溶酶体在2,2’,4,4’-四溴联苯醚诱导HepG2细胞凋亡中的作用  

Possible Involvement of Lysosomes in 2, 2’, 4, 4’-Tetrabromodiphenyl Ether Mediated Apoptosis in HepG2 Cells

作　　者：刘晓晖[1] 黄丹[1] 王若希[1] 邵静[1] 闫金松[2] 

机构地区：[1]大连医科大学公共卫生学院劳动卫生与环境卫生教研室,辽宁116044 [2]大连医科大学附属第二医院血液科,辽宁116027

出　　处：《环境与职业医学》2013年第7期510-514,共5页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金资助项目(编号:81273031);大连医科大学引进人才科研启动基金(编号:201069)

摘　　要：[目的]探讨2,2’,4,4’-四溴联苯醚(BDE 47)致人肝癌HepG2细胞凋亡机制。[方法]不同浓度BDE 47(0.00、6.25、12.50、25.00、50.00、100.00μmol/L)染毒HepG2细胞24h,采用四氮唑盐比色法(MTT法)检测细胞存活率;用2’,7’-二氢二氯荧光素(DCFH-DA)探针检测活性氧水平;用吖啶橙(AO)探针及罗丹明(Rh123)荧光探针分别检测溶酶体膜通透性和线粒体膜电势,并通过溶酶体组织蛋白酶B特异性抑制剂环氧酶琥珀酰肽甲基酯(CA-074)验证溶酶体在BDE 47细胞毒性的作用。[结果]与对照组比较,50.00、100.00μmol/L BDE 47染毒组HepG2细胞存活率明显降低(P<0.01);各BDE 47染毒组HepG2细胞凋亡率明显升高(P<0.01),呈现剂量-效应关系(R2=0.981);各BDE 47染毒组HepG2细胞活性氧含量明显升高(P<0.01),≥12.50μmol/L BDE 47染毒组HepG2细胞内溶酶体膜通透性明显升高(P<0.05;P<0.01),各BDE 47染毒组HepG2细胞内线粒体膜电势明显降低(P<0.01),上述3项指标与染毒浓度均呈剂量-效应关系(R2=0.918,R2=0.636,R2=0.678)。25μmol/L CA-074能够明显干预50μmol/L BDE 47对细胞的毒性作用使细胞存活率升高,细胞调亡减少(均P<0.05)。[结论]BDE 47可能通过溶酶体介导线粒体途径诱导HepG2细胞凋亡。[Objective]To explore the possible involvement of lysosomes in 2, 2’, 4, 4’-tetrabromodiphenyl ether （BDE 47） induced apoptosis in HepG2 cells.[Methods]HepG2 cells were exposed to different levels of BDE 47 （0.00, 6.25, 12.50, 25.00, 50.00, 100.00μmol/L） for 24h, and the effects on cytotoxicity, reactive oxygen species （ROS）, lysosomal and mitochondrial membrane stability were examined.[Results]The data demonstrated that the 50.00 and 100.00μmol/L BDE 47 statistically reduced cell viability （P 〈0.01）, compared with the control group. The BDE 47 exposure induced apoptosis in HepG2 cells significantly （P 〈0.01） and in a dose-dependent manner （R 2 =0.981） and statistically increased ROS generation in HepG2 cells （P 〈0.01, R 2 =0.918）. The treatment with 12.50μmol/L and higher doses of BDE 47 also led to elevated lysosomal membrane permeabilization （P 0.05; P 〈0.01） along with mitochondrial membrane potential disturbance （P 〈0.01）, both in a dose-effect manner （R 2 =0.918, R 2 =0.636, R 2 =0.678）. The toxic effect of 50 μmol/L BDE 47 was significantly alleviated by 25 μmol/L CA-074 （P 〈0.05）, a specific inhibitor for cathepsin B, for which increased cell viability and decreased apoptosis were demonstrated. [Conclusion]The findings indicate that the apoptotic event induced by BDE 47 in HepG2 might be initiated through a lysosomalmitochondrial pathway.

关 键 词：2 2’ 4 4’-四溴联苯醚 细胞凋亡 溶酶体 线粒体 组织蛋白酶B HEPG2细胞 

分 类 号：R114[医药卫生—卫生毒理学]