Hirudin Ameliorates Renal Interstitial Fibrosis via Regulating TGF-β1/Smad and NF-κB Signaling in UUO Rat Model
Hirudin Ameliorates Renal Interstitial Fibrosis via Regulating TGF-β1/Smad and NF-κB Signaling in UUO Rat Model
复制标题
水蛭素通过调节 UUO 大鼠模型中的 TGF-β1/Smad 和 NF-κB 信号传导改善肾间质纤维化
DOI：
10.1155/2020/7291075
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发表时间：
2020-06-27
影响因子：
--
通讯作者：
Yang, Hongtao
中科院分区：
文献类型：
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作者：
Yang, Kang;Fan, Boya;Yang, Hongtao
目的水蛭素是从药用水蛭中提取的一种具有抗凝血作用的天然凝血酶抑制剂，是一种由65个氨基酸组成的多肽结构。据报道，它具有抗炎和抗纤维化特性。本研究探讨水蛭素对单侧输尿管梗阻（UUO）所致肾间质纤维化（RIF）的保护作用。方法大鼠随机分为假手术组、单纯UUO组、UUO +水蛭素组（10、20、40 IU/kg/d，连续14 d）。在实验期结束时，处死动物。HE染色及Masson染色观察肾组织病理变化。采用免疫组化和/或Western blotting法检测肾组织中III型胶原（Col III）、纤维连接蛋白（FN）、α-平滑肌肌动蛋白（α-SMA）、蛋白酶激活受体1（PAR-1）以及TGF-β1/Smad和NF-κB通路蛋白的表达。结果HE和Masson染色显示水蛭素治疗组大鼠肾组织损伤程度和细胞外基质（ECM）沉积均较UUO组减轻。免疫组化和WB结果显示水蛭素治疗后Col III、FN、α-SMA、PAR-1以及肿瘤坏死因子-α和白细胞介素-6等炎症标志物的蛋白表达均降低。水蛭素还能降低UUO模型中转化生长因子β1（TGF-β1）、磷酸化Smad 2和磷酸化Smad 3的表达。同时，我们发现水蛭素处理后抑制核因子-κB（NF-κB）信号传导，下调P65、磷酸化P65和磷酸化i κBα的蛋白表达，并增加iκBα。结论水蛭素可改善UUO大鼠肾损伤，抑制炎症反应和细胞外基质的积聚，其机制可能与抑制TGF-β1/Smad和NF-κB信号通路有关。
Purpose Hirudin, a polypeptide structure containing 65 amino acids, is a potent natural thrombin inhibitor with anticoagulant property extracted from Hirudo medicinalis. It has been reported to have anti-inflammatory and antifibrotic property. Here we explored the renoprotective effect of hirudin on unilateral ureteral obstruction (UUO) induced renal interstitial fibrosis (RIF). Methods Rats were randomly divided into five groups: sham group, UUO alone group, and three UUO + hirudin-treatment groups (10, 20, or 40 IU/kg/d, for 14 continuous days). At the end of the experiment period, animals were sacrificed. Pathologic changes in renal specimens were observed using hematoxylin and eosin (HE) staining and Masson staining. The expressions of collagen III (Col III), fibronectin (FN), α-smooth muscle actin (α-SMA), protease-activated receptor 1 (PAR-1), and proteins in the TGF-β1/Smad and NF-κB pathways in renal tissues were examined by immunohistochemistry and/or Western blotting. Results HE and Masson staining showed that hirudin-treated UUO rats had lower extent of renal injury and deposition of extracellular matrix (ECM) in renal interstitium than those in the UUO group. The results of immunohistochemistry and WB indicated decreased protein expressions of Col III, FN, α-SMA, PAR-1, and inflammatory markers such as tumor necrosis factor-α and interleukin-6 after hirudin treatment. Furthermore, hirudin reduced the expressions of transforming growth factor β1 (TGF-β1), phosphorylated-Smad2, and phosphorylated-Smad3 in the UUO model. In parallel, we found inhibited nuclear factor-κB (NF-κB) signaling after hirudin treatment, with downregulated protein expressions of P65, phosphorylated-P65, and phosphorylated-iκBα and increased iκBα. Conclusion Hirudin improves kidney injury and suppresses inflammatory response and ECM accumulation in UUO rats; its underlying mechanism may be associated with the inhibition of TGF-β1/Smad and NF-κB signaling.