Faculty of Education, Ehime University, Ehime, Japan.
Department of Food and Nutritional Environment, Kinjo Gakuin University, Aichi, Japan.
Drug Discov Ther. 2024 Jul 9;18(3):143-149. doi: 10.5582/ddt.2024.01016. Epub 2024 Jun 6.
This study aims to investigate the antiallergic effects of Shiikuwasha (Citrus depressa Hayata) leaf and peel extracts by examining the regulation of degranulation and inflammatory cytokine production from rat basophilic leukemia (RBL-2H3) cells and antigen-specific antibody production in sensitized mouse spleen lymphocytes. In vivo antiallergic activity was evaluated using the passive cutaneous anaphylaxis (PCA) reaction model. Extracts of Shiikuwasha leaves and peel were prepared using 80% methanol and dissolved in dimethylsulfoxide. The dinitrophenyl-human serum albumin-induced β-hexosaminidase levels in immunoglobulin (Ig) E-sensitized RBL-2H3 cells were assessed using enzymatic assays. Cytokine production was measured by enzyme-linked immunosorbent assay. Antibody production capacity was evaluated using lymphocytes isolated from spleens of type I allergy model mice. Lymphocytes were cultured for 72 h with Shiikuwasha extracts, and ovalbumin-specific IgE, IgG1, and IgG2a levels were measured. Shiikuwasha leaf and peel extract significantly reduced β-hexosaminidase release and suppressed interleukin-4 and tumor necrosis factor-α production from RBL-2H3 cells. Ovalbumin-specific IgE and IgG1 production decreased in Shiikuwasha extract-treated lymphocytes. These extracts also significantly suppressed the PCA reaction. Shiikuwasha leaf and peel extract reduce degranulation in RBL-2H3 cells and antibody production in spleen-derived lymphocytes and therefore exhibit antiallergic effects.
本研究旨在通过考察 Shiikuwasha(枳实)叶和果皮提取物对大鼠嗜碱性白血病(RBL-2H3)细胞脱颗粒和炎症细胞因子产生的调节作用,以及对致敏小鼠脾淋巴细胞中抗原特异性抗体产生的影响,来研究其抗过敏作用。采用被动皮肤过敏反应(PCA)模型评估体内抗过敏活性。使用 80%甲醇制备 Shiikuwasha 叶和果皮提取物,并溶解在二甲基亚砜中。采用酶法测定二硝基苯人血清白蛋白诱导的免疫球蛋白(Ig)E 致敏 RBL-2H3 细胞β-己糖胺酶水平。通过酶联免疫吸附试验(ELISA)测定细胞因子产生。采用从 I 型过敏模型小鼠脾脏分离的淋巴细胞评估抗体产生能力。将淋巴细胞与 Shiikuwasha 提取物共培养 72 h,测定卵清蛋白特异性 IgE、IgG1 和 IgG2a 水平。Shiikuwasha 叶和果皮提取物可显著降低β-己糖胺酶的释放,并抑制 RBL-2H3 细胞中白细胞介素-4 和肿瘤坏死因子-α的产生。卵清蛋白特异性 IgE 和 IgG1 的产生在 Shiikuwasha 提取物处理的淋巴细胞中减少。这些提取物还显著抑制 PCA 反应。Shiikuwasha 叶和果皮提取物可减少 RBL-2H3 细胞的脱颗粒和脾源性淋巴细胞中抗体的产生,因此具有抗过敏作用。