机构地区:[1]南京晓庄学院,南京210017 [2]上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室,上海201306 [3]上海交通大学医学院上海市免疫学研究所,上海200025
出 处:《现代免疫学》2011年第5期389-394,共6页Current Immunology
基 金:上海市科委重点攻关项目(013212101);江苏省教育厅高校自然科学基金项目(08KJD180013)
摘 要:本研究旨在应用环磷酰胺诱导建立的免疫抑制性异育银鲫的实验模型,以研究壳聚糖和益生菌壳聚糖和益生菌对异育银鲫免疫功能的影响。实验分别设立基础饲料、生理盐水为对照,基础饲料、环磷酰胺为阴性对照组,实验饲料加注环磷酰胺为实验组。饲养异育银鲫(体重123.2±4.8)g 30 d,在实验的第15天按10 mg/kg.bw的剂量给实验组和阴性对照组的每尾鱼注射环磷酰胺液,空白对照组每尾鱼注射等量生理盐水,每隔5 d注射1次,共注射3次。第3次注射后的第6天,分别取各组鱼的外周血和头肾为实验标本材料。以瑞氏染色计数和分析外周血白细胞数量及分类,白细胞吞噬白色葡萄球菌的活性。应用硝酸还原酶法测定NO含量,比浊法测定头肾溶菌酶活力。采用IFN-αELISA试剂盒和ABC-HRP方法测定IFN-α、ELISA法测定IgM含量。采用靶细胞乳酸脱氢酶释放法测定外周血NK细胞杀伤活性以及采用3H-TdR掺入法测定外周血淋巴细胞转化能力。实验结果表明:阴性对照组各项免疫指标均显著低于空白对照组;添加的壳聚糖和益生菌能够不同程度地恢复免疫抑制实验组的大多数免疫指标。三种壳聚糖的免疫调节能力各不相同,分子量居中的壳聚糖A的调节能力较强;分子量较大的壳聚糖B的调节能力较弱;壳聚糖C的调节能力并没有随添加量的增加而提高。因此,本研究的初步结论为:壳聚糖的免疫调节能力受分子量的影响较大,并不与添加量成正比,而适量的壳聚糖或益生菌能够不同程度地消除鱼类的免疫抑制,具有使其拓展成为鱼类免疫功能调节剂的前景。The experimental model of the immunosuppressed gibel carp induced by cyclophoshamide was established to investi gate the effect of chitosan and probiotics in this immunosuppressive model. The tested diets used consisted of the basal diet supplemented with 0.2% probiotics, 0.5% chitosan A, 0.5% chitosan B, 0.5% chitosan C and 0.75% chitosan C respective- ly, while the normal control group was injected with 0.9% NaCl solution and fed on the basal diet. The negative control group was injected with cyclophosphamide solution and fed on the basal diet. The tested groups were injected with cyclophosphamide solution and fed on the tested diets. Every group was duplicated. Gibel carp (123.2±4.8)g were fed for 30 days. The negative control and tested groups were intraperitoneally injected of 10 mg/kg bw cyclophosphamide while the normal control group was intraperitoneally injected with the same dosage of 0.9 % NaCI solution every 5 days for three times after 15 days of the feeding period. A lot of tests were performed in the experiments, such as number, classification and phagocytosis of peripheral blood leucocytes; the lysozyme activity of head kidney, proliferation of peripheral blood lymphocytes, IgM and IFN-α content, activity of natural killer cell and NO content in head kidney. The experimental results showed that immune function of the negative control group was lower than that of the normal control group evidently and chitosans and probiotics could improve immune function of tested groups in varying degree. The immunosuppressive activities of three chitosan on the immunosuppressive fish varied a great deal, i.e. higher in chitosan A with middle molecular weight; weaker in chitosan B with large molecular weight. The ability of chitosan C would not enhance with supplemented amount. From these observations, it is evident that, the chitosan together with probiotics an renew the immune function of immunosuppressive fish and their ablity of chitosan to improve immune function would be influenced by the molecular we
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