Abstract
The aim of this study was to elucidate the anti-allergic effects of polymethoxyflavonoids in combination with milk proteins and the mechanism of inhibition. Three polymethoxyflavonoids and two milk proteins were exposed to the rat basophilic leukemia cell line RBL-2H3. β-hexosaminidase was used as an indicator of degranulation inhibition. The mechanism of inhibition was examined by measuring intracellular Ca2+ levels and western blot method. In the degranulation inhibition test with polymethoxyflavonoids and milk proteins alone, nobiletin was the strongest inhibitor in the polymethoxyflavonoid group and lactoferrin in the milk protein group. Next, co-stimulation with nobiletin and lactoferrin showed stronger synergistic degranulation inhibition than treatment with nobiletin or lactoferrin alone. Western blot analysis showed that co-stimulation with nobiletin and lactoferrin significantly downregulated the induction of phospholipase Cγ 1 phosphorylation. The degranulation response in RBL-2H3 cells was synergistically suppressed by co-stimulation of nobiletin and lactoferrin acting on both Ca2+-dependent and Ca2+-independent pathways.
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This work was supported by JSPS KAKENHI Grant Number 19K05880. We thank Wendy Hempstock, PhD, from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.
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KM, TO, and MT designed the research; KM, SY, LR and HK performed the experiments; KM, MT, SY, AK and HK analyzed the data; all authors wrote the draft and final manuscript; MT had primary responsibility for the final content; and all authors read and approved the final manuscript.
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Miyake, K., Tanaka, M., Yokoyama, S. et al. Degranulation of RBL-2H3 rat basophilic leukemia cells is synergistically inhibited by combined treatment with nobiletin and lactoferrin. Cytotechnology (2024). https://doi.org/10.1007/s10616-024-00625-2
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DOI: https://doi.org/10.1007/s10616-024-00625-2