本文關(guān)鍵詞：中樞AMPK在應(yīng)激肉雞食欲調(diào)控中的作用研究　出處：《山東農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 皮質(zhì)酮 肉雞 食欲 下丘腦 AMPK信號(hào)通路

【摘要】：本課題圍繞著“應(yīng)激-AMPK-食欲”研究主題展開了以下研究,主要是探討AMPK在應(yīng)激肉雞的食欲調(diào)控中的作用。應(yīng)激對(duì)肉仔雞下丘腦食欲及AMPK信號(hào)通路的影響。試驗(yàn)選取1日齡體重相近AA肉仔雞144只,隨機(jī)分為3個(gè)處理(6個(gè)重復(fù)/8只雞)。試驗(yàn)分為對(duì)照組、應(yīng)激組、應(yīng)激抑制組。7日齡進(jìn)行試驗(yàn)處理,處理時(shí)間持續(xù)7天。試驗(yàn)結(jié)果顯示:日糧中添加皮質(zhì)酮顯著降低了14日齡肉仔雞體增重;皮質(zhì)酮處理顯著降低心臟指數(shù)、脾臟指數(shù)和法氏囊指數(shù),相反,顯著提高了肝臟指數(shù)和腹脂率;皮質(zhì)酮組血漿中皮質(zhì)酮、葡萄糖、甘油三酯、肝糖原和胸肌糖原的含量與對(duì)照組相比顯著升高;與對(duì)照組相比,日糧中添加皮質(zhì)酮顯著上調(diào)14日齡肉仔雞下丘腦中mTOR、GR、AMPKα2、NPY、LKB1、AMPKα1、FAS基因表達(dá)量,ACC蛋白的磷酸化水平在飼喂皮質(zhì)酮時(shí)顯著提高,同時(shí)AMPK總蛋白及磷酸化水平也顯著提高。由此可見,應(yīng)激導(dǎo)致肉雞體重降低,糖元異生過程加強(qiáng);應(yīng)激反應(yīng)通過提高促食神經(jīng)肽NPY基因表達(dá)促進(jìn)食欲,同時(shí)激活了中樞LKB1-AMPK-ACC信號(hào)通路。中樞注射Compound C對(duì)應(yīng)激肉雞食欲和能量代謝的影響。試驗(yàn)選取體重相近的AA肉仔雞48只,7日齡進(jìn)行腦室埋管,隨機(jī)分為4個(gè)試驗(yàn)組(6個(gè)重復(fù)/2只雞):DEX+Compound C組、DEX組、Compound C組、對(duì)照組。8日齡早上8:00 DEX處理組皮下注射地塞米松,對(duì)照組注射等劑量的生理鹽水,連續(xù)注射3天。10日齡注射后空腹3小時(shí),Compound C組注射Compound C、對(duì)照組注射等劑量的DMSO,每隔30分鐘統(tǒng)計(jì)一次采食量,連續(xù)統(tǒng)計(jì)120分鐘。根據(jù)采食量數(shù)據(jù),重復(fù)上述試驗(yàn),腦室注射Compound C后繼續(xù)空腹1小時(shí),采集樣品。試驗(yàn)結(jié)果顯示:DEX顯著降低了肉仔雞8日齡和9日齡日增重;與對(duì)照組相比,地塞米松處理顯著上調(diào)了肉雞下丘腦AMPKα2、LKB1、NPY以及m TOR基因表達(dá),但后期中樞注射Compound C明顯降低肉仔雞下丘腦mTOR基因表達(dá);與對(duì)照組相比,腦室注射Compound C顯著降低了應(yīng)激狀態(tài)下肉雞下丘腦AMPK總蛋白及磷酸化水平水平;與對(duì)照組相比,地塞米松處理顯著降低了mTOR蛋白的磷酸化水平,后期中樞注射Compound C同樣明顯降低mTOR蛋白的磷酸化水平。由此可知,應(yīng)激狀態(tài)下中樞注射Compound C明顯的降低采食量持續(xù)到1小時(shí);應(yīng)激激活下丘腦AMPK,但后期中樞注射Compound C明顯抑制AMPK活性,并且通過AMPK-mTOR信號(hào)通路調(diào)控機(jī)體能量代謝。日糧不同能量水平對(duì)肉雞食欲及中樞AMPK的影響。選取1日齡的雄性AA肉仔雞216只,隨機(jī)分為3個(gè)處理組(6個(gè)重復(fù)/12只雞):低能日糧組(2900 kcal/kg)、正常日糧組(3200 kcal/kg)、高能日糧組(3500 kcal/kg);飼喂至21日齡各個(gè)重復(fù)取2只采集血漿及下丘腦樣品。試驗(yàn)結(jié)果顯示:低能日糧組平均日采食量明顯高于其他兩組,高能日糧組平均日增重明顯高于其他兩組;與對(duì)照組相比,高能日糧組和低能日糧組血漿中UREA的含量明顯升高,其中低能日糧組含量最高;與高能日糧組相比,低能日糧處理顯著升高了肉仔雞血漿中TG含量;此外,高能日糧和低能日糧均顯著降低了21日齡肉仔雞脾臟指數(shù);高能日糧處理顯著下調(diào)肉仔雞下丘腦LKB1、NPY、CCK、GR基因表達(dá);與對(duì)照組相比,低能日糧顯著上調(diào)肉仔雞下丘腦AMPKα2基因表達(dá);與高能日糧組相比時(shí),低能日糧處理顯著上調(diào)21日齡肉仔雞下丘腦AMPKα1基因表達(dá)。綜上所述,飼喂高能日糧時(shí),肉雞中樞AMPK信號(hào)通路被抑制,NPY表達(dá)量下降;飼喂低能日糧時(shí),激活中樞AMPK,通過AMPK信號(hào)通路調(diào)控機(jī)體能量平衡。日糧不同能量水平對(duì)應(yīng)激肉雞食欲及AMPK信號(hào)通路的影響。選取體重相近1日齡雄性AA肉仔雞216只,隨機(jī)分為6個(gè)處理組(3個(gè)重復(fù)/12只):低能日糧*DEX組,正常日糧*DEX組,高能日糧*DEX組;低能日糧*Con組,正常日糧*Con組,高能日糧*Con組。14日齡與28日齡早上8:00 DEX組皮下注射地塞米松,對(duì)照組注射等劑量的生理鹽水,連續(xù)注射3天。試驗(yàn)結(jié)果顯示:在相對(duì)生長(zhǎng)高峰期,無論飼喂何種日糧,糖皮質(zhì)激素處理均顯著提高肉雞下丘腦LKB1、AMPKα1和GR基因表達(dá)量。在絕對(duì)生長(zhǎng)高峰期,糖皮質(zhì)激素使正常日糧組和高能日糧組肉雞下丘腦LKB1、NPY基因表達(dá)量顯著升高;同時(shí),DEX處理顯著降低能日糧組肉雞下丘腦ACC總蛋白及磷酸化水平,顯著提高正常日糧組肉雞下丘腦AMPK磷酸化蛋白水平。綜上所述,相對(duì)生長(zhǎng)高峰期,DEX對(duì)LKB1-AMPK信號(hào)通路的影響不受日糧類型的影響,促食神經(jīng)肽的表達(dá)受日糧類型的影響;絕對(duì)生長(zhǎng)高峰期,DEX對(duì)LKB1-AMPK-ACC信號(hào)通路、促食神經(jīng)肽的影響均受日糧類型的影響。綜上所述,應(yīng)激顯著影響肉仔雞食欲,并且激活中樞LKB1-AMPK-ACC信號(hào)通路;中樞AMPK信號(hào)通路確實(shí)參與了應(yīng)激對(duì)家禽食欲及能量代謝的調(diào)控。外周GCs使整個(gè)生長(zhǎng)階段食欲均受日糧類型的影響,而中樞AMPK信號(hào)通路的變化依賴于生長(zhǎng)階段。
[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.

【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S831

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本文編號(hào)：1374327