甘氨鹅脱氧胆酸盐对人正常肝细胞HL-7702的影响  

Effects of glycochenodeoxcholate on the human normol hepatocyte HL-7702

作　　者：陈念平[1] 周军[1] 秦勇[1] 廖博懿[1] 邱志东[1] 

机构地区：[1]广东医学院附属医院肝胆外科,广东湛江524001

出　　处：《广东医学》2010年第14期1773-1775,共3页Guangdong Medical Journal

基　　金：广东省科技计划项目(编号:2007B031405001)

摘　　要：目的观察甘氨鹅脱氧胆酸盐(glycochenodeoxcholate,GCDC)对人正常肝细胞HL-7702凋亡诱导作用,探讨GCDC诱导HL-7702细胞凋亡的机制。方法终浓度分别为50、100、150、200、250μmol/L的GCDC处理HL-7702细胞4、8、12、24h,光学显微镜镜下观察细胞形态学改变、用Am-Blue细胞活性检测和乳酸脱氢酶活性的测定及AnnexinV-FITC/PI双染色法检测GCDC对HL-7702细胞凋亡的影响,用荧光指示剂Fluo-3/AM测定HL-7702细胞内钙离子浓度。结果 (1)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,包括核固缩、核碎裂和凋亡小体形成等。(2)Am-Blue吸光光度法、GENMED乳酸脱氢酶(LDH)总活性比色法及AnnexinV-FITC/PI双染色法定量检测结果均显示,GCDC均可诱导HL-7702细胞凋亡且呈剂量及时间依赖性。(3)GCDC能使HL-7702细胞内钙离子浓度增加且具有浓度和时间依赖性。结论 GCDC可能通过增加HL-7702细胞内钙离子浓度诱导人正常肝细胞HL-7702凋亡,其作用具有浓度和时间依赖性。Objective To investigate the roles of glycochenodeoxcholate in apoptosis of human normol hepatocyte HL - 7702. Methods HL - 7702 cells were incubated in various concentrations of GCDC （final concentrations of 50 μmol/L, 100 Ixmol/L, 150 μmol/L, 200 μmol/L and 250 ixmoL/L, respectively） for 4 h, 8 h, 12 h and 24 h. The apoptosis of HL - 7702 cells was detected by light microscope, while the apoptotic ratio and intracellular calcium concentra- tions of HL - 7702 were determined by Annexin V - FITC/PI double staining and Fluo - 3/AM load technique, respectively. Results （ 1 ） After incubation with 150 I^mol/L GCDC for 24 h, HL - 7702 cells appeared typical morphological changes of apoptosis, including nuclear pyknosis, nuclear fragmentation and apoptotic body formation. （2） SunBioTM Am -Blue Absorptiophotometry detection, GENMED lactate dehydrogenase （LDH） total activity quantitative colorimetric detection and AnnexinV -FITC/PI double -staining results showed that 50,100,150,200,250 p, mol/1 GCDC could induce apoptosis of HL - 7702 in a dose and time dependent manner. GCDC - induced apoptosis was revealed in HL - 7702 cells in a time - dependent and concentration - dependent manner according to Am - Blue Absorpticphotometry, lactate dehydrogenase （LDH） total activity quantitative colorimetry and Annexin V -FITC/PI double -staining. （3） GCDC increased intracellular calcium concentration in HL -7702 cells in a concentration and time dependent manner. Conclusion GCDC can increase HL -7702 intracellular calcium concentration that leads to cellular apoptosis in a concentration and time de- pendent manner.

关 键 词：甘氨鹅脱氧胆酸盐 细胞凋亡 HL-7702细胞 钙离子 

分 类 号：R85.5[医药卫生—航空、航天与航海医学]