苦荞麦总黄酮对软脂酸诱导的脐静脉内皮细胞NOX4蛋白表达的影响  

Effect of Total Flavonoids from Fagopyrum tataricum on Expression of NOX4 Protein in EA. hy926 Cell Induced by Palmitic Acid

作　　者：冯晓帆[1] 柳春[1] 赵丹玉[1] 刘恋[1] 王艳杰[1] 洪涌涛 刘博通[1] 

机构地区：[1]辽宁中医药大学基础医学院,沈阳110847

出　　处：《中国实验方剂学杂志》2016年第9期106-110,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：辽宁省科学技术计划项目(2013226012)

摘　　要：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 investigate effect of total flavonoids from Fagopyrum tataricum on expression of nicotinamide adenine dinucleotidephosphateoxidase-4（ NOX4） in EA. hy926 cell induced by palmitic acid,and to discuss mechanism of total flavonoids from F. tataricum in signal transduction pathway of insulin resistance.Method： EA. hy 926 cells were divided into blank group, insulin resistance model group, total flavonoids of F. tataricum group（ 31. 25,62. 5,125 mg·L-1） and metformin group. Every groups were combined insulin with 50 nmol·L-1. Compared with the blank group,other groups were combined 600 μmol·L-1palmitic acid.Total flavonoids of F. tataricum group was combined 125 mg·L-1total flavonoids of F. tataricum. Metformin group was combined 2 mmol·L-1metformin. The content of asymmetric dimethylarginine（ ADMA） was tested by double antibody sandwich method. The content of NO was tested by nitrate reductase method. NOX4 protein expression levels were determined by Western blot. Result： Compared with model group,contents of ADMA in the middle and high dose groups of total flavonoids were significantly reduced, contents of NO were increased significantly. Compared with model group,expression levels of NOX4 protein were reduced significantly in high dose groups of total flavonoids. Conclusion： Total flavonoids from F. tataricum can promote synthesis of NO by inhibiting synthesis of NOX4 and ADMA in EA. hy926 cell induced by palmitic acid,then it may restrain insulin resistance.

关 键 词：原型尼克酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4) 苦荞麦 总黄酮 脐静脉内皮细胞 软脂酸 胰岛素抵抗 

分 类 号：R945[医药卫生—微生物与生化药学] R285.5[医药卫生—药剂学]