大豆异黄酮对褐牙鲆免疫细胞功能的体外研究

In Vitro Effects of Soybean Isoflavones on Immune Function of Bastard Halibut(Paralichthys olivaceus)

在超净台内剖取、研磨褐牙鲆头肾,用Ortuńo的方法分离头肾巨噬细胞;无菌抗凝尾静脉抽血分离外周血白细胞。用含有0、0.1、0.5、1.0、1.5mg/mL和2.0mg/mL大豆异黄酮的细胞培养液分别体外培养这些细胞,24h后分别测定细胞的氧呼吸爆发活力、增殖活力及吞噬活力等免疫指标。试验结果表明,在体外培养下,大豆异黄酮显著影响褐牙鲆头肾巨噬细胞和外周血白细胞免疫力(P<0.05),且随着培养液中大豆异黄酮质量浓度的增加,上述细胞免疫力先增后降。当培养液中大豆异黄酮质量浓度为0.5mg/mL时,褐牙鲆头肾巨噬细胞和外周血白细胞的氧呼吸爆发活力、增殖活力以及吞噬活力显著高于其他处理组(P<0.05);而当大豆异黄酮质量浓度达到2mg/mL时,显著抑制褐牙鲆头肾巨噬细胞和外周血白细胞免疫力(P<0.05)。在本试验条件下,体外细胞培养中添加0.5mg/mL大豆异黄酮促进了褐牙鲆头肾巨噬细胞和外周血白细胞的免疫力;2mg/mL大豆异黄酮则明显抑制细胞免疫力。

In vitro,the head kidney macrophages and peripheral blood leukocytes were isolated from bastard halibut（Paralichthys olivaceus）by Ortuńo method and cultured in medium with soybean isoflavone at a dose of 0,0.1,0.5,1.0,1.5 mg/mL and 2.0 mg/mL for 24 hours.Then the respiratory burst activity,cell proliferation activity and phagocytic activity were determined to evaluate the effects of soybean isoflavones on the immunity of head kidney macrophages and peripheral blood leukocytes in bastard halibut.The results showed that in vitro soy isoflavones concentration in the medium had significant effects on the immunity in head kidney macrophages and peripheral blood leukocytes in bastard halibut（P〈0.05）and the immunity of head kidney macrophages and peripheral blood leukocytes were shown to be increased first and then decreased with the increase in the concentration of soybean isoflavone in the culture medium.There were significantly higher respiratory burst activity,cell proliferation activity,and phagocyticactivity in the head kidney macrophages and peripheral blood leukocytes in the group with 0.5 mg/mL soybean isoflavone in culture medium than those in other groups（P〈0.05）.However,significantly lower immunity in the head kidney macrophages and peripheral blood leukocytes was observed in the maximal dose of soybean isoflavone concentration（2 mg/mL）in culture medium（P〈0.05）.Under the experimental conditions,the lower concentration of soybean isoflavone（0.5 mg/mL）in the culture medium led to enhance the immunity of head kidney macrophages and peripheral blood leukocytes in bastard halibut while the immunity was inhibited by the maximal soybean isoflavone（2.0 mg/mL）in the culture medium.