ROCK1 inhibitor stabilizes E-cadherin and improves barrier function in experimental necrotizing enterocolitis

Abstract

Necrotizing enterocolitis (NEC) is a devastating gastrointestinal disease of newborns. Although incompletely understood, NEC is associated with intestinal barrier dysfunction. E-cadherin, an adherens junction, is a protein complex integral in maintaining normal barrier homeostasis. Rho-associated protein kinase-1 (ROCK1) is a kinase that regulates the E-cadherin complex, and p120-catenin is a subunit of the E-cadherin complex that has been implicated in stabilizing the cadherin complex at the plasma membrane. We hypothesized that E-cadherin is decreased in NEC and that inhibition of ROCK1 would protect against adherens junction disruption. To investigate this, a multimodal approach was used: In vitro Caco-2 model of NEC (LPS/TNFα), rap pup model (hypoxia + bacteria-containing formula), and human intestinal samples. E-cadherin was decreased in NEC compared with controls, with relocalization from the cell border to an intracellular location. ROCK1 exhibited a time-dependent response to disease, with increased early expression in NEC and decreased expression at later time points and disease severity. Administration of ROCK1 inhibitor (RI) resulted in preservation of E-cadherin expression at the cell border, preservation of intestinal villi on histological examination, and decreased apoptosis. ROCK1 upregulation in NEC led to decreased association of E-cadherin to p120 and increased intestinal permeability. RI helped maintain the stability of the E-cadherin-p120 complex, leading to improved barrier integrity and protection from experimental NEC. NEW & NOTEWORTHY This paper is the first to describe the effect of ROCK1 on E-cadherin expression in the intestinal epithelium and the protective effects of ROCK inhibitor on E-cadherin stability in necrotizing enterocolitis.

Low doses of imidacloprid induce disruption of intercellular adhesion and initiate proinflammatory changes in Caco-2 cells

“…Therefore, the abnormalities in E-cadherin adhesion can initiate the intercellular communication disturbance and disrupt epithelial barrier. Furthermore, the role of E-cadherin expression in the maintaining of intestinal epithelium integrity was demonstrated in the model of necrotizing enterocolitis while the disturbance in barrier function was accompanied by disruption of tight junctions (Buonpane et al, 2020).…”

Rho-Proteins and Downstream Pathways as Potential Targets in Sepsis and Septic Shock: What Have We Learned from Basic Research

“…Although scarcely investigated, septic shock and experimental models of sepsis can result in a dysfunctional gastrointestinal system [159,160]. The regulation of the epithelial barrier and calcium sensitization by Rho proteins have been involved in both physiological and inflammation-associated disorders in the intestinal system [161][162][163][164][165][166][167]. The enhanced activity of RhoA was described as a determinant step for diminished epithelial resistance, leading to reduced levels of occludin and E-cadherin found in Caco-2 cells previously incubated with LPS [68].…”

Bifidobacterium bifidum Enhances the Intestinal Epithelial Tight Junction Barrier and Protects against Intestinal Inflammation by Targeting the Toll-like Receptor-2 Pathway in an NF-κB-Independent Manner

“…Intestinal epithelial tight junction (TJ) barrier dysfunction has been implicated as an important pathogenic factor of various inflammatory conditions of the gut, including inflammatory bowel disease (IBD), necrotizing enterocolitis (NEC), and celiac disease [ 1 , 2 , 3 , 4 ]. Clinical studies in IBD patients have shown that a persistent increase in intestinal permeability following clinical remission is predictive of poor clinical outcome and an early recurrence of the disease [ 5 , 6 ], while normalization of intestinal permeability correlated with a sustained long-term clinical remission.…”