Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, North Carolina 27157, USA.
Stem Cells Transl Med. 2012 May;1(5):373-83. doi: 10.5966/sctm.2011-0048. Epub 2012 May 3.
New therapeutic strategies for chronic kidney disease (CKD) are necessary to offset the rising incidence of CKD and donor shortage. Erythropoietin (EPO), a cytokine produced by fibroblast-like cells in the kidney, has recently emerged as a renoprotective factor with anti-inflammatory, antioxidant properties. This study (a) determined whether human renal cultures (human primary kidney cells [hPKC]) can be enriched in EPO-positive cells (hPKC(F+)) by using magnetic-bead sorting; (b) characterized hPKC(F+) following cell separation; and (c) established that intrarenal delivery of enriched hPKC(F+) cells would be more beneficial in treatment of renal injury, inflammation, and oxidative stress than unsorted hPKC cultures in a chronic kidney injury model. Fluorescence-activated cell sorting analysis revealed higher expression of EPO (36%) and CD73 (27%) in hPKC(F+) as compared with hPKC. After induction of renal injury, intrarenal delivery of hPKC(F+) or hPKC significantly reduced serum creatinine, interstitial fibrosis in the medulla, and abundance of CD68-positive cells in the cortex and medulla (p < .05). However, only hPKC(F+) attenuated interstitial fibrosis in the renal cortex and decreased urinary albumin (3.5-fold) and urinary tubular injury marker kidney injury molecule 1 (16-fold). hPKC(F+) also significantly reduced levels of renal cortical monocyte chemotactic protein 1 (1.8-fold) and oxidative DNA marker 8-hydroxy-deoxyguanosine (8-OHdG) (2.4-fold). After 12 weeks, we detected few injected cells, which were localized mostly to the cortical interstitium. Although cell therapy with either hPKC(F+) or hPKC improved renal function, the hPKC(F+) subpopulation provides greater renoprotection, perhaps through attenuation of inflammation and oxidative stress. We conclude that hPKC(F+) may be used as components of cell-based therapies for degenerative kidney diseases.
治疗慢性肾脏病（CKD）的新策略对于抵消 CKD 的发病率上升和供体短缺是必要的。促红细胞生成素（EPO）是一种由肾脏中的成纤维样细胞产生的细胞因子，最近已成为一种具有抗炎、抗氧化特性的肾保护因子。本研究：（a）通过磁珠分选确定人肾培养物（人原代肾细胞 [hPKC]）是否可以富集 EPO 阳性细胞（hPKC(F+)）；（b）在细胞分离后对 hPKC(F+)进行特征描述；（c）建立在慢性肾损伤模型中，与未分选的 hPKC 培养物相比，富含 hPKC(F+)细胞的肾内给药在治疗肾损伤、炎症和氧化应激方面将更有益。荧光激活细胞分选分析显示 hPKC(F+)中 EPO（36%）和 CD73（27%）的表达高于 hPKC。在诱导肾损伤后，hPKC(F+)或 hPKC 的肾内给药均显著降低血清肌酐、髓质间质纤维化以及皮质和髓质中 CD68 阳性细胞的丰度（p <.05）。然而，只有 hPKC(F+)可减轻肾皮质间质纤维化，并降低尿白蛋白（3.5 倍）和尿肾小管损伤标志物肾损伤分子 1（16 倍）。hPKC(F+)还显著降低肾皮质单核细胞趋化蛋白 1（1.8 倍）和氧化 DNA 标志物 8-羟基脱氧鸟苷（8-OHdG）（2.4 倍）的水平。12 周后，我们检测到很少的注射细胞，这些细胞主要定位于皮质间质。虽然 hPKC(F+)或 hPKC 的细胞治疗均改善了肾功能，但 hPKC(F+)亚群提供了更大的肾脏保护作用，这可能是通过减轻炎症和氧化应激。我们得出结论，hPKC(F+)可作为基于细胞的退行性肾脏疾病治疗的组成部分。
