瘦素与奶牛泌乳早期能量平衡和生殖机能的关系

为阐明奶牛血浆瘦素、能量状况和生殖机能之间的关系,随机选取年龄、胎次、泌乳量相近的奶牛60头,分别在产后1、14、28、42、56和70 d时颈静脉采血,根据血糖浓度在6个时间点分为低血糖组(L)和对照组(CK).同时调查和检测奶牛的生产性能(体重、泌乳量和干物质摄入量)、繁殖性能(首次配种间隔、情期受胎率、发情率和配种指数)、血液生化指标(血糖、β-羟丁酸和游离脂肪酸)和内分泌指标(胰岛素、胰高血糖素、瘦素、雌二醇和孕酮).结果显示:奶牛泌乳早期存在不同程度的能量负平衡,产后14 d最高.能量负平衡(negative energy balance,NEB)能使奶牛产后干物质摄入量(DMI)、体重(BW)和泌乳量(MY)减少,首次配种间隔延长,情期受胎率和发情率降低,配种指数增高,血浆胰岛素(Ins)、胰高血糖素(Gn)、瘦素(leptin)、雌二醇(E2)和孕酮(P4)浓度降低,呈现低血糖(Glu)、高血浆游离脂肪酸(NEFA)和β-羟丁酸(BHBA)生化特性;能量负平衡时,奶牛血浆瘦素与血浆Glu、E2、P4之间呈显著正相关,与BW损失、血浆NEFA之间呈明显负相关;能量平衡时,奶牛血浆瘦素与DMI、Glu、Ins之间呈显著正相关,与Bw损失呈显著负相关,与血浆E2和P4之间相关不显著.这些结果提示,奶牛泌乳早期易发能量负平衡不仅与干物质摄入量、体重、泌乳量密切相关,而且与机体内分泌未完全协调发挥作用有关.能量负平衡明显降低泌乳早期奶牛的生产性能和繁殖性能.血浆瘦素与生产性能指标、血浆生化指标的相关性分析证实瘦素在奶牛能量平衡、生产性能和繁殖性能之间起到一个平衡纽带的作用.     

半滑舌鳎两种leptin基因的体外重组表达和生物活性分析

根据半滑舌鳎lepa和lepb编码氨基酸序列合成开放阅读框ORF肽段。利用原核表达载体pEQ30成功构建了半滑舌鳎lepa/pQE30和lepb/pQE30重组质粒,分别转化至大肠杆菌M15后,经IPTG诱导获得了N端含6个组氨酸的半滑舌鳎LepA和LepB重组蛋白。获得的重组蛋白大小均为16 ku,37°C下用0.5 mmol/L的IPTG诱导4 h后目的蛋白表达量最高,主要以包涵体形式存在。检测半滑舌鳎LepA和LepB重组蛋白浓度分别为0.3和0.25 mg/mL。Western blot免疫印迹和质谱分析表明,获得的LepA和LepB重组蛋白均具有免疫活性且序列正确。Ni2+-NTA亲和层析柱纯化可获得高纯度的半滑舌鳎LepA和LepB重组蛋白。离体孵育实验表明,获得的半滑舌鳎LepA和LepB重组蛋白能显著抑制半滑舌鳎下丘脑lepa、lepb和gnrh3 mRNA的表达水平,表明获得的重组蛋白具有明显的生物活性。研究结果可为探究leptin在半滑舌鳎生长发育中的调控机制提供重要参考。     

不同品种初生肉仔鸡卵黄囊耗用规律及其激素调节的研究

选用北京油鸡和AA鸡的公雏研究不同品种初生肉仔鸡卵黄囊耗用规律及其激素调节。每个品种276只健康公雏按体重随机分为6个重复，每个重复46只。分别在孵出后第0、1、3、5、7、9、11天称重后，每个品种选取36只雏鸡进行屠宰。结果表明：（1）北京油鸡的初生重和生长速度均显著低于AA鸡；（2）2个品种鸡的卵黄整重、卵黄囊风干物质含量和总能值均随日龄增长呈指数方式递减，但北京油鸡的指数函数系数均高于AA鸡；（3）卵黄囊瘦素和胰岛素的水平随日龄和品种的不同而变化。在出壳后前3天，北京油鸡和AA鸡的卵黄囊瘦素和胰岛素水平均随日龄的增长而增加。且在孵出后第0和3天，北京油鸡的卵黄囊瘦素和胰岛素水平显著高于相同日龄的AA鸡（P≤0．0413）。试验结果提示，2个品种的初生肉仔鸡卵黄囊耗用规律基本一致，但北京油鸡的耗用速度比AA鸡快，部分原因可能是北京油鸡卵黄囊瘦素和胰岛素水平更高。     

小鼠胚胎附植前后瘦素水平与胃瘦素长型受体相关性研究

为了对小鼠胚胎附植前后瘦素与脂肪细胞变化的相关性进行研究,本研究采用ELISA方法对小鼠生殖周期不同阶段的血清中的瘦素浓度进行了检测,皮下脂肪组织冰冻切片用苏丹(Ⅲ)染色计量脂肪细胞数量和大小的变化,用免疫组织化学染色法对胃瘦素长型受体进行定位分析。结果显示:妊娠期血清瘦素浓度高于间情期,且瘦素浓度从间情期到妊娠4日龄,呈上升趋势,妊娠5日龄稍下降,然后随妊娠日龄逐渐增加,与单位面积内脂肪细胞数量的变化趋势一致。妊娠7日龄,单位面积内脂肪细胞比间情期少,单个细胞面积大。胃底腺中下部部分胞质和细胞核有棕褐色阳性颗粒且阳性率随妊娠日龄的增加而增加。表明小鼠胚泡附植前后脂肪组织越多血清瘦素水平越高,瘦素及其长型受体可能通过影响胃的消化吸收达到调节机体能量平衡的目的。     

瘦素在动物生殖调控中的作用（续）

由以上可以得出：瘦素对动物的初情期启动有重要作用，是发情周期得以维持正常的代谢信号，可以通过激活卵巢，引起性激素分泌增加，对卵巢内分泌功能有一定促进作用，在早期胚胎发育中也起重要作用，对新生儿生长发育起调节作用，也可促进乳腺发育和乳汁的分泌。     

Prediction of incipient pasture‐associated laminitis from hyperinsulinaemia,hyperleptinaemia and generalised and localised obesity in a cohort of ponies

Reasons for performing study: The ability to predict ponies at increased risk of laminitic episodes, when exposed to nutrient dense pasture, would facilitate management to avoid disease. Objectives: To identify variables and clinically useful cut‐off values with reproducible diagnostic accuracy for the prediction of ponies that subsequently developed laminitis when exposed to nutrient dense pasture. Methods: A cohort of predominantly Welsh and Dartmoor ponies from a closed herd was evaluated in March 2006 (n = 74) and March 2007 (n = 57). Ponies were categorised as never laminitic or previously laminitic according to reported laminitic history and as clinically laminitic (CL) if laminitis was observed within 3 months following evaluation. Body condition score (BCS), cresty neck score (CNS), girth and neck circumferences (NC), withers height, blood pressure and hoof surface temperature, and plasma insulin, glucose, triglyceride, leptin, cortisol, ACTH, uric acid and TNF‐α concentrations were measured. Analysis of sensitivity, specificity and receiver operating characteristic curves was used to evaluate the diagnostic accuracy for a variable to predict CL ponies. Results: Variables with diagnostic accuracy for the prediction of CL ponies included insulin, leptin, BCS, CNS, and NC:height ratio. Specific cut‐off values of insulin (>32 mu/l), leptin (>7.3 ng/ml), BCS (≥7), CNS (≥4) and NC:height ratio (>0.71) had reproducible diagnostic accuracy for the prediction of laminitis. Combining tests did not result in higher diagnostic accuracy than individual tests of insulin or leptin during either evaluation. Conclusions: Tests of insulin and leptin concentrations and measures of generalised (BCS) and localised (CNS or NC:height ratio) obesity were beneficial in the prediction of laminitic episodes. Potential relevance: These results highlight the importance of monitoring and reducing insulin concentration, and generalised and regional obesity in ponies to reduce risk of laminitis.     

陆川猪和大白猪LEPR基因对产仔数的效应分析

为了比较LEPR基因对猪产仔数效应大小,采用PCR-SSCP技术在陆川猪和大白猪群中进行了LEPR基因的基因型频率检测及不同基因型的总产仔数和产活仔数效应分析。结果表明,LEPR基因对陆川猪头胎和头4胎平均总产仔数（TNB）及产活仔数（NBA）影响差异显著(P<0.05),对大白猪头胎的总产仔数及产活仔数影响差异显著(P<0.05),而对大白猪头4胎平均总产仔数及产活仔数影响差异不显著(P>0.05)。     

瘦素在妊娠小鼠子宫着床点与非着床点中的表达研究

目的:探讨瘦素在妊娠小鼠子宫着床点与非着床点中的表达规律。方法:性成熟雌性NIH小鼠与成熟雄鼠自然交配,在妊娠5,6 d分别脱臼处死小鼠取其子宫,然后进行以下实验:1.分别提取总RNA,检测胚胎着床点和非着床点子宫瘦素mRNA表达规律;2.分别进行石蜡包埋,切片,然后进行免疫组化试验,检测瘦素蛋白的表达规律。结果:妊娠5,6 d小鼠子宫着床点与非着床点中均有瘦素mRNA和蛋白表达,蛋白主要定位于腔上皮和腺上皮,着床点mRNA水平比非着床点显著增强。结论:瘦素在着床点与非着床点子宫组织中的表达差异提示瘦素在胚胎着床中的生理作用。     

Adipokines: a review of biological and analytical principles and an update in dogs,cats, and horses

Abstract: In addition to its role as an energy storage depot, adipose tissue is now recognized as a complex endocrine organ. Adipose tissue releases a variety of factors, termed adipokines, that regulate energy metabolism, cardiovascular function, reproductive status, and immune function. Some of the better‐studied adipokines include leptin, adiponectin, and components of the renin‐angiotensin system such as angiotensinogen. The function of more recently discovered adipokines such as resistin are under intense scrutiny. Abnormal production or regulation of adipokines occurs in obese individuals and is implicated in the development of a variety of associated co‐morbidities including metabolic syndrome, type 2 diabetes, atherosclerosis, heart disease, and cancer in people, although evaluation in domestic species is just beginning. Adipokines are now being examined as potential biomarkers for risk assessment for development of complications related to obesity. This article summarizes the function and regulation of some better‐characterized adipokines. It also reviews the current information on the characterization of adipokines in some domestic species in which rates of obesity and obesity‐related disorders are increasing, such as the dog, cat, and horse.