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Evaluation of Chemical-induced Mitochondrial Damage in HepG2 Cells Using Energy Charge
YAKHAK HOEJI 2018;62(6):394-400
Published online December 30, 2018
© 2018 The Pharmaceutical Society of Korea.

Jang Su Jeon and Sang Kyum Kim#

College of Pharmacy, Chungnam National University, Daejeon, 305-763, Korea
Correspondence to: Sang Kyum Kim College of Pharmacy, Chungnam National University, Daejeon, 305-763, Korea Tel.: +82-42-821-5930 Fax.: +82-42-823-6566 E-mail: sangkim@cnu.ac.kr
Received October 19, 2018; Revised November 9, 2018; Accepted November 29, 2018.
Abstract
In the presence study, we evaluated chemical-induced mitochondrial damage in HepG2 cells using energy charge, calculated by (ATP+0.5*ADP)/(ATP+ADP+AMP). ATP and its metabolites, ADP and AMP, were determined using LC-MS/MS. The cellular ATP, ADP and AMP were identified by comparison of mass spectra and retention time with their authentic standards. To establish sampling method, effects of extraction solvent composition on determination of ATP, ADP and AMP were evaluated. The stability of ATP, ADP and AMP was improved by using the sampling solution consisted of acetonitrile:methanol:water (4:4:2), relative to 10% perchloric acid solution. The energy charge of HepG2 cells treated with 0.02 μM rotenone, 25 μM troglitazone or 20 μM flutamide was decreased to 52, 83, or 84%, respectively, relative to control cells. The ratio of AMP to ATP was 13-, 2- or 2-fold higher in HepG2 cells treated with rotenone, troglitazone or flutamide, respectively, compared with control cells. In conclusion, the method established in this study can provide information of cellular energy status to predict chemical-induced mitochondrial damage.
Keywords : mitochondria damage, chemical-induced liver injury, energy status, ATP


June 2019, 63 (3)
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