Purified fish skin collagen hydrolysate attenuates TNF-α induced barrier dysfunction in-vitro and DSS induced colitis in-vivo model

Inflammatory mediators are key components in establishing pathogenesis in inflammatory bowel disease. Balanced expression of anti-inflammatory and pro-inflammatory cytokines is an important cue in maintaining gut native and adaptive immunity. In the present study, purified hydrolysate fraction of fish skin collagen from Clarias batrachus and Pangasius pangasius was evaluated as a treatment agent against TNF-α induced barrier dysfunction in Caco-2 cell line model and DSS induced colitis in mice model. Cell adhesion on purified hydrolysate fraction coated surfaces was found to be enhanced with increasing concentration in both Clarias batrachus and Pangasius pangasius. Alkaline phosphatase activity was enhanced in a concentration-dependent manner. The paracellular permeability assay demonstrated that Pangasius pangasius purified hydrolysate fraction had countered TNF-α induced barrier dysfunction. Analysis of the tight junction proteins (occludin, zonulae occluden, and claudin) by RT PCR, immunofluorescence, and western blot, further confirmed the effectiveness of Pangasius pangasius purified hydrolysate fraction against TNF-α. The Pangasius pangasius purified hydrolysate fraction was further evaluated for efficacy in DSS-induced colitis mice model. Two concentration of Pangasius pangasius purified hydrolysate was chosen based on in-vitro experiments, 80 μg/kg and 200 μg/kg BW of Balb/C male mice administered through intra-rectal route along with fish skin collagen 80 μg/kg BW. Pangasius pangasius purified hydrolysate fraction treatment improved the clinical signs of colitis such as body weight, rectal bleeding, colon length, and stool consistency caused by DSS administration. Immunofluorescence of colon tissue section showed that Pangasius pangasius purified hydrolysate fraction enhanced the expression of occludin protein. This study hints at the use of Pangasius pangasius purified hydrolysate fraction as a potential nutraceutical or treatment agent in healing ulcers of the mucosa.