WO2023213303A1 - 提高雌性家畜胎盘组织抗氧化能力和营养转运能力的方法 - Google Patents

提高雌性家畜胎盘组织抗氧化能力和营养转运能力的方法 Download PDF

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WO2023213303A1
WO2023213303A1 PCT/CN2023/092310 CN2023092310W WO2023213303A1 WO 2023213303 A1 WO2023213303 A1 WO 2023213303A1 CN 2023092310 W CN2023092310 W CN 2023092310W WO 2023213303 A1 WO2023213303 A1 WO 2023213303A1
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melatonin
pregnancy
day
nutrient transport
antioxidant
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PCT/CN2023/092310
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French (fr)
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刘国世
王立凯
李广栋
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中国农业大学
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/02—Breeding vertebrates

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  • the present invention relates to the field of animal husbandry and veterinary medicine, and specifically, to a method for improving the antioxidant and nutrient transport capabilities of placental tissue by treating female livestock in late pregnancy with melatonin.
  • Melatonin is a hormone secreted by the pineal gland. Current research shows that melatonin is involved in various biological processes such as regulating biological rhythms, regulating sleep and immune cells.
  • the purpose of the present invention is to provide a method for processing A method to improve the antioxidant and nutrient transport capacity of placental tissue in female livestock during late pregnancy.
  • the present invention provides the use of melatonin in improving the antioxidant capacity and/or nutrient transport capacity of placental tissue in female livestock.
  • increasing the antioxidant capacity of the placental tissue includes increasing the level of the antioxidant gene glutathione-S-transferase M3.
  • increasing the antioxidant capacity of the placental tissue further includes increasing the level of the antioxidant gene microsomal glutathione-S-transferase.
  • improving the nutrient transport capacity includes increasing the expression level of the nutrient transport-related gene Slc38A2.
  • improving the nutrient transport capacity further includes increasing the expression level of the nutrient transport-related gene Slc7A8.
  • the present invention provides a method for improving the antioxidant capacity and/or nutrient transport capacity of placental tissue of female livestock, which method includes feeding melatonin to the female livestock during the period from late pregnancy to calving.
  • the dosage of melatonin is 1.5-2.5 mg/kg/day, for example, it can be 1.5 mg/kg/day, 1.6 mg/kg/day, 1.7 mg/kg/day, 1.8 mg/kg/day, 1.9mg/kg/day, 2.0mg/kg/day, 2.1mg/kg/day, 2.2mg/kg/day, 2.3mg/kg/day, 2.4mg/kg/day or 2.5mg /kg/day.
  • the dosage of melatonin is 1.8-2.2 mg/kg/day.
  • the dosage of melatonin is 2.0 mg/kg/day. Based on the average weight of female livestock such as sows, calculated as 200kg, the amount of melatonin fed to each sow per day is 0.4g.
  • the feeding dose of melatonin will affect the antioxidant and nutrient transport capabilities of placental tissue. If the feeding dose is too high, the nutrient transport capability will be inhibited, while if the feeding dose is too low, the effect will be poor.
  • melatonin in the form of powder, and melatonin can be added to sow feed; preferably, the number of times of melatonin feeding per day is a single time; more preferably,
  • the melatonin feeding time is every morning, which can be between seven and eight o'clock. body Melatonin levels are low during the day. A single feeding in the morning can effectively increase melatonin levels during the day and help melatonin utilization.
  • the third trimester is pregnancy day 90 ⁇ 5, for example, it can be pregnancy day 85, 86, 87, 88, 89, 90, or 91 , day 92, day 93, day 94 or day 95.
  • the third trimester of pregnancy is the 90th ⁇ 2nd day of pregnancy.
  • the third trimester of pregnancy is the 90th day of pregnancy.
  • the period of high incidence of oxidative stress is the late pregnancy. If the melatonin feeding time is too long, it will easily increase the cost, and if it is too short, it will have no effect.
  • the melatonin feeding cycle is 30 ⁇ 5 days, for example, it can be 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days.
  • the melatonin feeding cycle is 30 ⁇ 2 days.
  • the melatonin feeding cycle is 30 days.
  • the female livestock includes: pigs, cattle, horses, sheep, rabbits, dogs and cats.
  • the female livestock is a sow.
  • the present invention unexpectedly discovered that treating late-pregnancy sows with melatonin can increase the concentration of melatonin in the sow's peripheral blood and colostrum, and at the same time improve the antioxidant and nutrient transport capabilities of the placental tissue.
  • the inventors through extensive efforts and long-term experiments, obtained the optimal time period for feeding melatonin to pregnant sows, as well as the most suitable amount of melatonin, which is beneficial to improving the performance of sows.
  • the antioxidant and nutrient transport capabilities of placental tissue are of great significance.
  • the method of the present invention has simple implementation steps, good effects, and strong generalizability.
  • the present invention adopts the method of feed addition and utilizes melatonin to treat sows in late pregnancy until the sows give birth.
  • the treatment method of the present invention is simple, easy to operate, will not cause stress reactions in sows, and is highly feasible. In addition, it can significantly improve the antioxidant and nutrient transport capabilities of sow placenta tissue, and It provides a useful reference for further research on improving the antioxidant capacity and nutrient transport capacity of female livestock placenta tissue.
  • Figure 1 shows the changes in MT concentration in peripheral blood and colostrum after feeding MT compared with the control group.
  • Figure 2 shows the changes in PRL, COR, and E2 contents in peripheral blood after feeding MT compared with the control group.
  • Figure 3 shows the expression of antioxidant-related genes CAT, GSH, GSTM3, and MGST1 in placenta tissue after feeding MT compared with the control group.
  • Figure 4 shows the expression of placental tissue nutrient transport-related genes Slc38A1, Slc38A2, and Slc38A4 after feeding MT compared with the control group.
  • the start time point of melatonin treatment was the late pregnancy of sows (D90 ⁇ 5), and the end time point was ⁇ 3 days after sow farrowing, with a total cycle of 30 ⁇ 5 days;
  • the feeding dose of each pig was 1.5 ⁇ 2.5mg/kg (the weight of the sow is calculated based on 200kg, and the daily feeding amount of each pig is 0.3 ⁇ 0.5g);
  • the melatonin feeding time point every day is from 7 to 8 o'clock in the morning; the feeding method is feed addition.
  • the blood collection time point is two days before the sow's expected delivery date (approximately D112), and the blood collection method is neck blood collection, which is used to analyze melatonin and hormone changes; the colostrum collection time point is after the sow starts giving birth.
  • qPCR was used to detect the expression of antioxidant and nutrient transport-related genes in placenta tissue.
  • melatonin group Ten healthy sows in late pregnancy were selected and randomly divided into melatonin group (MT, 5) and control group (5).
  • the control group was fed normally.
  • the melatonin group was treated with melatonin from the 90th day of pregnancy until sows farrowed.
  • Sow blood was collected two days before gestation, and sow colostrum was collected at farrowing.
  • the LC-MS method was used to detect the concentration of melatonin in peripheral blood and colostrum. The measurement results are shown in Figure 1.
  • control group VS melatonin group All data are expressed as mean ⁇ standard error. Different letters indicate significant differences within the group (p ⁇ 0.05). Same letters indicate that the differences within the group are not significant (p> 0.05).
  • melatonin group (MT, 3 pigs) and control group (3 pigs).
  • the control group was fed normally.
  • the melatonin group was treated with melatonin from the 90th day of pregnancy until sows farrowed.
  • the blood of sows was collected two days before pregnancy, and the concentration of reproductive hormones in peripheral blood was detected. The measurement results are shown in Figure 2.
  • control group VS melatonin group All data are expressed as mean ⁇ standard error. Different letters indicate significant differences within the group (p ⁇ 0.05). Same letters indicate that the differences within the group are not significant (p> 0.05).
  • A: Relative expression level of catalase: n 5 (unit: head);
  • control group VS melatonin group All data are expressed as mean ⁇ standard error. Different letters indicate significant differences within the group (p ⁇ 0.05). Same letters indicate that the differences within the group are not significant (p> 0.05).
  • melatonin group Ten healthy sows in late pregnancy were selected and randomly divided into melatonin group (MT, 5) and control group (5).
  • the control group was fed normally.
  • the melatonin group was treated with melatonin from the 90th day of pregnancy until sows farrowed.
  • the placenta tissue was collected, and the expression of nutrient transport-related genes in the tissue was detected by qPCR. The statistical results are shown in Figure 4.
  • control group VS melatonin group All data are expressed as mean ⁇ standard error. Different letters indicate significant differences within the group (p ⁇ 0.05). Same letters indicate that the differences within the group are not significant (p> 0.05).

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Zoology (AREA)
  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

一种提高雌性家畜胎盘组织抗氧化能力和/或营养转运能力的方法，包括用褪黑素处理妊娠后期雌性家畜。通过动物实验证明，利用褪黑素处理妊娠后期母猪，可提高母猪外周血和初乳中褪黑素浓度，同时提高胎盘组织的抗氧化能力和营养转运能力。该方法简单有效，可行性强。

Description

提高雌性家畜胎盘组织抗氧化能力和营养转运能力的方法 技术领域
本发明涉及畜牧兽医领域，具体地，涉及通过褪黑素处理妊娠后期雌性家畜提高胎盘组织抗氧化和营养转运能力的方法。
背景技术
长期实践证明，在猪的妊娠过程中，由于胎儿的快速生长极易导致母体代谢失衡，引发严重氧化应激反应。胎盘作为连接母体和胎儿的“枢纽”，在胎儿的营养供应、代谢废物排除和气体交换过程中发挥极为重要的作用。研究表明，妊娠期氧化应激易破坏胎盘组织的功能，导致不良妊娠结局，如流产、宫内发育迟缓和早产等，严重制约养殖业的发展。因此，合理提高母猪胎盘组织的抗氧化和营养转运能力，对于养猪业的发展至关重要。
目前，在养猪业中，为提高母猪的抗氧化能力和营养转运能力，养殖户一般根据母猪的实际体况进行科学合理地饲喂，提高母猪日粮能量，保证能量供应。此外，还需给母猪提供适宜的生存环境，避免高温、高湿等恶劣环境影响猪的生产性能。但在实际生产中，以上方案处理要求高(饲养规模化、养殖户具备相关养殖技术等)、实施工作繁琐，不易于具体实施和普遍推广。
褪黑素(Melatonin，MT)是一种由松果体分泌的激素，目前研究表明褪黑素参与调节生物节律、调节睡眠和免疫细胞等多种生物学过程。
因此，鉴于现有技术中存在的问题，需要寻求一种简单、有效提高母猪胎盘组织抗氧化能力和营养转运能力的方法，这对于养猪业发展至关重要。
发明内容
为了解决现有技术中存在的问题，本发明的目的是提供一种通过处理 妊娠后期雌性家畜来提高胎盘组织抗氧化和营养转运能力的方法。
为了实现本发明目的，本发明的技术方案如下：
第一方面，本发明提供褪黑素在提高雌性家畜胎盘组织抗氧化能力和/或营养转运能力中的用途。
在一些实施方案中，提高所述胎盘组织抗氧化能力包括提高抗氧化基因谷胱甘肽-S-转移酶M3的水平。
在一些实施方案中，提高所述胎盘组织抗氧化能力还包括提高抗氧化基因微粒体谷胱甘肽-S-转移酶的水平。
在一些实施方案中，提高所述营养转运能力包括提高营养转运相关基因Slc38A2的表达水平。
在一些实施方案中，提高所述营养转运能力还包括提高营养转运相关基因Slc7A8的表达水平。
第二方面，本发明提供提高雌性家畜胎盘组织抗氧化能力和/或营养转运能力的方法，所述方法包括在雌性家畜妊娠后期至产仔期间，给所述雌性家畜饲喂褪黑素。
在一些实施方案中，所述褪黑素的施用量为1.5～2.5mg/kg/天，例如，可以为1.5mg/kg/天、1.6mg/kg/天、1.7mg/kg/天、1.8mg/kg/天、1.9mg/kg/天、2.0mg/kg/天、2.1mg/kg/天、2.2mg/kg/天、2.3mg/kg/天、2.4mg/kg/天或2.5mg/kg/天。
优选地，所述褪黑素的施用量为1.8～2.2mg/kg/天。
更优选地，所述褪黑素的施用量为2.0mg/kg/天。以雌性家畜例如母猪的平均体重按照200kg计算，每头母猪每天饲喂褪黑素的量为0.4g。
褪黑素的饲喂剂量会影响胎盘组织的抗氧化和营养转运能力，饲喂剂量过高抑制营养转运能力，过低则产生的效果不佳。
在一些实施方案中，本发明的方法中，褪黑素为粉剂的形式，可将褪黑素添加在母猪饲料中饲喂；优选地，每天褪黑素饲喂次数为单次；更优选地，褪黑素饲喂时间点为每天早上饲喂，可以在七点至八点之间。机体 褪黑素在白天水平较低，采用早上单次饲喂的方式可有效提高白天褪黑素水平，有助于褪黑素的利用。
在一些实施方案中，所述妊娠后期为妊娠第90±5天，例如，可以为妊娠第85天、第86天、第87天、第88天、第89天、第90天、第91天、第92天、第93天、第94天或第95天。
优选地，所述妊娠后期为妊娠第90±2天。
更优选地，所述妊娠后期为妊娠第90天。
氧化应激高发阶段为妊娠后期，褪黑素饲喂时间过长易造成成本增加，过短则没有效果。
在一些实施方案中，褪黑素的饲喂周期为30±5天，例如，可以为25天、26天、27天、28天、29天、30天、31天、32天、33天、34天、35天。
优选地，褪黑素的饲喂周期为30±2天。
更优选地，褪黑素的饲喂周期为30天。
在本发明的技术方案中，所述雌性家畜包括：猪、牛、马、羊、兔、狗和猫。
优选地，在本发明的技术方案中，所述雌性家畜为母猪。
本发明的有益效果在于：
本发明意外地发现利用褪黑素处理妊娠后期母猪，可提高母猪外周血和初乳中褪黑素浓度，同时能够提高胎盘组织抗氧化和营养转运能力。基于这一惊喜的发现，本发明人通过大量努力和长期的多次实验，获得了对妊娠母猪饲喂褪黑素的最优时间段，以及褪黑素的最适用量，对提高母猪胎盘组织抗氧化和营养转运能力具有重要的意义。本发明的方法实施步骤简单且效果好，同时可推广性较强。
本发明采用饲料添加的方式，利用褪黑素来处理妊娠后期的母猪，直至母猪分娩。本发明的处理方式简单，易操作，不会造成母猪的应激反应，可行性强。此外，可以显著提高母猪胎盘组织抗氧化和营养转运能力，并 为进一步开展提高雌性家畜胎盘组织抗氧化能力和营养转运能力的相关研究提供有益的参考。
附图说明
图1示出与对照组相比饲喂MT后外周血和初乳中MT浓度变化。
图2示出与对照组相比饲喂MT后外周血中PRL、COR、E2含量变化。
图3示出与对照组相比饲喂MT后胎盘组织抗氧化相关基因CAT、GSH、GSTM3、MGST1的表达。
图4示出与对照组相比饲喂MT后胎盘组织营养转运相关基因Slc38A1、Slc38A2、Slc38A4的表达。
具体实施方式
以下实施例用于说明本发明，但不用来限制本发明的范围。
以下实施例中使用的试剂和材料均为市售商品。
实验方法
1.褪黑素处理方案
本研究中褪黑素处理开始时间点为母猪妊娠后期(D90±5)，结束时间点为母猪分娩产仔±3天，周期共30±5天；每头猪饲喂剂量为1.5～2.5mg/kg(母猪体重按照200kg计算，每头猪每天饲喂量为0.3～0.5g)；每天中褪黑素饲喂时间点为早上七点至八点；饲喂方式为饲料添加。
2.血液、初乳样本采集
血液采集时间点为母猪预产期前两天(约D112)，血液采集方式为颈部采血，用于分析褪黑素和激素变化；初乳采集时间点为母猪开始生产后。
3.褪黑素含量测定
本研究中褪黑素浓度测定方式为LC-MS检测。
4.胎盘组织抗氧化和营养转运相关基因表达检测
利用qPCR的方式检测胎盘组织中抗氧化和营养转运相关基因的表达。
实验例1
1.饲喂褪黑素后外周血和初乳中褪黑素的变化规律
选择处于妊娠后期的健康母猪10只，随机分为褪黑素组(MT，5只)和对照组(5只)。对照组正常饲喂。褪黑素组从妊娠第90天开始采用褪黑素处理，直至母猪分娩。于妊娠前两天采集母猪血液，并于分娩时收集母猪初乳。利用LC-MS法检测外周血和初乳中褪黑素浓度，测定结果见图1。
图1中，
A：外周血中MT浓度变化：
(54.92±5.782VS 170.8±18.47)n＝5(单位：头)；
B：初乳中MT浓度变化：
(61.00±6.675VS 135.5±12.20)n＝3(单位：头)。
注：以上数据表示为：对照组VS褪黑素组，所有数据采用平均值±标准误表示，字母不同者表示组内差异显著(p<0.05)，字母相同表示差异组内不显著(p>0.05)。
通过图1可知，采用2mg/kg的褪黑素处理妊娠后期的母猪，可显著提高母猪外周血中褪黑素浓度(54.92±5.782VS 170.8±18.47)(p<0.05)；对初乳中褪黑素浓度进行检测发现，饲喂褪黑素组(MT)母猪初乳中褪黑素浓度显著高于对照组(61.00±6.675VS 135.5±12.20)(p<0.05)。2.饲喂褪黑素后母猪体内生殖激素变化规律
选择处于妊娠后期的健康母猪6只，随机分为褪黑素组(MT，3只)和对照组(3只)。对照组正常饲喂。褪黑素组从妊娠第90天开始采用褪黑素处理，直至母猪分娩。于妊娠前两天采集母猪血液，检测外周血中生殖激素浓度，测定结果见图2。
图2中，
A：PRL含量变化：
(30.33±1.378VS 71.78±0.6307)n＝3(单位：头)；
B：COR含量变化：
(71.48±7.792VS 89.99±22.60)n＝3(单位：头)；
C：E2含量变化：
(931.3±81.52VS 862.8±256.9)n＝3(单位：头)。
注：以上数据表示为：对照组VS褪黑素组，所有数据采用平均值±标准误表示，字母不同者表示组内差异显著(p<0.05)，字母相同表示差异组内不显著(p>0.05)。
通过图2可知，采用2mg/kg的褪黑素处理妊娠后期的母猪，检测生殖激素发现，褪黑素处理可显著增加外周血中PRL的水平(30.33±1.378VS 71.78±0.6307)(p<0.05)。
3.胎盘组织抗氧化相关基因表达检测
选择处于妊娠后期的健康母猪10只，随机分为褪黑素组(MT，5只)和对照组(5只)。对照组正常饲喂。褪黑素组从妊娠第90天开始采用褪黑素处理，直至母猪分娩。妊娠母猪分娩完成后，收集胎盘组织，qPCR检测组织中抗氧化相关基因的表达，统计结果见图3。
图3中，
A：过氧化氢酶相对表达水平：n＝5(单位：头)；
B：谷胱甘肽相对表达水平：n＝5(单位：头)；
C：谷胱甘肽-S-转移酶M3相对表达水平：n＝5(单位：头)；
D：微粒体谷胱甘肽-S-转移酶1：n＝5(单位：头)。
注：以上数据表示为：对照组VS褪黑素组，所有数据采用平均值±标准误表示，字母不同者表示组内差异显著(p<0.05)，字母相同表示差异组内不显著(p>0.05)。
结果表明，采用2mg/kg的褪黑素处理妊娠后期的母猪可显著提高胎盘组织中抗氧化基因谷胱甘肽-S-转移酶M3和微粒体谷胱甘肽-S-转移酶的表达水平(p<0.05)。
4.胎盘组织营养转运相关基因表达检测
选择处于妊娠后期的健康母猪10只，随机分为褪黑素组(MT，5只)和对照组(5只)。对照组正常饲喂。褪黑素组从妊娠第90天开始采用褪黑素处理，直至母猪分娩。妊娠母猪分娩完成后，收集胎盘组织，qPCR检测组织中营养转运相关基因的表达，统计结果见图4。
图4中，
A：Slc38A1相对表达水平：n＝5(单位：头)；
B：Slc38A2相对表达水平：n＝5(单位：头)；
C：Slc38A4相对表达水平：n＝5(单位：头)；
D：Slc7A8相对表达水平：n＝5(单位：头)。
注：以上数据表示为：对照组VS褪黑素组，所有数据采用平均值±标准误表示，字母不同者表示组内差异显著(p<0.05)，字母相同表示差异组内不显著(p>0.05)。
结果表明，采用2mg/kg的褪黑素处理妊娠后期的母猪可显著提高胎盘组织中营养转运基因Slc38A2和Slc7A8的表达水平(p<0.05)。
以上结果表明，通过饲料添加的方式，采用2mg/kg的褪黑素处理处于妊娠后期的母猪，可显著提高母猪外周血和初乳中褪黑素浓度；褪黑素饲喂可提高外周血中PRL的水平，刺激母猪泌乳；同时，褪黑素处理可显著增加母猪胎盘组织中抗氧化基因谷胱甘肽-S-转移酶M3和微粒体谷胱甘肽-S-转移酶以及营养转运相关基因Slc38A2和Slc7A8的表达水平。因此，褪黑素可以提高妊娠后期雌性家畜的胎盘组织抗氧化能力和营养转运能力。
虽然，上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述，但在本发明基础上，可以对之作一些修改或改进，这对本领域技术人员而言是显而易见的。因此，在不偏离本发明精神的基础上所做的这些修改或改进，均属于本发明要求保护的范围。

Claims (10)

  1. 褪黑素在提高雌性家畜胎盘组织抗氧化能力和/或营养转运能力中的用途。
  2. 根据权利要求1所述的用途，其特征在于，提高所述胎盘组织抗氧化能力包括提高抗氧化基因谷胱甘肽-S-转移酶M3的水平。
  3. 根据权利要求1或2所述的用途，其特征在于，提高所述胎盘组织抗氧化能力包括提高抗氧化基因微粒体谷胱甘肽-S-转移酶的水平。
  4. 根据权利要求1所述的用途，其特征在于，提高所述营养转运能力包括提高营养转运相关基因Slc38A2的表达水平。
  5. 根据权利要求1或4所述的用途，其特征在于，提高所述营养转运能力包括提高营养转运相关基因Slc7A8的表达水平。
  6. 提高雌性家畜胎盘组织抗氧化能力和/或营养转运能力的方法，其特征在于，所述方法包括在雌性家畜妊娠后期至产仔期间，给所述雌性家畜饲喂褪黑素。
  7. 根据权利要求6所述的方法，其特征在于，所述褪黑素的施用量为1.5～2.5mg/kg/天；
    优选地，所述褪黑素的施用量为1.8～2.2mg/kg/天；
    更优选地，所述褪黑素的施用量为2.0mg/kg/天。
  8. 根据权利要求6或7所述的方法，其特征在于，所述妊娠后期为妊娠第90±5天；
    优选地，所述妊娠后期为妊娠第90±2天；
    更优选地，所述妊娠后期为妊娠第90天。
  9. 根据权利要求6或7所述的方法，其特征在于，褪黑素的饲喂周期为30±5天；优选地，褪黑素的饲喂周期为30±2天。
  10. 根据权利要求1-5任一项所述的用途或权利要求6-9任一项所述的方法，其特征在于，所述雌性家畜包括：猪、牛、马、羊、兔、狗和猫；
    优选地，所述雌性家畜为猪。
PCT/CN2023/092310 2022-05-05 2023-05-05 提高雌性家畜胎盘组织抗氧化能力和营养转运能力的方法 WO2023213303A1 (zh)

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