Hirudin Regulates Vascular Function in Chronic Renal Failure through Modulating Macrophage Polarization

Abstract

Excessive inflammation is responsible for arteriovenous fistula (AVF) failure, which determines the therapeutic effect of chronic renal failure (CRF). Macrophage polarization is of great significance in the inflammatory response. Hirudin (Hiru) has been reported to possess a definite anti-inflammatory effect. This study is to uncover the impacts of Hiru on classically (M1)/alternatively (M2) macrophage polarization in the CRF rat model and rat vascular smooth muscle cells (VSMCs). After the CRF rat model was administrated with different concentrations of Hiru, blood urea nitrogen (BUN) and serum creatinine (Scr) levels were tested. H&E staining was to detect vascular injury, and IHC assay was to analyze inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1) expressions in vascular tissues. Levels of inflammatory factors were examined by ELISA. Besides, western blot was to estimate the levels of marker proteins related to macrophage, proliferation, and apoptosis. CCK-8 was to measure cell viability. We discovered that Hiru alleviated renal function injury and vascular injury, exacerbated VSMC hyperplasia, and stimulated the differentiation and activation of M1 macrophage towards M2 macrophage in vivo. Moreover, after treatment with lipopolysaccharide (LPS)/IFN-gamma (IFN-γ), the increased M1/M2 ratio and enhanced levels of inflammatory factors were observed. Furthermore, Hiru boosted the proliferation and ameliorated the inflammatory response and apoptosis of rat VSMCs during the process of coincubation of M1-conditioned medium (CM). Collectively, Hiru played a protective role against vascular injury in CRF directly or through its influence on M1 macrophage polarization and inflammation.

Hirudin in the Treatment of Chronic Kidney Disease

“…A study found that hirudin alleviated renal function and vascular injury in chronic renal failure rats induced by adenine, promoted SMC hyperplasia, and stimulated the differentiation of M1 macrophages towards M2 macrophages. In vitro, hirudin promoted the proliferation of rat VSMCs and improved the inflammatory response and apoptosis during co-incubation in an M1-conditioned medium [ 83 ].…”

“…Metastatic kidney cancer inhibiting F-actin formation and reducing metastatic growth [82] Arteriovenous fistula stenosis Promoting SMC hyperplasia; stimulating the differentiation of M1 macrophage towards M2 macrophage Regulates endothelial cell permeability and apoptosis, inhibiting RhoA/ROCK signaling [83,84] IgA nephrology Reducing cell apoptosis; inflammatory response and keeping balance of T cells [85] Using network pharmacology and molecular docking to reveal the mechanism of hirudin in the treatment of DN, the results indicated hirudin may regulate various signaling pathways including the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (AKT) signaling pathway and the vascular endothelial-derived growth factor (VEGF) signaling pathway. Further animal experimental validation showed hirudin decreased the expression of caspase-3 related to apoptosis, and inhibited the PI3K-AKT signaling pathway in the kidney of DN rats.…”