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目的探讨尿毒血清对人肾上管上皮细胞侏(HK-2)增殖和胶原蛋白分泌的影响。方法在HK-2细胞培养液中加入正常人血清及不同浓度的尿毒血清,采用四氮甲基唑蓝(MTT)比色法和流式细胞术测定HK-2细胞增殖活性的变化;采用消化法检测细胞培养上清中羟脯氨酸的含量,换算成总胶原分泌量。结果5%￣20%浓度尿毒血清组A490值均高于正常对照组,10%￣20%浓度尿毒血清组G1期细胞均低于正常对照组,P1指数均高于正常对照组,但不同浓度尿毒血清组组间差异无统计意义;各浓度尿毒血清组羟脯氨酸和胶原蛋白含量较正常对照组增高,且随着尿毒血清浓度的升高而增高。结论慢性肾衰竭患者体内蓄积的尿毒症素可能通过促进肾小管上皮细胞增殖,促进细胞外基质的分泌,从而加速肾小管—间质纤维化进程,导致肾功能进行性恶化。
Objective To investigate the effect of uremic serum on the proliferation and collagen secretion of human renal epithelial cells (HK-2). Methods Normal human serum and different concentrations of uremic serum were added into HK-2 cell culture medium. The proliferation of HK-2 cells was assayed by MTT assay and flow cytometry. Law test cell culture supernatant hydroxyproline content, converted into total collagen secretion. Results The A490 values of 5% ~ 20% uremic serum group were higher than that of the normal control group. The cells in G1 phase of 10% ~ 20% uremic serum group were lower than the normal control group, and the P1 index were higher than the normal control group, There was no significant difference between the groups of uremic serum. The concentrations of hydroxyproline and collagen in each concentration of uremic serum were higher than that of the normal control group, and increased with the increase of serum concentration of uremia. Conclusion The accumulation of uremic toxins in patients with chronic renal failure may accelerate the process of tubulointerstitial fibrosis by promoting the proliferation of renal tubular epithelial cells and promoting the secretion of extracellular matrix, resulting in progressive worsening of renal function.