不同剂量环磷酰胺对雏鸡法氏囊免疫抑制的影响  

Effects of Cyclophosphamide at Different Doses on Immunosuppression of Chicken Bursa of Fabricius

作　　者：李冰心 李婉雁 曹楠 杨保和 袁明凤 付新亮 陈文斌 田允波 黄运茂 许丹宁 LI Bingxin;LI Wanyan;CAO Nan;YANG Baohe;YUAN Mingfeng;FU Xiniang;CHEN Wenbin;TIANYunbo;HUANG Yunmao;XU Danning(College of Animal Science&Technology,Zhongkai University of Agriculture and Engineering,Guangzhou Guangdong 510225;Yunnan Kuoduo Animal Husbandry Technology Co.,Ltd.,Yuxi Yunnan 653100)

机构地区：[1]仲恺农业工程学院动物科技学院,广东广州510225 [2]云南快大多畜牧科技有限公司,云南玉溪653100

出　　处：《广东畜牧兽医科技》2023年第4期7-12,53,共7页Guangdong Journal of Animal and Veterinary Science

基　　金：国家自然科学基金项目(NO.32102747和NO.32202764);广东省自然科学基金面上项目(2022A1515010677);湖南省自然科学基金面上项目(2021JJ30312)。

摘　　要：该研究旨在探究不同剂量环磷酰胺(CTX)对岭南黄鸡法氏囊免疫抑制的影响,为后续探究如何提高法氏囊免疫力提供动物模型参考。该试验将120只1日龄岭南黄鸡随机分成4组(对照组、CTX20组、CTX40组和CTX80组),每组3个重复,每个重复10只。所有组饲喂基础日粮,CTX20组、CTX40组和CTX80组雏鸡在19、20、21日龄时分别肌肉注射20、40、80 mg/kg BW的CTX,对照组注射等量的生理盐水,最后一次注射7天后,每组随机选取15只雏鸡无菌采集法氏囊和血清进行组织学观察、Tunel检测、实时荧光定量PCR和ELISA检测。组织学观察结果显示,CTX处理可以诱导法氏囊小结间空隙增大、髓质中细胞密度降低、髓质区纤维增生、细胞核溶解、核固缩、坏死细胞数量增加等现象。Tunel检测结果显示,CTX可以诱导法氏囊细胞凋亡(P<0.05),其中CTX40组内细胞凋亡率超过35%。实时荧光定量PCR检测结果显示,CTX处理可以诱导法氏囊内细胞凋亡相关基因Bcl-2和NF-κB mRNA表达量显著降低,TNF-α和Caspase3 mRNA表达量显著上升(P<0.05)。血清ELISA检测结果显示,与对照组相比,CTX处理可以诱导TNF-α和IFN-γ表达量显著上升,IL-2和IL-6表达量显著降低(P<0.05)。上述结果表明,该试验成功构建了雏鸡法氏囊免疫抑制模型,其中40 mg/kg CTX免疫抑制效果最优。The purpose of this study was to explore the effect of different doses of cyclophosphamide(CTX)on the immunosuppression of the bursa of Fabricius in Lingnan yellow chickensand to provide an animal model reference for further research on how to improve the immunity of the bursa of Fabricius.In this study,1201-day-old Lingnan yellow chickens were randomly divided into 4 groups(Control group,CTX20 group,CTX40 group,and CTX80 group)with 3 replicates per group and 10 in each replicate.All groups were fed with basic diet.The chickens in the CTX20,CTX40,and CTX80 groups were intramuscularly injected with 20,40,and 80 mg/kg BW of CTX at 19,20,and 21 days of age,and the control group was injected with the same amount of normal saline.Seven days after the last injection,15 chicks from each group were randomly selected to collect the bursa of Fabricius and serum for histological observation,Tunel,real-time fluorescence quantitative PCR,and ELISA detection.The histological results showed that CTX treatment could induce the increase of space between the nodules of the bursa of Fabricius,the decrease of cell density in the medullary tissue,fibroplasia in the medullary region,the dissolution of the nucleus,the pyknosis of the nucleus,and the increase of necrotic cells.The Tunel test results showed that CTX could induce apoptosis of bursa cells(P<0.05),and the apoptosis rate in the CTX40 group was more than 35%.The results of real-time fluorescence quantitative PCR showed that the apoptosis-related genes expressions of Bcl-2 and NF-κB were significantly decreased,and the mRNA expressions of TNF-αand Caspase3 were significantly increased in the bursa of Fabricius by CTX treatment(P<0.05).The results of serum ELISA showed that compared with the control group,the expression level of TNF-αand IFN-γwas significantly increased,and the expression level of IL-2 and IL-6 was significantly decreased(P<0.05).These results indicate that the immunosuppression model of the chicken bursae of Fabricius was successfully constructed in this s

关 键 词：环磷酰胺 岭南黄鸡 法氏囊 免疫抑制 

分 类 号：S859.7[农业科学—临床兽医学]