發(fā)布時間：2018-03-11 07:17

  本文選題：酸棗仁皂苷　切入點(diǎn)：肉雞　出處：《遼寧醫(yī)學(xué)院》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：目的通過研究酸棗仁皂苷對下丘腦原代神經(jīng)元細(xì)胞采食調(diào)控因子表達(dá)的影響,探明酸棗仁皂苷調(diào)節(jié)采食量的作用途徑。揭示肉雞飼糧中添加酸棗仁皂苷改善生產(chǎn)性能的作用機(jī)制。方法本實(shí)驗(yàn)所需種蛋和孵化條件由遼寧醫(yī)學(xué)院家禽養(yǎng)殖團(tuán)隊(duì)提供。提取低胚齡雞胚的下丘腦組織,分離神經(jīng)元細(xì)胞培養(yǎng)并鑒定。取發(fā)育良好的下丘腦神經(jīng)細(xì)胞,分別在培養(yǎng)液中添加不同濃度的酸棗仁皂苷、HSP70蛋白和CRH蛋白作為處理因子,在40.5℃高溫下培養(yǎng),用Real-time PCR和Western Blot的方法檢測下丘腦神經(jīng)細(xì)胞中NPY、Ag Rp、GABA A-α1、MC4R、POMC、CRH和HSP70基因的m RNA和蛋白表達(dá)量,應(yīng)用SPSS軟件進(jìn)行統(tǒng)計(jì)分析,闡明應(yīng)激對采食量調(diào)節(jié)因子的影響以及酸棗仁皂苷對其的調(diào)節(jié)作用。結(jié)果1.建立雞下丘腦神經(jīng)元細(xì)胞培養(yǎng)方法體系,并經(jīng)鑒定證實(shí)所培養(yǎng)的細(xì)胞為下丘腦神經(jīng)元細(xì)胞。2.經(jīng)酸棗仁處理后NPY、GABA A-α1、MC4R和Ag Rp基因m RNA的相對表達(dá)量極顯著的提高(P0.01),而POMC和CRH基因的m RNA相對表達(dá)量極顯著降低(P0.01)。3.經(jīng)HSP70處理后NPY和GABA A-α1m RNA的相對表達(dá)量極顯著提高(P0.01),MC4R和CRH基因m RNA的相對表達(dá)量極顯著降低(P0.01),Ag Rp m RNA表達(dá)量相對提高(P0.05),POMC m RNA表達(dá)量明顯下降(P0.05)。4.經(jīng)CRH處理后NPY、MC4R、POMC、CRH m RNA的相對表達(dá)量極顯著提高(P0.01),Ag Rp m RNA的相對表達(dá)量極顯著降低(P0.01)。5.用酸棗仁皂苷預(yù)處理后40.5℃培養(yǎng),極顯著低下丘腦神經(jīng)元細(xì)胞NPY、POMC、GABA A-α1m RNA的相對表達(dá)量(P0.01),其他基因MC4R、Ag Rp和HSP70 m RNA的相對表達(dá)量有所提高(P0.01)。6.添加HSP70預(yù)處理后40.5℃培養(yǎng),下丘腦神經(jīng)元細(xì)胞中NPY、POMC基因的m RNA相對表達(dá)量極顯著降低(P0.01),而GABA A-α1基因m RNA相對表達(dá)量有增高的趨勢,Ag Rp、MC4R m RNA相對表達(dá)量與對照間差異不顯著(P0.05)。7.39℃培養(yǎng)的下丘腦細(xì)胞樣本,NPY m RNA相對表達(dá)量與POMC m RNA和HSP70 m RNA相對表達(dá)量正相關(guān)均呈顯著正相關(guān)(r2=0.777,P=0.020;r2=0.806,P=0.0.015);40.5℃培養(yǎng),MC4R m RNA相對表達(dá)量與POMC m RNA相對表達(dá)量呈顯著負(fù)相關(guān)(r2=0.840,P=0.029)。結(jié)論1.獲得雞下丘腦神經(jīng)元原代細(xì)胞培養(yǎng)的成熟技術(shù)。2.酸棗仁皂苷和HSP70蛋白可調(diào)控下丘腦神經(jīng)元原代細(xì)胞采食相關(guān)基因的表達(dá)。3.CRH與40.5℃高溫處理,下丘腦神經(jīng)元細(xì)胞中NPY、POMC、GABA A-α1、MC4R、Ag Rp m RNA等基因的變化趨勢相同,實(shí)驗(yàn)條件下CRH可替代高溫處理。4.酸棗仁皂苷和HSP70均對40.5℃高溫培養(yǎng)的下丘腦神經(jīng)元原代細(xì)胞NPY、POMC、GABA A-α1、MC4R、Ag Rp等基因m RNA的表達(dá)量有恢復(fù)作用。5.NPY m RNA相對表達(dá)與POMC m RNA和HSP70 m RNA相對表達(dá)量呈顯著正相關(guān);MC4R m RNA相對表達(dá)量與POMC m RNA相對表達(dá)量呈顯著負(fù)相關(guān)。
[Abstract]:Objective to study the effect of Zizyphylloside on the expression of intake regulatory factors in primary hypothalamic neurons. The effect of jujube seed saponins on regulating feed intake was explored. The mechanism of adding jujube seed saponin to broiler diet to improve production performance was revealed. Methods the conditions of egg breeding and hatching required in this experiment were obtained from poultry breeding team of Liaoning Medical College. Extraction of hypothalamic tissue from chicken embryos of low embryo age, The hypothalamic nerve cells were cultured at 40.5 鈩，

本文編號：1597122