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目的探讨磷酸化视网膜母细胞瘤蛋白(phospho-retinoblastoma protein,p-Rb)在苯并[a]芘致神经元凋亡过程中的作用。方法用CCK-8试剂检测细胞活性;Annexin-V/PI双染法检测细胞凋亡率,免疫荧光细胞化学染色观察p-Rb表达,免疫印记法(Western-blot)检测p-Rb的表达情况。结果与对照组相比,各染毒组神经元活性降低(P<0.05),凋亡率增高(P<0.05),p-Rb表达量增高(P<0.05),2μmol/L组加入抑制剂后细胞活性增强(P<0.05),凋亡率下降(P<0.05),p-Rb阳性细胞数减少(P<0.05),p-Rb表达量降低(P<0.05)。结论苯并[a]芘可引起磷酸化Rb水平升高,是苯并[a]芘致神经元凋亡的可能机制。
Objective To investigate the role of phospho-retinoblastoma protein (p-Rb) in the apoptosis of neurons induced by benzo [a] pyrene. Methods The cell viability was detected by CCK-8 reagent. The apoptosis rate was detected by Annexin-V / PI double staining and the expression of p-Rb was detected by immunofluorescence staining. The expression of p-Rb was detected by Western-blot . Results Compared with the control group, the activity of neurons decreased (P <0.05), the apoptosis rate increased (P <0.05) and the expression of p-Rb increased (P <0.05) (P <0.05). The apoptosis rate decreased (P <0.05), the number of p-Rb positive cells decreased (P <0.05) and the expression of p-Rb decreased (P <0.05). Conclusions Benzo [a] pyrene can cause the increase of phosphorylated Rb, which is a possible mechanism of neuronal apoptosis induced by benzo [a] pyrene.