Hirudin Ameliorates Renal Interstitial Fibrosis via Regulating TGF-1/Smad and NF-B Signaling in UUO Rat Model.

Purpose: Hirudin, a polypeptide structure containing 65 amino acids, is a potent natural thrombin inhibitor with anticoagulant property extracted from . 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), -smooth muscle actin (-SMA), protease-activated receptor 1 (PAR-1), and proteins in the TGF-1/Smad and NF-B 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, -SMA, PAR-1, and inflammatory markers such as tumor necrosis factor- and interleukin-6 after hirudin treatment. Furthermore, hirudin reduced the expressions of transforming growth factor 1 (TGF-1), phosphorylated-Smad2, and phosphorylated-Smad3 in the UUO model. In parallel, we found inhibited nuclear factor-B (NF-B) signaling after hirudin treatment, with downregulated protein expressions of P65, phosphorylated-P65, and phosphorylated-iB and increased iB.

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-1/Smad and NF-B signaling.