蕨麻多糖对晚期糖基化终产物诱导内皮细胞损伤的抑制作用及机制研究

Inhibitory Effects and Mechanism of Potentilla Anserine Polysaccharide in Advanced Glycation End-products Induced Human Umbilical Vein Endothelial Cells Injury

目的:探讨蕨麻多糖对晚期糖基化终产物(AGEs)诱导人脐静脉内皮细胞(HUVECs)损伤的保护作用及其机制。方法:以不同浓度蕨麻多糖与体外培养的HUVECs预先共孵育2 h,再以AGEs对细胞进行干预。MTT法检测细胞增殖活力,生化方法检测超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量、一氧化氮(NO)分泌水平,酶联免疫法检测内皮素1(ET-1)水平,化学荧光法检测活性氧簇(ROS)水平,Western Blot检测各组细胞内皮型一氧化氮合酶(e NOS)、诱导型一氧化氮合酶(i NOS)、糖基化终产物受体(RAGE)、NAPDH氧化酶亚单位p22^(phox)蛋白表达水平。结果:与对照组比较,模型组细胞增殖活力、SOD活性、NO含量和e NOS蛋白表达显著降低,MDA、ET-1、ROS水平及i NOS、RAGE、p22^(phox)蛋白表达水平均显著增加(P <0. 01)。与模型组比较,蕨麻多糖各浓度组细胞增殖活力、SOD活性、NO含量和e NOS蛋白表达增加,MDA、ET-1、ROS水平及i NOS、RAGE、p22^(phox)蛋白表达水平均显著降低(P <0. 01)。结论:蕨麻多糖能抑制AGEs诱导的人脐静脉内皮细胞氧化应激反应,可能与通过下调RAGE/NADPH途径、抑制NADPH氧化酶、减少ROS生成、改善内皮细胞功能有关。

Objective:To explore the effects and mechanism of potentilla anserine polysaccharide in advanced glycation end-products(AGEs)induced human umbilical vein endothelial cells(HUVECs)injury in vitro.Methods:Potentilla anserine polysaccharides at three different concentrations were incubated with HUVECs in vitro for 2 h,and then AGEs were added and incubated with the cells for 24 h.The cell proliferation activity was detected by MTT assay.Superoxide dismutase(SOD)activity,malondialdehyde(MDA)content and nitric oxide(NO)secretion level were measured by biochemical methods.Endothelin-1(ET-1)content was detected by enzyme-linked immunosorbent assay(ELISA).Reactive oxygen species(ROS)level was measured by chemical fluorescence method.The protein expression of endothelial nitric oxide synthase(e NOS),inducible nitric oxide synthase(i NOS),receptor for advanced glycation end-products(RAGE)and NAPDH oxidase subunit p22phox were detected by Western blot.Results:Compared with those in the control group,the cell proliferation activity,SOD activity,NO content and e NOS protein expression in the model group significantly decreased,and MDA content,ET-1 level,ROS level,and i NOS protein,RAGE protein and p22phox protein expression levels in the model group increased significantly(P<0.01).Compared with the model group,Potentilla anserina polysaccharide at different concentrations significantly increased the cell proliferation activity,SOD activity,NO content and e NOS protein expression,and decreased MDA content,ET-1 level,ROS level,and i NOS protein,RAGE protein and p22phox protein expression levels(P<0.01).Conclusion:Potentilla anserina polysaccharide can inhibit AGEs-induced oxidative stress response in HUVECs,and the possible mechanism may be related to the down-regulation of RAGE/NADPH pathway,the inhibition of NADPH oxidase,the reduction of ROS production and the improvement of endothelial cell function.