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目的观察不同类型脂肪酸对HepG2细胞胰岛素抵抗的作用及探讨可能的作用机制。方法 250μmol/L的棕榈酸(PA)、花生四烯酸(AA)以及二十碳五烯酸(EPA)分别与HepG2细胞共同培养24 h,葡萄糖氧化酶法测定细胞上清液中葡萄糖含量,酶联免疫吸附法测定炎性因子白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量,以及实时聚合酶链反应(Real-time PCR)检测Toll样受体4(TLR4)mRNA的表达。结果 PA组培养液中葡萄糖含量显著高于空白对照组(P<0.05),EPA组可增加葡萄糖消耗量,与PA组及AA组比较差异具有统计学意义(P<0.05)。各脂肪酸干预组细胞上清液中炎性因子IL-6和TNF-α含量均增加,EPA组增加量最低,PA组和AA组上清液中IL-6含量与空白组比较差异有统计学意义(P<0.05)。各脂肪酸干预组与空白组比较TLR4 mRNA表达量均增加(P均<0.05),EPA组增加量最低。结论高浓度PA降低了HepG2细胞对胰岛素的敏感性,能够引起胰岛素抵抗,AA具有降低胰岛素敏感性的趋势,而EPA则具有增加胰岛素敏感性的趋势,TLR4信号转导通路及其启动的炎症反应可能是其重要的作用机制。
Objective To observe the effect of different types of fatty acids on insulin resistance in HepG2 cells and to explore the possible mechanism. Methods 250μmol / L palmitic acid (PA), arachidonic acid (AA) and eicosapentaenoic acid (EPA) were co-cultured with HepG2 cells for 24 h. Glucose oxidase method was used to determine the glucose content in the cell supernatant. The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay and Toll-like receptor 4 (TLR4) mRNA expression. Results The glucose content of PA group was significantly higher than that of blank control group (P <0.05). EPA group increased glucose consumption, with statistical significance (P <0.05) compared with PA group and AA group. The levels of IL-6 and TNF-αin the supernatant of each fatty acid intervention group were increased, while those in the EPA group were the lowest. The levels of IL-6 in the supernatant of the PA and AA groups were statistically different from the blank group Significance (P <0.05). Compared with the blank group, the expression of TLR4 mRNA increased in all the fatty acid intervention groups (all P <0.05), and the lowest in the EPA group. CONCLUSION: High concentration PA can reduce the sensitivity of HepG2 cells to insulin, lead to insulin resistance, reduce the sensitivity of AA to insulin, while EPA can increase the sensitivity of insulin. TLR4 signal transduction pathway and its inflammatory response It may be its important mechanism of action.