發(fā)布時間：2019-06-14 12:16

【摘要】：為探討CB1受體信號通路在應激調(diào)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(jié)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(zhì)激素DEX處理,研究應激時CB1受體及相關(guān)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(jié)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(zhì)激素可以激活CB1受體和AMPK信號通路,從而調(diào)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(jù)結(jié)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(jié)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(zhì)激素可通過激活CB1受體信號通路和AMPK信號通路,調(diào)控家禽食欲。CB1受體可通過調(diào)控AMPK及其下游的食欲調(diào)控因子,調(diào)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(nóng)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】

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本文編號：2499377