Date of Award

11-18-2025

Degree Type

Thesis

Degree Name

MPhil in Biological and Biomedical Sciences

First Advisor

Dr. Amber Pall

Second Advisor

Dr. Irfan Khan

Third Advisor

Dr. Najeeha Talat Iqbal

Department

Biological and Biomedical Sciences

Abstract

Background Inflammatory Bowel Disease (IBD) is an autoimmune disorder comprises of ulcerative colitis (UC) and Chron’s disease (CD), characterized by immune dysregulation, microbial hemostasis and barrier dysfunction. Natural products with comprise bioactive constituents, balance multiple aspects of the disease. Fermentation of the natural product enhance the bioavailability and bioactive compounds, and it has been traditionally used, however, the effect of fermented soybean and flaxseed has not yet been pharmacologically validated. Hence, it was aimed to explore the pharmacological aspect of fermented and non-fermented soybean (SB) and flaxseed (FS) alone as well as in combination against acetic acid induced colitis mice model, as it was not studied previously. Methodology Fermented and non-fermented SB and FS was at first assessed for its effect in in-vitro epithelial cells lines induced by hydrogen peroxide H2O2. The fermented formulation were tested for cytotoxicity and to validate for safest concentration in vitro assays (spectrophotometric, XTT and DCFDA) was performed. Secondly, in-vivo analysis was performed in both male and female (BALB/c) mice at 150 and 300mg/kg dose. The colonic tissue was analyzed for Disease Activity Index (DAI), macroscopic damage and expression of inflammatory cytokines, followed by mucosal protective genes expression. Additionally, bacterial abundance of probiotic (Lactobacillus) was assessed before and after treatment to examine the effect of fermented formulation on commensal gut microbiota.
Results The overall findings indicated that fermented formulation with probiotic alone has profound antioxidant potential as it reduced the DAI score, macroscopic damage, downregulated the pro inflammatory cytokines (TNF-α, IL-6, IL17A, IL17F, IL21, IL23) gene expression upregulating the mucosal protective gene (TGF-β, MUC-2 and MUC-4). Simultaneously the fermented formulation especially the combination group has significantly upregulated Lactobacillus gene expression. The combination group of both SB and FS fermented with probiotic alone has shown 18 synergistic effect in mitigating the disease severity, suggesting that this particular formulation might be use against IBD.
Conclusion Amongst all the fermented formulation, fermentation with probiotic alone has shown promising therapeutic potential for IBD as it helped in suppressing the inflammation, balanced the commensal bacteria. The present study potentially confirms that fermentation enhances the bioactive compounds of the natural product, contributing in antioxidant, anti-inflammatory and immunomodulatory effect against IBD model. However, future studies require further microbial profiling, histological analysis and to confirm which bioactive compound help in mitigating.

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Last Page

117

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