高糖对人肾小球内皮细胞膜表面多糖-蛋白复合物厚度及核心蛋白Syndecan-1和Glypican-1表达的影响  

Effect of High Concentration of Glucose on Thickness of Glycocalyx and Expression of Syndecan-1 and Glypican-1 in Cultured Human Renal Glomerular Endothelial Cells

作　　者：邱红渝[1] 樊文星[1] 黄颂敏[1] 刘芳[1] 唐万欣[1] 左川[1] 

机构地区：[1]四川大学华西医院肾脏内科,成都610041

出　　处：《四川大学学报（医学版）》2010年第6期980-985,共6页Journal of Sichuan University(Medical Sciences)

摘　　要：目的观察高糖刺激对体外培养人肾小球内皮细胞(HRGECs)膜表面多糖-蛋白复合物厚度及对HRGECs表面核心蛋白Syndecan-1和Glypican-1表达的影响。方法体外培养HRGECs,随机分为3组,分别为:高糖组(30 mmol/L D-葡萄糖)、正常糖浓度组(5 mmol/L D-葡萄糖+25 mmol/L甘露醇)、甘露醇组(30 mmol/L甘露醇),分别培养72 h后,用特异性异硫氰酸盐荧光标记的麦胚凝集素(WGA-FITC)进行免疫荧光染色,共聚焦激光显微镜(CLSM)进行三维(3D)重建,观察HRGECs表面多糖-蛋白复合物厚度的变化。利用倒置荧光显微镜观察各组HRGECs膜表面核心蛋白Syndecan-1和Glypican-1免疫荧光染色情况。实时荧光定量PCR法检测各组细胞Syndecan-1和Glypican-1 mRNA的含量,Western blot检测Syndecan-1和Glypican-1蛋白的表达。结果与正常糖浓度组相比,高糖组HRGECs表面多糖-蛋白复合物厚度减少36.8%(P<0.05)。高糖组HRGECs表面Syndecan-1和Glypican-1免疫荧光染色弱于正常糖浓度组和甘露醇组。高糖组HRGECs Syndecan-1和Glypican-1表达弱于正常糖浓度组和甘露醇组(P<0.05),且未检测到高糖组Glypican-1蛋白的表达。结论高糖可导致HRGECs表面多糖-蛋白复合物的缺失(厚度减少)和HRGECs膜表面核心蛋白Sydecan-1和Glypican-1的表达减弱。Objective To investigate the influence of high concentration of glucose on the thickness of Glycocalyx and expression of core protein Sydecan-1 and Glypican-1 in cultured human renal glomerular endothelial cells （HRGECs）. Methods HRGECs in culture were randomly divided into 3 groups, high concentration of glucose （30 mmol/L D-glucose, high glucose group）, normal concentration of glucose as controlled group （5 mmol/L D- glucose＋25 mmol/L mannitol, normal control group）, and mannitol group （30 mmol/L mannitol） respectively. After 72 hours, confoeal laser scanning microscopy （CLSM） was used to observe and characterize the fully hydrated glyeoalyx of HRGECs. Real time quantitative PCR and Western blot were applied to detect the mRNA levels and protein expression of Syndecan-1 and Glypican-1, and the fluorescence microscope were used to observe the immunofluorescenee change of Syndeean-1 and Glypican-1. Results Compared with normal control group, the thickness of Glycoealyx on the surface of HRGECs in high glucose group decreased to 36. 8% （P〈0. 05）. Immunofluorescence shows the depositions of Syndecan-1 and Glypican-1 were weakened in high glucose group. The mRNA and protein expression of Syndecan-1 and Glypican-1 were significantly decreased （P〈 0. 05） compared with normal control group and mannitol group. Conclusion High concentration of glucose can reduce thickness of Glycocalyx on the surface of human glomerular endothelial cell. At the same time, high glucose can decrease the expression of core protein Sydecan-1 and Glypican-1 of HRGECs.

关 键 词：人肾小球内皮细胞 高糖 多糖-蛋白复合物 核心蛋白 SYNDECAN-1 Glypican-1 

分 类 号：R587.2[医药卫生—内分泌]