猪血多肽对建鲤抗氧化能力和免疫功能的影响

Effects of Peptides from Swine Blood on Antioxidant Capacity and Immune Function of Jian Carp(Cyprinus carpio var.Jian)

本试验旨在从体外和体内水平评估猪血多肽(peptides from swine blood,PSB)对建鲤(Cyprinus carpio var.Jian)抗氧化能力和免疫功能的影响。在体外试验中,制备建鲤外周血白细胞和头肾巨噬细胞,然后暴露在含有不同浓度(0、10、20、40、80μg/m L)猪血多肽的培养基中作用24 h,采用WST-1法测定外周血白细胞的增殖能力,氯化硝基四氮唑蓝(NBT)还原法和格里斯(Griess)试剂显色法分别测定头肾巨噬细胞氮呼吸爆发和氧呼吸爆发活性;在体内试验中,分别将0、0.5、1.0、2.0和5.0 g/kg猪血多肽添加到基础饲料中饲喂建鲤,在恒温循环水系统中进行8周的饲养试验,分别在饲喂第1、2、4、8周采集血样,检测血清中生化指标、抗氧化指标及免疫指标的变化。结果表明:在体外试验中,猪血多肽浓度为20、40和80μg/m L时,可以显著或极显著提高建鲤外周血白细胞的增殖能力(P<0.05或P<0.01);猪血多肽浓度为40和80μg/m L时,可以显著或极显著增强建鲤头肾巨噬细胞氮呼吸爆发和氧呼吸爆发活性(P<0.05或P<0.01)。在体内试验中,用添加5.0 g/kg猪血多肽的饲料饲喂建鲤后,第4和8周血清中3种生化指标[白蛋白(ALB)、总蛋白(TP)、碱性磷酸酶(AKP)]含量/活性、3种抗氧化酶[超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)]活性和3种免疫因子[白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、补体4(C4)]含量均显著升高(P<0.05);此外,第8周血清中补体3(C3)含量亦显著升高(P<0.05)。由此得出,猪血多肽能够增强建鲤的抗氧化能力和免疫功能。

This experiment was conducted to study the effects of peptides from swine blood （ PSB） on antioxi-dant capacity and immune function of Jian carp （ Cyprinus carpio var. Jian） at in vitro and in vivo levels. In vitro experiment, the head-kidney macrophages and peripheral blood leukocytes, isolated from Jian carp, were cultured and treated for 24 hours with different concentrations （0, 10, 20, 40 and 80 μg/mL） of PSB. To e-valuate the immunostimulating effects of PSB, the viability of peripheral blood leukocytes was measured by WST-1 method, and the respiratory burst activities of oxygen and nitrogen of head kidney macrophages were investigated by nitroblue tetrazolium （ NBT） reduction method and Griess reagent colorimetric method, respec-tively. In vivo experiment, PSB was added to the basal diet at the concentrations of 0, 0. 5, 1. 0, 2. 0 and 5.0 g/kg, fed to Jian carp in constant temperature cycle water system for 8 weeks. The blood samples were collected four times around the time points of the 1st week, the 2nd week, the 4th week and the 8th week, then detected the changes of serum biochemical indices, antioxidant indices and immune indices. The results showed as follows： in vitro experiment, the PSB at the concentrations of 20, 40 and 80 μg/mL could signifi-cantly enhance the viability of peripheral blood leucocytes （ P〈0.05 or P〈0.01）; PSB at the concentrations of 40 and 80 μg/mL could significantly enhance the respiratory burst activities of oxygen and nitrogen （ P〈0.05 or P〈0.01） . In vivo experiment, after Jian carp were fed the diet with PSB at the concentration of 5.0 g/kg, the contents/activity of 3 biochemical indices [ albumin （ ALB） , total protein （ TP） and alkaline phosphatase （ AKP） ] , the activities of 3 antioxidant enzymes [ superoxide dismutase （ SOD） , catalase （ CAT） and gluta-thione peroxidase （ GSH-Px） ] and the contents of 3 immune factors [ interleukin-1β （ IL-1β） , tumor necrosis factor-α （ TNF-α） and complement 4 （ C4） ] in serum at the 4th week and the 8th week were significantly in-creased （ P〈0.05）; moreover, the content of complement 4 （ C4） in serum at the 8th week was significantly increased aslo （ P〈0.05） . Therefor, the PSB can improve the antioxidant capacity and immune function of Jian carp.