探讨氨甲酰化促红细胞生成素(CEPO)对6-羟基多巴胺(6-hydroxydopamine,6-OHDA)诱导PC12细胞损伤及凋亡的保护作用及其可能机制。方法借助CCK8、流式细胞(Flow cytometry,FCM)、Western-blotting和逆转录PCR(RT-PCR)技术检测PC12模型细胞相关指标的变化,实验数据以SPSS15软件统计分析。结果 CCK8结果显示6-O-HDA处理能够显著降低PC12模型细胞的存活率,而CEPO处理对其变化显示抑制作用;FCM技术探察结果显示,6-OHDA处理能够明显诱导PC12模型细胞的损伤与凋亡,而CEPO预处理能够减轻PC12模型细胞的损伤与凋亡(P<0.05);借助Western-blotting技术探察显示6-OHDA+CEPO处理组的PC12细胞的Cleaved caspase-3蛋白表达量显著低于6-OHDA处理组(P<0.05);RT-PCR结果显示,6-OHDA处理明显降低PC12模型细胞的Bcl-2 mRNA的表达(P<0.05)以及上调PC12模型细胞的Bax mRNA的表达(P<0.01),而CEPO处理明显上调Bcl-2 mRNA的表达水平(P<0.05)以及抑制Bax mRNA的表达(P<0.05)。结论CEPO处理能够显著抑制6-OHDA所诱导的PC12细胞的损伤及凋亡,其作用机制可能与其上调Bcl-2,并下调Bax和Caspase-3的表达有关。
Objective To investigate the effect and possible mechanisms of carbamylated erythropoietin（CEPO） in inhibiting the injury of PC12 cells induced by 6-hydroxydopamine（6-OHDA）.Methods PC12 cells were divided into the three groups： control group,6-OHDA group and 6-OHDA ＋ CEPO group.The PC12 cells viability was measured by CCK8 assay.Flow cytometry（FCM） was used to determine apoptosis rate of PC12 cells.The expression of cleaved caspase-3 was detected by western-blotting.The expression of Bcl-2 and Bax mRNA in PC12 cells were measured by Reverse transcriptase polymerase chain reaction（RT-PCR）.Results CCK8 assay showed that the cell viability of PC12 cells treated with 200μmol 6-OHDA decreased to 56.70±7.86%,while 40U CEPO treatment increased the cell viability to（87.9±5.3）%（P〈0.05）.Flow cytometry demonstrated that CEPO treatment significantly inhibited the apoptosis of PC12 cells induced by 6-OHDA（P〈0.05）.RT-PCR results showed that the expression of Bcl-2 and Bax mRNA in PC12 cells were significantly upregulated and downregulated by CEPO as compared with 6-OHDA treated group（P〈0.05）.Western-blotting showed CEPO treatment markedly induced the downregulation of cleaved caspase-3 expression compared to 6-OHDA treatment alone（P〈0.05）.Conclusion CEPO protects PC12 cells from injury and apoptosis induced by 6-OHDA.The protective effect of CEPO might be executed by upregulation of Bcl-2 and downregulation of Bax and Caspase-3 expression in PC12 cells.
出处
《解剖学研究》
CAS
2013年第1期12-16,共5页
Anatomy Research
基金
国家自然科学基金(31070941
30770679)
国家科委973项目(2010CB530000)分题(2010CB530004)
