缬沙坦对高糖环境下大鼠肾小球系膜细胞氧化应激水平的影响

Effect of Valsartan on Rat Glomerular Mesangial Cells Oxidative Stress Induced by High Glucose

关键词:  糖尿病肾病  缬沙坦  NADPH氧化酶  氧化应激

Key words:  diabetic nephropathy  valsartan  NADPH oxidase  oxidative stress

摘要:

目的 观察缬沙坦对高糖环境下大鼠肾小球系膜细胞氧化应激水平的影响。方法 高糖(20 mmol·L^-1)体外培养大鼠肾小球系膜细胞，不同浓度缬沙坦(10^-7，10^-6，10^-5和10^-4 mol·L^-1)干预，以RT-PCR技术检测细胞内还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶亚基p22^phox的mRNA表达的变化，并以DCFH-DA和HE标记细胞内活性氧(ROS)，流式细胞术检测细胞内ROS相对浓度。结果 高糖时间相关性诱导系膜细胞超氧阴离子(·O₂^-)与过氧化氢(H₂O₂)明显升高，并引起p22phox的mRNA表达显著上调，缬沙坦对高糖引起系膜细胞p22^phox的mRNA表达增加有下调作用，明显降低细胞内·O₂^-和H₂O₂浓度，并呈浓度依赖性。结论 缬沙坦抑制高糖引起肾小球系膜细胞氧化应激可能和下调p22phox的mRNA表达有关。

Abstract:

OBJECTIVE To observe the effect of valsartan on rat glomerular mesangial cells oxidative stress induced by high glucose. METHODS Mesangial cell was cultured in high glucose(20 mmol·L^-1) and intervened by valsartan of different concentration(10^-7, 10^-6, 10^-5 and 10^-4 mol·L^-1). The mRNA of p22^phox, which was the primary subunit of NADPH oxidase, was detected by semi-quantitative RT-PCR. The relative concentration of reactive oxygen species (ROS) in cells was measured by flow cytometry, using DCFH-DA and HE label, in order to observe the effect of valsartan on glomerular mesangial cells oxidative stress induced by high glucose. RESULTS The expression of p22phox mRNA as well as ·O₂^- and H₂O₂ was up-regulated by glucose concentration and showed time dependent effect, and it was inhibited by valsartan, the inhibitory effect had dose dependent effect. CONCLUSION Valsartan inhibits the up-regulated of oxidative stress induced by high glucose, and it could be concerned with down regulation of the expression of p22^phox mRNA.