Department of Oral Biochemistry, Asahi University School of Dentistry, Mizuho, Gifu 501-0296, Japan; Department of Operative Dentistry, Asahi University School of Dentistry, Mizuho, Gifu 501-0296, Gifu 500-8471, Japan.
Chemistry Laboratory, Department of Business Administration, Asahi University School of Business Administration, Mizuho, Gifu 501-0296, Japan.
J Oral Biosci. 2022 Sep;64(3):366-375. doi: 10.1016/j.job.2022.05.007. Epub 2022 Jun 3.
In this report, we attempt to clarify the immune modulatory effects of Brazilian green propolis (BGP) and its major component, artepillin C, on the cytokine production of anti-CD3 antibody-stimulated mouse spleen cells. We also estimate the physiological mechanism affecting artepillin C's upon the cells.
Male C3H/HeN mouse spleen cells stimulated by antiCD3 monoclonal antibody were co-cultured with BGP, artepillin C, and HC030031, a transient receptor potential ankyrin 1 (TRPA1) Ca channel antagonist. The synthesis of interferon (IFN)-γ, interleukin (IL)-6, IL-17, IL-4, IL-10, and IL-2 was assayed by enzyme-linked immunoassay. The expression of IL-2 mRNA and the protein product were examined by reverse transcription-quantitative polymerase chain reaction and Western blot analyses, respectively.
The production of IL-2 was markedly enhanced, while that of IL-4 and IL-10 was not significantly affected; by contrast, the production of IFN-γ, IL-6, and IL-17 was significantly reduced in the antibody-stimulated spleen cells treated with BGP at a non-cytostatic concentration. These effects were reproduced in the cells treated with artepillin C. The expression of IL-2 mRNA was unaffected; however, that of the protein was significantly enhanced in the artepillin C-treated cells compared to untreated control cells. The enhancement of protein expression and the production of IL-2 by artepillin C was significantly alleviated by adding HC030031.
Artepillin C is an important regulator of cytokine synthesis from activated spleen cells. The agent specifically augmented the expression of IL-2 via the Ca-permeable cation channel, TRPA1, at least in part, at the translational or secretion levels.
本报告试图阐明巴西绿蜂胶（BGP）及其主要成分 Artepillin C 对抗 CD3 抗体刺激的小鼠脾细胞细胞因子产生的免疫调节作用。我们还评估了生理机制对 Artepillin C 作用于细胞的影响。
用抗 CD3 单克隆抗体刺激雄性 C3H/HeN 小鼠脾细胞，与 BGP、Artepillin C 和 HC030031（瞬时受体电位锚蛋白 1（TRPA1）钙通道拮抗剂）共同培养。通过酶联免疫吸附试验测定干扰素（IFN）-γ、白细胞介素（IL）-6、IL-17、IL-4、IL-10 和 IL-2 的合成。通过逆转录定量聚合酶链反应和 Western blot 分析分别检测 IL-2 mRNA 的表达和蛋白产物。
BGP 在非细胞毒性浓度下处理的抗体刺激脾细胞中，IL-2 的产生明显增强，而 IL-4 和 IL-10 的产生没有明显影响；相比之下，IFN-γ、IL-6 和 IL-17 的产生则显著减少。该作用在 Artepillin C 处理的细胞中重现。IL-2 mRNA 的表达不受影响；然而，Artepillin C 处理的细胞中蛋白的表达明显增强，与未处理的对照细胞相比。HC030031 的加入显著减轻了 Artepillin C 处理细胞中蛋白表达的增强和 IL-2 的产生。
Artepillin C 是激活脾细胞细胞因子合成的重要调节剂。该药物通过钙通透阳离子通道 TRPA1 特异性地增强了 IL-2 的表达，至少部分是在翻译或分泌水平上。
