乙醇对小鼠胰岛瘤细胞凋亡的影响及其分子机制  

The study on apoptosis and its molecular mechanism in mouse insulinoma cells induced by ethanol

作　　者：郝丽萍[1] 胡学锋[1] 庞红[1] 曲巍 杨年红[1] 应晨江[1] 孙秀发[1] 

机构地区：[1]华中科技大学同济医学院公共卫生学院营养与食品卫生学系,湖北武汉430030

出　　处：《毒理学杂志》2006年第3期138-140,共3页Journal of Toxicology

基　　金：国家自然科学基金资助(30170807;30300282)

摘　　要：目的观察乙醇对小鼠胰岛瘤细胞(NIT-1细胞)凋亡的影响及Bcl-2、Bax和Caspase-3表达的变化,探讨乙醇诱发小鼠胰岛瘤细胞凋亡的分子机制。方法体外培养的小鼠NIT-1细胞,不同剂量乙醇(0、50、100、2004、00 mmol/L)作用24 h,单细胞凝胶电泳实验(SCGE),AnnexinV/PI双标记法流式细胞仪检测细胞凋亡情况。不同剂量的乙醇作用6、122、4 h,RT-PCR法检测与凋亡有关的基因Bcl-2,Bax和Caspase-3 mRNA表达水平。结果SCGE实验结果显示,低剂量乙醇损伤不明显,高剂量(2004、00 mmol/L)组DNA迁移率、DNA损伤程度分级、DNA平均迁移长度与对照组比较,差异均有显著性。AnnexinⅤ/PI检测结果表明,2004、00 mmol/L乙醇组NIT-1细胞凋亡率升高,与对照组相比,差异有非常显著性。乙醇作用6 h,随着乙醇剂量增加,Bcl-2/Bax mRNA表达先升高后降低;乙醇作用12 h,400 mmol/L剂量Bcl-2/Bax比值下降;作用24 h,各剂量组Bcl-2/Bax比值均下降。Caspase-3 mRNA表达水平与乙醇剂量和作用时间相关。400 mmol/L剂量组作用24 h,Caspase-3 mRNA表达增加。结论乙醇可诱发体外培养的NIT-1细胞凋亡,凋亡的发生很可能与Bcl-2家族和Caspase-3激活有关。Objective To investigate the effect of ethanol on apoptosis in mouse insulinoma cells（NIT-1 ceils） and to evaluate the pathway associated with the regulation of Bcl-2, Bax and Caspase-3.Methods After NIT-1 cells were treated with ethanol at concentrations of 0, 50, 100, 200 and 400 mmol/L for 24 h, single ceil gel electrophoresis （SCGE） and AnnexinV/PI methods were performed to detect the apoptosis. After NIT-1 cells were exposed to various concentrations of ethanol （same as above） for 6, 12 and 24 h, the levels of Bcl-2, Bax and Caspase-3 mRNA expression were determined by RT-PCR method. Results SCGE results showed that the low concentration of ethanol induced no obviously DNA damage in NIT-1 ceils, but the rate of DNA migration, degree of DNA damage and average length of DNA migration of 200 and 400 mmol/L ethanol-treated groups were significantly changed compared to those of untreated group. The results of Annexin V/PI assay indicated that the apoptosis rates of NIT-1 cells in 200 and 400 mmol/L ethanol- treated groups were higher than that of untreated group. At 6 h, the ration of Bcl-2/Bax mRNA expression was increased first, then decreased with the increasing of ethanol dosage. At 12 h and 24 h, the ration was decreased. By treatment at 400 mmol/L, the decrease was dramatically compared to control group. Ethanol increased the caspase-3 mRNA expression at high concentrations and long exposure time. At 24 h, the caspase-3 mRNA expression was increased significantly by treatment at 400 mmol/L. Conclusion Ethanol induced apoptosis in NIT-1 ceils and the potential molecular mechanism probably relates with the Bcl-2 family and Caspse-3.

关 键 词：乙醇 NIT-1细胞 凋亡 单细胞凝胶电泳 

分 类 号：R152.41[医药卫生—营养与食品卫生学]