杜仲叶绿原酸提取物联合西红花苷对肝癌细胞胆固醇代谢的影响

发布时间：2018-06-15 10:58

本文选题：脂类代谢紊乱 + 肝癌细胞　；参考：《山东医药》2017年08期

【摘要】：目的观察杜仲叶绿原酸提取物(CAEF)联合西红花苷对人肝癌细胞Hep G2胆固醇代谢的影响,并探讨其可能的作用机制。方法将人肝癌细胞Hep G2随机分为空白对照组、油酸诱导组、辛伐他汀组、CAEF组、联合用药组。油酸诱导组、辛伐他汀组、CAEF组、联合用药组分别加入油酸诱导脂肪变性模型;同时,辛伐他汀组加入10μmol/L的辛伐他汀10μL,CAEF组加入25 mg/L的CAEF 10μL,联合用药组加入含1μmol/L西红花苷、30μmol/L绿原酸、10μmol/L京尼平苷、10μmol/L槲皮素的混合液10μL;空白对照组仅加入等体积培养基。取各组干预48 h细胞,行油红O染色,检测细胞内中性脂滴含量。取干预24、48 h细胞上清液,检测TC、TG含量。采用RT-PCR法检测胆固醇代谢相关基因(ABCA1、SREBP2、CYP7A1、LXRα、AMPK2α、HMGCR)表达,免疫细胞化学法和Western blotting法检测HMGCR蛋白表达。结果辛伐他汀组、CAEF组、联合用药组干预48 h,细胞内中性脂滴含量明显降低,其作用强度依次为联合用药组CAEF组辛伐他汀组(P均0.05);与对照组比较,CAEF组、联合用药组干预24、48 h时细胞外液TC、TG含量明显增加,细胞外液TC、TG含量依次为联合用药组CAEF组辛伐他汀组(P均0.05)。CAEF组、联合用药组ABCA1、CYP7A1、AMPK2αmRNA表达明显升高,SREBP2、HMGCR mRNA表达明显降低(P均0.05);此外,联合用药组LXRαmRNA表达明显降低(P0.05)。辛伐他汀组、CAEF组、联合用药组HMGCR蛋白表达明显降低,以联合用药组降低最明显(P均0.05)。结论 CAEF联合西红花苷可协同抑制肝癌细胞脂质积聚,其作用机制可能与调控肝癌细胞胆固醇代谢相关基因和蛋白的表达有关。
[Abstract]:Objective to investigate the effect of Lv Yuan acid extract from Eucommia ulmoides leaves and crocetin on cholesterol metabolism in HepG2 cells and its possible mechanism. Methods HepG2 cells were randomly divided into control group, oleic acid induced group, simvastatin group and combined treatment group. Oleic acid induction group, simvastatin group, CAEF group, combined treatment group were added oleic acid induced fatty degeneration model, at the same time, In simvastatin group, 10 渭 mol / L simvastatin 10 渭 L CAEF was added to 25 mg / L CAEF10 渭 L, and 1 渭 mol / L Crocin 30 渭 mol / L Lv Yuan was added to 10 渭 mol / L quercetin mixture of 10 渭 mol / L geniposide 10 渭 mol / L and 10 渭 mol / L quercetin in blank control group. After 48 h of intervention, the cells were stained with oil red O and the content of neutral lipid droplets in the cells was detected. The supernatant of the cells was taken after 24 hours of intervention and the content of TCU TG was detected. The expression of LXR 伪 AMPK2 伪 and HMGCRs were detected by RT-PCR, and the expression of HMGCR protein was detected by immunocytochemistry and Western blotting. Results in the simvastatin group and CAEF group, the content of intracellular neutral lipid droplets was significantly decreased after 48 h intervention in the combined treatment group, and the effect intensity was in turn as follows: the simvastatin group in the combined treatment group (P 0.05), and the CAEF group in comparison with the control group. In the combined group, the content of TCN TG in extracellular fluid increased significantly at 24 h after intervention, and the content of TCN TG in extracellular fluid in CAEF group was 0.05%. In CAEF group, the expression of CYP7A1AMPK2 伪 mRNA in simvastatin group was significantly higher than that in CAEF group, and the expression of HMGCR mRNA in SREBP2HMGCR was significantly decreased (P 0.05) in combination group, and the expression of CYP7A1AMPK2 伪 mRNA in CAEF group was significantly lower than that in CAEF group (P < 0.05). The expression of LXR 伪 mRNA in combined treatment group was significantly decreased (P 0.05). The expression of HMGCR protein in simvastatin group and CAEF group was significantly lower than that in combined treatment group (P < 0.05). Conclusion CAEF combined with crocetin can synergistically inhibit lipid accumulation in hepatoma cells, and its mechanism may be related to the regulation of the expression of genes and proteins related to cholesterol metabolism in hepatoma cells.
【作者单位】：遵义医学院珠海校区;广东省第二人民医院;贵州省人民医院;
【基金】：贵州省中药现代化科技产业研究开发专项项目(黔科合ZY字[2013]3018号) 珠海市优势学科建设计划
【分类号】：R735.7

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