摘要
目的研究Cr（Ⅵ）对果蝇S2细胞凋亡的影响,初步探讨其可能的凋亡机制。方法将果蝇S2细胞分别暴露于含Cr（Ⅵ）终浓度为0（对照）、50、100、200、400、800μmol/L的新鲜培养基培养12 h。检测果蝇S2细胞的凋亡情况和凋亡相关基因Debcl、Buffy、Drice、P53 mRNA的表达水平及Drice的相对活化程度。结果 Cr（Ⅵ）处理浓度为400μmol/L时可观察到S2细胞染色质浓染、出现凋亡小体,当Cr（Ⅵ）浓度达到800μmol/L时,可明显观察到细胞崩解及坏死;与对照组比较,各剂量Cr（Ⅵ）染毒组果蝇S2细胞的早期凋亡率均较高,差异有统计学意义（P〈0.05）。当Cr（Ⅵ）浓度为50~400μmol/L时,果蝇S2细胞的早期凋亡率随着Cr（Ⅵ）浓度的上升而升高;当Cr（Ⅵ）浓度达到800μmol/L时,果蝇S2细胞的早期凋亡率下降。与对照组比较,400μmol/L Cr（Ⅵ）染毒组果蝇S2细胞Debcl、Drice mRNA的表达水平均较低,P53 mRNA的表达水平均较高,差异有统计学意义（P〈0.05）;而Buffy mRNA的表达水平无明显改变,Drice相对活化程度降低（P〈0.01）。结论天冬氨酸蛋白水解酶（Caspases）并未参与高浓度下Cr（Ⅵ）诱导果蝇S2细胞的凋亡过程,其凋亡机制可能与P53基因相关。
Objective To investigate the apoptosis mechanisms of Drosophila S2 cells exposed to chromium（ Ⅵ） [ Cr（ Ⅵ）].Methods Drosophila S2 cells were cultured by medium containing different concentrations（0, 50, 100, 200, 400, 800 μmol/L）of Cr（Ⅵ） for 12 h. Real-time PCR was used to evaluate expression of apoptosis-related genes including Debcl, Buffy, Drice and p53. A spectrophotometer was used to detect the relative degree of activation of Drice. Results Obvious apoptotic morphological changes such as heavy chromatin staining and apoptotic bodies were observed at 400 μmol/L when Cr（ Ⅵ）.Compared with the control group, Drosophila S2 cells exposed to Cr（Ⅵ） showed higher rate of early apoptosis,the difference was statistically significant. When Cr（ Ⅵ） exposure dose increased from 50 to 400 μmol/L,S2 cell early apoptosis rate also increased gradually（P0.05）. When the Cr（Ⅵ） concentration was 800 μmol/L,S2 cell early apoptosis rate decreased and S2 cell late apoptosis rate increased.With 400 μmol/L Cr（Ⅵ） treatment,Debcl m RNA expression decreased（P0.01）,Buffy m RNA expression had no significant change,p53 m RNA expression increased（P0.05）,Drice m RNA expression decreased（P0.01）,the relative degree of activation of Drice decreased（ P〈0. 01）. Conclusion Caspases does not participate in apoptosis in Drosophila S2 cells exposed to higher Cr（Ⅵ）; Cr（Ⅵ） may induce apoptosis in Drosophila S2 cells through p53 gene.
出处
《环境与健康杂志》
CAS
北大核心
2015年第7期583-586,F0003,共5页
Journal of Environment and Health
基金
国家自然科学基金(31260528)
贵州省卫生厅优秀医学青年科技人才基金(gzwkj2010-2-003)
