不同血液净化方法对人肾小管上皮细胞分泌细胞外基质的影响

Effects of different blood purification on ECM excretion of human renal tubular epithelial cells

目的探讨不同血液净化方法前后尿毒症患者血清对人近端肾小管上皮细胞株(HK-2)分泌细胞外基质(ECM)的影响。方法选择慢性肾衰竭维持性透析(MHD)的患者24例,随机分成3组,分别接受常规血液透析(HD)、高通量透析(HFHD)、血液透析滤过(HDF)后稀释法治疗,提取患者透析前后的血清。将透析前后血清分别混匀后培养HK-2细胞。应用碱水解法、ELISA法检测HK-2细胞培养上清中胶原、纤连蛋白(FN)等ECM的蛋白水平。结果HDF组透析后血清使HK-2细胞胶原蛋白和FN分泌减少,与透析前比较,差异有统计学意义(P<0.01);而HD、HFHD组透析后与透析前比较,差异无统计学意义(P>0.05)。HDF组透析前后胶原蛋白及FN分泌的差值与HD、HFHD组比较,差异有统计学意义(P<0.01),而HD组透析前后的差值与HFHD组比较,差异无统计学意义(P>0.05)。结论HDF可能通过清除致肾间质ECM异常积聚的尿毒症毒素和减少透析用水对机体的影响,从而减少HK-2细胞合成胶原和FN等ECM,对肾间质纤维化的发生、发展起一定的抑制作用,而HD、HFHD在此方面的作用不显著。

Objective To investigate the effects of uremic serum on extracellular matrix （ECM） of human renal tubular epithelial cells before and after the application of different methods of blood purification. Methods Twenty-four maintenance hemodialysis （MHD） patients were randomly divided into 3 groups that received conventional hemodialysis（HD） , high-flux hemodialysis（HFHD） and postdilution hemodiafiltration （HDF） treatment respectively. Uremic serum samples from patients before and after hemodialysis were collected and mixed to culture HK-2 cells respectively. Alkaline hydrolysis technique and ELISA were used to determine the concentrations of collagen and fibronectin （ FN ） in cell culture supernatants. Results After dialysis, uremic serum of HDF caused a significant decline in HK-2＇ s collagen and FN excretion, compared with the pre-dialysis value （ P 〈 0.01 ） ; whereas in HD and HFHD groups, the pre- and pest-dialysis difference was not significant （ P 〉 0. 05 ）. Difference of collagen and FN excretion in HDF group was significant to that in HD and HFHD groups （P 〈 0.01 ） ; while between group HD and HFHD, the difference was not significant （P 〉 0.05 ）. Conclusion HDF can undermine the synthesis of ECM like collagen and FN, and restrains renal interstitial fibrosis（RIF） occurrence and development via elimination of uremic serum that causes abnormal ECM accumulation and decrease of dialysis water influence on human body, whereas HD and HFHD are not effective in these regards.