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Rintu Das, Bhaswati Trafadar, Prosenjit Das, Srabani Kar, Shinjini Mitra, Garima Hore, Silpak Biswas and Ena Ray Banerjee
Inflammatory Bowel Diseases (IBD) is a chronic disorder of the gastrointestinal (GI) tract characterized by body weight loss, hemorrhage, lower abdominal pain and diarrhea. Indiscriminate use of antibiotics and modern life style disrupts microbial ecosystem of the human GI tract which leads to IBD. No disease modifying treatment exists for IBD which are both inflammatory and degenerative in nature. This study aimed at evaluating anti-inflammatory and pro-regenerative potential of probiotics in preclinical IBD. For in vitro study, inflammation was induced on RAW264.7 cell treated with 3% DSS (w/v), 1μg/ml LPS and 600ng/ml PMA. Novel combinations of probiotics were administrated on inflamed cell to measure their anti-inflammatory role. Cell viability and NO release assay were also done. For in vivo study, composite IBD phenotype was developed by 3% DSS by oral gavage (40μl) for 7 days and our probiotic combo was administered over 7 days (at 108 CFU/200μl) of DSS induction in Balb/c mice. Body weight, NO and ascorbic acid production by cells harvested from key lymphoid organs post-mortem, and histological studies were done to assess inflammation. We found cell viability increased from 30% to 92.5 % and NO concentration was reduced 2.22 fold after administration of probiotics. Mouse body weight (BW) reduced by 30.2% and 35.4% on 7th and 14th day respectively following DSS induction. Oral probiotics increased mouse BW in by 47%. NO concentration decreased 1.19 fold and 2.17 fold in colon and spleen tissue and ascorbic acid concentration increased 1.82 fold and 5.50 fold in colon and spleen tissue respectively. Clonogenic potential of intestine and spleen was decreased 2.38 fold and 2.34 fold in DSS treated mouse on 7th and 14th day respectively but it was increased 1.29 fold and 1.36 fold on 7th and 14th day respectively after probiotics administration in DSS treated mice. This indicates greater clonogenicity in intestine and spleen of probiotic treated mice as opposed to the reduced colony count of progenitor’s post-DSS only mice. Overall, orally administered probiotics showed anti-inflammatory and pro-regenerative action to revive the cells and tissues of DSS treated mice in which all symptoms of IBD were detected earlier. This study validates the use of combination of probiotic microbial strains supplemented in food (curd, yoghurt) as nutraceuticals and possible therapeutic as well as prophylactic to combat inflammation and degeneration in IBD.