三-(2,3-二溴丙基)异氰脲酸酯通过线粒体信号通路诱导HepG2细胞凋亡  

Tri-(2, 3-dibromopropyl) isocyanurate induced apoptosis in HepG2 cells via mitochondrial pathway

作　　者：綦峥 杨明 胡扬[2] 袁洪亮 李金龙 QI Zheng;YANG Ming;HU Yang;YUAN Hong-liang;LI Jin-long(Engineering Research Center for Medicine,Harbin University of Commerce,Harbin Heilongjiang 150076,China;College of Pharmacy,Harbin University of Commerce,Harbin Heilongjiang 150076,China)

机构地区：[1]哈尔滨商业大学药物工程技术研究中心,黑龙江哈尔滨150076 [2]哈尔滨商业大学药学院,黑龙江哈尔滨150076

出　　处：《毒理学杂志》2020年第6期460-465,共6页Journal of Toxicology

基　　金：国家自然科学基金(41702368);黑龙江省自然科学基金(LH2019D007)。

摘　　要：目的为确定环境外源性污染物三-(2,3-二溴丙基)异氰脲酸酯(TBC)对人肝癌细胞株HepG2的细胞毒性和诱导凋亡作用,并探讨TBC诱导HepG2细胞凋亡的线粒体途径。方法采用MTT法、流式细胞术和Western blot法检测TBC对HepG2细胞的毒性作用、细胞凋亡率,并对线粒体信号通路关键蛋白Bcl-2、Bax、Bcl-xL、Cyt-C、Caspase-3和Caspase-9的表达进行检测。结果不同浓度(0、12.5、25和50)μg/ml的TBC对HepG2细胞的增殖有抑制作用,通过流式细胞仪的PI单染法检测产生了明显的凋亡峰,表明DNA被抑制了合成。随着TBC给药浓度的增加,凋亡峰越明显,凋亡率也呈上升趋势;经TBC染毒后,细胞内Bcl-xL和Bcl-2蛋白表达量逐渐降低,Bax蛋白表达量逐渐升高,给药组Bcl/Bax蛋白表达量比值降低。细胞内Cyt-C蛋白和Caspase-9蛋白表达量增加,并且随着TBC浓度的增加而增强,呈现剂量线性关系。结论研究首次采用体外实验的方法对环境外源性污染物TBC的肝细胞毒性进行阐述,结果表明TBC可通过线粒体信号通路诱导HepG2细胞凋亡。Objective To determine the cytotoxicity and apoptosis-inducing effects of exogenous tri-(2,3-dibromopropyl) isocyanurate(TBC) on human hepatocyte cell line HepG2, and explore the mitochondrial pathway of TBC-induced apoptosis in HepG2 cells. Methods Toxic effects of TBC on HepG2 cells and apoptosis rate were detected by MTT assay, flow cytometry and Western blot assay, and the expressions of key proteins Bcl-2, Bax,Bcl-xL, Cyt-C, Caspase-3 and Caspase-9 in mitochondrial signaling pathway were detected. Results Different concentrations(0, 12.5, 25, 50) μg/ml of TBC inhibited the proliferation of HepG2 cells. Detection of obvious apoptotic peaks by flow cytometry PI single staining, which indicated that DNA synthesis inhibited. With the increase of the concentration of TBC, the apoptotic peak was more obvious, and the apoptotic rate was rising. Infected by TBC, the expressions of Bcl-xL and Bcl-2 protein in cells down regulated gradually, and the expression of Bax protein gradually up regulated. The ratio of Bcl/Bax protein expression was reduced in the treatment groups. The expressions of Cyt-C protein and Caspase-9 protein in cells were increased with the up regulation of TBC concentration, which appeared a dose linear relationship. Conclusion This experimental study was the first to use the method of in vitro experiments to explain the hepatotoxicity of environmental exogenous pollutants TBC, TBC induces apoptosis in HepG2 cells via mitochondrial signaling pathway.

关 键 词：三-(2 3-二溴丙基)异氰脲酸酯 细胞毒性 细胞凋亡 线粒体信号通路 

分 类 号：R994.3[医药卫生—毒理学]