Joshi Sunil, Wang Wei, Khan Saeed R
Department of Pathology, Immunology & Laboratory Medicine, College of Medicine, University of Florida, Gainesville, Florida, United States of America.
Department of Urology, College of Medicine, University of Florida, Gainesville, Florida, United States of America.
PLoS One. 2017 Nov 1;12(11):e0185009. doi: 10.1371/journal.pone.0185009. eCollection 2017.
Hyperoxaluria associated with renal deposition of calcium oxalate (CaOx) crystals causes renal injury and inflammation leading to number of diseases including chronic kidney disease (CKD). It is however, not been possible to separate the renal consequences of hyperoxaluria from that of CaOx crystal deposition. We decided to utilize ethylene glycol (EG) model where hyperoxaluria and CaOx crystal deposition can be separated in time. To test our hypothesis, male rats were made hyperoxaluric by administering EG, rats were euthanized and kidneys were extracted on day 14, when occasional crystal is seen in the kidneys and day 28, when all animals have developed renal CaOx crystal deposits. Total RNA was extracted for microarray analysis and genome wide analysis of differentially expressed genes was performed to investigate differences between hyperoxaluria and crystal induced alterations in the kidneys. Immunohistochemical and Hematoxylin and Eosin (H&E) staining was also done for macromolecules with significant role in stone formation. All EG fed rats became hyperoxaluric by day 7, showed a few crystal deposits on day 14, and had heavy crystal deposition by day 28. There were significant changes in the expression of genes encoding for NADPH Oxidases; macromolecular crystallization modulators; genes involved in inflammasome activation; and osteogenic marker genes. Results demonstrate major differences between hyperoxaluria and CaOx crystal induced changes in the kidneys. Injury and inflammation are mainly associated with crystal deposition indicating significant role played by crystal retention.
与草酸钙（CaOx）晶体在肾脏沉积相关的高草酸尿症会导致肾损伤和炎症，进而引发包括慢性肾脏病（CKD）在内的多种疾病。然而，一直无法将高草酸尿症的肾脏后果与CaOx晶体沉积的后果区分开来。我们决定利用乙二醇（EG）模型，在该模型中高草酸尿症和CaOx晶体沉积可以在时间上分离。为了验证我们的假设，给雄性大鼠施用EG使其产生高草酸尿症，在第14天（此时在肾脏中偶尔可见晶体）和第28天（此时所有动物都已形成肾脏CaOx晶体沉积物）对大鼠实施安乐死并取出肾脏。提取总RNA用于微阵列分析，并对差异表达基因进行全基因组分析，以研究高草酸尿症和晶体诱导的肾脏变化之间的差异。还对在结石形成中起重要作用的大分子进行了免疫组织化学和苏木精-伊红（H&E）染色。所有喂食EG的大鼠在第7天时都出现了高草酸尿症，在第14天时显示有少量晶体沉积物，到第28天时则有大量晶体沉积。编码NADPH氧化酶、大分子结晶调节剂、参与炎性小体激活的基因以及成骨标记基因的表达有显著变化。结果表明高草酸尿症和CaOx晶体诱导的肾脏变化之间存在主要差异。损伤和炎症主要与晶体沉积有关，表明晶体滞留起着重要作用。
