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目的观察NaAsO2在诱导人胚肝细胞株(L-02)细胞凋亡过程中对细胞色素-C(Cytochrome C;Cyto-c)释放和半胱天冬酶-3(Caspase-3)表达的影响。方法分别以0(对照组)、50、100和150μmol/L的NaAsO2染毒L-02肝细胞24 h后,采用流式细胞仪检测细胞凋亡率;Western blot法检测Cyto-c和Caspase-3在NaAsO2诱导L-02肝细胞凋亡过程中的变化。结果流式细胞术检测到0、50、100和150μmol/L浓度NaAsO2组细胞凋亡率为6.3%±0.40%、12.1%±0.89%、20.15%±0.50%和28.23%±4.77%。与对照组相比细胞凋亡率明显增高,差异具有统计学意义(P<0.05或P<0.01)。Western blot法检测Cyto-c的释放量分别是1.10%±0.35%,1.73%±0.73%、1.83%±0.67%和2.43%±0.74%,随染砷剂量的增加而增加,仅高剂量组差异具有统计学意义(P<0.05);Caspase-3分别为0.41%±0.45%,0.74%±0.14%、0.76%±0.04%和1.02%±0.18%,随染砷剂量的增加而增加,差异具有统计学意义(P<0.05或P<0.01);结论在该试验条件下,NaAsO2可诱导L-02肝细胞发生凋亡,Cyto-c和Caspase-3在细胞凋亡过程中发挥着重要作用。
Objective To investigate the effects of NaAsO2 on the release of Cytochrome C (Cyto-C) and the expression of Caspase-3 during the induction of apoptosis in human embryonic hepatocyte line (L-02) . Methods L-02 hepatocytes were treated with 0 (control), 50, 100 and 150 μmol / L NaAsO2 for 24 h, respectively. The apoptosis rate of L-02 hepatocytes was detected by flow cytometry. The expressions of Cyto-c and Caspase- 3 in NaAsO2-induced apoptosis in L-02 hepatocytes. Results The apoptotic rate of NaAsO2 group was 6.3% ± 0.40%, 12.1% ± 0.89%, 20.15% ± 0.50% and 28.23% ± 4.77% by flow cytometry at 0, 50, 100 and 150μmol / L, respectively. Compared with the control group, the apoptosis rate was significantly increased, the difference was statistically significant (P <0.05 or P <0.01). Western blot showed that the release of Cyto-c was 1.10% ± 0.35%, 1.73% ± 0.73%, 1.83% ± 0.67% and 2.43% ± 0.74%, respectively, and increased with the increase of arsenic dose. (P <0.05). Caspase-3 was 0.41% ± 0.45%, 0.74% ± 0.14%, 0.76% ± 0.04% and 1.02% ± 0.18% respectively with the increase of arsenic dose Statistically significant (P <0.05 or P <0.01). Conclusions Under this experimental condition, NaAsO2 induces apoptosis in L-02 hepatocytes and Cyto-c and Caspase-3 play an important role in apoptosis.