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Archive > Volume 10 Issue 2 > 2018,10(2):198-204. DOI:10.1016/j.chmed.2018.03.005 Prev Next

高濃度綠原酸對非酒精性肝炎的干預作用及其機制研究

Original articles

Effect of high exposure of chlorogenic acid on lipid accumulation and oxidative stress in oleic acid-treated HepG2 cells

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    摘要:
    目的：本文通過油酸（oleic acid，OA）處理人肝癌細胞系HepG2體外模擬肝細胞非酒精性脂肪肝（non-alcoholic fatty liver disease ，NAFLD）的病理狀態(tài)，研究了高濃度綠原酸（chlorogenic acid ，CGA）對NAFLD的干預作用，包括對肝細胞脂堆積、糖攝取和氧化應激的影響。 方法：用0.5 mmol/L OA處理HepG2 細胞24h建立體外非酒精性肝炎的模型。正常組和OA誘導組的細胞用CGA（0, 0.5, 1, and 2 mmol/L）處理24h后有油紅O染色和生化分析試劑盒檢測胞內的脂堆積情況、活性氧( reactive oxygen species, ROS)含量和糖攝取能力。我們還通過Western blot和RT-qPCR檢測了脂合成相關指標（SREBP-1c和PNPLA3）和氧化應激相關蛋白（CYP2E1和CYP4A）的相對表達量。 結果： CGA組相對于OA誘導組其細胞內脂滴含量呈上升趨勢，SREBP-1c和PNPLA3的mRNA和蛋白表達量呈上升趨勢，CGA還可以促進HepG2對熒光葡萄糖的攝取，降低胞內ROS含量，下調CYP450酶2E1和4A的表達。 結論：本文的研究結果提示，高濃度CGA可以通過上調SREBP-1c和PNPLA3的mRNA和蛋白的表達促進脂合成進而導致細胞內脂滴含量增加；CGA還可以促進HepG2對熒光葡萄糖的攝取，下調CYP450酶2E1和4A的表達進而降低胞內ROS含量，減輕氧化損傷。

    Abstract:
    Objective To evaluate the effect of high concentration of chlorogenic acid (CGA) on non-alcoholic fatty liver disease (NAFLD) in normal and oleic acid (OA) treated HepG2 cells, as well as the underlying mechanism involved in the fat accumulation, oxidative stress and insulin resistance (IR) induced by CGA treatment. Methods OA (0.5 mmol/L) induced hepatic steatosis was established in HepG2 cells as an in vitro model of NAFLD. The normal and OA-treated HepG2 cells were treated by CGA (0, 0.5, 1, and 2 mmol/L) for 24 h, then cellular lipid droplets, reactive oxygen species (ROS), and glucose uptake were evaluated by Oil Red O staining and cellular biochemical assays, respectively. Signaling pathways involved in?adipogenesis including SREBP-1c and PNPLA3, oxidative stress, and IR including CYP2E1 and CYP4A, were investigated by Western blot and RT-qPCR. Results CGA (0.5, 1, and 2 mmol/L) treatment increased the cellular lipid droplets and the expression of SREBP-1c and PNPLA3 in the tested cells. Additionally, 2-NBDG uptake was significantly increased, whereas the cellular ROS and protein levels of CYP2E1 and CYP4A were significantly decreased in OA-treated cells. Conclusion Our results suggest that high concentrations of CGA ameliorate OA-induced oxidative damage and IR likely by inhibiting the expression of CYP2E1 and CYP4A, promote lipid accumulation by inducing the expression of SREBP-1c and PNPLA3 in the tested cells.

    關鍵詞:
    綠原酸;糖攝取;HepG2;脂堆積;氧化應激

    Keywords:


    Project Supported:
    Sci-Tech Support Plan of Hubei Province (grant no.2015BCA273); United fund for innovation and entrepreneurship of Ministry of education (201601031006).

    Clc Number:

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