苦荞麦总黄酮对软脂酸诱导EA.hy926细胞IRS-2合成的影响

Effect of Total Flavonoids of Tartary buckwheat on Palmitic Acid Induced IRS-2 Synthesis in EA.hy926 Cells

摘要:

目的：研究苦荞麦总黄酮对于软脂酸诱导EA.hy926细胞胰岛素受体底物2（IRS-2）合成的影响。方法：将体外培养的EA.hy926细胞分为正常对照组、模型组、苦荞麦总黄酮组、二甲双胍组，各组均加入10%FBS的DMEM完全培养基和终浓度为50 nmol·L^-1的胰岛素，除正常对照组其余各组加入终质量浓度为600 μmol·L^-1的软脂酸建立胰岛素抵抗细胞模型；苦荞麦总黄酮组加入终浓度为125 mg·L^-1的苦荞麦总黄酮；二甲双胍组加入终浓度为2 mmol·L^-1的二甲双胍。采用RT-PCR以及western blot法分别测定各组细胞IRS-2 mRNA和蛋白的表达。结果：模型组细胞IRS-2 mRNA和蛋白表达与正常组相比明显下降（P<0.05），正常对照组IRS-2 mRNA的相对表达量为（1.203±0.040），蛋白相对表达量为（1.164±0.034），模型组为（0.611±0.022）和（0.580±0.006），苦荞麦总黄酮组与二甲双胍组细胞IRS-2 mRNA和蛋白表达与模型组相比明显增加（P<0.05），治疗组间无明显差异（P>0.05）。苦荞麦总黄酮组IRS-2 mRNA的相对表达量为（0.904±0.157），蛋白相对表达量为（0.845±0.029），二甲双胍组为（0.890±0.054）和（0.841±0.006）。结论：苦荞麦总黄酮对于软脂酸诱导下EA.hy926细胞IRS-2合成具有明显促进作用。

Abstract:

Objective: To observe the effects of total flavonoids of tartary buckwheat on IRS-2 in EA.hy926 cells in the presence of palmitic acid. Method: EA.hy 926 cells were cultured in vitro and divided into control group, insulin resistance induced by palmitic acid group, total flavonoids of tartary buckwheat group and metformin group randomly. Each group combined 10% FBS ADMA complete medium and insulin (50 nmol·L^-1), compared with control group, other groups contain palmitic acid (600 μmol·L^-1), and total flavonoids of tartary buckwheat group contain total flavonoids of tartary buckwheat flavonoids (125 mg·L^-1), metformin group contain metformin (2 mmol·L^-1). The IRS-2 mRNA and protein expression levels were determined by RT-PCR and Western blot respectively. Result: Compared with control groups, the expression levels of IRS-2 mRNA and protein were significantly lower in insulin resistance group (P<0.05).the expresssion levels of IRS-2 mRNA and protein of control group were (1.203±0.040 )and (1.164±0.034), and insulin resistance induced by palmitic acid group were (0.611±0.022) and (0.580±0.006), Compared with insulin resistance group, the IRS-2 mRNA and protein expression were markedly increased in both total flavonoids of tartary buckwheat group and metformin group (P<0.05), and there is no significant difference between two groups. the expression levels of IRS-2 mRNA and protein of total flavonoids of tartary buckwheat group were (0.904±0.157) and (0.845±0.029), and metformin group randomly were (0.890±0.054) and (0.841±0.006). Conclusion: Total flavonoids of tartary buckwheat effectively promotes the expression of IRS-2 mRNA and protein in endothelial cells under palmitic acid stimulation.