Degranulation of RBL-2H3 rat basophilic leukemia cells is synergistically inhibited by combined treatment with nobiletin and lactoferrin

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Cytotechnology Pub Date : 2024-04-04 DOI:10.1007/s10616-024-00625-2

Kaho Miyake, Mamoru Tanaka, Sayaka Yokoyama, Lu Rui, Ayaka Koida, Hana Kozai, Takeaki Okamoto

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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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联合使用金霉素和乳铁蛋白可协同抑制 RBL-2H3 大鼠嗜碱性粒细胞的脱颗粒作用

本研究旨在阐明多甲氧基黄酮与牛奶蛋白结合的抗过敏作用及其抑制机制。将三种多甲氧基黄酮和两种牛奶蛋白暴露于大鼠嗜碱性粒细胞白血病细胞系 RBL-2H3。通过测量细胞内 Ca2+ 水平和 Western 印迹法研究了抑制机制。在单独使用多甲氧基黄酮和牛奶蛋白的脱颗粒抑制试验中，多甲氧基黄酮组中金雀花素的抑制作用最强，牛奶蛋白组中乳铁蛋白的抑制作用最强。其次，与单独使用金没药和乳铁蛋白相比，金没药和乳铁蛋白的联合刺激显示出更强的协同脱颗粒抑制作用。Western 印迹分析表明，金胆碱和乳铁蛋白共同刺激可显著降低磷脂酶 Cγ 1 磷酸化的诱导。金没药素和乳铁蛋白共同刺激钙离子依赖性和钙离子非依赖性途径，可协同抑制 RBL-2H3 细胞的脱颗粒反应。

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来源期刊

Cytotechnology 生物-生物工程与应用微生物

CiteScore

4.10

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： The scope of the Journal includes: 1. The derivation, genetic modification and characterization of cell lines, genetic and phenotypic regulation, control of cellular metabolism, cell physiology and biochemistry related to cell function, performance and expression of cell products. 2. Cell culture techniques, substrates, environmental requirements and optimization, cloning, hybridization and molecular biology, including genomic and proteomic tools. 3. Cell culture systems, processes, reactors, scale-up, and industrial production. Descriptions of the design or construction of equipment, media or quality control procedures, that are ancillary to cellular research. 4. The application of animal/human cells in research in the field of stem cell research including maintenance of stemness, differentiation, genetics, and senescence, cancer research, research in immunology, as well as applications in tissue engineering and gene therapy. 5. The use of cell cultures as a substrate for bioassays, biomedical applications and in particular as a replacement for animal models.