Antioxidant and Immunoprotective Effects of Moroccan Honey, Bee Pollen, and Propolis Against Cyclophosphamide‐Induced Immunosuppression in Mice: In Vitro, In Vivo, and In Silico Insights

Abstract

The immune system plays a crucial role in protecting the body against infections, and its suppression can lead to severe complications during chemotherapy. Bee products are rich in bioactive phenolic compounds with antioxidant and immunomodulatory properties. This study aimed to characterize the phenolic composition of Moroccan honey, bee pollen, and propolis and to evaluate their potential, individually and in combination, to counteract CTX-induced immunosuppression in mice. Phenolic compounds were analyzed by HPLC-DAD, total phenolics, and flavonoids were quantified using colorimetric assays, and antioxidant activity was determined by DPPH, RP, and TAC methods. In vivo, CTX-induced immunocompromised mice were distributed into six groups and treated orally for 21 days with honey (1 g/kg), bee pollen (100 mg/kg), propolis (100 mg/kg), or their mixture at a volume ratio of 1:1:1 (v/v/v). Results showed that propolis had the highest total phenolic and flavonoid content and exhibited the strongest antioxidant activity. All bee products significantly improved leukocyte counts, thymus and spleen indices, IgG/IgM levels, and delayed hypersensitivity response compared with the CTX non-treated group, with propolis, bee pollen, and the mixture showing the most pronounced effects. Molecular docking revealed strong interactions of major phenolics (apigenin, kaempferol, isorhamnetin, pinocembrin) with immune-related proteins (STAT3, JNK, and SYK), suggesting a multi-target mechanism of immunoprotection. These findings highlight Moroccan bee products as promising natural adjuvants for supporting immune function during chemotherapy.