Leptin对动物繁殖机能的影响及其应用前景

Leptin是由脂肪组织分泌的一种“厌食”激素。自1995年发现以来，就成为最活跃的研究领域。它可通过下丘脑或直接作用于卵巢，对动物青春期的发育、生殖器官的发育以及性激素的分泌都有重要的调节作用。本文综述了Leptin的生物学特性、作用机理及其在繁殖方面的作用，井指出了它在畜牧生产中应用前景。     

阿勒泰羊生长速度与主要血液生化指标含量及尾脂Leptin基因表达关系

试验通过研究不同生长速度阿勒泰羊主要血液生化指标含量和尾脂中瘦素（Leptin）基因的表达水平，探讨阿勒泰羊生长速度与脂肪沉积之间的关系。选取300只出生时间相近的阿勒泰羊公羔，正常饲喂至6月龄，记录日增重，取平均日增重排前10%和后10%的公羔各30只，分为生长快速组（HBW）和生长慢速组（LBW），从生长快速组随机选12只分为抑制生长组（饲喂低能量饲粮以抑制其生长，HBW-75%，n=6）和生长快速对照组（饲喂标准饲粮，HBW-100%，n=6），生长慢速组随机选12只分为促进生长组（饲喂高能量饲粮以促进生长，LBW-125%，n=6）和生长慢速对照组（饲喂标准饲粮，LBW-100%，n=6）；预饲期7 d，正饲期30 d，试验结束后称重并采集血液，屠宰后采集尾部脂肪。检测血液TG、CK、TSH、T3、T4、S.S、GH、INS、PG、Leptin含量或活性及尾部脂肪Leptin mRNA和蛋白表达量，并观察尾部脂肪细胞形态学变化。结果显示，HBW-100%和LBW-100%组初体重与末体重均差异显著（P<0.05），HBW-75%和HBW-100%组初体重与末体重均无显著差异（P>0.05）；LBW-100%和LBW-125%组初体重差异显著（P<0.05），末体重无显著差异（P>0.05）。LBW-100%组的T4、S.S、Leptin、TSH、T3含量显著低于HBW-100%组（P<0.05）；LBW-125%组T3含量显著高于LBW-100%组（P<0.05）；HBW-75%组GH、PG和Leptin含量显著低于HBW-100%组（P<0.05）。LBW-125%组尾部脂肪细胞面积大于LBW-100%组，HBW-75%组小于HBW-100%组（P<0.05）。Leptin mRNA和蛋白表达量检测结果表明，HBW-100%组的Leptin mRNA表达量与LBW-100%组无显著差异（P>0.05），Leptin蛋白表达量极显著高于LBW-100%组（P<0.01）。LBW-125%组Leptin mRNA和蛋白的表达量均显著高于LBW-100%组（P<0.05）。HBW-75%组Leptin mRNA的表达量极显著低于HBW-100%组（P<0.01），Leptin蛋白表达量显著低于HBW-100%组（P<0.05）。综上所述，生长快速组与生长慢速组中，Leptin的表达量与体重和体脂有一定关联；在生长慢速组促进生长后体重和体脂显著增加，Leptin的表达量呈上升趋势；在生长快速组抑制生长后体重和体脂减少，Leptin的表达呈下降趋势。改变生长速度对Leptin的表达量会产生影响，从而改变机体的脂肪沉积状况。     

Assessment of Uterine Vascular Perfusion During the Estrous Cycle of Mares in Connection to Circulating Leptin and Nitric Oxide Concentrations

The objective of this study was to find difference in vascular perfusion of uterine horns or uterine body throughout the estrous cycle and their relation to circulating nitric oxide and leptin concentrations. Five cyclic mares were subjected to transrectal Doppler ultrasonography and blood sampling for 18 days. Area of color and power Doppler modes was measured in pixels. Day (P = .0001) of the estrous cycle and ovulation (P = .0001) influenced uterine blood flow. Uterine body blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day 0 (day of ovulation), and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until days 12 and 10, respectively. Conversely to the contralateral uterine horn, ipsilateral uterine horn blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day −1, and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until day 10. Nitric oxide concentrations (P = .0001) attained two major peaks; the first on day −3 and the other persisted from day 2 until day 5. Leptin concentrations increased (P > .001) with a maximum value on day 0 and then decreased until a minimum value on day 9. In conclusion, during the estrous cycle, ipsilateral uterine horn and uterine body blood vessels had similar blood flow. Both leptin and nitric oxide played a role during follicle growth, ovulation, and corpus luteum development and modulated uterine blood flow before and after ovulation.     

Leptin对猪成肌细胞中PGC-1α和UCPs mRNA表达的影响

采用链酶蛋白酶消化法获得猪成肌细胞、肌细胞生成素(MyoG)和生肌决定因子(MyoD)的RT-PCR和Desmin免疫组化染色鉴定猪成肌细胞后,分别研究外源Leptin对猪成肌细胞产热相关基因过氧化体增殖活化受体gamma辅助活化因子α(PGC-1α)和解偶联蛋白家族(UCPs)mRNA表达的影响。对产热相关基因的RT-PCR检测结果表明,30和100nmol/L Leptin均可促进成肌细胞中PGC-1α mRNA表达,100 nmol/L Leptin作用效果显著高于30 nmol/L,且作用24h促进效果明显优于12h作用效果(P<0.05)。30 nmol/L Leptin可明显提高成肌细胞中UCP2 mRNA的表达,且作用24h促进效果明显优于12h处理效果(P<0.05),100 nmol/L Leptin作用效果显著低于30n mol/L Leptin作用效果,但显著高于对照组(P<0.05)。30和100 nmol/L Leptin均不影响UCP3 mRNA的表达(P<0.05)。结果提示,Leptin可能通过上调PGC-1α和UCP2转录,促进成肌细胞能量消耗。     

Biology of leptin in the pig

The recently discovered protein, leptin, which is secreted by fat cells in response to changes in body weight or energy, has been implicated in regulation of feed intake, energy expenditure and the neuroendocrine axis in rodents and humans. Leptin was first identified as the gene product found deficient in the obese ob/ob mouse. Administration of leptin to ob/ob mice led to improved reproduction as well as reduced feed intake and weight loss. The porcine leptin receptor has been cloned and is a member of the class 1 cytokine family of receptors. Leptin has been implicated in the regulation of immune function and the anorexia associated with disease. The leptin receptor is localized in the brain and pituitary of the pig. The leptin response to acute inflammation is uncoupled from anorexia and is differentially regulated among swine genotypes. In vitro studies demonstrated that the leptin gene is expressed by porcine preadipocytes and leptin gene expression is highly dependent on dexamethasone induced preadipocyte differentiation. Hormonally driven preadipocyte recruitment and subsequent fat cell size may regulate leptin gene expression in the pig. Expression of CCAAT-enhancer binding protein (C/EBP) mediates insulin dependent preadipocyte leptin gene expression during lipid accretion. In contrast, insulin independent leptin gene expression may be maintained by C/EBP auto-activation and phosphorylation/dephosphorylation. Adipogenic hormones may increase adipose tissue leptin gene expression in the fetus indirectly by inducing preadipocyte recruitment and subsequent differentiation. Central administration of leptin to pigs suppressed feed intake and stimulated growth hormone (GH) secretion. Serum leptin concentrations increased with age and estradiol-induced leptin mRNA expression in fat was age and weight dependent in prepuberal gilts. This occurred at the time of expected puberty in intact contemporaries and was associated with greater LH secretion. Further work demonstrated that leptin acts directly on pituitary cells to enhance LH and GH secretion, and brain tissue to stimulate gonadotropin releasing hormone secretion. Thus, development of nutritional schemes and (or) gene therapy to manipulate leptin secretion will lead to practical methods of controlling appetite, growth and reproduction in farm animals, thereby increasing efficiency of lean meat production.     

Effects of a high-protein, low-energy diet in finishing lambs: 2. Weight change, organ mass, body composition, carcass traits, fatty acid composition of lean and adipose tissue, and taste panel evaluation

An experiment was conducted to evaluate the effects of a high protein – low energy diet on body composition of feedlot lambs The objective was to assess the capability of such a diet to reduce body fat while preserving lean body mass. Three dietary regimens were compared: a typical feedlot diet (CON), barley straw at libitum (STR), and barley straw supplemented with fish meal (SFM). The latter two diets caused an energy deficit in the experimental animals. Significant treatment effects on mass of metabolic organs, body composition, and carcass data were observed. Conventional analysis of sensory data and fatty acid profiles of adipose and muscle tissue identified few significant dietary effects, yet discriminant analysis clearly separated all treatment groups. It was concluded that protein supplementation of ruminants in the catabolic state does not conserve protein mass. Given the high cost of protein supplementation, the post hoc modification of body composition of feedlot lambs is not a viable option. It is suggested that the production of feedlot lambs with a desirable degree of fatness should employ a feeding strategy dynamically adapted to the changes in protein – energy balance required by the growing ruminant.     

烟酸对高脂血症兔瘦素、过氧化体增殖物激活型受体γ和CD36表达的影响

目的探讨烟酸对高脂血症兔血清瘦素及皮下脂肪组织瘦素、过氧化体增殖物激活型受体γ及CD36 mRNA表达的影响。方法12只健康雄性新西兰兔给予高胆固醇饮食饲养8周后,随机分为高脂组和烟酸组:高脂组继续饲以高胆固醇饲料6周;烟酸组在饲以高胆固醇饲料的基础上给予烟酸0.2g/(kg.d),共6周。另选择普通饮食14周兔(n=6)作为对照组。实验结束后,取腹股沟处皮下脂肪组织称重并冻存,用酶联免疫吸附法测定血清及脂肪细胞培养基瘦素水平;半定量逆转录聚合酶链反应测定脂肪组织瘦素、过氧化体增殖物激活型受体γ及CD36mRNA的表达。此外,在体外观察不同浓度的烟酸对高脂兔脂肪细胞瘦素、过氧化体增殖物激活型受体γ及CD36 mRNA的表达。结果高脂组兔血清及脂肪组织瘦素水平明显高于正常对照组,烟酸治疗6周后可降低血清及脂肪组织瘦素水平。逆转录聚合酶链反应表明烟酸组较高脂组瘦素mRNA表达降低,瘦素mRNA与过氧化体增殖物激活型受体γmRNA和CD36 mRNA的表达呈负相关。体外实验亦表明,烟酸呈剂量依赖性地降低脂肪细胞瘦素mRNA表达,并剂量依赖性地上调过氧化体增殖物激活型受体γ及CD36 mRNA的表达。结论烟酸治疗能降低高脂血症兔血清及脂肪分泌瘦素水平,上调过氧化体增殖物激活型受体γ及CD36 mRNA的表达。     

The Effect of Different Physical Exercise on Plasma Leptin,Cortisol, and Some Energetic Parameters Concentrations in Purebred Arabian Horses

Leptin is a hormone that coordinates food intake, energy expenditure, and metabolic rate; it is, however, unknown how exercise influences plasma leptin concentration in horses. The aim of the study was to evaluate the exercise-induced changes in plasma leptin concentration in purebred Arabian horses competing in races and endurance rides. A total number of 26 horses (12 purebred Arabian racehorses aged 3–5 years and 14 endurance horses aged 7–14 years) were studied during performing routine physical exercise. From each horse, blood samples were collected at rest and immediately after the exercise session. Concentrations of plasma leptin, cortisol, lactic acid, uric acid, triacylglycerols, glycerol and free fatty acids were determined. Exercise-induced increase in cortisol and free fatty acid values were higher in endurance horses than in racehorses. Neither endurance nor race exercises influenced the plasma leptin concentration. Further research is needed to elucidate the effects of different types of exercise on leptin synthesis and secretion in horses.     

Leptin的生殖调控机制研究进展

Ｌｅｐｔｉｎ是一种具有多种生殖生理效应的活性多肽。研究表明，它促进生殖系统发育，参与卵母细胞动物极的决定，影响内细胞团的建立和滋养层的形成。在妊娠中参与附植，母体妊娠状态，胎盘功能和胎儿生长发育的调节，另外还影响多种生殖激素的释放。     

侧脑室注射Leptin对大鼠下丘脑前阿黑皮素mRNA表达的影响

给体重 180～ 2 0 0 g的雌性 SD大鼠侧脑室中注射重组瘦素 (L eptin) 1μg,采用 RT- PCR测定了其下丘脑前阿黑皮素 (POMC) m RNA和 L eptin受体 (OB- R) m RNA的表达水平。结果表明 ,注射重组 L eptin后 ,OB- R基因和 POMC基因表达量均上升 ,与生理盐水注射组、空白对照组相比差异显著 (P<0 .0 5 ,P<0 .0 1)。由此认为 ,L eptin在大鼠下丘脑 POMC的调控功能中起着重要作用