Nephrology, Dialysis and Kidney Transplantation Unit, Maggiore della Carità Hospital-University of Eastern Piedmont, Novara, Italy.
Department of Medical Sciences and Center for Experimental Medical Research (CeRMS), Nephrology, Dialysis and Kidney Transplantation Center, University of Torino, Torino, Italy.
Nephrol Dial Transplant. 2018 Jul 1;33(7):1110-1121. doi: 10.1093/ndt/gfx328.
The renal assist device (RAD) is a blood purification system containing viable renal tubular epithelial cells (TECs) that has been proposed for the treatment of acute kidney injury (AKI) and multiple organ failure. Perfluorocarbons (PFCs) are oxygen carriers used for organ preservation in transplantation. The aim of this study was to investigate the effect of PFCs on hypoxia- and sepsis-induced TEC injury and on renal CD133+ progenitor differentiation in a microenvironment similar to the RAD.
TECs were seeded in a polysulphone hollow fibre under hypoxia or cultured with plasma from 10 patients with sepsis-associated AKI in the presence or absence of PFCs and were tested for cytotoxicity (XTT assay), apoptosis (terminal deoxynucleotidyl transferase dUTP nick end labeling assay, caspases, enzyme-linked immunosorbent assay, Fas/Fas Ligand pathway activation), mitochondrial activity, cell polarity [transepithelial electrical resistance (TEER)] and adenosine triphosphate production. The effect of PFCs on proliferation and differentiation of human CD133+ progenitors was also studied.
In the presence of PFCs, TECs seeded into the polysulphone hollow fibre showed increased viability and expression of insulin-like growth factor 1, hepatocyte growth factor and macrophage-stimulating protein. Plasma from septic patients induced TEC apoptosis, disruption of oxidative metabolism, alteration of cell polarity and albumin uptake, down-regulation of the tight junction protein ZO-1 and the endocytic receptor megalin on the TEC surface. These detrimental effects were significantly reduced by PFCs. Moreover, PFCs induced CD133+ renal progenitor cell proliferation and differentiation towards an epithelial/tubular-like phenotype.
PFCs improved the viability and metabolic function of TECs seeded within a polysulphone hollow fibre and subjected to plasma from septic AKI patients. Additionally, PFCs promoted differentiation towards a tubular/epithelial phenotype of CD133+ renal progenitor cells.
肾辅助装置（RAD）是一种含有有活力的肾小管上皮细胞（TEC）的血液净化系统，被提议用于治疗急性肾损伤（AKI）和多器官衰竭。全氟化碳（PFC）是用于移植器官保存的氧载体。本研究的目的是在类似于 RAD 的微环境中，研究 PFC 对缺氧和脓毒症诱导的 TEC 损伤以及肾 CD133+祖细胞分化的影响。
将 TEC 接种在聚砜中空纤维下，在缺氧或存在或不存在 PFC 的情况下，用来自 10 名脓毒症相关 AKI 患者的血浆培养，并通过 XTT 测定法、末端脱氧核苷酸转移酶 dUTP 缺口末端标记法、半胱天冬酶、酶联免疫吸附试验、Fas/Fas 配体途径激活来检测细胞毒性（XTT 测定法）、细胞凋亡、线粒体活性、细胞极性[跨上皮电阻（TEER）]和三磷酸腺苷（ATP）产生。还研究了 PFC 对人 CD133+祖细胞增殖和分化的影响。
在 PFC 的存在下，接种到聚砜中空纤维中的 TEC 表现出更高的活力和胰岛素样生长因子 1、肝细胞生长因子和巨噬细胞刺激蛋白的表达。脓毒症患者的血浆诱导 TEC 凋亡、氧化代谢破坏、细胞极性和白蛋白摄取改变、紧密连接蛋白 ZO-1 和 TEC 表面内吞受体 megalin 的下调。这些有害影响被 PFC 显著降低。此外，PFC 诱导 CD133+肾祖细胞增殖并向上皮/管状样表型分化。
PFC 提高了接种在聚砜中空纤维内并暴露于脓毒症 AKI 患者血浆中的 TEC 的活力和代谢功能。此外，PFC 促进 CD133+肾祖细胞向管状/上皮样表型分化。
