ABSTRACT

Puromycin treatment can cause glomerular injury to the kidney, leading to proteinuria. However, the pathogenesis of acute kidney injury and subsequent regeneration after puromycin administration in animal models remain unclear. In this work, we examined the characteristics of kidney injury and subsequent regeneration following puromycin treatment in adult zebrafish. We intraperitoneally injected 100 μg of puromycin into zebrafish; sacrificed them at 1, 3, 5, 7, or 14 days post-injection (dpi); and examined the morphological, functional, and molecular changes in the kidney. Puromycin-treated zebrafish presented more rapid clearance of rhodamine dextran than control animals. Morphological changes were observed immediately after the puromycin injection (1–7 dpi) and had recovered by 14 dpi. The mRNA production of lhx1a, a renal progenitor marker, increased during recovery from kidney injury. Levels of NFκB, TNFα, Nampt, and p-ERK increased significantly during nephron injury and regeneration, and Sirt1, FOXO1, pax2, and wt1b showed an increasing tendency. However, TGF-β1 and smad5 production did not show any changes after puromycin treatment. This study provides evidence that puromycin-induced injury in adult zebrafish kidneys is a potential tool for evaluating the mechanism of nephron injury and subsequent regeneration.

摘要

嘌呤霉素治疗可引起肾小球损伤，导致蛋白尿。然而，动物模型中给予嘌呤霉素后急性肾损伤的发病机制和随后的再生仍不清楚。在这项工作中，我们检查了成年斑马鱼嘌呤霉素治疗后肾损伤和随后再生的特征。我们给斑马鱼腹腔注射了 100 μg 嘌呤霉素；在注射后 1、3、5、7 或 14 天 (dpi) 处死它们；并检查了肾脏的形态、功能和分子变化。嘌呤霉素处理的斑马鱼比对照动物表现出更快的罗丹明葡聚糖清除率。嘌呤霉素注射后（1-7 dpi）立即观察到形态学变化，并在 14 dpi 时恢复。lhx1a的mRNA产生，一种肾祖细胞标记物，在肾损伤恢复期间增加。肾单位损伤和再生过程中NFκB、TNFα、Nampt和p-ERK水平显着升高，Sirt1、FOXO1、pax2和wt1b呈升高趋势。然而，TGF-β1 和 smad5 的产生在嘌呤霉素处理后没有显示任何变化。这项研究提供的证据表明，嘌呤霉素对成年斑马鱼肾脏的损伤是评估肾单位损伤机制和随后再生的潜在工具。