Food derived raffinose attenuates cyclophosphamide‐induced immunosuppression in mice with associated changes in gut microbiota, short‐chain fatty acids and intestinal barrier integrity

Abstract BACKGROUND Cyclophosphamide (CTX)‐induced immunosuppression is accompanied by systemic immune impairment and intestinal dysbiosis. This study evaluated whether raffinose (Raf), a food‐derived prebiotic trisaccharide, is associated with improved immune and intestinal outcomes during CTX challenge. RESULTS Forty male ICR mice were assigned to control, CTX, and low‐, medium‐ or high‐dose Raf pretreatment groups. Raf was administered orally at 100, 200 or 400 mg kg −1 body weight for 21 days, followed by oral CTX at 80 mg kg −1 body weight for 3 days. Compared with CTX alone, Raf pretreatment was associated with reduced body‐weight loss, less spleen and thymus atrophy, improved peripheral blood parameters, higher serum IgG and IgM concentrations and improved spleen enzyme activities. Raf was also associated with reduced hepatic oxidative injury, increased abundance and mRNA expression of selected intestinal tight‐junction markers, increased colonic short‐chain fatty acids and changes in gut microbial composition, including Lactobacillus and Desulfobacterota . However, the study did not assess epithelial permeability, short‐chain fatty acid receptors, upstream inflammatory signaling or causal microbiota‐transfer/manipulation experiments. CONCLUSION Raf pretreatment was associated with improved immune, hepatic, intestinal barrier‐related, metabolite and microbiota outcomes in CTX‐treated mice. These findings support a proposed working model rather than a confirmed causal gut microbiota–short‐chain fatty acid–barrier mechanism. The prophylactic design, absence of a positive control and small microbiota sample size limit translation and dose interpretation. © 2026 Society of Chemical Industry.