Department of Nephrology, The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine, Nanning, Guangxi 530023, China.
College of Graduate School, Guangxi University of Traditional Chinese Medicine, Nanning, Guangxi 530001, China.
Int Immunopharmacol. 2020 Apr;81:106249. doi: 10.1016/j.intimp.2020.106249. Epub 2020 Feb 14.
Renal interstitial fibrosis (RIF) often occurs in many chronic kidney diseases (CKD). Hirudin now is applied to treat fibrosis in some organs. In this study, we verified the treatment effects of hirudin on RIF in vivo and in vitro with the underlying mechanism. The RIF in vivo was the unilateral ureteral obstruction (UUO) model and RIF in vitro was the renal tubular epithelial cells induced by TGF-β. The renal pathological changes and renal fibrosis were observed by hematoxylin and eosin (H&E) staining and Masson staining. The α-SMA in renal tissues was detected by immunohistochemistry. The inflammatory factors were analyzed by the ELISA assay. The cell apoptosis was observed by TUNEL assay. The related proteins of fibrosis, epithelial-mesenchymal transition (EMT) and apoptosis were assessed by western blot analysis. The experimental data demonstrated that hirudin decreased fibrosis, EMT, inflammation and cell apoptosis in renal tissues of UUO rats and TGF-β-induced renal tubular epithelial cells. Furthermore, hirudin also reduced the expression of collgen-I, FN, α-SMA, N-cad, slug, E-cad, IL-1β, IL-6 and TNF-α in mice serum and TGF-β-induced renal tubular epithelial cells. The apoptosis related proteins (pro-caspase3, pro-caspase9, bcl2 and bax) expression was also down-regulated in renal tissues of UUO rats. In conclusion, hirudin depressed the fibrosis in renal tissues and renal tubular epithelial cells by inhibiting the inflammation, regulating the related proteins of fibrosis and ETM and decreasing the apoptosis of renal tubular epithelial cells. These findings may offer an effective treatment method for RIF.
肾间质纤维化（RIF）常发生于多种慢性肾脏病（CKD）中。目前，水蛭素被用于治疗某些器官的纤维化。本研究通过体内和体外实验验证了水蛭素对 RIF 的治疗作用及其潜在机制。体内 RIF 模型为单侧输尿管梗阻（UUO）模型，体外 RIF 模型为 TGF-β诱导的肾小管上皮细胞。通过苏木精-伊红（H&E）染色和 Masson 染色观察肾脏病理变化和肾纤维化，免疫组化检测肾组织中α-SMA 的表达，ELISA 检测炎症因子，TUNEL 检测细胞凋亡，Western blot 分析纤维化、上皮间质转化（EMT）和凋亡相关蛋白。实验数据表明，水蛭素可减轻 UUO 大鼠肾脏组织和 TGF-β诱导的肾小管上皮细胞中的纤维化、EMT、炎症和细胞凋亡。此外，水蛭素还降低了 UUO 大鼠血清和 TGF-β诱导的肾小管上皮细胞中胶原-I、FN、α-SMA、N-cad、slug、E-cad、IL-1β、IL-6 和 TNF-α的表达。UUO 大鼠肾脏组织中凋亡相关蛋白（pro-caspase3、pro-caspase9、bcl2 和 bax）的表达也下调。综上所述，水蛭素通过抑制炎症、调节纤维化和 EMT 相关蛋白以及减少肾小管上皮细胞凋亡，抑制肾脏组织和肾小管上皮细胞的纤维化。这些发现可能为 RIF 提供一种有效的治疗方法。
