银杏内酯对PC12细胞缺糖损伤凋亡相关基因的影响

Effective of ginkgolides on expression of apoptosis related gene during PC12 cells glucose deprivation

目的:考察银杏内酯对缺糖引起的PC12细胞损伤的保护作用及对凋亡相关基因表达的影响,探讨银杏内酯对中枢神经系统的保护机制。方法:缺糖培养引起PC12细胞损伤,分别加入100,10,1,0.1mg.L-1的银杏内酯,MTT法检测给药前后与正常组的细胞活力变化,实时荧光定量PCR(Fluorescence Quantitative PCR,FQ-PCR)法分析各组抑制凋亡基因(Bcl-2),凋亡诱导基因(Bax)及原癌基因(c-myc)的表达情况。结果:缺糖培养早期12h时,银杏内酯给药使Bcl-2表达量增高,Bax和c-myc表达量降低;缺糖培养24h时,银杏内酯仅能引起c-myc的表达量下降。结论:在细胞缺糖损伤早期,银杏内酯通过调节凋亡相关基因Bcl-2,Bax和c-myc产生细胞应激保护效应,在损伤晚期主要通过调节c-myc的表达量产生应激保护效应。

Objective： To investigate the protective effects of ginkgolides on glucose deprivation-induced apoptosis in PC12 cells and the mechanism underlying the protective effect. Method： PC12 cells were treated under glucose deprivation, and the proliferation was determined by tetrazolium （M13＂） assay. Furthermore, the mRNA levels of Bcl-2, Bax, c-myc were measured by Fluorescence Quantitative PCR （FQ-PCR）. Result： Ginkgolides could markedly inhibit the injury of glucose deprivation on the PC12 cells and increase the cell proliferation compared with the model groups （P 〈 0.01 ）. Ginkgolides could up-regulate Bcl-2 and down-regulate Bax and c-myc at 12 h, respectively. There were no significant differences in the Bcl-2 and Bax levels in both groups at 24 h, and ginkgolides only reduced the elevation of c-myc from 4. 32-fold to 2. 87-fold at this time. Conclusion： Ginkgolides are able to protect the injured PC12 cells against cell apoptosis. During the early period of glucose deprivation, Bcl-2, Bax and c-myc were regulated to inhibit cell apoptosis by ginkgolides. After that, ginkgolides seems inhibit the apoptosis through attenuating the elevation of c-myc.