Dual blockade of protease-activated receptor 1 and 2 additively ameliorates diabetic kidney disease

Abstract

Protease-activated receptors (PARs) are coagulation protease targets, and they increase expression of inflammatory cytokines and chemokines in various diseases. Of all PARs, previous reports have shown that PAR1 or PAR2 inhibition is protective against diabetic glomerular injury. However, how PAR1 and PAR2 cooperatively contribute to diabetic kidney disease (DKD) pathogenesis and whether dual blockade of PARs is more effective in DKD remain elusive. To address this issue, male type I diabetic Akita mice heterozygous for endothelial nitric oxide synthase were used as a model of DKD. Mice (4 mo old) were divided into four treatment groups and administered vehicle, PAR1 antagonist (E5555, 60 mg·kg−1·day−1), PAR2 antagonist (FSLLRY, 3 mg·kg−1·day−1), or E5555 + FSLLRY for 4 wk. The results showed that the urinary albumin creatinine ratio was significantly reduced when both PAR1 and PAR2 were blocked with E5555 + FSLLRY compared with the vehicle-treated group. Dual blockade of PAR1 and PAR2 by E5555 + FSLLRY additively ameliorated histological injury, including mesangial expansion, glomerular macrophage infiltration, and collagen type IV deposition. Marked reduction of inflammation- and fibrosis-related gene expression in the kidney was also observed. In vitro, PAR1 and PAR2 agonists additively increased mRNA expression of macrophage chemoattractant protein 1 or plasminogen activator inhibitor-1 in human endothelial cells. Changes induced by the PAR1 agonist were blocked by a NF-κB inhibitor, whereas those of the PAR2 agonist were blocked by MAPK and/or NF-κB inhibitors. These findings suggest that PAR1 and PAR2 additively contribute to DKD pathogenesis and that dual blockade of both could be a novel therapeutic option for treatment of patients with DKD.

Lack of miRNA-17 family mediates high glucose-induced PAR-1 up-regulation in glomerular mesangial cells

“…Moreover, our recent study demonstrated that a natural product sarsasapogenin alleviated DN in rats by downregulating PAR-1 (Tang et al, 2020). Additionally, dual blockade of PAR-1 and PAR-2 additively ameliorates DN in male type 1 diabetic Akita mice (Mitsui et al, 2020). Taken together, PAR-1 plays an important role in the development of DN.…”

Nicotinamide Attenuates the Progression of Renal Failure in a Mouse Model of Adenine-Induced Chronic Kidney Disease

“…Immunohistochemistry for F4/80 were performed as previously described [ 44 ]. We used rat-anti-mouse F4/80 monoclonal antibody (1:200, Clone A3-1, Bio-Rad, Hercules, CA, USA) as the primary antibody.…”

Coagulation, Protease-Activated Receptors, and Diabetic Kidney Disease: Lessons from eNOS-Deficient Mice

“…We have demonstrated that PAR1 and PAR2 cooperatively contribute to DKD pathogenesis (Mitsui et al 2020). In this study, male type I diabetic Akita mice heterozygous for eNOS (Ins2 Akita/+ ; eNOS +/-) were used as a model of DKD.…”

“…We focused on the pro-inflammatory effects of PAR1 and PAR2 agonists on human endothelial cells (Mitsui et al 2020). The results showed that stimulation with both PAR1 and PAR2 agonists synergistically increased the expression levels of MCP1 and PAI1 mRNA.…”