Remediation of Hemorrhagic Shock-Induced Intestinal Barrier Dysfunction by Treatment with Diphenyldihaloketones EF24 and CLEFMA

Abstract

Gut is very sensitive to hypoperfusion and hypoxia, and deranged gastrointestinal barrier is implicated in systemic failure of various organs. We recently demonstrated that diphenyldihaloketone EF24 [3,5-bis(2-fluorobenzylidene)piperidin-4-one] improves survival in a rat model of hemorrhagic shock. In this study, we tested EF24 and its other analog CLEFMA (4-[3,5-bis(2-chlorobenzylidene)-4-oxo-piperidine-1-yl]-4-oxo-2-butenoic acid) for their effect on intestinal barrier dysfunction in hypovolemic shock. Hypovolemia was induced in rats by withdrawing 50% of blood. EF24 or CLEFMA (0.4 mg/kg i.p.) treatment was provided, without volume resuscitation, after 1 hour of hemorrhage. Ileum was collected 5 hours after the treatment to investigate the expression of tight junction proteins (zonula occludens, claudin, and occludin) and epithelial injury markers [myeloperoxidase, ileal lipid-binding protein (ILBP), CD163, and plasma citrulline]. The ileal permeability for dextran-fluoroisothiocyanate and Evan's blue dye was determined. EF24 and CLEFMA reduced the hypovolemiainduced plasma citrulline levels and the ileal expression of myeloperoxidase, ILBP, and CD163. The drugs also restored the basal expression levels of zonula occludens, claudin, and occludin, which were substantially deranged by hypovolemia. In ischemic ileum, the expression of phospho(tyrosine)-zonula occludens-1 was reduced, which was reinstated by EF24 and CLEFMA. In contrast, the drug treatments maintained the hypovolemia-induced expression of phospho(threonine)-occludin, but reduced that of phospho(tyrosine)-occludin. Both EF24 and CLEFMA treatments reduced the intestinal permeability enhanced by hypovolemia. EF24 and CLEFMA attenuate hypovolemic gut pathology and protect barrier function by restoring the status of tight junction proteins. These effects were observed in unresuscitated shock, implying the benefit of EF24 and CLEFMA in prehospital care of shock.

Hemorrhagic shock-induced cerebral bioenergetic imbalance is corrected by pharmacologic treatment with EF24 in a rat model

“…Gut barrier dysfunction is known to trigger SIRS and cerebral damage (Grenz et al 2012, Hsieh et al 2011, Moore et al 2004, Rhodes et al 1973, Zhou et al 2012). This hypothesis is also supported by our recent report where we showed that EF24 prevents gut barrier dysfunction and this effect was associated with rapid accumulation of EF24 in the intestinal tissue (Yadav et al 2014a). Interestingly, brain tissue accumulates negligible amounts of administered EF24 (Lagisetty et al 2012).…”

“…The leading cause of SIRS is the loss of gut barrier function because of ischemic injury to the gut epithelium (Moore et al 2004, Grenz et al 2012, Rhodes et al 1973) which has also been implicated in brain dysfunction (Hsieh et al 2011, Zhou et al 2012). In previous reports, we described that an anti-inflammatory molecule EF24 exerts a pro-survival effect in a rat model of 50% hemorrhagic shock (Yadav et al 2014a, Yadav et al 2013) by preventing gut injury. In this work, we investigated the effect of EF24 on brain energy crisis caused by hemorrhagic shock in the same model.…”

“…EF24 in small volume was as effective in restoring cerebral energy stores as was a combination of EF24 and LEH. Given our previous observations that the majority of administered EF24 localizes in intestinal tissue (Lagisetty et al 2012, Yadav et al 2014a) and negligible amounts accumulated in the brain, it appears that the cerebral metabolic derangement after blood loss is a combined consequence of hypovolemia-induced perfusion/oxygen deficit and inflammatory processes triggered by access of gut contents to systemic space, secondary to the dysfunction of intestinal barrier.…”

“…Recently, we have shown that treatment of severely hypovolemic rats with EF24, 3,5-bis(2-fluorobenzylidene)piperidin-4-one, protected gut barrier function and reduced systemic inflammatory response (Yadav et al 2014a). The primary goal of this study was to evaluate the status of brain energetics in response to a treatment with EF24 in shock model of 45% hemorrhage in rats.…”

Nanovesicular liposome-encapsulated hemoglobin (LEH) prevents multi-organ injuries in a rat model of hemorrhagic shock

“…Any untoward effect of a resuscitation fluid on the complement system has a potential to further complicate the pathology of shock, because the complement system also contributes to the SIRS (Szebeni et al, 2003). By virtue of their non-phospholipid nature, the HDAS-lipids reduce the complement activation induced by the liposome particles (Nag et al, 2013; Yadav et al, 2014). …”