低氧、高糖对肾小球内皮细胞膜表面多糖-蛋白复合物厚度影响的分析

Effect of hypoxia and high glucose on the thickness of polysaccharide-protein complex on human glomerular endothelial cell surface

目的探讨低氧、高糖对人肾小球内皮细胞(HRGECs)膜表面多糖-蛋白复合物厚度的影响。方法体外培养HRGECs,随机分为低氧高糖组、常氧高糖组和正常对照(Con)组,对各组HRGECs进行免疫荧光染色,观察膜表面多糖-蛋白复合物厚度情况。结果 Con组HRGECs膜表面多糖-蛋白复合物免疫荧光染色最强,常氧高糖组荧光染色减弱,而低氧高糖组荧光染色减弱更明显;Con组、常氧高糖组和低氧高糖组膜表面多糖-蛋白复合物厚度比较,差异有统计学意义[(3.892±0.183)vs(2.913±0.137)vs(2.062±0.152)nm,P<0.05];Con组HRGECs培养24h、48h和72h的一氧化氮(NO)分泌水平均高于其他两组,且低氧高糖组各时点NO分泌水平最低(P<0.05)。结论低氧、高糖可使HRGECs膜表面多糖-蛋白复合物厚度减少,且导致NO分泌减少。

Objective To investigate the effect of hypoxia and high glucose on the thickness of polysaccharide-protein complex on human glomerular endothelial cell（HRGEC）surface. MethodsHRGECs by in vitro culture were randomly divided into three groups：hypoxia and high glucose group（n=20）,normoxia and high glucose group（n=20）and normal control group（Con,n=20）.The thickness of polysaccharide-protein complex on the HRGEC surface among three groups was compared through immunofluorescent staining. Results Immunofluorescent staining of polysaccharide-protein complex was strongest in Con group,with a decreased trend in normoxia and high glucose group,followed by hypoxia and high glucose group.The thickness of polysaccharide-protein complex in Con group was higher than in normoxia and high glucose group and in hypoxia and high glucose group [（3.892±0.183）vs（2.913±0.137）vs（2.062±0.152）nm,P〈0.05].The levels of nitric oxide（NO）secreted by HRGECs cultured for 24 h,48hand 72 hwere highest in Con group and lowest in hypoxia and high glucose group（P〈0.05）. Conclusion Hypoxia and high glucose can reduce the thickness of polysaccharide-protein complex on HRGECs surface and inhibit the secretion of NO.