Title: [Study on the role and possible mechanism of hemeoxygenase-1/carbon monoxide system in protection of quercetin against ethanol-induced hepatocytes oxidative injury].
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.目的: 研究I型血红素加氧酶(HO-1)/一氧化碳(CO)介导的槲皮素对大鼠原代肝细胞酒精性氧化损伤保护作用及其潜在机制。 方法: 二步胶原酶技术分离培养大鼠原代肝细胞。在乙醇染毒同时,加入槲皮素(100 μmol/L)或/和血红蛋白(100 μmol/L),或不同剂量的CO释放分子(CORM-2,5~50 μmol/L)共同作用,作用结束后检测细胞培养上清液中乳酸脱氢酶(LDH)、天冬氨酸转氨酶(AST)活性,细胞丙二醛(MDA)和谷胱甘肽(GSH)水平。乙醇染毒肝细胞与槲皮素、CORM-2、血红蛋白、锌原卟啉Ⅸ单独或联合作用,检测细胞微粒体内CYP2E1的活性。数据统计分析采用方差分析(ANOVA)和组间比较(SNK-检验)。 结果: 乙醇染毒同时加入100 μmol/L的槲皮素,乙醇诱导的AST与LDH的释放明显减少,GSH消耗与MDA升高程度也明显下降;而血红蛋白(CO阻断剂)不仅使槲皮素的保护效应丧失,反而进一步加重了乙醇所致的脂质过氧化。CORM-2能减少乙醇诱导的肝细胞AST与LDH释放,GSH消耗与MDA生成也明显减轻,且这种保护作用呈剂量依赖性。乙醇导致细胞CYP2E1活性显著升高,槲皮素或CORM-2能抑制CYP2E1的活性,血红蛋白或锌原卟啉Ⅸ则消除了槲皮素的这种抑制效应,使CYP2E1活性升高。当槲皮素、CORM-2和锌原卟啉Ⅸ与乙醇共同孵育肝细胞时,CYP2E1活性并未升高,与乙醇组比较反而明显下降,P值均< 0.05,差异均有统计学意义。 结论: HO-1的代谢产物CO介导了槲皮素对肝细胞酒精性氧化损伤的保护作用,这种保护效应可能与其抑制CYP2E1活性有关。.
Publication Year: 2020
Publication Date: 2020-06-20
Language: en
Type: article
Indexed In: ['pubmed']
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Cited By Count: 1
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