Inhibition of peptidylarginine deiminase attenuates inflammation and improves survival in a rat model of hemorrhagic shock

Abstract

Background We have recently shown that inhibition of peptidylarginine deiminase (PAD) improves survival in a rodent model of lethal cecal ligation and puncture (CLP). The roles of PAD inhibitors in hemorrhagic shock (HS), however, are largely unknown. The goal of this study was to investigate the effects of YW3-56, a novel PAD inhibitor, on survival following severe HS. Methods Mouse macrophages were exposed to hypoxic conditions followed by reoxygenation in the presence or absence of YW3-56. Enzyme-linked immunosorbent assay (ELISA) was performed to measure levels of secreted tumor necrosis factor α (TNF-α) and interleukin (IL)-6 in the culture medium. Cell viability was determined by methyl thiazolyl tetrazolium assay. In the survival experiment, anesthetized male Wistar-Kyoto rats (n=10/group) were subjected to 55% blood loss, treated with or without YW3-56 (10 mg/kg, intraperitoneally). Survival was monitored for 12 h. In the non-survival experiment, morphorlogic changes of the lungs were examined. Levels of circulating cytokine-induced neutrophil chemoattractant 1 (CINC-1) and myeloperoxidase (MPO) in the lungs were measured by ELISA. Expression of lung intercellular adhesion molecules-1 (ICAM-1) was also determined by Western blotting. Results Hypoxia/reoxygenation (H/R) insult induced TNF-α and IL-6 secretion from macrophages, which was significantly attenuated by YW3-56 treatment. YW3-56 treatment also increased cell viability when macrophages were exposed to H/R up to 6/15h, and improved survival rate from 20% to 60% in lethal HS rat model. Compared to the sham groups, pulmonary MPO activity and ICAM-1 expression in the HS group were significantly increased, and acute lung injury was associated with a higher degree of CINC-1 levels in serum. Intraperitoneal delivery of YW3-56 significantly reduced pulmonary MPO and ICAM-1 expression and attenuated acute lung injury (ALI). Conclusions Our results demonstrate for the first time that administration of YW3-56, a novel PAD inhibitor, can improve survival in a rat model of hemorrhagic shock as well as in a cell culture model of H/R. The survival advantage is associated with an attenuation of local and systemic pro-inflammatory cytokines and the protection against ALI following hemorrhage. Thus, PAD inhibition may represent a novel and promising therapeutic strategy for severe HS.

Comparative analysis of isoform-specific and non-selective histone deacetylase inhibitors in attenuating the intestinal damage after hemorrhagic shock

“…During HS, there is inadequate perfusion of the bowel which leads to epithelial damage; cellular injury during hypoperfusion and subsequent reperfusion can release proinflammatory cytokines in the circulation and trigger multiorgan failure 25. After HS, we observed that rats in the vehicle group had significantly increased levels of proinflammatory cytokines IL-1β and TNF-α, and CINC-1 in the intestine and serum, which is consistent with previous studies 26–28…”

Role of Peptidylarginine Deiminase and Neutrophil Extracellular Traps in Injuries: Future Novel Diagnostics and Therapeutic Targets

“…The survival outcome in the study by Biron et al (53) is similar to the other sepsis models (47) and the study by He et al (51). However, both He et al (51) and Pan et al (47) used a pan-PAD inhibitor instead of KO animals, which notably also inhibits PAD2. Given the conflicting findings, the pathophysiology of the two-hit hemorrhage and sepsis model and PADs remains to be further clarified.…”

Roles of PADI4 in the expression of cytokines involved in inflammation and adhesion in differentiated NB4 cells treated with ATRA

“…In a previous study, PADI4 activation was associated with the exacerbation of kidney ischemiareperfusion injury by enhancing renal tubular inflammatory responses and neutrophil infiltration (27). In addition, PADI4 silencing in a rat model of hemorrhagic shock can attenuate local inflammatory responses (28). Other studies indicate that PADI4 can be involved in the prevention of bacterial infection by promoting the formation of neutrophil extracellular traps (29,30).…”