Experimental Protocol for Detecting Mitochondrial Function in Hepatocytes Exposed to Organochlorine Pesticides

被引:3
作者
Liu, Qian [1 ,2 ,3 ]
Jiang, Zhaoyan [3 ]
Gu, Aihua [1 ,2 ]
机构
[1] Nanjing Med Univ, Inst Toxicol, State Key Lab Reprod Med, Nanjing, Peoples R China
[2] Nanjing Med Univ, Sch Publ Hlth, Key Lab Modern Toxicol, Minist Educ, Nanjing, Peoples R China
[3] Tongji Univ, Shanghai East Hosp, Ctr Gallbladder Dis, Inst Gallstone Dis,Sch Med, Shanghai, Peoples R China
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2020年 / 163期
基金
中国国家自然科学基金;
关键词
PERSISTENT ORGANIC POLLUTANTS;
D O I
10.3791/56800
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
This paper presents detailed methods on detecting hepatic mitochondrial function for a better understanding the cause of metabolic disorders caused by environmental organochlorine pesticides (OCPs) in hepatocytes. HepG2 cells were exposed to beta-hexachlorocyclohexane (beta-HCH) for 24 h at an equivalent dose of internal exposure in general population. Ultrastructure in hepatocytes was examined by transmission electron microscopy (TEM) to show the damage of mitochondria. Mitochondrial function was further evaluated by mitochondrial fluorescence intensity, adenosine 5'-triphosphate (ATP) levels, oxygen consumption rate (OCR) and mitochondrial membrane potential (MMP) in HepG2 cells incubated with beta-HCH. The mitochondria fluorescence intensity after stained by mitochondrial green fluorescent probe was observed with a fluorescence microscopy. The luciferin-luciferase reaction was used to determine ATP levels. The MMP was detected by the cationic dye JC-1 and analyzed under flow cytometry. OCR was measured with an extracellular flux analyzer. In summary, these protocols were used in detecting mitochondrial function in hepatocytes with to investigate mitochondria damages.
引用
收藏
页码:1 / 12
页数:12
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