Kidney tubular epithelium is restored without replacement with bone marrow-derived cells during repair after ischemic injury
Kidney tubular epithelium is restored without replacement with bone marrow-derived cells during repair after ischemic injury
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DOI：
10.1111/j.1523-1755.2005.00629.x
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发表时间：
2005-11-01
影响因子：
19.6
通讯作者：
Bonventre, JV
中科院分区：
文献类型：
--
作者：
Duffield, JS;Bonventre, JV
肾脏具有恢复近端小管的结构和功能完整性的能力，近端小管在长时间暴露于缺血后经历广泛的上皮细胞死亡。为了研究成体骨髓来源的干细胞可能在损伤后的肾重塑中发挥的作用，我们采用了缺血/再灌注（I/R）损伤的鼠模型，其中已经表征了损伤、功能障碍、修复、肾小管细胞增殖和功能恢复的程度[Park KM等人，J Biol Chem 276：11870-11876，2001]。我们使用表达细菌LacZ基因或增强型绿色荧光蛋白（eGFP）基因的小鼠的骨髓，或从雄性小鼠移植到雌性小鼠的骨髓，产生嵌合小鼠。在每种情况下，在移植后6周确认嵌合体的建立。通过用微动脉瘤夹闭塞肾动脉和静脉30分钟在嵌合小鼠中诱导I/R损伤。在eGFP嵌合体中功能恢复后，虽然有许多间质细胞，但没有来自骨髓细胞的管状细胞。在细菌β-半乳糖苷酶（β-gal）嵌合小鼠中，我们通过5-bomo-4-chloro-3-indolyl-beta-D-galactopyranoside（X-gal）染色发现了哺乳动物（内源性）β-gal的证据，但在小管细胞中没有发现细菌β-gal。荧光原位杂交（FISH）检测Y染色体在缺血后的性别不匹配的嵌合体肾脏显示Y染色体阳性，仅在间质细胞的细胞核，当仔细检查时，通过去卷积显微镜。在我们的I/R损伤模型中，存活的受损肾小管细胞大量增殖，表明受损肾小管被存活肾小管细胞的子细胞重新填充。I/R损伤后间质细胞和血管细胞的表型分析显示，少量的管周内皮细胞来自骨髓细胞，可能在修复过程中发挥作用。
The kidney has the ability to restore the structural and functional integrity of the proximal tubule, which undergoes extensive epithelial cell death after prolonged exposure to ischemia. In order to study the role that adult bone marrow-derived stem cells might play in kidney remodeling after injury, we employed a murine model of ischemia/reperfusion (I/R) injury in which the degree of injury, dysfunction, repair, tubular cell proliferation and functional recovery have been characterized [Park KM, et al, J Biol Chem 276:11870-11876, 2001]. We generated chimeric mice using marrow from mice expressing the bacterial LacZ gene, or the enhanced green fluorescence protein (eGFP) gene, or from male mice transplanted into female mice. The establishment of chimerism was confirmed at 6 weeks following transplantation in each case. I/R injury was induced in chimeric mice by occluding the renal arteries and veins with microaneurysm clamps for 30 minutes. After functional recovery in the eGFP chimeras, although there were many interstitial cells, no tubular cells were derived from bone marrow cells. In the bacterial beta-galactosidase (beta-gal) chimeric mice we found evidence of mammalian (endogenous) beta-gal by 5-bomo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining, but not bacterial beta-gal in tubule cells. Detection of the Y chromosome by fluorescence in situ hybridization (FISH) in the postischemic kidneys of gender-mismatched chimeras revealed Y chromosome positivity only in the nuclei of interstitial cells, when scrutinized by deconvolution microscopy. In our model of I/R injury there was a large amount of proliferation of surviving, injured tubular cells indicating that the injured tubule is repopulated by daughter cells of surviving tubular cells. Analysis of the phenotype of interstitial and vascular cells following I/R injury revealed small numbers of peritubular endothelial cells to be derived from bone marrow cells that may serve in the repair process.