Effect of Dihydroartemisinin on Oxidative Damage and Energy Metabolism of HepG2 Cells and Its Synergistic Effect with Sorafenib
10.13422/j.cnki.syfjx.20211202
- VernacularTitle:双氢青蒿素对HepG2细胞氧化损伤和能量代谢的影响及其与索拉非尼的协同作用
- Author:
Zhao CUI
1
;
Shuo LI
1
;
Hua-jing WANG
1
;
Ji MA
1
;
Ting-ting QIN
1
;
Hang SHI
1
;
Lan-fang LI
1
;
Gui-hua YU
1
;
Cang-hai LI
1
;
Ting-liang JIANG
1
Author Information
1. Institute of Chinese Materia Medica，China Academy of Chinese Medical Sciences，Beijing 100700，China
- Publication Type:Research Article
- Keywords:
dihydroartemisinin;
sorafenib;
synergism;
aerobic glycolysis;
mitochondrial oxidative phosphorylation;
reactive oxygen species （ROS）;
heme oxygenase-1 （HO-1）;
glutamate-cysteine ligase catalytic subunit （GCLC）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(12):24-32
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the inhibitory effect of dihydroartemisinin （DHA） on the proliferation of HepG2 cells， elucidate the mechanism from the perspectives of oxidative damage and energy metabolism， and discuss the possibility of combined use of DHA with sorafenib （Sora）. Method:Cell counting kit-8 （CCK-8） assay was used to obtain the 50% inhibitory concentration （IC50） of DHA and Sora on HepG2 and SW480 cells and Chou-Talalay method was used to obtain the combination index （CI） of DHA and Sora. HepG2 cells were classified into the control group， DHA group （10 µmol·L-1）， Sora group （5 µmol·L-1）， and DHA + Sora group （DHA 10 µmol·L-1， Sora 5 µmol·L-1） and then incubated with corresponding drugs for 8-12 h. Seahorse XF glycolytic rate assay kit and cell mito stress test kit were employed to respectively detect the glycolysis function of cells and oxidative phosphorylation function of mitochondria. DCFH-DA and lipid peroxidation MDA assay kit were separately used to analyze the intracellular levels of reactive oxygen species （ROS） and malondialdehyde （MDA）. Western blot was applied to determine the intracellular levels of heme oxygenase-1 （HO-1） and glutamate-cysteine ligase catalytic subunit （GCLC）. Result:Compared with the control group， DHA alone inhibited the ATP synthesis in mitochondrial oxidative phosphorylation and glycolysis （P<0.01）， increased the levels of intracellular ROS and MDA （P<0.05）， and decreased the levels of HO-1 and GCLC （P<0.05） in HepG2 cells. DHA and Sora had synergistic inhibitory effect on proliferation of HepG2 and SW480 cells， with CI < 0.90. The DHA + Sora group showed stronger suppression of ATP synthesis in mitochondrial oxidative phosphorylation and glycolysis （P<0.01）， higher levels of intracellular ROS and MDA （P<0.01）， and lower levels of intracellular antioxidation-related proteins HO-1 and GCLC in HepG2 cells （P<0.01） than the DHA group. Conclusion:DHA may increase the level of MDA by reducing HO-1 and GCLC and increasing ROS in HepG2 cells， which results in mitochondria oxidative damage， restricts cell glycolysis and mitochondrial oxidative phosphorylation， and thus finally inhibits the proliferation of HepG2 cells. DHA and Sora have synergistic inhibitory effect on the proliferation of HepG2 and SW480 cells， and the mechanism may be related to the synergistic oxidative damage that affects the mitochondrial electron transport chain and suppresses cell energy metabolism.
