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5-Aza-2'-dc预处理致敏百草枯对V79细胞活性氧及Bcl-2/Bax表达的影响
王 晔
(昆明医科大学附属延安医院急诊医学科,云南 昆明 650051)
摘要:
[摘要]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.
关键词:  [关键词]5-氮杂-2'-脱氧胞苷  百草枯  V79细胞  活性氧  Bcl-2/Bax
DOI:
分类号:
基金项目:[基金项目]云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目(2013FB190)
Influences of Pretreating with 5-Aza-2'-dc on the Reactive Oxygen Level and the Expression of Protein Bcl-2 and Bax in Paraquat Sensitized V79 Cells
WANG Ye
(Dept. of Emergency,Affiliated Yan’an Hospital of Kunming Medical University,Kunming Yunnan 650051,China)
Abstract:
[Abstract]Objective To study the reactive oxygen level and the expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax after treatment of DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-Aza-2'-dc) and paraquat in V79 cells.Methods Cultured V79 cells were divided into 5-Aza-2'-dc treatment group (group A), paraquat treatment group(group B),5-Aza-2'-dc and paraquat treatment group (group C, V79 cells were pretreated with 5-Aza-2'-dc for 12h followed by exposure to paraquat for 12h)and control group (group D). Reactive oxygen level in V79 cells was measured by DCFH-DA flow cytometry and expression of Bcl-2 and Bax was detected by Western blot.Results Reactive oxygen levels and expression levels of Bcl-2 and Bax in V79 cells were significantly different(P<0.05) in 5-Aza-2'-dc and paraquat treatment group(group C),compared with 5-Aza-2'-dc treatment group (group A), paraquat treatment group(group B) and control group(group D). Expression levels of Bcl-2 and the ratio of Bcl-2 and Bax were lower while reactive oxygen levels and expression levels of Bax were higher in group C than in groups A, B and D. Conclusion 5-Aza-2'-dc regulates DNA methylation by the imbalancing the reactive oxygen metabolism and apoptosis, thus up-regulating the toxic effect of paraquat on V79 cells.
Key words:  [Key words]5-Aza-2'-dc  Paraquat  V79 cells  Reactive oxygen level  Bcl-2/Bax