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, confocal laser scanning microscopy (CLSM) was used to observe and characterize the fully hydrated glycoalyx 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 immunofluorescence change of Syndecan-1 and Glypican-1.
Results: Compared with normal control group, the thickness of Glycocalyx 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.