Department of Pediatric Nephrology, Shengjing Hospital, China Medical University, Shenyang 110004, China.
Ren Fail. 2010 Jun;32(5):603-11. doi: 10.3109/08860221003778049.
To investigate the effect of paired box 2 (PAX2) gene re-expression in renal tubular epithelial cells on renal interstitial fibrosis in rats with obstructive nephropathy.
Wistar rats were randomly divided into two groups: sham-operated group (sham group) and unilateral ureteral obstruction group (UUO group), 40 in each group. After the surgery, rats were killed at 3, 5, 7, and 14 days (n = 10 each) to remove and collect kidneys. Morphological changes of the kidney were determined by histopathology. Protein and mRNA expression of PAX2 in the kidney were determined by immunohistochemistry, Western blot and real-time polymerase chain reaction (PCR).
The new major findings of this study include the following: (1) Hematoxylin and Eosin (HE) and Masson staining revealed obvious renal interstitial fibrosis in UUO group; (2) increased PAX2 expression was found in renal tubular epithelial cells in UUO group by immunohistochemistry, but not in sham group; (3) by Western blot, a significant increase in PAX2 protein level in UUO group was detected 3d after the surgery when compared to sham group (p < 0.05), which continued to increase with prolonged obstruction; the same pattern was also found in PAX2 mRNA expression by real-time PCR in UUO group after the surgery; (4) PAX2 protein level was positively correlated with the severity of renal tubular damage (r = 0.937 and p < 0.05) and the level of renal interstitial fibrosis (r = 0.987 and p < 0.05).
In rats with obstructive nephropathy, embryonic development gene PAX2 is re-expressed in the renal tubules, which may participate in the pathogenesis of renal tubular damage and renal interstitial fibrosis.
探讨配对盒基因 2(PAX2)在肾小管上皮细胞中的重新表达对梗阻性肾病大鼠肾间质纤维化的影响。
将 Wistar 大鼠随机分为两组:假手术组(假手术组)和单侧输尿管梗阻组(UUO 组),每组 40 只。手术后,每组各取 10 只大鼠分别于 3、5、7、14 天处死并取肾。通过组织病理学观察肾脏形态学变化。采用免疫组织化学、Western blot 和实时聚合酶链反应(PCR)检测肾脏中 PAX2 的蛋白和 mRNA 表达。
本研究的新主要发现包括:(1)苏木精-伊红(HE)和 Masson 染色显示 UUO 组出现明显的肾间质纤维化;(2)免疫组织化学显示 UUO 组肾小管上皮细胞中 PAX2 表达增加,但假手术组未见;(3)Western blot 检测显示,与假手术组相比,UUO 组术后 3d 时 PAX2 蛋白水平显著升高(p<0.05),随着梗阻时间的延长而持续增加;实时 PCR 也显示 UUO 组术后 PAX2 mRNA 表达呈相同模式;(4)PAX2 蛋白水平与肾小管损伤严重程度呈正相关(r=0.937,p<0.05),与肾间质纤维化程度呈正相关(r=0.987,p<0.05)。
在梗阻性肾病大鼠中,胚胎发育基因 PAX2 在肾小管中重新表达,可能参与肾小管损伤和肾间质纤维化的发病机制。