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目的研究西红花苷对Aβ_(25-35)诱导的大鼠肾上腺嗜铬细胞瘤细胞(PC12)凋亡的影响。方法体外培养的PC12细胞,分为5组:正常对照组、阿尔茨海默病(AD)模型组、西红花苷(低、中、高)剂量组。CCK-8法检测细胞存活率,Hoechst 33258检测细胞凋亡状况,流式细胞仪检测细胞凋亡率,Western blot实验检测凋亡相关蛋白Bcl-2、Caspase-3的表达。结果与模型组比较,西红花苷各剂量组细胞的存活率均明显升高(q值分别为3.455、6.975、7.078,P值分别<0.05、<0.01、<0.01),凋亡率显著性下降(q值分别为4.609、13.460、13.679,P值均<0.01),Bcl-2蛋白表达增加(q值分别为7.642、14.577、16.983,P值均<0.01),Caspase-3表达减少(q值分别为4.236、8.210、7.555,P值分别<0.05、<0.01、<0.01)。与西红花苷低剂量组比较,西红花苷中、高剂量组细胞存活率均升高(q值分别为3.544、5.462,P值均<0.05),细胞凋亡率呈显著性减低(q值分别为8.849、9.064,P值均<0.01),但中、高剂量组之间无统计学差异。结论西红花苷对Aβ25-35诱导的PC12细胞损伤具有明显的保护作用,其机制可能与增加Bcl-2的表达、抑制Caspase-3依赖性凋亡信号通路有关。
Objective To study the effect of crocin on Aβ 25-35 induced apoptosis in adrenal pheochromocytoma cells (PC12). Methods PC12 cells cultured in vitro were divided into 5 groups: normal control group, Alzheimer’s disease (AD) model group and crocin (low, medium and high) dose groups. The cell viability was detected by CCK-8 assay. The apoptosis of Hoechst 33258 cells was detected by flow cytometry. The expressions of Bcl-2 and Caspase-3 were detected by Western blot. Results Compared with the model group, the survival rates of crocetin in each dose group were significantly increased (q = 3.455,6.975,7.078, P <0.05, <0.01, <0.01, respectively) (Q values were 7.642, 14.577 and 16.983, P <0.01 respectively), and the expression of Caspase-3 decreased (q values were 4.609, 13.460 and 13.679, respectively, P <0.01) Values were 4.236,8.210,7.555, P values were <0.05, <0.01, <0.01). Compared with crocin low dose group, crocetin medium and high dose group cell survival rates were increased (q values were 3.544,5.462, P values were <0.05), the apoptotic rate was significantly reduced q values were 8.849,9.064, P values were <0.01), but no significant difference between the medium and high dose groups. Conclusion Crocin obviously protects PC12 cells induced by Aβ25-35, and its mechanism may be related to the increase of Bcl-2 expression and the inhibition of Caspase-3-dependent apoptosis signaling pathway.