糖化白蛋白诱导人近端肾小管上皮细胞产生损伤与炎性反应的表达

发布时间：2019-01-04 08:09

【摘要】：目的探讨在糖化白蛋白(glycated albumin,GA)刺激下,人近端肾小管上皮细胞(HK-2细胞系)中肾损伤分子-1(kidney injury molecule-1,KIM-1)、中性粒细胞明胶酶相关载脂蛋白(neutrophil gelatinase-associated lipocalin,NGAL)及炎性反应相关细胞因子的表达情况。方法分别采用不同浓度的GA与白蛋白(albumin,Alb)刺激HK-2细胞12 h、24 h及48 h,通过real-time PCR和酶联免疫吸附法(enzyme-linked immunesorbent assay,ELISA)分别检测NGAL、KIM-1的mRNA表达与蛋白释放水平,并运用流式液相多重蛋白定量技术检测血管内皮生长因子(vascular endothelial growth factor,VEGF)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-8(interleukin-8,IL-8)和可溶性细胞间黏附分子-1(soluble intercellular cell adhesion molecule-1,sICAM-1)等细胞因子的浓度。结果与对照组与Alb组比较,GA刺激HK-2细胞12 h、24 h和48 h均能显著上调KIM-1和NGAL的mRNA表达(P0.05)以及KIM-1和NGAL蛋白的释放(P0.05);GA组sICAM-1、VEGF与IL-8水平在各时间点均高于Alb组和对照组(P0.05)。结论 GA能上调KIM-1和NGAL mRNA的表达与蛋白的释放,并促进炎性细胞因子的分泌从而对肾小管造成损伤,提示GA可反映糖尿病肾脏受累情况。
[Abstract]:Objective to investigate the expression of renal injury molecule-1 (kidney injury molecule-1,KIM-1 in human proximal tubular epithelial cells (HK-2 cell lines) stimulated by glycosylated albumin (glycated albumin,GA). Expression of neutrophil gelatinase-associated apolipoprotein (neutrophil gelatinase-associated lipocalin,NGAL) and inflammatory response-related cytokines. Methods HK-2 cells were stimulated with different concentrations of GA and albumin (albumin,Alb) for 24 h and 48 h, respectively. NGAL, was detected by real-time PCR and enzyme-linked immunesorbent assay,ELISA. MRNA expression and protein release level of KIM-1 were detected by flow liquid phase multiplex protein quantitative technique. Vascular endothelial growth factor (vascular endothelial growth factor,VEGF), tumor necrosis factor- 伪 (TNF- 伪) and tumor necrosis factor 伪 (TNF- 伪) were detected by flow cytometry. Concentrations of cytokines such as interleukin-8 (interleukin-8,IL-8) and soluble intercellular adhesion molecule-1 (soluble intercellular cell adhesion molecule-1,sICAM-1. Results compared with control group and Alb group, GA stimulated HK-2 cells significantly up-regulated mRNA expression of KIM-1 and NGAL (P0.05) and release of KIM-1 and NGAL protein (P0.05). The levels of sICAM-1,VEGF and IL-8 in GA group were higher than those in Alb group and control group at each time point (P0.05). Conclusion GA can up-regulate the expression of KIM-1 and NGAL mRNA and release of protein, and promote the secretion of inflammatory cytokines, thus causing damage to renal tubules, suggesting that GA can reflect the renal involvement in diabetic patients.
【作者单位】：北京大学第一医院检验科;
【分类号】：R587.2;R692.9

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