甘氨脱氧胆酸盐诱导人正常肝细胞株HL-7702凋亡过程中caspase3,9活性与mRNA表达水平变化

传良敏; 黄文方; 曾娅莉; 洪华; 邓君; 杨永长

甘氨脱氧胆酸盐诱导人正常肝细胞株HL-7702凋亡过程中caspase3,9活性与mRNA表达水平变化

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- 出版日期: 2008-01-20

Changes in caspase-3, 9 activity and mRNA expression of normal human hepatocyte HL-7702 cell in the process of apoptosis induced by glycodeoxycholate.

摘要

摘要: 目的观察甘氨脱氧胆酸盐（glycodeoxycholate,GCDC）诱导人正常肝细胞株HL-7702凋亡过程中caspase3,9活性与mRNA表达的变化,探讨GCDC诱导肝细胞凋亡的机制。方法体外培养HL-7702细胞,不同浓度的GCDC为处理因素,用Annexin V-FITC/PI双染色法结合流式细胞技术仪检测细胞凋亡率,比色法测定caspase3,9活性,RT-PCR检测caspase3,9mRNA表达水平。结果100μM250μM的GCDC处理HL-7702细胞24h后,其细胞凋亡率、caspase3,9活性与mRNA表达水平明显升高,并与GCDC呈浓度依赖性。结论caspase3,9参与GCDC诱导HL-7702细胞凋亡调控,GCDC通过活化caspase9,进一步活化caspase3诱导肝细胞凋亡。
- 甘氨脱氧胆酸盐 /
- 细胞凋亡 /
- caspase
Abstract: Objective To investigate the apoptosis and changes of caspase-3, 9 activity and mRNA expression induced by glycodeoxycholate (GCDC) in cultured normal human hepatocyte HL-7702 cell in vitro and to explore the possible mechanism of hepatocyte apoptosis.Methods HL-7702 cells were cultured with GCDC which the final concentration was 100μM、150μM、200μM、250μM respectively.The apoptosis ratio of HL-7702 cell was determined by Annexin V-FITC/PI double staining.Colorimetric method was used to measure caspase-3, 9 activity.The expression of caspase-3, 9 mRNA was determined by RT-PCR.Results When hepatocyte were incubated with GCDC for 24 hours, the apoptotic ratio of hepatocyte、caspase-3, 9 activity and mRNA expression of HL-7702 cells increased remarkably and had concentration-dependent with GCDC.Conclusion The caspase-3, 9 were concerned with the hepatocyte apoptosis induced by GCDC.The mechanism might be related to up-regulation of caspase9 activity which subsequently transforms caspase-3 into its active form.
- glycodeoxycholate /
- apoptosis /
- caspase

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参考文献(11)

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