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)。④TLR4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。
Objective To discuss the effect of astragalosideⅣ(ASⅣ)on lipid metabolism in HepG2 cell 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-Ⅳgroups,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.Results①In high-dose AS-Ⅳgroup,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-Ⅳgroup).②In the medium-and high-dose AS-Ⅳgroups,the expression levels of TLR4,MYD88 and TRAF6 mRNA were significantly decreased(not significantly in lowdose group).③TLR4,MyD88,TRAF6 protein expression was significantly decreased in the high-and mediumdose AS-Ⅳgroups(not significantly in low-dose AS-Ⅳgroup).④TLR4 inhibitor TAK-242,MyD88 inhibitor ST2825 and TRAF6 inhibitor C25-140 were able to inhibit the action of AS-Ⅳ.Conclusion AS-Ⅳcan 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.
作者
杨柳
熊小平
马国斌
李昱
胡林
丁海强
曾圣强
王洪
YANG Liu;XIONG Xiao-ping;MA Guo-bin(First Affiliated Hospital of Nanchang Medical College,Jiangxi Provincial People's Hospital,Nanchang 330006,China)
出处
《中国现代药物应用》
2022年第22期184-189,共6页
Chinese Journal of Modern Drug Application
基金
江西省中医药科技计划项目(项目编号:2019A162)
南昌医学院第一附属医院江西省人民医院博士启动基金(项目编号:19-236)。
