脱氧雪腐镰刀菌烯醇对PC12细胞凋亡的影响

[目的] 本试验旨在研究脱氧雪腐镰刀菌烯醇（deoxynivalenol，DON）诱导PC12细胞凋亡及对相关基因Bcl-2、Bax、Bid mRNA和蛋白cleaved-Caspase 3、cleaved-Caspase 9表达水平的影响。[方法] 用不同质量浓度的DON（0、125、250、500、1 000和2 000 ng？mL-1）对PC12细胞染毒24 h，采用透射电镜观察、流式细胞术、荧光定量PCR、Western-blot等方法，研究DON对PC12细胞的超微结构、凋亡率、凋亡相关基因及蛋白表达的影响。[结果] 不同浓度的DON均可诱导细胞凋亡，各试验组细胞凋亡率均极显著高于对照组（P<0.01），并随着染毒剂量的增加而升高，在1 000 ng？mL-1时达到峰值；与对照组相比，各试验组Bax mRNA表达水平和Bax/Bcl-2值均极显著增加（P<0.01），Bcl-2 mRNA表达水平在DON浓度超过500 ng？mL-1时极显著下降（P<0.01），而Bid mRNA表达水平在DON浓度超过250 ng？mL-1时极显著下降（P<0.01）；Western-blot检测结果显示，各试验组cleaved-Caspase 9蛋白表达量均极显著高于对照组（P<0.01），而cleaved-Caspase 3蛋白表达量在DON浓度超过500 ng？mL-1时极显著增加。[结论] DON能够诱导PC12细胞发生凋亡，凋亡相关基因Bcl-2、Bax、Bid及蛋白Caspase3、Caspase9等参与了DON诱导PC12细胞凋亡的调控。
[Objectives] The experiment aimed at the effect of DON on Bcl-2,Bax,Bid mRNA and the cleaved-Caspase 3,cleaved-Caspase 9 protein in PC12 cells. [Methods] PC12 cells were exposed to deoxynivalenol(DON) at different concentrations(0,125,250,500,1 000,2 000 ng？mL-1) for 24 h. The changes of cell ultrastructure,cell apoptosis ratio,apoptosis-related genes expression and apoptosis-related protein expression were determined by transmission electron microscopy,flow cytometry,qPCR and Western-blot,respectively. [Results] Different concentrations of DON could induce cell apoptosis,compared with control group,the cell apoptosis ratio in experimental groups significantly increased(P<0.01),and the changes with the increase of DON concentrations,peak at 1 000 ng？mL-1. Compared with control group,the Bax mRNA and Bax/Bcl-2 ratio was significantly increased(P<0.01),the Bcl-2 mRNA was significantly decreased(P<0.01) when the concentration of DON above 500 ng？mL-1,and the Bid mRNA was significantly decreased(P<0.01) when the concentration of DON above 250 ng？mL-1. Western-blot results showed that the cleaved-Caspase 9 in experimental groups were significantly increased(P<0.01),however,the cleaved-Caspase3 significantly increased(P<0.01) when the concentration of DON was above 500 ng？mL-1. [Conclusions] DON could induce PC12 cell apoptosis,and Bcl-2,Bax,Bid,Caspase 3 and Caspase 9 participated in the regulation of cell apoptosis