Study on the role and possible mechanism of hemeoxygenase-1/carbon monoxide system in protection of quercetin against ethanol-induced hepatocytes oxidative injury
10.3760/cma.j.cn501113-20200522-00269
- VernacularTitle:HO-1/CO在槲皮素拮抗酒精性肝细胞氧化损伤中的作用及其可能机制研究
- Author:
Wei HOU
1
;
Pei QIU
;
Nianjun CHEN
;
Ping YAO
;
Shuang LIU
;
Hua QIN
Author Information
1. 华中科技大学同济医学院附属同济医院消化内科，武汉 430030
- Keywords:
Oxidative damage;
Quercetin;
Carbon monoxide;
Heme oxygenase-1;
Cytochrome P450 2E1
- From:
Chinese Journal of Hepatology
2020;28(6):504-508
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the protective effect and potential mechanism of heme oxygenase (HO-1)/carbon monoxide (CO)-mediated quercetin on alcoholic oxidative damage of primary rat hepatocytes.Methods:Primary rat hepatocytes were isolated and cultured by two-step collagenase technique. Ethanol exposed primary rat hepatocytes were simultaneously added with quercetin (100 μmol/L) and/or hemoglobin (100 μmol/L) or different doses of CO-releasing molecules (CORM-2, 5-50 μmol/L) for their combined action. After polling, LDH, AST activities and MDA and GSH levels were measured in the supernatant of cell culture. The alone or combined effects of quercetin, CORM-2, hemoglobin and zinc protoporphyrin IX exposed to ethanol were detected by the activity of CYP2E1 in liver microsomes. Statistical analysis of data was performed by analysis of variance (ANOVA) and intergroup comparison was done by SNK-test.Results:Simultaneous addition of 100 μmol/L quercetin had significantly reduced ethanol-induced AST and LDH release, and GSH consumption and MDA elevation extent. Moreover, quercetin had not only lost the hemoglobin (CO blocker) protective effect but also had further exacerbated ethanol-induced lipid peroxidation. CORM-2 had reduced ethanol-induced AST and LDH release, and GSH consumption and MDA production in liver cells, and thus had dose-dependent protective effect. Ethanol had increased significantly CYP2E1 activity. Quercetin or CORM-2 had inhibited CYP2E1 activity, while hemoglobin or protoporphyrin IX had eliminated quercetin inhibitory effect and had increased the CYP2E1 activity. Quercetin, and CYP2E1 activity was constant as compared to ethanol group when CORM-2, zinc protoporphyrin IX and ethanol were incubated with hepatocytes, but the CYP2E1 activity was significantly decreased ( P < 0.05), and the differences were statistically significant. Conclusion:CO/HO-1 metabolite mediates the protective effect of quercetin on alcoholic oxidative damage of hepatocytes, which may be related to the inhibition of CYP2E1 activity.
