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鸡血管生成素样蛋白4重组蛋白对肉鸡肝脏和胸肌脂肪代谢的影响
引用本文：侯彦茹,赵旭,黄华山,张金红,井长伟.鸡血管生成素样蛋白4重组蛋白对肉鸡肝脏和胸肌脂肪代谢的影响[J].动物营养学报,2022,34(2):1260-1267.
作者姓名：侯彦茹  赵旭  黄华山  张金红  井长伟
作者单位：临沂大学农林科学学院,临沂 276000;山东隆大生物工程有限公司,临沂 276400;山东亚太中慧集团有限公司,潍坊 262400
基金项目：临沂大学高层次人才科研启动项目(LYDX2019BS036);临沂大学2020年大学生创新创业训练计划项目(X202010452116)。
摘    要：本试验旨在探究鸡血管生成素样蛋白4(ANGPTL4)重组蛋白对肉鸡肝脏和胸肌脂肪代谢的影响。分体内动物试验和体外细胞试验2部分进行。体内动物试验选用35日龄健康禁食状态下爱拔益加肉公鸡36只,随机分为6组,每组6个重复,每个重复1只鸡。对照组翅静脉注射灭菌的生理盐水,试验组分别翅静脉注射20、100、500、2 500、12 500 ng/kg BW鸡ANGPTL4重组蛋白,注射剂量均为550μL,试验期30 min。体外细胞试验设3个组,分别是生理盐水组、组氨酸-小分子泛素样修饰蛋白(His-SUMO)标签组和鸡ANGPTL4重组蛋白组,分别在细胞培养基中添加灭菌的生理盐水、His-SUMO标签蛋白(其含量与鸡ANGPTL4重组蛋白组中标签蛋白含量一致)和鸡ANGPTL4重组蛋白(250 pg/mL),添加剂量均为5μL,5%CO2、37℃孵育24 h。结果发现:1)与对照组相比,20、100、500、2 500 ng/kg BW鸡ANGPTL4重组蛋白组的肝脏脂肪酸合成酶(FAS)mRNA相对表达量和500 ng/kg BW鸡ANGPTL4重组蛋白组的肝脏FAS活性均显著提高(P<0.05)。随着鸡ANGPTL4重组蛋白注射剂量增加,肝脏FAS mRNA相对表达量和活性均呈现一次线性和二次曲线增加的效应(P<0.05)。2)与对照组相比,100、500、2 500 ng/kg BW鸡ANGPTL4重组蛋白组的肝脏苹果酸酶(ME) mRNA相对表达量和12 500 ng/kg BW鸡ANGPTL4重组蛋白组的肝脏乙酰辅酶A羧化酶(ACC) mRNA相对表达量均显著降低(P<0.05)。随着鸡ANGPTL4重组蛋白注射剂量增加,肝脏ACC mRNA相对表达量呈现一次线性和二次曲线降低的效应(P<0.05)。不同注射剂量的鸡ANGPTL4重组蛋白对肉鸡肝脏ME和ACC活性均无显著影响(P>0.05)。3)与对照组相比,500 ng/kg BW鸡ANGPTL4重组蛋白组的胸肌脂蛋白脂酶(LPL)活性显著增加(P<0.05)。4)鸡ANGPTL4重组蛋白组的成肌细胞甘油三酯(TG)含量显著高于生理盐水组和His-SUMO标签组(P<0.05)。综上所述,鸡ANGPTL4重组蛋白具有调控肉鸡肝脏脂肪合成相关基因mRNA表达和酶活性以及促进肉鸡胸肌脂肪沉积的作用,以500 ng/kg BW注射剂量效果较好。
关 键 词：鸡ANGPTL4重组蛋白  肉鸡  肝脏  胸肌  脂肪代谢

Effects of Chicken Angiopoietin-Like Protein 4 Recombinant Protein on Fat Metabolism in Liver and Breast Muscle of Broilers
HOU Yanru,ZHAO Xu,HUANG Huashan,ZHANG Jinhong,JING Changwei.Effects of Chicken Angiopoietin-Like Protein 4 Recombinant Protein on Fat Metabolism in Liver and Breast Muscle of Broilers[J].Acta Zoonutrimenta Sinica,2022,34(2):1260-1267.
Authors:HOU Yanru  ZHAO Xu  HUANG Huashan  ZHANG Jinhong  JING Changwei
Institution:(College of Agriculture and Forestry Science,Linyi University,Linyi 276000,China;Shandong Longda Bio-Products Co.,Ltd.,Linyi 276400,China;Shandong Yatai Zhonghui Group Co.,Ltd.,Weifang 262400,China)
Abstract:This experiment aimed to explore the effects of chicken angiopoietin-like protein 4(ANGPTL4)recombinant protein on fat metabolism in liver and breast muscle of broilers. It was divided into 2 parts:animal experiment in vivo and cell experiment in vitro. In the animal experiment in vivo,thirty-six healthy 35-day-old Arbor Acres broilers under fasting condition were randomly divided into 6 groups with 6 replicates per group and 1 broiler per replicate. Broilers in the control group were intravenously injected with sterile normal saline,and others in experimental groups were intravenously injected 20,100,500,2 500 and 12 500 ng/kg BW chicken ANGPTL4 recombinant protein,respectively. The injection dose was 550μL,and the experiment period was 30 minutes. Three groups were established for the cell experiment in vitro,including the normal saline group,histidine-small ubiquitin-like modifier(His-SUMO)tag group and chicken ANGPTL4 recombinant protein group,which supplemented 5μL sterile normal saline,His-SUMO tag protein(its content is consistent with that of chicken ANGPTL4 recombinant protein group)and chicken ANGPTL4 recombinant protein(250 pg/mL),respectively,with 5% CO2 and 37 ℃ incubated 24 h. The results showed as follows:1)compared with the control group,the liver fatty acid synthase(FAS)mRNA expression level of 20,100,500 and2 500 ng/kg BW chicken ANGPTL4 recombinant protein groups and liver FAS activity of 500 ng/kg BW chicken ANGPTL4 recombinant protein group were significantly increased(P<0.05). With the chicken ANGPTL4 recombinant protein injection dose increased,the liver FAS mRNA expression level and activity showed linear and quadratic increase effects(P<0.05). 2)Compared with the control group,the liver malic enzyme(ME)mRNA expression level of 100,500 and 2 500 ng/kg BW chicken ANGPTL4 recombinant protein groups and liver acetyl-CoA carboxylase(ACC)mRNA expression level of 12 500 ng/kg BW chicken ANGPTL4 recombinant protein group were significantly decreased(P<0.05). With the chicken ANGPTL4 recombinant protein injection dose increased,the liver ACC mRNA expression level and activity showed linear and quadratic decrease effects(P<0.05). Different injection dose of chicken ANGPTL4 recombinant protein had no significant effects on activities of ME and ACC in liver of broilers(P>0.05). 3)Compared with the control group,the breast muscle lipoprotein lipase(LPL)activity of 500 ng/kg BW chicken ANGPTL4 recombinant protein group was significantly increased(P<0.05). 4)The myoblasts triglyceride(TG)content of chicken ANGPTL4 recombinant protein group was significantly higher than that of normal saline group and HisSUMO tag group(P<0.05). In summary,chicken ANGPTL4 recombinant protein can regulate the mRNA expression and activity of fat synthesis related genes and enzyme in liver of broilers and promote fat deposition in breast muscle of broilers,and the 500 ng/kg BW injection dose has the better effect.Chinese Journal of Animal Nutrition,2022,34(2):1260-1267]
Keywords:chicken ANGPTL4 recombinant protein  broilers  liver  breast muscle  fat metabolism
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