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

Effects of glycated albumin on inducing injury and inflammation in human proximal tubular epithelial cells

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(P<0.05)以及KIM-1和NGAL蛋白的释放(P<0.05);GA组sICAM-1、VEGF与IL-8水平在各时间点均高于Alb组和对照组(P<0.05)。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。

Objective To investigate the effects of glycated albumin （GA） on the expression of kidney injury molecule-1 （KIM-1）,neutrophil gelatinase-associated lipocalin （NGAL） and inflammatory cytokines in human proximal tubular epithelial cells （HK-2 cells）.Methods The HK-2 cells were treated with the different concentrations of GA and albumin （Alb） for 12h,24h and 48h.The mRNA expressions of KIM-1 and NGAL were detected by real-time PCR.The release of KIM-1 and NGAL in the supernatants were detected by ELISA.The concentrations of VEGF,TNF-α,IL-8 and sICAM-1 were detected by cytometric beads array method.Results Compared with control and Alb groups,the mRNA levels of KIM-1 and NGAL in HK-2 cells were significantly up-regulated at 12 h,24 h and 48 h after GA treatment （P〈0.05）;the protein levels of KIM-1 and NGAL released in supernatants of GA-treated cells were significantly higher than those in control group and Alb groups at the same time points （P〈0.05）;GA groups also had significantly higher levels of sICAM-1,VEGF and IL-8 than control group at each time point （P〈0.05）.Conclusion GA can up-regulate the expression and release of KIM-1 and NGAL and promote the secretion of inflammatory cytokines which could cause damage to renal tubules,suggesting that GA may reflect the diabetic renal involvement.