N-乙酰半胱氨酸和caspase-3抑制剂对镉诱导293细胞凋亡的拮抗作用  

Antagonistic effect of N-acetyl cysteine and caspase-3 inhibitor on apoptosis of 293 cells induced by cadmium

作　　者：王雨[1] 徐兆发[1] 喻道军[1] 

机构地区：[1]中国医科大学公共卫生学院环境卫生教研室,辽宁沈阳110001

出　　处：《中国工业医学杂志》2006年第6期321-323,361,共4页Chinese Journal of Industrial Medicine

基　　金：国家自然科学基金资助项目(30371200)

摘　　要：目的研究N-乙酰半胱氨酸(-Nacetyl cysteine,NAC)和caspase-3抑制剂(Z-DEVD-fmk)对镉致293细胞凋亡的影响,探讨氧化损伤和caspase途径在镉致细胞凋亡中的作用。方法体外培养的转化人胚肾293细胞分别以0、20、40、80、120、200μmol/L的氯化镉(CdCl2)处理0、3、6、12、24 h,NAC和Z-DEVD-fmk预处理组分别在镉处理前以2.5、5.0、10.0 mmol/L的NAC和0.1、1.0、10μmol/L的Z-DEVD-fmk预处理24 h和1 h,然后再以40μmol/LCdCl2处理24 h,MTT法测细胞相对存活率,流式细胞术Annexin-V-PI双染法检测细胞凋亡百分率。结果随着镉处理浓度的增加,CdCl2对细胞活力的抑制作用增强,细胞的相对存活率明显降低,与0μmol/L镉处理组比较,40、80、120、200μmol/L镉处理组293细胞的凋亡百分率显著升高(P<0.01);200μmol/L镉处理组与120μmol/L CdCl2组比较293细胞的凋亡百分率显著下降(P<0.01)。与40μmol/L镉处理组比较,5.0 mmol/L和10 mmol/L NAC预处理组细胞凋亡百分率显著降低,0.1、1.0、10μmol/L Z-DEVD-fmk预处理可使镉致293细胞凋亡百分率显著降低。结论NAC和Z-DEMD-fmk对镉诱导293细胞凋亡有明显的抑制作用,镉诱导细胞凋亡与氧化损伤和caspase途径密切相关。Objective To study the effects of N-acetyl cysteine and caspas-3 inhibitor on apoptosis of transformed human embryonic kidney 293 cells induced by cadmium tot exploring the role of oxidative damage and caspase pathway played in the apoptosis. MeUlod Transformed human embryonic kidney 293 cells wen incubated with 0, 20, 40, 80, 120, 200 μmol/L CdCl2 respectively for 0, 3, 6, 12 and 24 h. The cells in groups of NAC and caspase-3 inhibitor （Z-DEVD-fmk） were pre-treated with 2.5, 5.0, 10.0 mmol/L NAC and 0.1, 1.0, 10 ttmol/L Z-DEV1）-fmk for 24 h and 1 h respectively before 24 h incubation with 40 μmol/L CdCI2. Using MTT method for measuring the relative survival rate and flow cytometry annexin-V/Pi double stain method for measuring the apoptosis percentage of test cells. Result The results showed that the cellular relative survival rate was time and dose dependent upon the dose of cadmium. Compared with 0 μmol/L cadmium, the apeptosis rate increased obviously in the incubating cells with 40, 80, 120, 200 μmol/L cadmium （ P 〈 0.01）. Both NAC （an antioxidant agent） and Z-DEVD-fmk （a selective inhibitors of caspase-3）, might significantly suppress cadmium-induced cell apoptosis （ P 〈 0.01 ）. Conclusion Antioxidant agent and selective inhibitor of caspase-3 may suppress the apoptosis induced by Cd and oxidative stress and caspase-3 might be involved in Cd-induced apoptosis.

关 键 词：CASPASE-3抑制剂 N-乙酰半胱氨酸 镉 凋亡 

分 类 号：R994.6[医药卫生—毒理学]