论文部分内容阅读
目的研究DNA甲基化酶抑制剂5-氮杂-2’-脱氧胞苷(5-Aza-2’-dc)与百草枯联合对V79细胞作用后的活性氧含量及抗凋亡Bcl-2蛋白和促凋亡Bax蛋白表达的影响.方法 V79细胞经传代培养后分为5-Aza-2’-dc作用组(A组)、百草枯作用组(B组)、5-Aza-2’-dc加百草枯作用组(C组,即先应用5-Aza-2’-dc对V79细胞进行预处理12h,然后给予百草枯作用12h)、对照组(D组).采用2,7二氯荧光黄双乙酸盐(DCFH-DA)荧光探针装载,流式细胞仪测定各实验组V79细胞内活性氧含量,Western blot检测各实验组V79细胞内抗凋亡Bcl-2蛋白和促凋亡Bax蛋白的表达情况.结果 5-Aza-2’-dc与百草枯联合作用组(C组)V79细胞内活性氧含量、抗凋亡Bcl-2蛋白、促凋亡Bax蛋白表达与5-Aza-2’-dc组(A组)、百草枯组(B组)和对照组(D组)比较有统计学差异(P<0.05);C组V79细胞内抗凋亡Bcl-2蛋白表达、抗凋亡Bcl-2蛋白/促凋亡Bax蛋白比值降低,活性氧含量和促凋亡Bax蛋白表达升高.结论 5-Aza-2’-dc调控DNA甲基化可能通过活性氧代谢失衡、细胞调亡来促进百草枯对V79细胞的毒性损伤.
Objective To investigate the effects of DNA methylation inhibitor 5-Aza-2’-deoxycytidine (5-Aza-2’-dc) combined with paraquat on V79 cells and the effect of anti-apoptotic Bcl-2 Aza-2’-dc group (group A), paraquat group (group B), 5-Aza-2 ’ -dc plus paraquat (group C, pretreatment of V79 cells for 12h with 5-Aza-2’-dc and then with paraquat for 12h) and control group (group D) DCFH-DA fluorescent probe was used to load and the reactive oxygen species (ROS) in V79 cells of each experimental group were measured by flow cytometry. The anti-apoptotic Bcl-2 protein and the pro-apoptotic rate of V79 cells in each experimental group were detected by Western blot Bax protein expression in V79 cells after combined with 5-Aza-2’-dc and paraquat.Results The levels of reactive oxygen species (ROS), anti-apoptotic Bcl-2 protein and proapoptotic Bax protein in V79 cells were significantly decreased compared with 5-Aza (P <0.05). The anti-apoptotic Bcl-2 protein expression in V79 cells in group C was significantly higher than that in group C (P <0.05) Anti-apoptotic Bcl-2 protein / pro-apoptotic Bax protein ratio decreased, reactive oxygen species and pro-apoptotic B ax protein.Conclusion The regulation of DNA methylation by 5-Aza-2’-dc may promote paraquat toxicity to V79 cells through the imbalance of active oxygen metabolism and cell apoptosis.