Abstract
Objective To investigate the effect of erythropoietin (EPO) on the expression of aquaporin-2 (AQP2) in the kidney of young SD rats after release of bilateral ureter obstruction (BUO-R). Methods Thirty-two young SD rats were equally divided into 4 groups randomly (BUO group, BUO-R group, BUO-R+ EPO group and Sham group, 8 rats in each group). The BUO model was built through bilateral ureteral ligation.EPO (500 U/kg) was given to BUO-R+ EPO rats at 2 h after release of BUO, and then repeated 6 h, 12 h, 24 h and 36 h thereafter and the same volume of 9 g/L saline was simultaneously given to BUO-R rats.The Sham group was prepared in parallel by laparotomy and free dissection of bilateral ureters but not ligated.Both side kidneys were harvested 48 h (72 h for Sham group) after release of BUO to examine the effect of EPO on the expression of AQP2 in inner medulla by immunohistochemistry, Real-time PCR and Western blot.The urine samples were collected by using metabolic cage before death. Results The osmotic pressure of BUO-R+ EPO group was higher than that of BUO-R group, but lower than that of Sham group (P<0.05). Immunohistochemistry showed that the collecting duct wall thinned and lumen enlarged.After the pictures were analysized by using Image-Pro Plus software, it showed that the expression of AQP2 in collecting duct in BUO group was significantly down-regulated compared with that in Sham group, whereas, it was slightly weaker in BUO-R group and BUO-R+ EPO group than Sham group(P<0.05). These results were further confirmed by adopting Western blot, and the relative quantity of AQP2 in BUO group was also the lowest of the four groups (P<0.05). Real-time PCR showed that the level of AQP2 mRNA in Sham group was (24.30±1.03) folds of BUO group, (10.60±1.05) folds of BUO-R group and (5.70±1.01) folds of BUO-R+ EPO group, respectively. Conclusion EPO could promote not only the recovery of AQP2 mRNA and protein expression but also the recovery of AQP2 function in young BUO-R rats. Key words: Erythropoietin; Ureter obstruction; Aquaporin-2; Kidney
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