低氧环境下蛋内注射CuSO4对藏鸡和肉鸡鸡胚发育的影响  

Effects of in ovo injection of CuSO4 on the development of Tibetan chicken and broiler chicken embryos hatched under hypoxia conditions

作　　者：姚宏 郑玉才[1] 李志雄 王云浩 谈永萍[1] 徐亚欧[1] 王爱林 董传豪 冯卫东[2] 饶开晴[1] YAO Hong;ZHENG Yucai;LI Zhixiong;WANG Yunhao;TAN Yongping;XU Ya'ou;WANG Ailin;DONG Chuanhao;FENG Weidong;RAO Kaiqing(College of Life Science and Technology,Southwest Minzu University,Chengdu 610041,China;Ganzi Prefectural Livestock Research Institute,Kangding 626000,China)

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041 [2]甘孜州畜牧业科学研究所,四川康定626000

出　　处：《南京农业大学学报》2020年第6期1079-1086,共8页Journal of Nanjing Agricultural University

基　　金：四川省科技支撑计划“十三五育种攻关”项目(2016NYZ0043)。

摘　　要：[目的]本文旨在研究低氧环境下改变种蛋内铜离子(Cu^2+)含量对藏鸡、科宝肉鸡两品种鸡胚发育、血管生成和机体抗氧化性能的影响及机制。[方法]选择质量相近的藏鸡、科宝肉鸡种蛋各132枚,随机分为3组。孵化前蛋内注射不同Cu^2+浓度的CuSO4注射液100μL:对照组(仅注射100μL 90μg·mL^-1 NaCl溶液)、低剂量组(75μg·mL^-1 Cu^2+),高剂量组(300μg·mL^-1 Cu^2+),低氧环境下孵化。12胚龄时固定和采集尿囊膜,采用Image-Pro Plus 6.0图像分析软件对鸡胚尿囊膜血管面积、条数进行测量;记录出雏率,采集1日龄血清及各组织并测定其质量;使用试剂盒检测与抗氧化相关的指标;采用RT-qPCR检测肝脏和尿囊膜中血管内皮生长因子(VEGF)基因的表达。[结果]肉鸡出雏率随蛋内注射Cu^2+浓度增加而升高;与对照组相比,低剂量组肉鸡出雏质量/蛋质量、腿体比以及高剂量组肝体比均显著提高(P<0.05),藏鸡低剂量组肝体比显著升高(P<0.05)。与对照组相比,肉鸡高剂量组的尿囊膜血管面积和中血管数显著增加(P<0.05),高剂量组的藏鸡尿囊膜上大、中、小血管形成以及血管面积增加(P<0.05),低剂量组的藏鸡尿囊膜中、小血管数量显著增加(P<0.05)。藏鸡、肉鸡试验组血清中铜锌超氧化物歧化酶(Cu/Zn-SOD)活性均显著高于对照组(P<0.05),丙二醛(MDA)含量显著低于对照组(P<0.05),而谷胱甘肽过氧化物酶(GSH-Px)活性不受Cu^2+的影响。注射高剂量的CuSO4能显著上调藏鸡、肉鸡肝脏和尿囊膜中VEGF基因的表达量(P<0.05),且藏鸡尿囊膜和肝脏中VEGF基因的表达量均显著高于肉鸡(P<0.05)。[结论]低氧环境下种蛋内注射CuSO4可提高藏鸡和肉鸡机体的抗氧化能力,促进两品种鸡胚血管生成以应对低氧环境;肉鸡机体抗氧化能力强于藏鸡,而藏鸡的血管生成能力强于肉鸡。[Objectives]The present experiment was conducted to study the effects of changing the copper ion content on embryonic development,angiogenesis and anti-oxidant performance of Tibetan chickens and Cobb broilers under hypoxic conditions,and to explore the mechanism of these effects.[Methods]Each of 132 fertilized eggs with similar quality of Tibetan chickens and Cobb broilers were selected and randomly allocated into three groups.100μL of CuSO4 injection with different Cu^2+concentrations were in ovo injection before incubation.90μg·mL^-1 NaCl solution in the control group,75μg·mL^-1 Cu^2+in the low-dose group,and 300μg·mL^-1 Cu^2+in the high-dose group were hatched under hypoxia conditions.Chorioallantoic membrane(CAM)was fixed and collected on 12 embryonic age(E12).Image-Pro Plus 6.0 image analysis software was used to measure the blood vessel area and number of strips in CAM on E12;the hatching rate was observed at hatching,and each tissue sample was collected and measured on the quality on one-day-old;the anti-oxidation related indexes were detected using the kit;the expression of VEGF gene in liver and CAM was detected by RT-qPCR.[Results]The hatching rate of broiler chickens enhanced with the increase concentration of Cu^2+injected into the eggs.Compared with the control group,the hatching weight/egg weight,leg-body ratio of the low dose group and liver-body ratio of the high dose group significantly increased(P<0.05).The liver-body ratio of Tibetan chickens in the low-dose group was significantly higher than that of the control group(P<0.05).The vascular area and the number of middle blood vessels in CAM were significantly increased in broiler chicken group with high dose(P<0.05).The high dose of copper ion could promote the formation of large,medium and small blood vessels and increase the vascular area in CAM of Tibetan chickens(P<0.05).The number of small and medium blood vessels in CAM of Tibetan chickens in the low-dose group significantly increased(P<0.05).The activity of Cu,Zn-superoxide dismut

关 键 词：低氧 蛋内注射 铜离子 血管生成 抗氧化 

分 类 号：S831.1[农业科学—畜牧学]