本文關(guān)鍵詞：日糧硒缺乏對肉雞硒蛋白基因及胰島素信號相關(guān)基因表達(dá)的影響　出處：《四川農(nóng)業(yè)大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 肉雞 硒 硒蛋白基因 胰島素信號 表達(dá)

【摘要】：胰腺是肉雞硒缺乏最為敏感的器官,長期飼喂肉雞缺硒日糧可導(dǎo)致胰腺病變萎縮,意味著硒在胰腺功能維持中起著重要作用,但日糧缺硒對肉雞胰島素代謝是否有影響尚不清楚。胰腺是分泌胰島素的器官,肝臟和肌肉是兩個(gè)重要的胰島素效應(yīng)靶組織,硒主要通過硒蛋白在機(jī)體組織中發(fā)揮作用,然而日糧硒缺乏對肉雞上述組織中硒蛋白基因和胰島素代謝相關(guān)基因表達(dá)是否有影響尚不清楚。本試驗(yàn)以肉雞為研究材料,研究日糧硒缺乏對肉雞生產(chǎn)性能,生理生化指標(biāo)、組織硒蛋白基因和胰島素信號相關(guān)基因表達(dá)的影響,試驗(yàn)結(jié)果可為探索硒在胰島素代謝中的生物學(xué)機(jī)制提供線索。本試驗(yàn)采用四川嚴(yán)重缺硒地區(qū)(漢源山區(qū))生長的大豆、玉米為原料,配制基礎(chǔ)日糧(Se0.02mg Se/Kg),進(jìn)而研究日糧硒缺乏對肉雞胰島素合成與作用靶組織中硒蛋白基因表達(dá)的影響。試驗(yàn)采用120只一日齡健康科寶肉雞,隨機(jī)分成2個(gè)處理組(n=60)。分別飼喂缺硒日糧(Se, 0mg/kg)和對照日糧(Se,0.3 mg/kg),試驗(yàn)期5周。每周稱量肉雞體重和計(jì)算生長性能,并記錄滲出性素質(zhì)和其他典型硒缺乏癥。于試驗(yàn)?zāi)┢诘?5d,每個(gè)處理選取6只雞屠宰,采集血液、肝臟、胰腺和肌肉,用于測定硒含量、抗氧化性能、生化指標(biāo)測定,同時(shí)考察日糧硒缺乏對肝臟、脂肪和胰腺組織23各硒蛋白基因及17個(gè)胰島素信號相關(guān)基因表達(dá)量影響。實(shí)驗(yàn)結(jié)果如下：1.日糧硒缺乏對肉雞病理、生長性能及組織硒含量的影響日糧硒缺乏導(dǎo)致肉雞出現(xiàn)滲出性素質(zhì)、胰腺萎縮和其它硒缺乏癥,并伴隨高發(fā)病率(90%)和死亡率(62%)。同時(shí)日糧硒缺乏降低了肉雞生長性能,從14日齡到35日齡平均體重與對照組相比顯著降低(P0.05)。同對照組相比,缺硒組血清和肝臟中的硒含量降低了80%和73%(P0.01)。2.日糧硒缺乏對肉雞生理生化指標(biāo)的影響日糧硒缺乏導(dǎo)致肉雞血清葡萄糖濃度、白介素6(IL-6)和腫瘤壞死因子a(TNF-a)含量分別升高了233.3%、40%和23%(P0.05),血清胰島素濃度、甘油三酯(TG)和總膽固醇(TCH)含量分別降低了32%、61%和45%(P0.05)。此外,缺硒組肉雞血清谷胱甘肽過氧化物酶(GSH-PX)活性和總抗氧化能力(T-AOC)與對照日糧組相比降低了76%和57%(P0.05)；另外肝臟組織GSH-Px活性和超氧化物歧化酶(SOD)還降低了66%和24%(P0.05),丙二醛(MDA)含量升高了63%(P0.05)。在肌肉組織中僅有GSH-Px活性受到缺硒影響而降低了48%(P0.05);胰腺組織中GSH-Px和SOD活性受缺硒影響降低了32%(P0.05)。3.日糧硒缺乏對肉雞組織硒蛋白基因表達(dá)影響與對照組相比,日糧硒缺乏下調(diào)了(P0.05)肝臟組織中9個(gè)硒蛋白基因(Gpx1、 Gpx2、Gpx3、Gpx4、Diol、Seli、Selm、Selt和Selu的表達(dá)豐度),其中除Gpxl和Selt這2個(gè)基因?yàn)椴町愶@著外(P0.05),其余7個(gè)基因均為差異極顯著(P0.01),同時(shí),日糧缺硒還上調(diào)了肝臟組織中2個(gè)硒蛋白基因Txnrd1和SeIs的表達(dá)(P0.05)。日糧缺硒下調(diào)了肉雞肌肉組織中14個(gè)硒蛋白基因(Gpx2、Gpx3、Dio1、Dio3、Txnrd1、 Selk、Seli、Selm、Selo、Sels、Selt、Sepn1、Sepp1和Selx)的表達(dá)豐度(P0.05)。相比肝臟和肌肉組織,胰腺是受日糧缺硒影響改變硒蛋白基因表達(dá)量最多的組織,下調(diào)了18個(gè)硒蛋白基因(Gpx2、Gpx3、Gpx4、Txnrd1、Txnrd3、Selk、Seli、Selm、Selo、 Sels、Selu、Selt、Sep15、Sepn1、Sepp1、Sepw1、Selx和Sephs2)的表達(dá)豐度(P0.05),其中只有Txnrd1基因表現(xiàn)為顯著下調(diào)(P0.05),其余的17個(gè)基因均表現(xiàn)為極顯著下調(diào)(P0.01)。4.日糧硒缺乏對肉雞組織胰島素信號相關(guān)基因表達(dá)影響與對照組相比,日糧缺硒顯著下調(diào)了肉雞肝臟組織中2個(gè)胰島素信號相關(guān)蛋白基因Ins和Ptpn1的表達(dá)(P0.05)：極顯著地下調(diào)了Irs2、Pdx1和Slc2A2這3個(gè)基因的表達(dá)(P0.01),其中Irs2基因的表達(dá)量下調(diào)最大。同時(shí),日糧缺硒極顯著上調(diào)了3個(gè)基因Foxa2、Gcg和Irsl的表達(dá)(P0.01),其中Irsl基因表達(dá)量上調(diào)最高。日糧硒缺乏下調(diào)了肌肉組織中14個(gè)胰島信號相關(guān)基因表達(dá),其中4個(gè)基因(Irs1、Ins、 Neurod1和Slc2A2)為顯著下調(diào)(P0.05)；Akt1、Braf、Foxo1、Foxa2、Ir、Irs2、 Ptpn1、Pi3k和Ucp等9個(gè)基因?yàn)闃O顯著下調(diào)(P0.01)。日糧缺硒使胰腺組織中的6個(gè)胰島素信號相關(guān)基因表達(dá)下調(diào),其中Akt1、Foxa2、Enf1A和Hnf4A為極顯著下調(diào)(P0.01)。結(jié)論：日糧硒缺乏降低肉雞的生長性能,減少肉雞血清和肝臟中硒沉積,影響了肉雞血清和胰島素代謝相關(guān)組織的抗氧化活性。同時(shí),缺硒還導(dǎo)致肉雞出現(xiàn)低胰島素和高血糖癥,影響血液中甘油三酯和膽固醇的正常代謝。RT-PCR結(jié)果分析表明：日糧缺硒不同程度的影響肉雞肝臟、肌肉和胰腺組織中胰島素信號通路相關(guān)基因的表達(dá),在這三個(gè)組織中,肌肉組織中受影響基因最多,其次為肝臟和胰腺組織,多數(shù)受影響的基因表現(xiàn)為下調(diào)表達(dá)。與此同時(shí)三個(gè)組織中硒蛋白基因的普遍下調(diào)表達(dá),其中胰腺中下調(diào)的硒蛋白基因最多,其次為肌肉組織和肝臟組織,在三個(gè)組織中,只有肝臟中上調(diào)的硒蛋白基因。上述研究結(jié)果表明,日糧硒缺乏影響肉雞胰島素代謝相關(guān)組織中硒蛋白基因與胰島素信號相關(guān)基因的表達(dá),導(dǎo)致肉雞血脂代謝異常,出現(xiàn)低胰島素血癥和高葡萄糖血癥,這預(yù)示著受影響的硒蛋白基因在維持胰島素合成和正常代謝中發(fā)揮著重要作用。
[Abstract]:Chicken pancreas is the most sensitive organ selenium deficiency, long-term feeding broiler diets deficient in selenium can lead to pathological changes in the pancreas atrophy, mean Se in pancreatic function plays an important role in the maintenance of dietary selenium deficiency, but whether the effect of insulin metabolism in broilers is unclear. Pancreatic insulin secretion is the organ, liver and muscle is the two important effect of insulin target tissue selenium mainly through selenoproteins in the tissue play a role, but the lack of dietary selenium on selenoprotein gene in the tissues of the broiler and insulin metabolism related gene expression has any effect is not clear. In this experiment, broilers as research materials, research of dietary selenium deficiency on broiler performance. Physiological and biochemical indexes, the effects of expression of genes related to selenoprotein gene and insulin signaling, the test results may provide clues for exploring the biological mechanism of selenium in insulin metabolism in this experiment. The Sichuan severe selenium deficiency (Hanyuan mountain) the growth of soybean, corn as raw materials, preparation of the basal diet (Se0.02mg Se/Kg), and study the dietary effects of selenium deficiency on the expression of selenoprotein insulin synthesis and action of chicken genes in target tissues. In this experiment, 120 one day old healthy Kebao broiler were randomly divided into 2 treatments group (n=60). With selenium deficiency diet (Se, 0mg/kg) and control diet (Se, 0.3 mg/kg), the experimental period was 5 weeks. The weekly weighing weight and calculating the growth performance of broilers, and record the exudative diathesis and other typical selenium deficiency. At the end of the test section 35d, each select 6 chicken slaughter, blood, liver, pancreas and muscle, for the determination of selenium content, antioxidant properties, biochemical indexes, and the effects of dietary selenium deficiency on the liver, fat and 23 of the selenoprotein gene in pancreatic tissue and 17 insulin signal related gene expression effect. The results are as follows: 1. dietary selenium deficiency on broiler pathological effects of dietary selenium deficiency, growth performance and tissue selenium content resulting in broiler appear exudative diathesis, pancreatic atrophy and other selenium deficiency, and with high incidence rate (90%) and mortality (62%). At the same time, the lack of dietary selenium reduced growth of broiler the performance, from the age of 14 days to 35 days of age, average weight significantly reduced compared with the control group (P0.05). Compared with the control group, the content of selenium in selenium deficiency group in serum and liver were reduced by 80% and 73% (P0.01).2. dietary selenium deficiency on physiological and biochemical indexes of broiler dietary selenium deficiency in Broiler Serum the concentration of glucose and interleukin 6 (IL-6) and tumor necrosis factor A (TNF-a) content were increased by 233.3%, 40% and 23% (P0.05), serum insulin, triglyceride (TG) and total cholesterol (TCH) decreased by 32%, 61% and 45% (P0.05). In addition, the serum selenium deficiency group glutathione 鐢樿偨榪囨哀鍖栫墿閰，

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