Hirudin Ameliorates Renal Interstitial Fibrosis via Regulating TGF‐β1/Smad and NF‐κB Signaling in UUO Rat Model

Abstract

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.

Autoantibodies from Patients with Scleroderma Renal Crisis Promote PAR-1 Receptor Activation and IL-6 Production in Endothelial Cells

“…Other in vitro studies showed that the natural PAR-1 ligand, thrombin, provoked a time- and concentration-dependent IL-6 release in endothelial cells [ 58 ]. The application of hirudin, a natural thrombin inhibitor, resulted in a reduced expression of both PAR-1 and IL-6, as well as decreased accumulation of ECM in the renal interstitium [ 59 ].…”

Pharmacological Activities and Mechanisms of Hirudin and Its Derivatives - A Review

“…Therefore, blocking TGF-β 1 signaling pathway may be an effective strategy for fibrosis, such as RIF. Recently, Yang et al confirmed that hirudin (10, 20, 40 IU/kg) could alleviate the renal damage and the deposition of ECM in renal interstitium of unilateral ureteral obstruction (UUO) rats (Yang et al, 2020). The expressions of TGF-β 1 , phosphorylated-Smad2, and phosphorylated-Smad3 in the kidneys of UUO rats were decreased, which illustrated the inhibition of TGF-β 1 /Smad signaling.…”

Transcriptome-Based Network Analysis Reveals Hirudin Potentiates Anti-Renal Fibrosis Efficacy in UUO Rats

“…The corresponding drug was given to each group of rats on the same day after the operation. The dose of the drug administered was determined according to the animal/human surface area method of conversion and combined with the results of the previous experiments ( Yang K. et al, 2020 ). UUO hirudin treatment group with a dose of 40 IU/kg/d by tail vein injection, and the UUO group as well as Sham group with stroke-physiological saline solution (SPSS) by tail vein injection too.…”