Effects of Dietary Lysine on Growth Performance,Serum Concentrations of Insulin-Like Growth Factor-I （IGF-I） and IGF-I mRNA Expression in Growing Rabbits

The effects of dietary lysine on production performance,serum concentrations of metabolites,growth hormone （GH）,insulin-like growth factor-I （IGF-I） and IGF-I mRNA expression in growing rabbits were examined.One hundred weaned New Zealand rabbits were allocated to individual cages and randomly offered a diet containing 5.5 （L1）,6.5 （L2）,7.5 （L3）,8.5 （L4）,or 9.5 g （L5） lysine per kg diet.The results showed that the average daily gain （ADG） of the rabbits from L3,L4 or L5 was higher than those from L1 or L2 （P 〈 0.05）.The feed gain ratio （F/G） in the rabbits from L4 or L5 was lower than those from L1 or L2 （P 〈 0.05）.Dietary lysine did not affect serum concentrations of total protein （TP）,glucose,insulin （INS）,and growth hormone （GH） （P 〉 0.05）.The quadratic effects of lysine on the serum urea nitrogen （SUN） concentration was detected （P = 0.035）.Serum IGF-I concentrations had a trend to increase quadratically with the increasing dietary lysine （P = 0.07）.A significant correlation was found between serum IGF-I concentrations （x,ng mL-1） and ADG （y,g kg-1）： y = -0.017x2 ＋ 1.984x ＋ 20.87 （R2 = 0.8982,P = 0.003）.The relative abundance of hepatic and muscular IGF-I mRNA tended to increase with increasing dietary lysine levels （P = 0.053 and 0.082,respectively）.Providing the diets mainly consisted of corn,wheat bran and peanut vine,the most appropriate dietary lysine level for growing meat rabbits from weaning to 70 d old was found to be 8.5 g kg-1,and IGF-I may be an important factor controlling growth of weaned rabbits.     

绵羊肌肉IGF-I.基因表达的发育性变化研究

生长激素（GH）在动物体内主要有两种作用方式：（1）GH与肝脏等靶器官上的GHR结合,产生胰岛素样生长因子（IGF）,随后IGF以内分泌方式进入血液,与胰岛素样生长因子结合蛋白（IGFBP）结合后到达靶器官,作用于胰岛素样生长因子受体（IGFR）,从而调节动物生长发育。②GH直接作用于靶器官GHR,通过旁分泌或自分泌IGF直接影响细胞的代谢,从而调节机体的生长发育。GH可作用于骨骼肌GHR,产生旁分泌或自分泌的IGF-I调节肌肉的生长发育,或直接影响细胞的代谢。笔者采用实时定量PCR法,以处于生长发育早期的雄性哈萨克羊和雄性新疆细毛羊为研究对象,检测了IGF-I基因在肌肉中表达情况。     

庄河大骨鸡IGF-Ⅰ基因多样性与产肉性能关联的分析研究

为了探究庄河大骨鸡IGF-Ⅰ基因的多样性与产肉性能相关的遗传标记。以庄河大骨鸡为试验材料,利用PCR-RFLP分子遗传标记技术分析了IGF-Ⅰ基因5′非编码区域的多样性与产肉性能的关系。结果表明：该基因存在AA、AB、BB 3种基因型,基因型的频率分别为0.445、0.300、0.255,A基因和B基因频率分为0.595和0.405;BB型个体的活体重、屠体重、半净膛重、全净膛重、腿肌重、平均日增重和料肉比分别显著的高于AA型和AB型(P<0.05),其他性状差异不显著(P>0.05);AB型和AA型各性状间不差异(P>0.05)。IGF-Ⅰ基因的BB型对鸡的产肉性能有显著影响,但是,其是否可以作为庄河大骨鸡产肉性能的重要遗传标记还需做进一步研究。     

Serum and intratumoural GH and IGF-I concentrations: Prognostic factors in the outcome of canine mammary cancer

The biological implication of the growth hormone/insulin like growth factor-I (GH/IGF-I) axis in canine mammary tumours (CMT) has been recently demonstrated, however its clinical and prognostic implications are unknown. Our aim was to investigate its prognostic significance.Hormonal determinations were done by enzyme immunoassays techniques validated for canine species in serum and tumour tissue from 32 bitches with CMT and in serum and normal mammary tissue from 10 controls. Serum and tissular GH and IGF-I concentrations were significantly higher in the case of malignant tumour compared with benign and controls. GH and IGF-I elevated concentrations were significantly associated with tumour relapse and/or metastases during follow-up and in dogs with reduced survival times; however these parameters were not independent prognostic factors in multivariate analysis. This association demonstrates a link between high serum and intratumoural GH and IGF-I concentrations and a worse prognosis and opens the possibility to new anticancer endocrine therapies in dogs.     

鸡IGF-I基因5'非翻译区Trul Ⅰ、Tai Ⅰ多态位点的发现

研究采用测序和PCR-RFLP技术对京海黄鸡、AA鸡和尤溪麻鸡IGF-I基因5'非翻译区(EF488284)进行多态性检测.结果表明:在5'非翻译区共发生2个SNP突变位点.43 bp位置发生A→G的突变导致Turl Ⅰ酶切位点发生改变,67 bp位置发生A→G的突变导致Tai Ⅰ酶切位点发生改变.PCR-RFLP结果显示2个突变位点完全连锁,产生AA/CC、AB/CD和BB/DD 3种基因型.等位基因和基因型在3个品种中分布不一致,AA鸡为纯合型(AA/CC),在京海黄鸡和尤溪麻鸡中出现3种基因型,可以作为新的标记住点进一步研究.     

朗德鹅肌肉生成抑制因子基因（MSTN）的克隆及其表达量与日粮能量、血清IGF-I和GH的关系

 【目的】采用反转录－聚合酶链反应（RT-PCR）技术,克隆朗德鹅肌肉生成抑制因子基因（MSTN）,研究MSTN基因表达和日粮能量及其部分血清激素含量的相互关系,了解MSTN的功能,为水禽分子营养和分子育种提供基础资料。【方法】从朗德鹅（Anser anser）的腿肌中抽提总RNA,用两步法RT-PCR扩增出MSTN基因的cDNA编码序列,以pGEM-T Vector为载体,将该片段克隆到大肠杆菌（Escherichia coli）中。通过筛选阳性克隆,双酶切鉴定后测序;以MSTN基因的克隆为基础,以β-actin为内标,构建优化的半定量RT-PCR方法,研究高中低（A:13.38MJ•kg-1、B:12.13MJ•kg-1、C:10.87MJ•kg-1）3种不同能量对朗德鹅21日龄和70日龄2个时期,肌肉组织MSTN基因表达的差异;同时用放免法测定21、70日龄的血清GH和IGF-I浓度。【结果】克隆朗德鹅MSTN基因cDNA的部分序列,其片段大小为1 128bp,编码375个氨基酸组成的多肽,与已发表的鸡、鸭、鹅的MSTN 核苷酸相似性分别为94%、94%、99%;氨基酸的相似性分别为98%、97%、98%;21日龄时,朗德鹅公鹅、母鹅MSTN表达量差异不显著;70日龄时朗德鹅公鹅MSTN的表达量情况为能量A＞C＞B,朗德鹅母鹅MSTN的表达量情况为C＞B＞A;对于朗德鹅21～70日龄阶段的生长,MSTN基因的表达量和血清IGF-I的变化基本一致;与血清GH含量之间并不存在很大关联,GH和能量的高低呈正相关。【结论】日粮能量对21日龄后朗德鹅MSTN基因的表达有影响,且MSTN基因表达量和血清IGF-I具有相关性,和血清GH无较大相关性。     

Role of ghrelin in regulating rabbit ovarian function and the response to LH and IGF-I

The aim of these in vivo and in vitro studies was to examine the role of ghrelin in the control of plasma hormone concentrations, the proliferation, apoptosis and secretory activity of ovarian granulosa cells and the response of these cells to hormonal treatments. Female rabbits were injected with ghrelin (10 μg/animal/day for one week before ovulation induced by 25 IU PMSG and 0.25 IU LHRH). On the day of ovulation, blood samples were collected and analyzed for concentrations of progesterone (P₄), testosterone (T), estradiol (E₂), estrone-sulphate (ES), insulin-like growth factor I (IGF-I) and leptin (L) by RIA. Some control and ghrelin-treated animals were killed in the periovulatory period, their ovaries were weighed and granulosa cells were isolated and cultured for 2 d. Cell proliferation (expression of PCNA) and apoptosis (expression of TdT) were evaluated by immunocytochemistry and TUNEL respectively. Secretion of P₄, T, E₂, IGF-I, and prostaglandin F (PGF) by granulosa cells cultured with and without LH or IGF-I (1, 10 or 100 ng/ml medium) was assessed by RIA. The remaining control and treated animals were kept until parturition, while the number, viability and body weight of pups were recorded.     

半胱胺和大豆黄酮对东北细毛羊部分组织IGF-I mRNA表达量的影响

为了探讨半胱胺和大豆黄酮对东北细毛羊肝脏、肌肉和皮肤组织中IGF-I mRNA表达量的影响,选取体重27～28 kg,5～6月龄健康的东北细毛羊育成羊21只,随机分成7组,每组3个重复,每个重复1只,采用实时荧光定量PCR法检测肝脏、肌肉和皮肤组织中IGF-I mRNA相对表达量.结果表明:(1)半胱胺和大豆黄酮均能显著提高东北细毛羊的生长性能(P<0.05),其中以7.5 mg/kg BW半胱胺组和7.5 mg/kg BW半胱胺+3 mg/kg BW大豆黄酮组效果较佳;(2)半胱胺和大豆黄酮对东北细毛羊肝脏和肌肉中IGF-I mRNA表达量有显著的提高作用(P<0.05),但各试验组之间差异不显著(P>0.05);(3)对皮肤中表达量的影响:对照组与添加3和6 mg/kg BW大豆黄酮组和7.5 mg/kg BW半胱胺+3 mg/kg BW大豆黄酮组之间差异显著(P<0.05),与添加7.5和15 mg/kg BW半胱胺组和15 mg/kg BW半胱胺+6 mg/kg BW大豆黄酬组之间差异不显著(P>0.05).由结果可知,半胱胺和大豆黄酮均能提高肝脏和肌肉中IGF-I mRNA的表达量,但协同应用并没有促使IGF-I mRNA表达量在肝脏、肌肉和皮肤组织中的明显提高,综合分析以添加3 mg/kg BW大豆黄唰效果最佳.     

酶制剂对雏鹅组织中胰岛素样生长因子的影响

18只 7日龄的杂交雏鹅 ,随机分成试验和对照两组 ,饲以大麦基础日粮 ,试验组日粮中添加 0 .1%的Biokyowa酶制剂。测定 2 1日龄血液、肝脏和肌肉中的IGF -I含量。添加粗酶制剂后 ,肝脏和肌肉中的IGF -Ⅰ分别较对照高18.2 3% (P <0 .0 1)和 2 2 .79% (P <0 .0 1) ,而血浆中IGF -I水平却较对照低 9.0 4 % (P <0 .0 5 )。结果提示 :酶制剂能促使肝脏合成IGF -I,血液中与IGFBP结合的IGF -I大量释放 ,从而显著提高肌肉中的IGF -I水平 ,加速了肌肉的生长。