生物人工肾小管体外构建及对氨基酸、钠离子、肌酐转运功能的研究

目的探讨体外构建生物人工肾小管(RAD)的方法及5h内RAD在体外对肌酐(Cr)、葡萄糖(Glu)、钠离子(Na )和天冬氨酸、苏氨酸、丝氨酸、谷氨酸的转运功能。方法采用猪肾近曲小管上皮细胞株(LLC-PK1)体外培养,然后植入FH66血液滤过器内腔继续培养两周构建RAD。测定RAD对Na 、Cr、葡萄糖及4种氨基酸的转运功能,并与未植入LLC-PK1的滤过系统做对照,并观察哇巴因、根皮苷对转运的特异性抑制作用。结果RAD组肌酐滤过率明显低于对照组(P<0.01)。RAD组Na 、葡萄糖、氨基酸的滤过率在哇巴因及根皮苷作用后明显低于对照组(P<0.01);去除药物抑制因素后,滤过率又能恢复,与对照组比较差异有显著性意义。结论体外构建的RAD具有对氨基酸、葡萄糖、钠离子、肌酐的选择性转运功能,且能在5h内稳定维持。

Construction of bioartificial renal tubule assist device and determination of transport to creatinine,amino acids and sodium in vitro

Objective To investigate the method of constructing bioartificial renal tubule assist device(RAD) and its transport function to creatinine,glucose,sodium and asparagine,theonine,serine,glutamic acid in vitro.Methods The cultured LLC PK1 cells were seeded into the inner space of the hemofilter(FH66)and the hemofilter was incubated for two weeks.Then the filtration in vitro was simulated to detect the function of the RAD.The transport rates of sodium,creatinine,glucose and four kinds of amino acids were measured.Meanwhile,the effects of oubaine and phlorizin on the transport rates of sodium,glucose and amino acids were determined and compared with no LLC PK1 device.Results The creatinine transport rate of RAD was significantly decreased as compared with control group(P< 0 01).The transport of sodium,glucose and amino acids could be significantly inhibited by oubaine and phlorizin compared to control group(P< 0 01),then it could be recovered after oubaine and phlorizin were drived out.Conclusion The RAD had selective transport function of sodium,creatinine,glucose and amino acids.The function could maintain well during 5 hours.