黄芪多糖对棕榈酸损伤的人脐静脉内皮细胞内一氧化氮水平的影响

The effects of Astragalus polysaccharides on nitric oxide in human umbilical vein endothelial cells induced with palmitic acid

目的:探讨黄芪多糖对棕榈酸损伤的人脐静脉内皮细胞内一氧化氮(NO)水平的影响。方法:用棕榈酸诱导人脐静脉内皮细胞建立内皮细胞损伤模型,实验设为对照组、模型组、黄芪多糖组(400 mg/L、800 mg/L、1 600 mg/L)。MTT试验检测黄芪多糖对细胞活性的影响。Western blot检测细胞内AMPK、p-AMPK、p-Akt、eNOS、p-e NOS、Nox4、GAPDH蛋白的表达。Q-PCR试验检测Nox4 mRNA以及eNOS mRNA的表达。硝酸酶还原法检测细胞内NO含量。结果:黄芪多糖可以上调棕榈酸损伤的人脐静脉内皮细胞内p-AMPK、p-Akt、pe NOS蛋白以及eNOS mRNA的表达(P<0.05);下调Nox4蛋白及Nox4 mRNA的表达,增加细胞内NO的生成(P<0.05)。结论:黄芪多糖可以促进内皮细胞内NO的生成,其调控机制可能与其能活化细胞内AMPK、Akt蛋白,激活eNOS以及降低Nox4的表达有关。

Objective:To explore and investigate the effects of Astragalus polysaccharides on nitric oxide in human umbilical vein endothelial cells induced with palmitic acid.Methods:Endothelial cell injury model was established by using human umbilical vein endothelial cells induced with palmitic acid.The experiment consisted of control group,model group,astragalus polysaccharide group(400 mg/L,800 mg/L,1 600 mg/L).The effects of Astragalus polysaccharides on cell viability was tested by MTT assay.Western blotting assay was used to detect the expressions of AMPK,p-AMPK,p-Akt,eNOS,p-eNOS,Nox4,and GAPDH.The Q-PCR assay was used to detect the expressions of Nox4 mRNA and eNOS mRNA.Nitrate reduction assay was used to detect intracellulr NO levels.Results:Astragalus polysaccharide could up-regulate the expressions of p-AMPK,p-Akt,p-eNOS and eNOS mRNA in human umbilical vein endothelial cells induced with palmitic acid,down-regulate the expressions of Nox4 and Nox4 mRNA,and increase the production of NO in cells.Conclusion:Astragalus polysaccharides can promote the production of NO in endothelial cells,which can prevent and treat the damage of vascular endothelial cells.Its regulation mechanism may be associated with its ability to activate intracellular AMPK,Akt,active eNOS,and decrease the expression of Nox4.