CN116870091B - 一种抗肉鸡lps应激的中药复方及其制备和应用 - Google Patents
一种抗肉鸡lps应激的中药复方及其制备和应用 Download PDFInfo
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Abstract
本发明公开一种抗肉鸡LPS应激的中药复方及其制备和应用,涉及兽药技术领域。该中药复方由五味子,金银花,知母,甘草组成,质量比为0.8~1.5:1~2:1~1.5:1~2。该中药复方能够改善肉鸡LPS导致的生长性能降低,免疫器官指数下降。全身炎症细胞增多及氧化应激反应。给该中药复方后,下调血清炎性因子IL‑1β、TNF‑α、IL‑6水平,降低血清ALT、AST含量,上调肝脏中SOD活性,下调肝脏中MPO的活性,降低肠道通透性。对LPS导致的应激有良好的预防效果,为养殖业生产中肉鸡应激提供了一种环保、安全的天然药物来源,同时其还具备成本低廉、安全无耐药性、环保无残留、不含抗生素的优点。
Description
技术领域
本发明涉及兽药技术领域,更具体地,涉及一种能够抗肉鸡LPS应激的中药复方及其制备和应用。
背景技术
兽用西药在禽病的防治中应用广泛,发挥了巨大的作用,但也存在着耐药性、药物残留等风险,不仅影响禽产品质量,且严重威胁着人类健康。近年来随着研究深入,中药以其得天独厚的绿色环保、无药物残留、无毒副作用、安全可靠、药效持久,生产成本低、不需要花费太多时间净化的优点逐渐被重视起来。
综上,现有技术存在的问题是在肉鸡饲养阶段,容易受到外来应激原的侵袭,兽用西药的添加有药物残留的风险,这些药物残留在鸡体内,有可能会使人产生毒副作用,现在亟需一种天然、无毒副作用,而且能够有效抗肉鸡应激的中草药饲料添加剂。
发明内容
为了克服现有技术的缺点与不足,本发明的目的在于提供一种抗肉鸡LPS应激的中药复方及其制备和应用。该中药复方由五味子8~15g;金银花10~20g;知母10~15g,甘草10~20g组成,诸药调和,共奏益气敛阴、补肾宁心、清热解毒、生津润燥、抗菌消炎、增强免疫抗炎抗氧化功效。该中药复方有抗肉鸡LPS应激的功效,同时其还具备成本低廉、安全无耐药性、环保无残留、不含抗生素的优点。
本发明的目的通过下述技术方案实现:
一种抗肉鸡LPS应激的中药复方,由五味子,金银花,知母,甘草组成,质量比为0.8~1.5:1~2:1~1.5:1~2。
优选的,质量比为1:1.5:1.2:2。
所述的抗肉鸡LPS应激的中药复方的制备方法,包括如下步骤:
称取五味子、金银花、知母、甘草,烘干,粉碎,过筛,混匀,制得中药复方。
优选的,所述的烘干的温度为60℃~70℃;进一步为60℃;
优选的,所述的过筛为过80~150目筛。
所述的抗肉鸡LPS应激的中药复方在制备抗肉鸡LPS应激的饲料或饲料添加剂中的应用。
优选的,所述的抗肉鸡LPS应激的中药复方在饲料中的添加量为5~20g/kg;进一步为10~20g/kg;更进一步为20g/kg。
优选的,所述的LPS应激为脂多糖(LPS)诱导的免疫应激。
优选的,所述的抗肉鸡LPS应激为提高LPS应激下肉鸡生长性能,具体为:上调LPS应激下肉鸡平均日采食量和平均日增重,下调料重比。
优选的,所述的抗肉鸡LPS应激为提高LPS应激下肉鸡免疫机能,具体为:上调LPS应激下肉鸡免疫器官指数;进一步的,上调LPS应激下肉鸡胸腺指数、法氏囊指数和脾脏指数。
优选的,所述的抗肉鸡LPS应激为提高LPS应激下肉鸡抗炎作用,具体为:降低LPS应激下肉鸡炎症损伤;进一步为,下调LPS应激下肉鸡血清中IL-6、TNF-α和IL-1β水平。
优选的,所述的抗肉鸡LPS应激为提高LPS应激下肉鸡抗氧化作用,具体为:降低LPS应激下肉鸡肝脏氧化损伤;进一步为,下调LPS应激下肉鸡血清中ALT、AST水平,上调LPS应激下肉鸡肝脏中SOD水平,下调LPS应激下肉鸡肝脏中MPO水平。
优选的,所述的抗肉鸡LPS应激为降低LPS应激下肉鸡肠道通透性;具体为:下调LPS应激下肉鸡血清中DAO、D-LA水平。
其中,五味子有促进机体免疫功能与作用;抗氧化及抗衰老;护肝、诱导肝脏药物代谢酶;镇咳祛痰,抗肿瘤;降血糖;强心;增强机体适应能力;抗溃疡;抗肾病变;抗菌等作用。研究表明,五味子对金黄色葡萄球菌、大肠杆菌、沙门氏菌有不同程度的抑制作用,在中国兽药典中成方制剂中有关五味子配伍规律,五味子可与清热类以及补虚类中药配伍,既能清热解毒,又能滋补肺肾,生津敛汗。金银花的功效主要是清热解毒,亦可抗击炎症,增强机体免疫力,有护肝、抗肿瘤、消炎、解热等作用。金银花在动物生产中的应用非常广泛,对动物生产过程中的多种致病菌、病毒均有抑制作用,对预防和治疗动物的温病发热、风热感冒、咽喉炎症、肺炎、痢疾等症均有很好的效果。知母是临床常用的传统中药材,是常用中药组合物的组成部分。知母为百合科植物知母的干燥根茎。具有清热泄火,滋阴润燥的功效,可用于外感热病,高热烦渴,肺热燥咳,骨蒸潮热,内热消渴,肠燥便秘。甘草为豆科甘草属多年生草本植物,又名美草、蜜甘、蜜草、国老、灵通、甜草、粉草等。现代药理学研究发现,甘草具有保肝、抗炎、抗菌、抗病毒、抗胃溃疡、镇咳、抗疟疾、抗氧化、抗癌、免疫调节、降糖和抗血小板凝集等多种活性。
该中药复方中,诸药相和,共奏抗菌消炎、清热燥湿、凉血解毒、养阴生津、增强免疫力功效,对肉鸡具有抗应激的功效,同时其还具备成本低廉、安全无耐药性、环保无残留、不含抗生素的优点。
经反复试验,作为一种最优选的方案,称取各原料五味子10g、金银花15g、知母12g、甘草20g,使用上述中药复方对肉鸡进行实验,结果显示,食用添加了该中药复方的饲料的肉鸡实验组在平均日增重、平均日采食量、料重比、免疫器官指数、血清炎性因子IL-6、TNF-α和IL-1β,血清中ALT、AST和肝脏中MPO、SOD等方面的表现均优于未添加中药复方的饲料的LPS组,且该中药复方不添加抗生素,不会导致耐药性,不会使大量畜禽甚至人类暴露于耐药菌株引起的感染危险中,更不会给食品安全带来隐患,因此,该中药复方整体上优于促生长抗生素,能够为无抗、绿色健康的养殖模式提供新的思路。
本发明所述的中药复方对LPS诱导的肉鸡免疫应激有抑制作用。
本发明相对于现有技术具有如下的优点及效果:
(1)本发明利用LPS模拟肉鸡免疫应激,其中的中药复方仅由4种中药根据其配伍规律组方而成,不仅能够有效治疗症状,还能从多方面调理机体生理功能,提高机体的免疫力,达到标本兼治的效果。因此,通过中草药来抵抗肉鸡LPS应激反应中具有重要的理论和实践意义。
(2)本发明首次研究发现该中药复方能够改善肉鸡LPS导致的生长性能降低,免疫器官指数下降。全身炎症细胞增多及氧化应激反应。给该中药复方后,下调血清炎性因子IL-1β、TNF-α、IL-6水平,降低血清ALT、AST含量,上调肝脏中SOD活性,下调肝脏中MPO的活性,降低肠道通透性。对LPS导致的应激有良好的预防效果,为养殖业生产中肉鸡应激提供了一种环保、安全的天然药物来源。
具体实施方式
下面结合实施例对本发明作进一步详细的描述,但本发明的实施方式不限于此。除非另有说明,本发明实施例采用的原料为常规购买的原料。
实施例1
1、实验材料
将五味子、金银花、知母、甘草,在60℃下烘干,粉碎后过80~150目筛,按照1:1.5:1.2:2的比例混合而成所需中药复方。
2、试验方法
试验选用200只1日龄凤山文昌鸡,随机分为5组,每组4个重复,每个重复10羽鸡。1日龄起,空白对照组和LPS组(即模型组)饲喂基础日粮,低剂量给药组在基础日粮中添加5g/kg中药复方、中剂量给药组在基础日粮中添加10g/kg中药复方、高剂量给药组在基础日粮中添加20g/kg中药复方。分别在29、31、33、35日龄时注射脂多糖(LPS),在29、36日龄给各组鸡只称重,计算每组肉鸡平均空腹体重,自由饮水,记录采食量,保证通风良好、温度适宜。在36日龄时停止给药,计算每组试验肉鸡在应激期间的平均日采食量、平均日增重、料重比(F/G);采集血清、脾脏、胸腺、法氏囊,剥离周围脂肪并用滤纸吸干水分后,称重并记录,计算免疫器官指数【免疫器官指数(g/kg)=免疫器官鲜重(g)/活重(kg)】。采用酶联免疫吸附试验(ELASA)试剂盒检测血清IL-6、TNF-α、IL-1β的含量,血清中DAO、D-LA的含量,试剂盒购自上海酶联生物科技有限公司。用全自动生化分析仪分析36日龄各组肉鸡血清AST、ALT含量。采用试剂盒检测肝脏中SOD、MPO的含量,试剂盒购自南京建成生物工程研究所。
3、试验结果
用统计软件SPSS 22.0中单因素方差分析对试验数据进行多组样本间的差异显著性分析,所有数据均以“平均值±标准差”表示,得到试验结果如表1~6所示。
表1 29~36日龄(即应激期间)肉鸡平均日增重结果(g)(n=6)
注:#表示与空白对照组相比差异显著(P<0.05);*表示与LPS组相比差异显著(P<0.05)。
由表1可知,与空白对照组相比,LPS组肉鸡在应激期间的平均日采食量和平均日增重都显著下降,各个给药组都能不同程度的提高鸡只的平均日采食量和平均日增重,低剂量给药组能够显著升高鸡只的平均日增重和平均日采食量,高剂量给药组能够显著升高鸡只的平均日增重。
表2 36日龄肉鸡免疫器官指数结果(g/kg)(n=6)
注:#表示与空白对照组相比差异显著(P<0.05),##表示与空白对照组相比差异极显著(P<0.01);*表示与LPS组相比差异显著(P<0.05),**表示与LPS组相比差异极显著(P<0.01)。
脾脏、胸腺和法氏囊是禽类机体发挥免疫效应的器官,检测这些器官的免疫指数可以初步衡量机体的免疫状态及免疫系统的发育情况。由表2可知,与空白对照组相比,LPS组肉鸡的免疫器官指数都有显著降低,高剂量给药组与中剂量给药组肉鸡的胸腺指数和法氏囊指数显著升高,而低剂量给药组肉鸡胸腺指数极显著升高。在一定程度上反映了动物免疫机能的增强,有利于促进机体产生抗炎因子和提高免疫水平。
表3 36日龄肉鸡血清IL-6、TNF-α、IL-1β的含量(pg/mL)(n=6)
注:#表示与空白对照组相比差异显著(P<0.05),##表示与空白对照组相比差异极显著(P<0.01);*表示与LPS组相比差异显著(P<0.05),**表示与LPS组相比差异极显著(P<0.01)。
IL-6、TNF-α、IL-1β都是炎症发展过程中,炎症细胞和巨噬细胞产生的炎症介质,在机体内具有很强的炎症损伤作用。
由表3可知,与空白对照组相比,LPS组的IL-6含量显著升高,TNF-α和IL-1β有明显的升高。与LPS组相比,高剂量给药组肉鸡血清中IL-6和TNF-α含量极显著下降,IL-1β含量显著下降,中剂量给药组肉鸡血清中TNF-α和IL-1β含量极显著降低,低剂量给药组肉鸡血清中TNF-α含量极显著降低、IL-1β含量显著降低。
表4 36日龄肉鸡血清中ALT、AST的含量(U/L)(n=6)
注:#表示与空白对照组相比差异显著(P<0.05);*表示与LPS组相比差异显著(P<0.05)。
谷丙转氨酶(ALT)、谷草转氨酶(AST)都是在肝脏细胞浆中合成的,而肝脏是免疫应激损伤的靶组织,当机体免疫应激时导致肝细胞膜通透性增加,大量的AST、ALT从细胞释放进入血液中,导致血清中两种酶含量增高。因此,血清中ALT、AST含量的上升可以表明肝细胞受到损伤。由表4可知,LPS组肉鸡血清中ALT含量显著升高,高剂量给药组肉鸡血清中ALT和AST的含量显著下降,中剂量给药组和低剂量给药组肉鸡血清中AST含量显著下降。
表5 36日龄肉鸡肝脏中SOD、MPO的活性(U/g)(n=6)
注:#表示与空白对照组相比差异显著(P<0.05);*表示与LPS组相比差异显著(P<0.05)。
动物机体处在免疫应激状态时,自由基稳态平衡受到破坏,机体内会产生大量的自由基,进而导致氧化应激,氧化应激能够破坏细胞氧化还原稳态,破坏动物体内的氧化平衡状态,导致动物生长性能下降。超氧化物歧化酶(SOD)主要存在于细胞液和线粒体基质中,是防御生物体氧化损伤的一种十分重要的酶。髓过氧化物酶(MPO)是反映中性粒细胞浸润和聚集程度的一个指标,其活性变化是组织氧化损伤炎症发展的重要过程。由表5可知,与空白对照组相比,LPS组肉鸡肝脏中SOD含量明显降低,MPO含量显著升高。与LPS组相比,低剂量给药组和中剂量给药组肉鸡肝脏中SOD含量有上升趋势,低剂量给药组肉鸡肝脏中MPO含量显著下降,中、高剂量给药组肉鸡肝脏中MPO含量有下降趋势。
表6 36日龄肉鸡血清中DAO、D-LA的活性(nmol/L)(n=6)
注:#表示与空白对照组相比差异显著(P<0.05),##表示与空白对照组相比差异极显著(P<0.01);*表示与LPS组相比差异显著(P<0.05),**表示与LPS组相比差异极显著(P<0.01)。
二胺氧化酶(DAO)是由肠道上皮产生的细胞内酶,研究发现完整的肠道上皮黏膜中存在机体95%以上的DAO。D-乳酸(D-LA)由肠道细菌代谢和裂解产生。动物机体健康情况下D-LA在血中含量很低,当肠道黏膜屏障受损时DAO就会释放到全身血液中。因此,DAO、D-LA的含量可反映肠道屏障受损情况,是可以评价肠道通透性的敏感指标。由表6可知,与空白对照组相比,LPS组肉鸡血清中DAO含量显著上升,D-LA含量极显著上升;与LPS组相比,低剂量给药组肉鸡血清中DAO含量显著下降,中剂量给药组肉鸡血清中D-LA含量显著降低,高剂量给药组肉鸡血清中D-LA含量显著下降。
综上,食用添加了该中药复方的饲料的肉鸡实验组在平均日增重、平均日采食量、料重比、脾脏指数、法氏囊指数、胸腺指数、抗炎、抗氧化、降低肠道通透性等方面的表现均优于未食用添加该中药复方的饲料的LPS组,且该中药复方不添加抗生素,降低产生耐药性的风险,不会使大量畜禽甚至人类暴露于耐药菌株引起的感染危险中,更不会给食品安全带来隐患,因此,该中药复方能够为肉鸡抗炎、抗应激的养殖提供新的思路。
此外,经发明人多次实验,发现中药复方组按照五味子10g、金银花15g、知母12g、甘草20g称取各原料,在复方总用量相同的情况下,各药物用量微小的变动对实验结果影响甚小,故本发明实施例详细记载为五味子8~15g;金银花10~20g;知母10~15g,甘草10~20g的实验结果,并经过剂量筛选,认为中药复方按20g/kg与饲料配比混匀,可以使药物性价比发挥到最大。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (7)
1.一种抗肉鸡LPS应激的中药复方,其特征在于,所述中药复方由五味子、金银花、知母、甘草组成,质量比为1:1.5:1.2:2。
2.权利要求1所述的抗肉鸡LPS应激的中药复方的制备方法,其特征在于,包括如下步骤:
称取五味子、金银花、知母、甘草,烘干,粉碎,过筛,混匀,制得中药复方。
3.根据权利要求2所述的制备方法,其特征在于:
所述的烘干的温度为60℃~70℃;
所述的过筛为过80~150目筛。
4.权利要求1所述的抗肉鸡LPS应激的中药复方在制备抗肉鸡LPS应激的饲料或饲料添加剂中的应用。
5.根据权利要求4所述的应用,其特征在于:
所述的抗肉鸡LPS应激的中药复方在饲料中的添加量为5~20 g/kg。
6.根据权利要求5所述的应用,其特征在于:
所述的抗肉鸡LPS应激的中药复方在饲料中的添加量为10~20 g/kg。
7.根据权利要求6所述的应用,其特征在于:
所述的抗肉鸡LPS应激的中药复方在饲料中的添加量为20 g/kg。
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