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目的探讨槲皮素对大鼠肝细胞同型半胱氨酸(homocysteine,Hcy)代谢的影响。方法实验1:培养的大鼠肝细胞分成2组,对照组和5μmol/L槲皮素组,分别用DMEM培养液和含有5μmol/L槲皮素的DMEM培养液培养;实验2(治疗性模型):培养的细胞分成4组:对照组、5μmol/L槲皮素组、50mmol/L蛋氨酸(methionine,Met)组和5μmol/L槲皮素+50mmol/L Met组,分别用空白DMEM培养液,含有5μmol/L槲皮素、50mmol/L Met以及5μmol/L槲皮素+50mmol/L Met的DMEM培养液培养24h,随后收集培养液检测Hcy;实验3(预防性模型):培养的细胞分成4组:对照组、5μmol/L槲皮素组、50mmol/L Met组和5μmol/L槲皮素+50 mmol/L Met组。在前23h,5μmol/L槲皮素组、5μmol/L槲皮素+50mmol/L Met组用含有5μmol/L槲皮素的DMEM培养液处理,其余两组用空白培养液处理;在实验结束前的1h,所有组均换液,分别用空白培养液,含有5μmol/L槲皮素、50 mmol/L Met和5μmol/L槲皮素+50mmol/L Met的培养液处理细胞1h。实验结束,收集各组培养液和肝细胞,检测培养液Hcy水平,肝细胞内谷胱甘肽(GSH)含量以及胱硫醚-β合成酶(CBS)活性和表达。结果在槲皮素处理组,槲皮素和甲基化槲皮素的含量分别为(36.74±0.05)μg/L和(31.57±0.03)μg/L。在实验2和实验3中,Met处理后Hcy水平显著增加,在实验3中槲皮素处理能够降低Hcy水平。实验3中槲皮素组及其槲皮素+Met处理组GSH含量增加,CBS活性显著增加。结论在Met负载大鼠肝细胞,槲皮素能通过促进转硫化途径来降低Hcy水平。
Objective To investigate the effect of quercetin on the metabolism of homocysteine (Hcy) in rat hepatocytes. Methods Experiment 1: The cultured rat hepatocytes were divided into 2 groups, control group and 5μmol / L quercetin group, cultured in DMEM medium and DMEM medium containing 5μmol / L quercetin respectively. Experiment 2 (Therapeutic model ): The cultured cells were divided into 4 groups: control group, 5μmol / L quercetin group, 50mmol / L methionine (Met) group and 5μmol / L quercetin + 50mmol / L Met group, , And cultured in DMEM containing 5μmol / L quercetin, 50mmol / L Met and 5μmol / L quercetin + 50mmol / L Met for 24 h, then the culture medium was collected to detect Hcy. Experiment 3 (Preventive model): cultured cells The mice were divided into 4 groups: control group, 5μmol / L quercetin group, 50mmol / L Met group and 5μmol / L quercetin + 50mmol / L Met group. In the first 23h, 5μmol / L quercetin group, 5μmol / L quercetin + 50mmol / L Met group were treated with 5μmol / L quercetin DMEM medium, the other two groups were treated with blank medium; at the end of the experiment Before 1 h, all the groups were replaced by liquid, and the cells were treated with blank medium containing 5μmol / L quercetin, 50mmol / L Met and 5μmol / L quercetin + 50mmol / L Met for 1h respectively. At the end of the experiment, the culture medium and hepatocytes of each group were collected, and the level of Hcy, the content of glutathione (GSH) and the activity and expression of cystathionine-β synthase (CBS) in hepatocytes were detected. Results The contents of quercetin and methylated quercetin in the quercetin treatment group were (36.74 ± 0.05) μg / L and (31.57 ± 0.03) μg / L, respectively. In experiments 2 and 3, Hcy levels increased significantly after Met treatment and in experiment 3 quercetin treatment reduced Hcy levels. In quercetin group and quercetin + Met treatment group, GSH content increased and CBS activity increased significantly in experiment 3. Conclusions Quercetin can reduce the level of Hcy by promoting the transsulfuration pathway in Met-loaded rat hepatocytes.