甲基尿石素A对油酸诱导的Huh-7细胞脂质累积的改善作用及其机制研究
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篇名: | 甲基尿石素A对油酸诱导的Huh-7细胞脂质累积的改善作用及其机制研究 |
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摘要: | 目的:研究甲基尿石素A对油酸诱导的人肝癌Huh-7细胞脂质累积的改善作用及机制。方法:采用油酸诱导建立细胞脂质累积模型。取Huh-7细胞,分为对照组(培养基)和模型组(1 mmol/L油酸)、低剂量药物组(1 mmol/L油酸+10 μmol/L甲基尿石素A)和高剂量药物组(1 mmol/L油酸+20 μmol/L甲基尿石素A),采用油红O染色法观察细胞中脂质累积情况;采用三酰甘油(TG)酶法测定细胞内TG含量,采用聚合酶链式反应(PCR)法检测细胞中脂肪酸合成酶(FASN)、胆固醇调节元件结合蛋白1(SREBP-1)、过氧化物酶体增殖剂激活受体α(PPAR-α)、PPAR-γ的mRNA表达水平;采用Western blotting法检测细胞中FASN的蛋白表达水平。结果:采用油酸诱导后,细胞膜周围出现大量脂滴累积;细胞内脂质及TG含量均显著升高,FASN、SREBP-1、PPAR-γ的mRNA表达水平及FASN的蛋白表达水平均显著升高,PPAR-α的mRNA表达水平显著降低(P<0.01)。采用甲基尿石素A干预后,细胞膜周围的脂滴明显减少;细胞中脂质及TG含量均显著降低,FASN、SREBP-1、PPAR-γ的mRNA表达水平及FASN的蛋白表达水平均显著降低(P<0.05或P<0.01)。结论:甲基尿石素A对油酸诱导的Huh-7细胞脂肪累积有一定的改善作用,其机制可能与下调FASN、SREBP-1、PPAR-γ等相关因子的表达,抑制脂肪酸从头合成、促进脂质代谢有关。 |
ABSTRACT: | OBJECTIVE: To study the improvement effect and mechanism of methylated urolithin A on oleic acid-induced lipid accumulation in human liver cancer Huh-7 cells. METHODS: Oleic acid was adopted to induce lipid accumulation model cells. Huh-7 cells were divided into control group (culture medium), model group (1 mmol/L oleic acid), low-dose group (1 mmol/L oleic acid+10 μmol/L methylated urolithin A) and high-dose group (1 mmol/L oleic acid+20 μmol/L methylated urolithin A). Oil red O staining was used to observe lipid accumulation in cells. Triglyceride(TG) enzyme assay was applied to determine the TG content in cells. PCR was employed to detect the mRNA expression of FASN, SREBP-1, PPAR-α and PPAR-γ in cells. Western blotting was used to determine the protein expression of FASN in cells. RESULTS: After induced by oleic acid, a large amount of lipid droplet accumulated around the cells; the intracellular lipid and TG content, mRNA expression levels of FASN, SREBP-1 and PPAR-γ, protein expression levels of FASN were increased significantly, while mRNA expression level of PPAR-α was decreased significantly (P<0.01). After intervened with methylated urolithin A, lipid droplet around the cells decreased significantly; the contents of lipid and TG in cells were decreased significantly, while the mRNA expression levels of FASN, SREBP-1 and PPARγ and protein expression level of FASN were decreased significantly (P<0.05 or P<0.01). CONCLUSIONS: Methylated urolithin A can improve oleic acid-induced lipid accumulation in Huh-7 cells, the mechanism of which may be associated with inhibiting fat synthesis, promoting lipid metabolism and down-regulating the expression of metabolism-related factors as FASN, SREBP-1 and PPAR-γ. |
期刊: | 2019年第30卷第6期 |
作者: | 张聪,周俊旋,盛磊,马俊俏,李歆,郑国华,刘思丹,邱振鹏 |
AUTHORS: | ZHANG Cong,ZHOU Junxuan,SHENG Lei,MA Junqiao,LI Xin,ZHENG Guohua,LIU Sidan,QIU Zhengpeng |
关键字: | 甲基尿石素A;油酸;人肝癌Huh-7细胞;脂质累积;机制 |
KEYWORDS: | Methylated urolithin A; Oleic acid; Human liver cancer Huh-7 cells; Lipid accumulation; Mechanism |
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