外源性三碘甲腺原氨酸对金定鸭出雏前后生长及肝脏中脱碘酶表达的影响

发布时间：2018-04-13 20:05

  本文选题：鸭 + T3　； 参考：《中国农业科学院》2015年硕士论文

【摘要】：胚胎期骨骼肌的发生决定鸭产肉量,比较发现生长速度差异较大两品种鸭胚胎期生长速度不同,且生长速度较快品种鸭蛋清内三碘甲腺原氨酸(T3)含量显著高于后者。假设蛋清内T3含量与鸭生长速度呈正相关,增加蛋清内T3含量会促进鸭生长。为验证假说,设计蛋清内注射T3试验,监测鸭出雏前后生长、血清中甲状腺激素(thyroid hormones,THs)含量及THs稳态的主要调控因子--肝脏中脱碘酶(deiodinases)的表达情况;设计外源性T3干扰鸭胚原代肝细胞生长试验,检测细胞中脱碘酶的表达。探讨外源性T3对鸭出雏前后生长及肝脏中脱碘酶表达的影响。1蛋清内注射T3对鸭出雏前后生长及血清中THs含量的影响金定鸭种蛋200枚,随机分为两组:对照组蛋清内(Control group,Con)注射100μL不含T3的生理盐水,试验组蛋清内(Treatment group,Tre)注射100μL含250 ng T3的生理盐水,两组种蛋在同一条件下同期孵化。监测27胚龄和7日龄鸭体重和胸肌重的变化,并采集鸭的血液;放射性免疫分析法检测血清中T3和T4的含量;分析鸭出雏前后体重、胸肌重与血清T3、T4含量之间的相关性。结果表明,试验组鸭体重和胸肌重在27胚龄和7日龄均显著高于对照组(P=0.035,0.044;0.027,0.037),试验组血清中T3和T4的含量在7日龄均显著低于对照组(P=0.040,0.040);试验组鸭体重与T3含量呈极显著负线性相关(r=-0.821,P=0.000)。2蛋清内注射T3对鸭出雏前后肝脏中脱碘酶表达的影响试验设计与上一章相同,采集鸭27胚龄和7日龄时的肝脏。实时荧光定量PCR(RT-PCR)检测肝脏中D1和D3的mRNA表达,ELISA法检测肝脏中D1和D3的蛋白含量及酶活性。分析D1和D3的表达与血清T3、T4含量之间的相关性。结果表明,试验组肝脏D3的mRNA表达以及D1、D3的蛋白含量和酶活性在7日龄均显著低于对照组(P=0.049;0.017,0.042;0.013,0.039)。试验组血清中T4含量与D3的mRNA表达呈极显著正线性相关(r=0.913,P=0.001),T3含量与D1和D3蛋白含量及酶活性均呈显著正线性相关(r=0.790,P=0.011;r=0.746,P=0.021;r=0.757,P=0.018;r=0.744,P=0.022)。3外源性T3对鸭胚原代肝细胞中脱碘酶表达的影响分离并培养鸭15胚龄原代肝细胞,24 h后随机分为三组即对照组(Control group,Con)、低浓度T3干扰组(Low T3 group,LT3)和高浓度T3干扰组(High T3 group,HT3)。Con组细胞液中导入2μL不含T3的生理盐水;LT3和HT3组细胞液中分别导入终浓度为5 ng/mL和20 ng/mL的T3,均以2μL生理盐水为载体。干扰36 h后细胞换液并继续培养12 h。在干扰后0 h和12 h时收集细胞并用Cell Counting Kit(CCK-8)法检测细胞活性,RT-PCR检测细胞中D1和D3的mRNA表达,ELISA检测细胞D1和D3的蛋白含量及酶活性。结果显示,LT3组细胞活性及D1的mRNA表达在干扰后的0 h和12 h均显著高于对照组(P=0.026,0.006;0.000,0.000)。全文结论:蛋清内T3含量的差异与鸭的早期生长有一定的相关性,外源性T3(250 ng)促进了鸭出雏前后体重和胸肌重的增长,降低了鸭出雏后血清中T3和T4的含量;鸭出雏前后生长模式发生改变,伴随着出雏前D3 mRNA表达的下调及出雏后D1和D3的蛋白含量及酶活性的降低;外源性T3干扰鸭胚原代肝细胞生长试验验证了D1对T3的直接调控作用。
[Abstract]:The amount of meat duck embryo determining skeletal muscle, that growth rate differences between the two varieties of duck embryo growth rate and faster growth of different varieties of duck's egg white three triiodothyronine (T3) was significantly higher than that of the latter. The content of T3 in duck egg white that growth rate was positively correlated with increased T3 content in egg white will promote growth. In order to verify the hypothesis of duck egg white, design of T3 injected into the test, monitoring duck hatching and growth of thyroid hormone in serum (thyroid, hormones, THs) deiodinase regulating factor of liver and the content of THs in steady state (deiodinases) the expression of exogenous T3 interference; design of duck embryo primary liver cell growth test, expression of deiodinases were detected. To investigate the effect of exogenous T3 on deiodinase expression before and after hatching and growth of duck liver.1 in egg white duck hatching before and after the injection of T3 on growth and serum THs content The influence of Jinding duck eggs 200, were randomly divided into two groups: control group (Control, group, Con in egg white) saline injection 100 L without T3, the test group (Treatment group, Tre in egg white) saline injection 100 L containing 250 ng T3, two groups of eggs in the same a period of incubation. 27 monitoring conditions below and 7 day old duck embryo weight and breast muscle weight changes, and duck blood collection; analysis of the content of T3 and T4 in serum were determined by radioactive immune analysis; duck hatching and weight, breast muscle weight and serum T3, the correlation between T4 content. The results show that the weight the test group 27 and chest on duck embryo age and 7 days of age were significantly higher than those in control group (P=0.035,0.044; 0.027,0.037), the content of T3 and T4 in serum of the experimental group at 7 days of age were significantly lower than the control group (P=0.040,0.040); experimental group duck weight and T3 content showed significant negative correlation (r= -0.821, P=0.000.2 injection of T3 in egg white) Effect on duck deiodinase expression in the liver before and after the young experimental design and the same chapter, collecting duck 27 and 7 day old embryo liver. Real time fluorescence quantitative PCR (RT-PCR) to detect the expression of D1 and D3 in the liver of mRNA, protein content and enzyme activity of D1 and D3 detection in liver by ELISA method the expression and analysis of serum T3. D1 and D3, the correlation between the content of T4. The results show that the experimental group of liver D3 mRNA expression and D1, protein content and enzyme activity of D3 at 7 days of age were significantly lower than the control group (P=0.049; 0.017,0.042; 0.013,0.039). The content of T4 in serum and D3 in test group the expression of mRNA had a significantly positive linear correlation (r=0.913, P=0.001), showed a significant positive linear correlation with T3 content of D1 and D3 protein content and enzyme activity (r=0.790, P=0.011; r=0.746, P=0.021; r=0.757, P=0.018; r=0.744, P=0.022) separation effect of deiodinase expression in primary liver cells.3 exogenous T3 the duck embryo 骞跺煿鍏婚腑15鑳氶緞鍘熶唬鑲濈粏鑳，

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