高浓度的糖刺激人肾小球内皮细胞表达单核细胞趋化蛋白-1的影响

目的 :探讨高浓度的糖对培养的人肾小球内皮细胞 (HUGEC)表达单核细胞趋化蛋白 1(MCP 1)的影响 ,以及HUGEC的条件培养基对单核细胞 (MC)的趋化作用及抗MCP 1抗体对MC迁移的影响。方法 :采用原位杂交技术和细胞ELISA法 ,观察MCP 1的表达 ;用改良的Boyden小室微孔滤膜法 ,测定高浓度的糖刺激HUGEC后的条件培养基 ,对MC的趋化作用 ,以及抗MCP 1抗体对MC迁移的影响。结果 :(1)在低浓度的糖(含 5 .5mmol/LD 葡萄糖 )条件下培养的HUGECMCP 1mRNA呈弱表达。高浓度的糖 (含 2 5mmol/LD 葡萄糖 )刺激后 ,8h即出现MCP 1表达增强 ,于 16h达高峰。(2 )高浓度的糖刺激HUGEC后的条件培养基 ,对MC具有明显的趋化作用 ;抗MCP 1抗体可抑制其作用。结论 :在高浓度的糖诱导下 ,人HUGECMCP 1的表达增强 ,其条件培养基对MC具有趋化作用 ,从而可能招引单核细胞迁入内皮下间隙。

Expression of monocyte chemoattractant protein-1 in glomerular endothelial cells stimulated by high concentration of glucose

AIM: To study the expression of monocyte chemoattractant protein-1 (MCP-1) in cultured human glomerular endothelial cells (HUGECs) stimulated by high concentration of glucose. METHODS: The effect of high concentration of glucose on the expression of MCP-1mRNA by HUGECs was detected by in situ hybridization and cell ELISA. The monocyte migration induced by the conditioned media of cultured HUGECs was assayed by a modified Boyden Chamber micropore filtration membrane method. Effects of anti -MCP-1 antibody on monocyte migration was detected as well. RESULTS: The MCP-1 mRNA was only weakly expressed in HUGECs cultured under the condition of low concentration of glucose. Following a stimulation of high concentration of glucose (25 mmol/L), MCP-1 mRNA expression was upregulated from the 8th hour and reached the maximum at the 16th hour. Conditioned medium of cultured HUGECs stimulated with high concentration of glucose had marked chemotaxis for monocytes, which was inhibited by anti-MCP-1 antibody. CONCLUSION: HUGEC stimulated by high glucose may highly express MCP-1, and produce chemotatic factors for monocytes.