中枢AMPK在应激肉鸡食欲调控中的作用研究

发布时间：2018-01-03 14:35

本文关键词：中枢AMPK在应激肉鸡食欲调控中的作用研究　出处：《山东农业大学》2017年硕士论文　论文类型：学位论文

更多相关文章： 皮质酮 肉鸡食欲 下丘脑 AMPK信号通路

【摘要】：本课题围绕着“应激-AMPK-食欲”研究主题展开了以下研究,主要是探讨AMPK在应激肉鸡的食欲调控中的作用。应激对肉仔鸡下丘脑食欲及AMPK信号通路的影响。试验选取1日龄体重相近AA肉仔鸡144只,随机分为3个处理(6个重复/8只鸡)。试验分为对照组、应激组、应激抑制组。7日龄进行试验处理,处理时间持续7天。试验结果显示:日粮中添加皮质酮显著降低了14日龄肉仔鸡体增重;皮质酮处理显著降低心脏指数、脾脏指数和法氏囊指数,相反,显著提高了肝脏指数和腹脂率;皮质酮组血浆中皮质酮、葡萄糖、甘油三酯、肝糖原和胸肌糖原的含量与对照组相比显著升高;与对照组相比,日粮中添加皮质酮显著上调14日龄肉仔鸡下丘脑中mTOR、GR、AMPKα2、NPY、LKB1、AMPKα1、FAS基因表达量,ACC蛋白的磷酸化水平在饲喂皮质酮时显著提高,同时AMPK总蛋白及磷酸化水平也显著提高。由此可见,应激导致肉鸡体重降低,糖元异生过程加强;应激反应通过提高促食神经肽NPY基因表达促进食欲,同时激活了中枢LKB1-AMPK-ACC信号通路。中枢注射Compound C对应激肉鸡食欲和能量代谢的影响。试验选取体重相近的AA肉仔鸡48只,7日龄进行脑室埋管,随机分为4个试验组(6个重复/2只鸡):DEX+Compound C组、DEX组、Compound C组、对照组。8日龄早上8:00 DEX处理组皮下注射地塞米松,对照组注射等剂量的生理盐水,连续注射3天。10日龄注射后空腹3小时,Compound C组注射Compound C、对照组注射等剂量的DMSO,每隔30分钟统计一次采食量,连续统计120分钟。根据采食量数据,重复上述试验,脑室注射Compound C后继续空腹1小时,采集样品。试验结果显示:DEX显著降低了肉仔鸡8日龄和9日龄日增重;与对照组相比,地塞米松处理显著上调了肉鸡下丘脑AMPKα2、LKB1、NPY以及m TOR基因表达,但后期中枢注射Compound C明显降低肉仔鸡下丘脑mTOR基因表达;与对照组相比,脑室注射Compound C显著降低了应激状态下肉鸡下丘脑AMPK总蛋白及磷酸化水平水平;与对照组相比,地塞米松处理显著降低了mTOR蛋白的磷酸化水平,后期中枢注射Compound C同样明显降低mTOR蛋白的磷酸化水平。由此可知,应激状态下中枢注射Compound C明显的降低采食量持续到1小时;应激激活下丘脑AMPK,但后期中枢注射Compound C明显抑制AMPK活性,并且通过AMPK-mTOR信号通路调控机体能量代谢。日粮不同能量水平对肉鸡食欲及中枢AMPK的影响。选取1日龄的雄性AA肉仔鸡216只,随机分为3个处理组(6个重复/12只鸡):低能日粮组(2900 kcal/kg)、正常日粮组(3200 kcal/kg)、高能日粮组(3500 kcal/kg);饲喂至21日龄各个重复取2只采集血浆及下丘脑样品。试验结果显示:低能日粮组平均日采食量明显高于其他两组,高能日粮组平均日增重明显高于其他两组;与对照组相比,高能日粮组和低能日粮组血浆中UREA的含量明显升高,其中低能日粮组含量最高;与高能日粮组相比,低能日粮处理显著升高了肉仔鸡血浆中TG含量;此外,高能日粮和低能日粮均显著降低了21日龄肉仔鸡脾脏指数;高能日粮处理显著下调肉仔鸡下丘脑LKB1、NPY、CCK、GR基因表达;与对照组相比,低能日粮显著上调肉仔鸡下丘脑AMPKα2基因表达;与高能日粮组相比时,低能日粮处理显著上调21日龄肉仔鸡下丘脑AMPKα1基因表达。综上所述,饲喂高能日粮时,肉鸡中枢AMPK信号通路被抑制,NPY表达量下降;饲喂低能日粮时,激活中枢AMPK,通过AMPK信号通路调控机体能量平衡。日粮不同能量水平对应激肉鸡食欲及AMPK信号通路的影响。选取体重相近1日龄雄性AA肉仔鸡216只,随机分为6个处理组(3个重复/12只):低能日粮*DEX组,正常日粮*DEX组,高能日粮*DEX组;低能日粮*Con组,正常日粮*Con组,高能日粮*Con组。14日龄与28日龄早上8:00 DEX组皮下注射地塞米松,对照组注射等剂量的生理盐水,连续注射3天。试验结果显示:在相对生长高峰期,无论饲喂何种日粮,糖皮质激素处理均显著提高肉鸡下丘脑LKB1、AMPKα1和GR基因表达量。在绝对生长高峰期,糖皮质激素使正常日粮组和高能日粮组肉鸡下丘脑LKB1、NPY基因表达量显著升高;同时,DEX处理显著降低能日粮组肉鸡下丘脑ACC总蛋白及磷酸化水平,显著提高正常日粮组肉鸡下丘脑AMPK磷酸化蛋白水平。综上所述,相对生长高峰期,DEX对LKB1-AMPK信号通路的影响不受日粮类型的影响,促食神经肽的表达受日粮类型的影响;绝对生长高峰期,DEX对LKB1-AMPK-ACC信号通路、促食神经肽的影响均受日粮类型的影响。综上所述,应激显著影响肉仔鸡食欲,并且激活中枢LKB1-AMPK-ACC信号通路;中枢AMPK信号通路确实参与了应激对家禽食欲及能量代谢的调控。外周GCs使整个生长阶段食欲均受日粮类型的影响,而中枢AMPK信号通路的变化依赖于生长阶段。
[Abstract]:This paper focuses on "stress -AMPK- appetite" research themes of the following research, mainly to explore the role of AMPK in appetite regulation stressed broilers. In the effect of stress on Broilers hypothalamus appetite and AMPK pathway. The experiment selects 1 day old AA broilers with similar body weight 144 rats were randomly divided into 3 treatments (6 a repeat /8 chicken). The experiment was divided into control group, stress group, stress inhibition group.7 days test, the processing time for 7 days. The test results show: the addition of corticosterone in diets significantly reduced 14 day old broiler body weight; corticosterone treatment significantly decreased cardiac index, spleen index and method bursal index, on the contrary, significantly improve the index of liver and abdominal fat rate; corticosterone, corticosterone group in plasma glucose, triglyceride, liver glycogen and muscle glycogen content was significantly higher than that in control group; compared with the control group, add skin in the diet MTOR, testosterone increased significantly 14 days old broilers in hypothalamus GR, AMPK alpha 2, NPY, LKB1, AMPK, alpha 1, FAS gene expression, the phosphorylation level of ACC protein increased significantly in feeding corticosterone, AMPK total protein and phosphorylation levels increased significantly at the same time. Thus, the stress caused by the weight of broilers decreased gluconeogenesis process of glycogen enhanced; stress response by increasing the feeding of neuropeptide NPY gene expression promotes appetite, while activation of the central pathway of LKB1-AMPK-ACC. The effects of central injection of Compound C stress on appetite and energy metabolism. Similar weight 48 AA broiler chickens were selected, 7 days of age were randomly divided into ventricle buried tube. For the 4 test groups (6 replicates of /2 chickens): DEX+Compound C group, DEX group, Compound group, C group,.8 days morning 8:00 DEX treatment group subcutaneous injection of dexamethasone group, saline control group injected dose, continuous injection of 3 days.10 days After the injection of fasting for 3 hours, Compound C Compound C injection group, control group were injected with the same dose of DMSO, feed intake for 30 minutes once every 120 minutes. For statistics, statistical data according to the intake, repeat the test, intracerebroventricular injection of Compound C after 1 hours of fasting, collecting samples. The results showed that DEX significantly decreased the broilers at the age of 8 days and 9 days of age, weight gain; compared with the control group, dexamethasone treatment significantly increased the chicken hypothalamus AMPK alpha 2, LKB1, NPY and M Expression of TOR gene, but later the central injection of Compound C significantly decreased the expression of hypothalamic mTOR gene in broilers; compared with the control group, intracerebroventricular injection of Compound C decreased significantly the stress state of chicken hypothalamus AMPK proteins and their phosphorylation level; compared with the control group, dexamethasone treatment significantly reduced the phosphorylation level of mTOR protein, the late central injection of Compound C also significantly reduced Low level of mTOR protein phosphorylation. Therefore, stress the central injection of Compound C significantly reduced food intake continued to 1 hours; stress activates the hypothalamic AMPK, but later the central injection of Compound C significantly inhibited the activity of AMPK, and through the AMPK-mTOR signaling pathway of body energy metabolism. Effect of dietary energy level on broiler appetite and the central AMPK. The 1 day old male AA broiler 216 rats were randomly divided into 3 treatment groups (6 replicates of /12 chickens): low energy diet group (2900 kcal/kg), normal diet group (3200 kcal/kg), high energy diet group (3500 kcal/kg) were fed to 21 days; the age of each repeat 2 collecting plasma and hypothalamus samples. Experimental results show that the low energy diet group average daily feed intake was significantly higher than that of the other two groups, high energy diet group average daily gain was significantly higher than that of the other two groups; compared with the control group, high energy diets and low energy diets The concentration of UREA in plasma was significantly increased, which was the highest in low energy diets; compared with high energy diets, low energy diet treatment significantly increased the content of TG in broilers in the pulp; in addition, high energy diet and low energy diets were significantly decreased in the 21 day old broiler spleen index; high energy diets treatment significantly reduced broilers hypothalamus LKB1, NPY, CCK, GR gene expression; compared with the control group, low energy diets of broilers significantly increased hypothalamic AMPK alpha 2 gene expression; compared with high energy diets, low energy diets significantly increased 21 days old broilers hypothalamus AMPK alpha 1 gene expression. In summary, feeding high energy diets of broilers, central AMPK signal pathway was inhibited, NPY expression decreased; feeding in low energy diets, activation of central AMPK, the energy balance regulation of the AMPK signaling pathway. Different dietary energy levels of stress in broilers appetite and AMPK signaling pathway. Select the similar weight ring. 1 day old male AA broiler 216 rats were randomly divided into 6 treatment groups (3 replicates /12): low energy diets *DEX normal diet group, *DEX group, *DEX group and high energy diets; low energy diet *Con group, normal diet group *Con, high energy diets *Con group.14 days and 28 days of age at 8:00 DEX group subcutaneous injection of dexamethasone and saline control group injected dose, continuous injection of 3 days. The test results showed that the relative growth peak, no matter what the fed diets, glucocorticoid treatment significantly increased hypothalamic LKB1 AMPK broilers, alpha 1 and GR gene expression in the absolute growth peak of glucocorticoid to normal diet group and high-energy diet group broilers hypothalamus LKB1, NPY gene expression increased significantly; at the same time, DEX treatment significantly reduced to broiler diet group hypothalamic ACC total protein and phosphorylation level, significantly improve the normal diet of broiler chickens hypothalamus AMPK The phosphorylated protein level. To sum up, the relative growth peak, the effect of DEX on LKB1-AMPK signaling pathway is not affected by the type of diet, promoting the expression of neuropeptides affected by food diets; the absolute growth peak of DEX on LKB1-AMPK-ACC signaling pathway, promoting effect of food neuropeptides are influenced by the type of diet. In summary, stress effect of broilers appetite, and activation of central LKB1-AMPK-ACC signal pathway; central AMPK signaling pathway is indeed involved in the regulation of stress on poultry appetite and energy metabolism. The peripheral GCs makes the whole growth stage of appetite are influenced by the type of diet, and the changes of AMPK signal pathway depends on the growth stage.

【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S831

【参考文献】