Department of Microbiology and Immunology, Western University, London, Ontario, Canada; Matthew Mailing Center for Translational Transplant Studies, London Health Sciences Center, London, Ontario, Canada.
Matthew Mailing Center for Translational Transplant Studies, London Health Sciences Center, London, Ontario, Canada.
Nitric Oxide. 2018 Jun 1;76:16-28. doi: 10.1016/j.niox.2018.03.002. Epub 2018 Mar 6.
To assess the effects of slow-releasing HS donor GYY4137 on post-obstructive renal function and injury following unilateral ureteral obstruction (UUO) by using the UUO and reimplantation (UUO-R) model in rats and to elucidate potential mechanisms by using an in vitro model of epithelial-mesenchymal transition (EMT).
Male Lewis rats underwent UUO at the left ureterovesical junction. From post-operative day (POD) 1-13, rats received daily intraperitoneal (IP) injection of phosphate buffered saline (PBS, 1 mL) or GYY4137 (200 μmol/kg/day in 1 mL PBS, IP). On POD 14, the ureter was reimplanted back into the bladder, followed by a right nephrectomy. Urine and serum samples were collected to monitor renal function. On POD 30, the left kidney was removed and tissue sections were stained with H&E, TUNEL, CD68, CD206, myeloperoxidase, and Masson's trichrome to determine cortical thickness, apoptosis, inflammation, and fibrosis. In our in vitro model of EMT, NRK52E cells were treated with 10 ng/mL TGF-β1, 10 μM GYY4137 and/or 50 μM GYY4137. Western blot analysis was performed to determine the expression of E-cadherin, vimentin, Smad7 and TGF-β1 receptor II (TβRII).
GYY4137 led to a moderate decrease in post-obstructive serum creatinine, cystatin C and FENa. We also observed a trend towards a decrease in post-obstructive proteinuria following GYY4137 treatment. Histologically, we observed a significant decrease in apoptosis, inflammation, and fibrosis. Furthermore, our in vitro studies demonstrate that in the presence of TGF-β1, GYY4137 significantly decreases vimentin and TβRII and significantly increases E-cadherin and Smad7.
HS may help to accelerate the recovery of renal function post-obstruction and attenuates renal injury associated with UUO. It is possible that HS mitigates fibrosis by regulating the TGF-β1-mediated EMT pathway. Taken together, our data suggest that HS may be a potential novel therapy for improving renal function and limiting renal injury associated with obstructive uropathy.
通过单侧输尿管梗阻（UUO）和再植入（UUO-R）大鼠模型，评估缓激肽供体 GYY4137 对梗阻后肾功能和损伤的影响，并通过上皮-间充质转化（EMT）的体外模型阐明潜在机制。
雄性 Lewis 大鼠在左侧输尿管膀胱交界处行 UUO。术后第 1-13 天，大鼠每天接受腹腔内（IP）注射磷酸盐缓冲盐水（PBS，1 mL）或 GYY4137（200 μmol/kg/天，1 mL PBS，IP）。术后第 14 天，输尿管重新植入膀胱，随后进行右肾切除术。收集尿液和血清样本以监测肾功能。术后第 30 天，切除左侧肾脏，对组织切片进行 H&E、TUNEL、CD68、CD206、髓过氧化物酶和 Masson 三色染色，以确定皮质厚度、细胞凋亡、炎症和纤维化。在我们的 EMT 体外模型中，NRK52E 细胞用 10 ng/mL TGF-β1、10 μM GYY4137 和/或 50 μM GYY4137 处理。进行 Western blot 分析以确定 E-钙粘蛋白、波形蛋白、Smad7 和 TGF-β1 受体 II（TβRII）的表达。
GYY4137 导致梗阻后血清肌酐、胱抑素 C 和 FENa 中度降低。我们还观察到 GYY4137 治疗后梗阻后蛋白尿有减少趋势。组织学上，细胞凋亡、炎症和纤维化明显减少。此外，我们的体外研究表明，在 TGF-β1 存在的情况下，GYY4137 显著降低波形蛋白和 TβRII，并显著增加 E-钙粘蛋白和 Smad7。
HS 可能有助于加速梗阻后肾功能恢复，并减轻 UUO 相关的肾损伤。HS 通过调节 TGF-β1 介导的 EMT 途径减轻纤维化的可能性。总之，我们的数据表明，HS 可能是一种改善梗阻性尿路病相关肾功能和限制肾损伤的潜在新型治疗方法。
