洋甘菊总黄酮对HepG2细胞脂质沉积的改善作用研究
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篇名: 洋甘菊总黄酮对HepG2细胞脂质沉积的改善作用研究
TITLE: Study on improvement effect of total flavonoids from Matricaria recutita on lipid deposition in HepG 2 cells
摘要: 目的 探讨洋甘菊总黄酮对人肝癌HepG2细胞脂质异常的调节作用及其降脂机制。方法采用大孔树脂分离纯化得到高含量的洋甘菊总黄酮提取物。将HepG2细胞分为对照组(不给药)、模型组(不给药)、非诺贝特组(阳性对照,3.61μg/mL)和洋甘菊总黄酮低、中、高剂量组(100、150、200μg/mL)后,除对照组不造模外,其余组细胞采用1mmol/L油酸和棕榈酸混合液联合诱导建立脂质沉积细胞模型。干预24h后,检测细胞上清液中的游离脂肪酸(FFA)水平和细胞中的三酰甘油(TG)、FFA水平;采用油红O染液染色观察细胞中脂滴沉积情况,并检测脂质含量;采用DAPI染液染色观察细胞中二酰基甘油酰基转移酶2(DGAT2)蛋白表达情况,并检测DGAT2蛋白表达荧光强度;采用Westernblot法检测TG合成关键酶乙酰辅酶A羧化酶(ACC)、脂肪酸合成酶(FAS)、DGAT2的蛋白表达水平。结果经分离纯化后,洋甘菊总黄酮含量从6.72%提高到56.20%。细胞实验结果显示,与对照组比较,模型组细胞中的TG、FFA水平和细胞上清液中的FFA水平显著升高,细胞中脂质含量显著升高,DGAT2蛋白表达荧光强度显著升高,ACC、FAS、DGAT2蛋白表达水平均显著升高(P<0.01);细胞中可见大量脂滴堆积。与模型组比较,洋甘菊总黄酮低、中、高剂量组以上指标水平均显著逆转(P<0.01);细胞中脂滴堆积现象明显减少。结论洋甘菊总黄酮可以抑制脂质沉积模型HepG2细胞的TG合成,减少细胞脂质积累,防止细胞脂质损伤,其机制可能与其下调ACC/FAS/DGAT2通路表达有关。
ABSTRACT: O BJECTIVE To investigate the regulatory effect of total fla vonoids of Matricaria recutita on lipid abnormalities in human hepatoma HepG 2 cells and its lipid-lowering mechanism. METHODS The high-content total flavonoids extract from M. recutita was isolated and purified by macroporous resin. HepG 2 cells were divided into control group (without administration ), model group (without administration ),fenofibrate group (positive control ,3.61 μg/mL)and M. recutita total flavonoids low-dose , medium-dose and high-dose groups (100,150 and 200 μg/mL). Except for control group ,lipid deposition model of HepG 2 cells in other groups were established by 1 mmol/L mixture of oleic acid and palmitic acid. After 24 hours of intervention ,the levels of free fatty acids (FFA)in cell supernatant and triglyceride (TG)and FFA in cells were detected ;Oil red O staining was used to observe the deposition of lipid droplets in cells and detect the content of lipid ;DAPI staining was used to observe the protein expression of diacylglycerol acyltransferase 2(DGAT2)in cells ,and fluorescence intensity of protein expression of DGAT 2 were also detected ; protein expressions of key enzymes of TG synthesis as acetyl CoA carboxylase (ACC),fatty acid synthase (FAS)and DGAT 2 were detected by Western blot. RESULTS After separation and purification ,the content of total flavonoids from M. recutita increased from 6.72% to 56.20%. The results of cell experiment showed that compared with control group ,the levels of TG and FFA in cells and FFA in the cell supernatant increased significantly in the model group ,the content of lipid in cells increased significantly,the fluorescence intensity of protein expression of DGAT 2 increased significantly ,and the protein expressions of ACC,FAS and DGAT 2 increased significantly (P<0.01); large number of lipid dro plets were accumulated in the cells. Compared with model group ,the levels of above indexes in M. recutita total flavonoids low-dose , medium-dose andhigh-dose groups were significantly reversed (P<0.01);the accumulation of lipid droplets in cells decreased significantly. CONCLUSIONS M. recutita total flavonoids can inhibit the TG synthesis of lipid depos ition model HepG 2 cell,reduce the lipid accumulation of cells and prevent the lipid damage of cells. Its mechanism may be related to the down-regulation of the expression of ACC/FAS/DGAT 2 pathway.
期刊: 2022年第33卷第11期
作者: 阿曼古丽·艾则孜,马红梅,兰卫
AUTHORS: Amanguli·Aizezi,MA Hongmei ,LAN Wei
关键字: 洋甘菊总黄酮;脂质沉积;降脂作用
KEYWORDS: total flavonoids from Matricaria recutita ;lipid deposition ;lipid-lowering effect
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