【摘要】：為探討CB1受體信號通路在應激調控食欲過程中起到的作用,本課題進行了以下研究。本研究通過測定不同日齡家禽各組織中CB1受體的表達量,研究CB1受體的時空分布。選擇體重相近的7,21和35日齡的肉仔雞和7日齡蛋公雛各8只,采集下丘腦,肝臟,十二指腸,空腸,回腸。試驗結果顯示,在7,21和35日齡的肉仔雞和7日齡蛋公雛中,CB1受體在下丘腦中的表達量均遠高于其他外周組織,且下丘腦中CB1受體的表達量隨日齡的增加而顯著增加,在7日齡時,肉仔雞和蛋公雛各組織中CB1受體的表達量并不存在差異。本研究通過熱應激和糖皮質激素DEX處理,研究應激時CB1受體及相關基因表達的變化情況。熱應激試驗:選擇體重相近的28日齡雄性AA肉仔雞144只,分為四組(10h熱應激組,10h對照組,72h熱應激組,72h對照組),每組6個重復,每個重復6只雞。DEX試驗:選擇體重相近的28日齡雄性AA肉仔雞144只,分為四組(24h DEX組、24h對照組,72h DEX組,72h對照組),每組6個重復,每個重復6只雞。試驗結果顯示,應激(熱應激和DEX處理)可導致采食量和體增重的顯著降低,肛溫的顯著升高。熱應激時,肉仔雞下丘腦中GR mRNA表達量顯著增加,AMPK?1mRNA表達量顯著降低,AMPK磷酸化水平降低。DEX處理時,肉仔雞下丘腦中GR、AMPK??、LKB1、CB1受體mRNA的表達量顯著升高,AMPK磷酸化水平升高。說明,糖皮質激素可以激活CB1受體和AMPK信號通路,從而調控了食欲,能量代謝和平衡。本研究通過腦室注射CB1受體激活劑ACEA和拮抗劑SR141716,研究CB1受體對AMPK信號通路的影響。兩個試驗均選擇體重165g左右的7日齡肉仔雞40只進行腦室埋管,恢復至10日齡進行腦室注射,腦室注射SR141716分為DMSO組,0.1,1,2ugSR141716組,腦室注射ACEA分為DMSO組,0.5,1,2ug ACEA組,每半小時統(tǒng)計采食量。根據結果分別選擇1ug SR141716和2ug ACEA為最佳注射劑量,1h和3h為最佳采樣時間,進行重復試驗。試驗結果顯示,腦室注射SR141716使肉仔雞采食量下降,下丘腦中CB1受體、AMPK?2、CART mRNA表達量顯著升高,AgRP、POMC mRNA表達量顯著降低。說明,抑制CB1受體可通過降低促食欲基因AgRP的表達,升高抑食欲基因CART的表達,抑制家禽食欲。腦室注射ACEA可增加肉仔雞的采食量,降低了其下丘腦中CB1受體、POMC mRNA的表達水平,升高了AgRP的mRNA表達水平。說明,激活CB1受體可通過增加促食欲基因AgRP的表達,促進家禽食欲。本研究表明,家禽CB1受體在下丘腦中高度表達,并廣泛分布在肝臟、腸道等外周組織。糖皮質激素可通過激活CB1受體信號通路和AMPK信號通路,調控家禽食欲。CB1受體可通過調控AMPK及其下游的食欲調控因子,調控家禽食欲。
[Abstract]:In order to investigate the role of CB1 receptor signaling pathway in the regulation of appetite by stress, the following studies were carried out. In this study, the temporal and spatial distribution of CB1 receptor was studied by measuring the expression of CB1 receptor in poultry tissues of different ages. Eight broilers of 7, 21 and 35 days old and 8 7-day-old male chicks of 7, 21 and 35 days old were selected to collect hypothalamus, liver, duodenum, Jejunum and ileum. The results showed that the expression of CB1 receptor in Hypothalamus was much higher than that in other peripheral tissues in 7, 35-day-old broilers and 7-day-old egg chicks, and the expression of CB1 receptor in hypothalamus increased significantly with the increase of age. At 7-day-old broilers and egg-male chicks, there was no difference in the expression of CB1 receptors between broilers and egg chicks. In this study, the changes of CB1 receptor and related gene expression during stress were studied by heat stress and glucocorticoid DEX treatment. Heat stress test: 144 28-day-old male AA broilers with similar body weight were divided into four groups (10 h heat stress group, 10 h control group, 72 h heat stress group, 72 h control group), 6 repeats in each group, 6 chickens in each group. Dex test: 144 28-day-old male AA broilers with similar body weight were divided into four groups (24 h DEX group, 24 h control group, 72 h DEX group, 72 h control group), 6 chickens in each group. The results showed that stress (heat stress and DEX treatment) could significantly decrease feed intake and body weight gain, and increase anal temperature. During heat stress, the expression of GR mRNA was significantly increased, the expression of AMPK?1mRNA was significantly decreased, and the phosphorylation level of AMPK was decreased. The expression of GR,AMPK??,LKB1,CB1 receptor mRNA and AMPK phosphorylation were significantly increased and AMPK phosphorylation was increased in the hypothalamus of broilers treated with DEX. It is suggested that glucocorticoids can activate CB1 receptor and AMPK signaling pathway, thus regulating appetite, energy metabolism and balance. In this study, the effect of CB1 receptor on AMPK signaling pathway was studied by intracerebroventricular injection of CB1 receptor activating agent ACEA and antagonist SR141716,. In both experiments, 40 7-day-old broilers weighing about 165 g were selected for intraventricular catheterization and restored to 10 days of age for intraventricular injection. SR141716 was divided into DMSO group, 0.1, 1, 2ugSR141716 groups. Intraventricular injection of ACEA was divided into DMSO group, 0.5 g group and 2ug ACEA group, and the feed intake was counted every half hour. According to the results, 1ug SR141716 and 2ug ACEA were selected as the best injection dose, 1 h and 3 h as the best sampling time, respectively, and the repeated tests were carried out. The results showed that intraventricular injection of SR141716 decreased feed intake, increased the expression of CB1 receptor and AMPK?2,CART mRNA and decreased the expression of AgRP,POMC mRNA in hypothalamus. It is suggested that inhibition of CB1 receptor can inhibit appetite of poultry by decreasing the expression of appetite promoting gene AgRP and increasing the expression of appetite gene CART. Intraventricular injection of ACEA increased feed intake, decreased the expression of CB1 receptor, POMC mRNA and increased the expression of mRNA in AgRP. It is suggested that the activation of CB1 receptor can promote the appetite of poultry by increasing the expression of appetite promoting gene AgRP. This study showed that CB1 receptor was highly expressed in the Hypothalamus and widely distributed in the liver, intestine and other peripheral tissues. Glucocorticoids can regulate poultry appetite by activating CB1 receptor signaling pathway and AMPK signaling pathway. CB1 receptor can regulate poultry appetite by regulating AMPK and its downstream appetite regulators.
【學位授予單位】：山東農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S831

【共引文獻】

相關期刊論文 前4條

1 欒虹;王帥;吳崇明;郭鵬;;激活腺苷酸活化蛋白激酶天然產物對脂代謝調節(jié)作用研究進展[J];國際藥學研究雜志;2014年02期

2 魏鳳仙;胡驍飛;李紹鈺;徐彬;孫全友;林敏;;慢性濕度應激對肉仔雞生產性能及血液生理生化指標的影響[J];河南農業(yè)科學;2013年10期

3 朱雪英;徐倩;馮青;向榮;;HU-308緩釋涂層對大鼠成骨細胞生物學特性的影響[J];實用口腔醫(yī)學雜志;2014年05期

4 曹可;馮智輝;劉健康;;天然AMPK激活劑防治代謝綜合征的研究進展[J];生命科學研究;2014年06期

相關博士學位論文 前8條

1 孫作為;飼養(yǎng)密度和飼糧營養(yǎng)影響肉雞生長、胴體組成及腿部健康的研究[D];中國農業(yè)大學;2013年

2 胡榮寬;天然產物銀線草醇對哺乳動物細胞糖脂代謝的調控作用研究[D];中國科學技術大學;2013年

3 李由然;Bacillus lentus CICIM304異淀粉酶的基因克隆、鑒定與表達[D];江南大學;2013年

4 田萬強;牛AMPK家族7個基因SNP檢測及其與生長和肉質性狀的關聯分析[D];西北農林科技大學;2013年

5 莊靜靜;AMPK及LXR通路新型天然調節(jié)劑的發(fā)現及相應作用機制研究[D];華東理工大學;2013年

6 EL MOUJAHID EL MOSTAFA;候選基因多態(tài)性與肉雞生長，采食量和飼料轉比率的關聯分析[D];中國農業(yè)大學;2014年

7 焦洪超;棉粕日糧對蛋雞生產性能和肝臟脂肪代謝的影響及其機制研究[D];山東農業(yè)大學;2014年

8 劉磊;糖皮質激素在應激家禽食欲調控中的作用機制[D];山東農業(yè)大學;2014年

相關碩士學位論文 前10條

1 張雷;運輸應激和禁食應激對AA肉雞組織結構的影響及相關基因表達的研究[D];東北農業(yè)大學;2013年

2 劉凡;植物甾醇添加劑對日本鵪鶉IGF-1和腎上腺功能的影響[D];南京農業(yè)大學;2011年

3 陳躍平;孵化重和日糧蛋氨酸水平對肉雞生長、免疫及抗氧化機能的影響[D];南京農業(yè)大學;2012年

4 段婧;亞硒酸鈉通過Akt/AMPK/mTOR通路促進NB4細胞凋亡[D];北京協(xié)和醫(yī)學院;2013年

5 魏堯;MSTN基因突變體免疫對小鼠應激性肌損傷的影響[D];山西農業(yè)大學;2013年

6 張巖;肉雞宰前管理初步調查及宰前應激對雞肉品質影響[D];南京農業(yè)大學;2012年

7 駱婧文;皮質酮對肉雞骨骼發(fā)育影響機理的研究[D];南京農業(yè)大學;2013年

8 付文艷;白羽肉雞養(yǎng)殖福利狀況的調查及慢性應激對肉雞骨骼肌生長的影響[D];南京農業(yè)大學;2013年

9 巫慧娟;調控內源性大麻素系統(tǒng)對不同腦膠質瘤細胞的影響研究[D];廈門大學;2014年

10 李亞丹;秀麗隱桿線蟲14-3-3蛋白FTT-2及AMPK蛋白AAK-2對熱抗性調控作用的研究[D];廈門大學;2014年

，

本文編號：2499377