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)

[1]Greim H, Trulzsch D, Czygan P, et al.Mechanismof cholestasis.6.Bile acids in human livers with or without biliary obstruction[J].Gastroenterology, 1972, 63∶846~850.

[2]黄文方, 康格非, 杨明清, 等.甘氨脱氧胆酸诱导大鼠肝细胞凋亡[J].中华肝脏病杂志, 2000, 8 (5) ∶305~306.

[3]Kwo P, Patel T, Bronk SF, et al.Nuclear serine protease activitycontributes to bile acid-induced apoptosis in hepatocytes[J].Am JPhysiol, 1995, 268∶G613~G621.

[4]Li D, Sun H.Bile salt induces apoptosis of hepatocytes:the mecha-nism of hepatic function injury during obstructive jaundice[J].Zhon-ghua Wai Ke Za Zhi, 1998, 36 (10) ∶624~626, 117.

[5]Dseagher S, Martinou JC.Mitochondria as the central control point ofapoptosis[J].Trends Cell Biol, 2000, 10 (9) ∶369~377.

[6]Ashkenazi A, Dixit VM.Death receptors:signaling andmodulation[J].Science, 1998, 281 (5381) ∶1305~1308.

[7]Nakamura K, Bossy-Wetzel E, Burns K, et al.Changes in endo-plasmic reticulum luminal environment affect cell sensitivity to apop-tosis[J].J Cell Biol, 2000, 150 (4) ∶731~740.

[8]Zou H, Henzel WJ, Liu X, et al.Apaf-1, a human protein homol-ogous to C.elegans CED-4, participates in cytochrome c-depend-ent activation of caspase-3[J].Cell, 1997, 90 (3) ∶405~413.

[9]Li P, Nijhawan D, Budihardjo I, et al.Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an ap-optotic protease caspase[J].Cell, 1997, 91 (4) ∶479~489.

[10]Cohen GM.Caspase:the executioners of apoptosis[J].BiochemicalJournal, 1997, 326 (Pt) ∶1~16.

[11]Cynthia RL, Webster, Paul U, et al.cAMP inhibits bile acid-in-duced apoptosis by blocking caspase activation and cytochrome c re-lease[J].Am J Physiol Gastrointest Liver Physiol, 2002, 283∶G727~G738.

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