【摘要】 目的:探讨应用人肾近曲小管上皮细胞株(HK2)构建的生物人工肾小管辅助装置(RAD)体外对β2微球蛋白(β2-MG)的清除作用及机制。方法:A组以HK2体外培养后植入F60血液滤过器内腔构建的RAD后用含β2-MG的置换液体外模拟液体滤过,B组以上述条件构建的RAD用不含β2-MG的置换液模拟液体滤过,C组用含β2-MG的置换液对无细胞植入的F60滤器模拟液体滤过,对各组测定不同时间点的β2-MG滤过率,并应用免疫荧光法观察滤过后A、B组HK2细胞Megalin与β2-MG的蛋白表达。结果:A组β2-MG滤过率在各时间点均明显高于C组(P<0.05);C组β2-MG滤过率在第4h较同组前期各时间点明显降低(P<0.05)。免疫荧光显示A组Megalin与β2-MG同步表达,B组则仅见Megalin表达。结论:以HK2细胞构建的RAD可显著提高β2-MG的滤过率,其功能在4小时内可稳定维持,作用机制可能与Megalin的内吞作用相关。更多还原

【Abstract】 Objective:To determine the clearance and mechanism of beta2-microglobulin(β2-MG) by using bioartificial renal tubule assist device(RAD) which constructed by HK2 cell line in vitro.Methods:HK2 cells were seeded into the hemofilter(F60) for constructing RAD,then the filtration in vitro was simulated by replacement fluids with β2-MG.RADs of group B were taken the filtration with replacement fluids without β2-MG.The hemofilters without HK2 cell line were uesd in filtration by replacement fluids with β2-MG as group C.The reabsorption rate of three groups were measured.The expression of megalin and β2-MG protein of group A and B were detected by immunofluorescence after filtration.Results:The β2-MG reabsorption rate of group A was significantly higher as compared with group C(P<0.05).The reabsorption rate of β2-MG in group C at the fourth hour of filtration significantly decreased as compared with other time points of group C.Immunofluorescence detection indicated that HK2 cells of group A both have the expression of megalin and β2-MG,meanwhile only the expression of megalin was detected in the cells of group B.Conclusion:The RAD cuntructed by HK2 cell line can obviously increase the reabsorption function of β2-MG.The function could maintain well during 4 hours.The probable mechanism is associated with the endocytosis of megalin.更多还原