Abstract
Inflammation and compromised immune responses often increase colorectal cancer (CRC) risk. The immune-modulating effects of limonin on carcinogen/inflammation-induced colorectal cancer (CRC) were studied in mice. Male Balb/c mice were randomly assorted into three groups (n = 6): healthy control, non-treated CRC-induced (azoxymethane/dextran-sulfate-sodium AOM/DSS) control, and CRC-induced + 50 mg limonin/kg body weight. The CRC developments were monitored via macroscopic, histopathological, ELISA, and mRNA expression analyses. Limonin downregulated inflammation (TNF-α, tumor necrosis factor-α), enhanced the adaptive immune responses (CD8, CD4, and CD19), and upregulated antioxidant defense (Nrf2, SOD2) mRNA expressions. Limonin reduced serum malondialdehyde (MDA, lipid peroxidation biomarker), prostaglandin E2, and histopathology inflammation scores, while increasing reduced glutathione (GSH) in CRC-induced mice. Limonin significantly (p < 0.05) increased T cells (CD4 and CD8) and B cells (CD19) in spleen tissues. The CD335 (natural killer cells) were increased in the CRC-induced mice and limonin treatment restored them to normal levels suggesting reinstatement to normal colon conditions. Limonin apparently mitigated CRC development, by ameliorating adaptive immune responses (CD8, CD4, and CD19), reducing inflammation (serum prostaglandin E2; TNF-α, innate immune responses) and oxidative stress, and enhancing the endogenous anti-oxidation defense reactions (GSH) in CRC-induced mice.
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Acknowledgements
We thank the Ministry of Higher Education for the partial scholarship (MyBrain15), Universiti Putra Malaysia for the seed research Putra Grant (GP-IPS/2018/9623000) and the staff from Comparative Medicine and Technology (COMeT), Institute of Bioscience, Universiti Putra Malaysia (UPM), for the help provided in handling the animals throughout the study.
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This work was funded by the Ministry of Higher Education partial scholarship (MyBrain15), and Universiti Putra Malaysia postgraduate Putra Grant (GP-IPS/2018/9623000).
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NIMI executed the experiments, planning, data collection, and manuscript preparation, NMM is the veterinary clinician for animal studies, and SM is the grant recipient, main supervisor, project planning coordinator and manuscript writing/editing, principal researcher. IFM and NME assisted in planning the animal study. All the researchers approved the final manuscript. All the authors read and approved the manuscript and all data were generated in-house and that no paper mill was used.
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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The animal study protocol was approved by the Institutional Animal Care and Use Committee (IACUC), Universiti Putra Malaysia (UPM/IACUC/AUP-R069/2017).
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Ishak, N.I.M., Mohamed, S., Madzuki, I.N. et al. Limonin modulated immune and inflammatory responses to suppress colorectal adenocarcinoma in mice model. Naunyn-Schmiedeberg's Arch Pharmacol 394, 1907–1915 (2021). https://doi.org/10.1007/s00210-021-02101-6
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DOI: https://doi.org/10.1007/s00210-021-02101-6