PARK7 Diminishes Oxidative Stress-Induced Mucosal Damage in Celiac Disease

Abstract

Coeliac disease (CD) is a chronic, immune-mediated small intestinal enteropathy, accompanied with gluten-triggered oxidative damage of duodenal mucosa. Previously, our research group reported an increased mucosal level of the antioxidant protein Parkinson’s disease 7 (PARK7) in children with CD. In the present study, we investigated the role of increased PARK7 level on the epithelial cell and mucosal integrity of the small intestine. The presence of PARK7 was investigated using immunofluorescent staining on duodenal mucosa of children with CD and on FHs74Int duodenal epithelial cells. To investigate the role of oxidative stress, FHs74Int cells were treated with H2O2 in the absence or presence of Comp23, a PARK7-binding compound. Intracellular accumulation of reactive oxygen species (ROS) was determined by DCFDA-based assay. Cell viability was measured by MTT, LDH, and Annexin V apoptosis assays. Disruption of cytoskeleton and cell adhesion was investigated by immunofluorescence staining and by real-time RT PCR. Effect of PARK7 on mucosal permeability was investigated ex vivo using intestinal sacs derived from control and Comp-23-pretreated mice. Comp23 treatment reduced the H2O2-induced intracellular accumulation of ROS, thus preserving the integrity of the cytoskeleton and also the viability of the FHs74Int cells. Accordingly, Comp23 treatment increased the expression of antioxidants (NRF2, TRX1, GCLC, HMOX1, NQO1), cell-cycle regulators (TP53, CDKN1A, PCNA, BCL2, BAX), and cell adhesion molecules (ZO1, CDH1, VCL, ITGB5) of H2O2-treated cells. Pretreatment with Comp23 considerably decreased the small intestinal permeability. In this study, we demonstrate that PARK7-binding Comp23 reduces the oxidative damage of duodenal epithelial cells, via increased expression of NRF2- and P53-regulated genes. Our results suggest that PARK7 plays a significant role in the maintenance of mucosal integrity in CD.

Immunomodulatory role of Parkinson’s disease 7 in inflammatory bowel disease

“…6 ) . In line with our previous experiments on small intestinal sacs 13 , we found that Comp23 greatly decreased the permeability of H 2 O 2 -treated colon sacs (Fig. 6 ), confirming the determinative role of PARK7 in the maintenance of mucosal integrity.…”

“…However, there are a few studies investigating the role of PARK7 in gastrointestinal diseases. Indeed, recently our research group demonstrated the role of PARK7 in the maintenance of duodenal mucosal integrity of children with coeliac disease 12 , 13 , and Di Narzo 23 and Zhang et al 24 investigated the amount of PARK7 in the plasma and intestine of adult patients with IBD, respectively. PARK7 was also shown to play a role in the pathomechanism of premature piglet model of enteral feeding induced necrotizing enterocolitis 25 , and in ketoprofen-induced oxidative damage of intestinal mucosa 26 .…”

“…The above considerations, and also our previous studies demonstrating the role of PARK7 in the maintenance of small intestinal mucosal integrity 12 , 13 led us to study the role of PARK7 in the pathogenesis of IBD. In this study we report the presence, regulation and role of PARK7 in the pathomechanism of mucosal inflammation using tissue samples of therapy-naive children with IBD, and in vitro and in vivo experimental models of colitis.…”

Impact of Exposure to Chronic Light–Dark Phase Shifting Circadian Rhythm Disruption on Muscle Proteome in Periparturient Dairy Cows

“…In addition to the proteins represented as increased in abundance in PS cows at 21 days BEC in Table 2, PARK7 and PRDX6 were both elevated. These proteins function as antioxidants that prevent buildup of oxidative species [24]. PS cows also exhibited an increased abundance of HSP70 at 21 days PP.…”

Transient Agarose Spot (TAS) Assay: A New Method to Investigate Cell Migration

“…The mRNA expressions were determined by real-time PCR using Light-Cycler 480 SYBR Green I Master enzyme mix on a LightCycler 96 system (Roche Diagnostics, Indianapolis, IN, USA). PCR primers (Table 2) were designed as previously described [51,52] and synthetized by Integrated DNA Technologies (IDT, Coralville, IA, USA). Results were analyzed using LightCycler 96 software v1.1.0.1320 (Roche Diagnostics).…”