盐酸米诺环素联合栀子苷对NO诱导PC12细胞凋亡的影响

摘要

Aim:To investigate the effects of minocycline,geniposide alone or combined on the apoptosis of PC12 cells induced by nitric oxide and the possible mechanism.Methods: PC12 cells were divided into control group,minocycline group,geniposide group and the combined group.The four groups were all given 0.5 mmol/L SIN-1 to induce apoptosis,minocycline group was given 0.1 mmol/L minocycline 12 hours before the SIN-1 intervention,geniposide group was given 0.25 mmol/L geniposide,the combined group was given the 2 drugs,and control group was not given any drug.The cell survival was determined by CCK-8 method,the apoptosis was observed by Hoechest 33342 fluorescent staining,the apoptosis rate was measured by flow cytometry,the expressions of the apoptotic related proteins Caspase-3 and Bcl-2 were detected by Western blot.Results: Compared with control group,the cell viability improved,the apoptosis rate decreased,the expression of Bcl-2 upregulated,and that of Caspase-3 downregulated in minocycline group and geniposide group(P<0.05);minocycline and geniposide had synergistic effects on the above indicators(P<0.05).Conclusion: Minocycline and geniposide both can inhibit the apoptosis of PC12 cells,but the effect of the combined application is superior to the single application.The mechanism may be related to the inhibition of Caspase-3 dependent apoptosis signaling pathways.%目的:探讨盐酸米诺环素、栀子苷单独及联合应用对NO诱导PC12细胞凋亡的影响及其可能机制.方法:PC12细胞分为对照组、盐酸米诺环素组、栀子苷组和盐酸米诺环素+栀子苷组(联合应用组).4组均用终浓度为0.5 mmol/L的NO供体SIN-1诱导细胞凋亡,SIN-1干预前12 h盐酸米诺环素组加入终浓度为0.1 mmol/L的盐酸米诺环素,栀子苷组加入终浓度为0.25 mmol/L的栀子苷,联合应用组加入同上浓度的盐酸米诺环素和栀子苷,对照组不给药.CCK-8法测定细胞存活率,Hoechest 33342荧光染色观察细胞凋亡情况,流式细胞仪测定细胞凋亡率,Western blot测定凋亡相关蛋白Caspase-3和Bcl-2的表达.结果:与对照组相比,单用盐酸米诺环素或栀子苷均能提高NO诱导的PC12细胞存活率,降低细胞凋亡率,增加Bcl-2蛋白表达,减少Caspase-3蛋白表达(P均<0.05);二者联合应用对上述指标的影响具有协同效应(P<0.05).结论:盐酸米诺环素和栀子苷均能抑制PC12细胞凋亡,且二者联合应用作用优于单独应用,其机制可能与抑制Caspase-3依赖性凋亡信号通路有关.