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目的:观察苦荞麦总黄酮对软脂酸诱导的人脐静脉内皮(EA.hy926)细胞中还原型尼克酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4)表达等指标的影响,探讨苦荞麦总黄酮在胰岛素抵抗(IR)信号传导通路中的作用机制。方法:将EA.hy926细胞株常规培养、传代,加入终浓度50 nmol·L~(-1)的胰岛素,分为空白组、模型组、苦荞麦总黄酮组(31.25,62.5,125 mg·L~(-1))和二甲双胍组,除空白组外,其他各组先用600μmol·L~(-1)软脂酸造IR模型,苦荞麦总黄酮组进一步加入125 mg·L~(-1)苦荞麦总黄酮,二甲双胍组加入2 mmol·L~(-1)二甲双胍。利用双抗体夹心法测定非对称性二甲基精氨酸(ADMA)含量,硝酸还原酶法测定NO含量,Western blot测定NOX4蛋白表达。结果:苦荞麦总黄酮中、高剂量组与模型组比较,ADMA含量明显减少,NO含量明显增加,均有显著性差异;苦荞麦总黄酮组高剂量组与模型组比较,NOX4蛋白表达量显著减少。结论:苦荞麦总黄酮可通过抑制软脂酸诱导下产生IR的EA.hy926细胞中NOX4和ADMA的合成来促进NO的合成,进而抑制IR的发生。
Objective: To observe the effects of total flavones of buckwheat on the expression of reduced nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in palmitic acid-induced human umbilical vein endothelial cells (EA.hy926) The Mechanism of Action of Total Flavones in Insulin Resistance (IR) Signaling Pathway. Methods: EA.hy926 cells were cultured, passaged and added with 50 nmol·L -1 insulin at final concentration. The cells were divided into blank group, model group, total flavonoids of buckwheat group (31.25, 62.5, 125 mg · L ~ (-1)) and metformin group. Except for the blank group, all the other groups were treated with 600 μmol·L -1 palmitate at first, and the total flavonoids of buckwheat were further treated with 125 mg · L -1 ) Total flavonoids of buckwheat, metformin group added 2 mmol·L -1 metformin. The asymmetric dimethylarginine (ADMA) content was determined by sandwich enzyme-linked immunosorbent assay (ELISA) and the NO content was determined by nitrate reductase method. The expression of NOX4 protein was determined by Western blot. Results: Compared with the model group, the content of ADMA decreased significantly and the content of NO significantly increased in the middle and high dose of total flavonoids of Tartary buckwheat compared with the model group. The NOX4 protein expression of high dose group cut back. CONCLUSION: Total flavonoids from buckwheat can promote the synthesis of NO by inhibiting the synthesis of NOX4 and ADMA in EA.hy926 cells induced by palmitate, thereby inhibiting the occurrence of IR.