甘氨脱氧胆酸盐诱导人正常肝细胞HL-7702凋亡及其机制的研究  

Apoptosis mechanism of cultured human hepatocyte HL-7702 induced by glycodeoxycholate

作　　者：传良敏[1] 洪华[1] 黄文芳[1] 杨永长[1] 饶绍琴[1] 邓君[1] 

机构地区：[1]四川省医学科学院,四川省人民医院检验科,四川成都610072

出　　处：《西安交通大学学报（医学版）》2009年第1期36-39,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的观察甘氨脱氧胆酸盐(glycodeoxycholate,GCDC)对人正常肝细胞株HL-7702凋亡的影响,并探讨其凋亡机制。方法终浓度分别为100、150、200、250μmol/L的GCDC处理HL-7702细胞24h,光学显微镜观察GCDC对HL-7702细胞形态的影响,AnnexinV-FITC/PI双染色法检测HL-7702细胞的凋亡率,用荧光指示剂Fluo-3/AM测定HL-7702细胞内钙离子浓度,RT-PCR测定Bcl-2、Bax基因mRNA表达水平。结果终浓度为150μmol/L的GCDC处理HL-7702细胞24h后,细胞出现典型的凋亡细胞形态学改变;100、150、200、250μmol/L的GCDC均可诱导HL-7702细胞凋亡,呈剂量依赖性,凋亡率分别为(13.16±2.9)%(t=6.41)、(20.3±3.0)%(t=10.22)、(25.02±2.1)%(t=18.11)、(45.02±3.5)%(t=28.89),较对照组(2.2±0.6)%显著升高(P<0.05);GCDC能使HL-7702细胞内钙离子浓度增加,且具有浓度依赖性;GCDC使细胞内Bcl-2mRNA的表达下降、BaxmRNA的表达增加。结论GCDC可能是通过增加HL-7702细胞内钙离子浓度,并进一步下调Bcl-2、上调Bax表达水平,从而诱导细胞凋亡。Objective To investigate the apoptosis of cultured normal human hepatocyte HL-7702 cells induced by glycodeoxycholate （GCDC） and to explore its possible mechanism. Methods HL-7702 cells were incubated with various concentrations （100, 150, 200 and 250μmol/L） of GCDC. The changes of cellular morphology were observed under optical microscope. The apoptosis rate of HL-7702 was determined by Annexin V-FITC/PI double staining. The changes of HL-7702 cell intracelluar [Ca^2＋ ]i were determined with Fluo-3/AM load technique. The mRNA expression levels of Bcl-2/Bax in HL-7702 cells were analyzed by RT-PCR. Results Typical apoptotic morphological changes were observed after HL-7702 cells had been treated with 150μmol/L GCDC for 24 h; HL-7702 cells could be induced to undergo apoptosis in a concentration-dependent manner after 100, 150, 200, and 250 μmol/ LGCDC treatment for 24 hours. The apoptosisrates were （13.16±2.9）%, （20.3±3.0）%, （25.02±2.1）% and （45.02±3.5）%, which were markedly higher （P〈0.05） than those of the control group. The intracellular [Ca^2＋ ]i was increased in a concentration-dependent manner when HL-7702 cells were incubated with GCDC. The expression of Bcl-2 mRNA was decreased and the expression of Bax mRNA was increased. Conclusion GCDC can induce HL- 7702 cells apoptosis. The potential mechanism of apoptosis might be related to increasing intracellular [Ca^2＋ ]i, downregulating the expression of Bcl-2 mRNA and up-regulating the expression of Bax mRNA.

关 键 词：甘氨脱氧胆酸盐 细胞凋亡 钙离子 BCL-2 BAX 肝细胞 

分 类 号：R575[医药卫生—消化系统]