【摘要】 目的 探讨黄芪甲苷(AS-Ⅳ)对HepG2高脂细胞模型血脂代谢的影响以及Toll-like信号通路在其中的作用。方法 使用HepG2细胞构建高脂血症模型,并随机分为六组：AS-Ⅳ低、中、高(0.05、0.1、0.2 mmol/L)剂量组,对照组,抑制剂组, AS-Ⅳ+抑制剂组。分别进行实时荧光定量聚合酶链式反应(PCR)和蛋白质免疫印迹(WB)分析,检测Toll样受体4(TLR4)、髓样分化因子88(MyD88)、肿瘤坏死因子受体相关因子6(TRAF6)mRNA和蛋白表达。结果 (1)在AS-Ⅳ高剂量组中,甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白(LDL)的水平显著降低(在AS-Ⅳ低剂量组中降低不显著)。(2)在AS-Ⅳ中、高剂量组中, TLR4、MYD88和TRAF6mRNA的表达水平明显下降(在AS-Ⅳ低剂量组中降低不显著)。(3)在AS-Ⅳ高、中剂量组中, TLR4、MyD88、TRAF6蛋白的表达水平显著下降(在AS-Ⅳ低剂量组中降低不显著)。(4) TLR4抑制剂TAK-242、MyD88抑制剂ST2825和TRAF6抑制剂C25-140能够抑制AS-Ⅳ的作用。结论 AS-Ⅳ在HepG2高脂细胞中有降低血脂累积作用,可能通过减轻脂肪代谢障碍实现,其机理与Toll-like信号路径有关。更多还原

【Abstract】 Objective To discuss the effect of astragaloside Ⅳ(AS Ⅳ) on lipid metabolism in HepG2cell model and the role of Toll-like signaling pathway in it. Methods HepG2 cells were used to construct hyperlipidemia model and randomly divided into six groups: low-, medium-and high-dose(0.05, 0.1, 0.2 mmol/L)AS-Ⅳ groups, control group, inhibitor group and AS-Ⅳ+inhibitor group. The expressions of Toll like receptor 4(TLR4), myeloid differentiation factor 88(MyD88), tumor necrosis factor receptor-associated factor 6, tumor necrosis factor receptor associated factor 6(TRAF6) mRNA and protein were detected by real-time quantitative polymerase chain reaction(PCR) and Western blot(WB) assay. Results(1)In high-dose AS-Ⅳ group, the levels of triglycerides(TG), total cholesterol(TC) and low-density lipoprotein(LDL) were significantly reduced at high doses(not significantly reduced in the low-dose AS-Ⅳ group).(2)In the medium-and high-dose AS-Ⅳ groups,the expression levels of TLR4, MYD88 and TRAF6 mRNA were significantly decreased(not significantly in lowdose group).(3)TLR4, MyD88, TRAF6 protein expression was significantly decreased in the high-and mediumdose AS-Ⅳ groups(not significantly in low-dose AS-Ⅳ group).(4)TLR4 inhibitor TAK-242, MyD88 inhibitor ST2825 and TRAF6 inhibitor C25-140 were able to inhibit the action of AS-Ⅳ. Conclusion AS-Ⅳ can reduce the accumulation of blood lipids in HepG2 cells, which may be achieved by reducing the disorder of fat metabolism, and its mechanism is related to the Toll like signaling pathway.更多还原