尿毒血清对HK-2细胞增殖和胶原蛋白分泌的影响(英文)

Effects of uremic serum on cell proliferation and procollagen of HK-2

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 explore the effects of uremic serum on proliferation and collagen secretion of human renal tubular epithelial cells. Methods： Human renal tubular epithelial cell line （HK-2） was cultured in RPMI-1640 medium. Normal serum and uremic serum in different concentrations were added into the cultured medium respectively. The proliferation of HK-2 was measured with methylene blue assay （MTT method） and flow cytometry. Hydroxyproline was measured as an index of procollagen production by digestive technique. Results： A490 was increased in 5%-20% uremic serum groups and cells in G1 phase were decreased, but proliferation index （PI） was increased in 10%-20% uremic serum groups compared with that of normal control, but significant difference of cell proliferation indexes was not found among uremic serum groups. Hydroxyproline and procollagen content was increased in uremic serum groups compared with normal control, and increased in parallel with the increasing of uremic serum soneentration. Conclusions： Uremic toxin may accelerate renal tubulointerstital fibrosis by promoting renal tubular epithelial cells proliferation and procollagen secretory in patients with chronic renal failure.