CN116211925A - 一种含有金银花和黄芪的兽类药物组合物 - Google Patents
一种含有金银花和黄芪的兽类药物组合物 Download PDFInfo
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Abstract
本发明公开了一种含有金银花和黄芪的兽类药物组合物,包括以下组分:金银花、黄芪、甘草、白术、黄岑、五倍子、陈皮、有机硒壳聚糖复合物。其中,有机硒壳聚糖复合物的制备是将壳聚糖先进行叠氮化再与末端连接有炔基的有机硒化合物反应,得到含有1,2,3‑***基团的复合物,其中***环状基团和硒原子的共价官能化赋予了多糖同时转移单个电子或氢原子以配对自由基中的未配对电子的能力,提升了抗氧化性。所得到的组合物能够提高畜禽免疫力、预防疾病,且安全性高。
Description
技术领域
本发明涉及兽药技术领域,尤其涉及一种含有金银花和黄芪的兽类药物组合物。
背景技术
近年来,随着我国国民经济的蓬勃发展,食品安全以及健康问题日益受到人们的重视,优质、绿色、健康食品的需求量不断增加。抗生素对于各种动物疾病的预防和治疗都有着十分重要的意义,但长期不当使用抗生素会导致机体产生抗药性、菌群失调、机体的免疫力下降、不良反应及机体药物残留等一系列危害。中兽药(Chinese veterinarymedicine)是指以天然动植物和矿物质为原料炮制加工而成的饮片及其制剂,在中兽医药学理论的指导下可预防及治疗动物疾病,提升其生产性能。中兽药具有无毒性、无残留和不易产生抗药性等产品优势以及多组分、多功能、多靶点及标本兼治等防治疾病的优势。当前,动物疾病复杂多样,人们对动物源性食品高度重视,畜禽产品的安全质量问题也越来越受到人们的重视和关注,因此中兽药产品的开发与应用将会更加广泛。
免疫力是指机体能够识别并消灭侵入的异物(如细菌和病毒等)以及处理老化及病毒感染等细胞的能力。近年来,中医药改善机体免疫力这一功能已经得到了社会的普遍认可。中药主要通过调节机体免疫器官功能、体液免疫、细胞免疫及吞噬细胞功能等方面来发挥免疫功能。动物机体除受遗传因素所致的免疫抑制外,还受到各种外界因素的影响,使机体的免疫器官、细胞、细胞因子的形态、功能受损,导致细胞免疫和体液免疫达不到预期效果,造成动物免疫力下降,发病率增加。造成动物机体免疫抑制的外界因素主要有:(1)病原微生物感染;(2)霉菌毒素:如黄曲霉毒素、T-2毒素、呕吐毒素、玉米赤霉烯酮等霉菌毒素的危害作用是对免疫***的破坏及对免疫应答的强烈抑制。毒素或通过损伤免疫器官,或抑制免疫细胞功能和活性,或诱导细胞凋亡,或减少抗体产生等而实现免疫抑制;(3)药物滥用:众所周知,长时间、大剂量、不合理的使用药物常可造成动物机体的免疫抑制,如***等糖皮质激素类药物、氯霉素类药物、四环素类药物以及磺胺类药物等抗菌药的不合理使用就容易导致机体免疫抑制;(4)应激:应激是导致动物机体免疫抑制的又一诱因。研宄证实过冷过热、断奶、仔猪去势、长途贩运、疫苗免疫应激等会影响淋巴细胞的活性,从而引起明显的免疫抑制,造成免疫器官不同程度的损伤;(5)营养因素:动物日粮中营养物质、微量元素、维生素不仅仅满足动物生长发育之所需,还可对动物免疫***进行调节。均衡的营养可促进免疫器官的发育和功能的正常发挥;促进淋巴细胞分化增殖、受体活化与表达;增强单核巨噬细胞的吞噬功能;促使合成抗体和补体;(6)其他因素如重金属、化学物质污染。
由于畜禽发生免疫抑制病的诱因较多,且在畜禽生活的外界环境中普遍存在,因此畜禽极易患免疫抑制病,从而导致免疫失败、继发感染,甚至发生死亡。因此用具有免疫增强作用的中药来提高畜禽机体的免疫力,抵御外邪入侵,对于提高养殖效益十分必要。为提高畜禽机体的免疫力,抵御外邪入侵,减少疾病发生,提高养殖效益,开发中兽药制剂,既是保障动物健康所需,也是国家产业发展所需,同时也能有利于保障食品安全,提高人们的生活水平和质量。
壳聚糖为甲壳质脱乙酰基产物,具有抗肿瘤、抗炎等多种生物活性,且生物相容性好,无毒副作用。壳聚糖分子中含有3种主要活性官能团:氨基(C2位)、一级羟基(C3位)和二级羟基(C6位),对这些基团进行化学修饰可获得大量在不同领域有应用价值的原材料。目前研究证明壳聚糖及其衍生物在细胞水平和动物整体水平上具有一定的抗氧化作用,对壳聚糖进行硒化可以改善其溶解性低的缺陷,其易于被机体吸收,同时兼具硒与多糖的双重活性,具有显著的抗肿瘤、抗氧化和免疫调节等功能。
专利CN108272874 A公开了一种用于蛋鸡的中药制剂及其制备方法。所述中药制剂是对传统中药的创新,由以下重量份数中药原料制备而成:黄芩10~25份、金银花10~25份、大青叶15~30份、连翘5~15份、黄芪10~25份、柴胡10~25份、大黄5~15份、川芎5~15份、益母草5~15份。上述组合物采用粉碎,超声提取,煎煮提取,壳聚糖纯化等工艺,制备成口服液或颗粒剂,工艺简便高效,提高药物生物利用度及药材利用率,研究质量规范,符合现代中兽药发展方向。
专利CN101624425 B公开了一种有机法制备硒化多糖的方法,包括如下步骤:1)将蕨麻多糖或沙蒿多糖放入反应器中,加入N,N-二甲基甲酰胺,搅拌,得悬浊液A;2)往***中滴加氯化亚砜进行反应,对反应后的溶液减压蒸馏,得固体物质B;3)用N,N-二甲基胺将固体物质B溶解,然后加入到悬浊液A中反应,得反应液C;4)将反应液C用蒸馏水溶解,用氢氧化钠调节pH=7,离心,将上清液装入截流量为8000~14000Da的透析袋中,通自来水透析,再用去离子水透析;将透析袋内溶液减压浓缩,在浓缩液中加入乙醇,室温下静置,离心,收集下层沉淀物,冷冻干燥,得到硒化蕨麻多糖或沙蒿多糖。该发明的方法具有反应时间短、反应副产物少、反应过程简单且产品硒含量高等优点。
壳聚糖由于其溶解性较低,较难分散,因此不易被吸收,对壳聚糖进行硒化可以改善这一问题。然而,目前的硒化改性中生物合成始终存在产率低、硒含量普遍较低、富集周期长,纯化困难等问题,化学合成中则存在过度硒酯化破坏多糖的空间构象的问题,因此,研发出一种针对多糖的不同改性方法以应用到实际提高动物的免疫力中将具有十分重要的意义。
发明内容
有鉴于现有技术中的上述缺陷,本发明所要解决的技术问题是提供一种抗氧化应激能力强,同时能够提升免疫力的含有金银花和黄芪的兽类药物组合物。
化学改性中无机化改性存在方法简单,但活性较低的问题,然而有机化改性虽然易于被机体吸收,同时兼具硒与多糖的双重活性,但仍存在一些缺陷。对壳聚糖进行化学改性主要是多糖C6位半缩醛羟基被***根取代形成硒多糖。硒化改性会影响多糖的相对分子质量,一些研究发现硒化改性会导致多糖的分子链在酸性环境中发生降解。然而,有研究指出硒化改性由于硒的引入会增加天然多糖的相对分子质量。过度的硒化改性可能会破坏多糖的空间构象。利用多糖结构的多样性,寻找不同的化学改性方法合成结构新颖的有机硒多糖对拓展硒多糖的结构,提高其生物活性具有重要的意义。本发明中,发明人通过点击反应合成了有机硒壳聚糖复合物,壳聚糖首先被叠氮化再与末端连接有炔基的有机硒化合物反应,最终得到含有1,2,3-***基团的复合物,其中***环状基团和硒原子的共价官能化赋予了多糖同时转移单个电子或氢原子以配对自由基中的未配对电子的能力,提升了抗氧化性,这一性能的提升不仅有利于对抗应激带来的损伤,而且有助于与其他中药共同发挥作用提升畜禽免疫力。
本发明的技术方案:
一种含有金银花和黄芪的兽类药物组合物,包括以下重量份的组分:金银花5~10份,黄芪5~10份,甘草5~8份,白术5~8份,黄岑5~8份,五倍子5~8份,陈皮5~8份,有机硒壳聚糖复合物1~3份。
所述有机硒壳聚糖复合物的制备方法,包括以下步骤:
S1称取壳聚糖1~5重量份分散在24~30重量份的水/乙酸乙酯的混合溶液中(水与乙酸乙酯质量比=1:7),加入稀盐酸0.5~2重量份,混合均匀后加入到3~10重量份浓度为4.5~5mol/L的NaNO2水溶液中,降温至0~5℃下搅拌1~2h,滴加浓度为4.5~5mol/L的NaN3水溶液1~5重量份,在0~5℃下继续搅拌30~60min,反应结束后,过滤,滤饼加入NaOH水溶液调节pH至中性,随后在40℃下干燥2~4h,得到叠氮化壳聚糖;
S2称取2-甲基-4-(苯基硒基)-3-丁炔-2-醇1~2重量份,KOH 0.6~1重量份,己烷15~30重量份,混合均匀后升温至50℃搅拌,反应1~5h,反应结束后,减压浓缩至干得到炔丙基苯基硒;
S3称取CuI 0.5~1重量份,三乙胺7~10重量份,甲苯8~20重量份,二甲基甲酰胺9~10重量份,混合均匀后升温至40~50℃下搅拌30~60min,得到溶液A;称取步骤S2中的炔丙基苯基硒0.005~0.05重量份溶于8~10重量份的甲苯中,得到溶液B;将溶液B加入到溶液A中,再加入步骤S1中的叠氮化壳聚糖5~10重量份,升温至40~50℃下搅拌10~12h,反应结束后降至室温,过滤,将滤饼分散在5~10重量份的水中,并进行透析,透析袋截留分子量=12000~14000,透析24~48h后冷冻干燥,得到有机硒壳聚糖复合物。
本发明还提供了一种含有金银花和黄芪的兽类药物组合物的制备方法,包括如下步骤:
按上述配方称取金银花、黄芪、甘草、白术、黄岑、五倍子、陈皮,洗净后烘干,粉碎,过筛,再加入有机硒壳聚糖复合物,混合均匀后,即得含有金银花和黄芪的兽类药物组合物。
进一步的,所述烘干温度为30~40℃,时间为4~8h;
进一步的,所述过筛目数为200~400目。
与现有技术相比,本发明的有益效果:
(1)本发明中通过将金银花、黄芪、甘草、白术、黄岑、五倍子、陈皮组成配方制成的组合物具有扶正祛邪、固本培元功效,适用于提高畜禽免疫力、预防疾病;
(2)本发明中的兽药药物组合物具有组方科学、疗效确切、安全性高、制剂稳定、质量可控、制备工艺简单、适合工业化生产等特点;
(3)通过点击化学反应制备得到的有机硒壳聚糖复合物使得兽药组合物具有良好的抗氧化应激能力,有助于进一步提升动物免疫力,且有机硒的加入使得复合物活性更高,更易被吸收。
具体实施方式
下面,通过具体实施例对本发明的技术方案进行详细说明,但是应该明确提出这些实施例用于举例说明,但是不解释为限制本发明的范围。
本发明实施例中部分原料的参数如下:
金银花(拉丁学名:Lonicera japonica Thunb.,),药材金银花呈棒状,上粗下细,略弯曲。气清香,味淡、微苦。性寒,味甘。归肺经、心经、胃经,主治清热解毒、凉散风热。属清热药下属分类的清热解毒药。
黄芪,本品为豆科植物蒙古黄芪Astragalus membranaceus(Fisch.)Bge.var.mongholicus(Bge.)Hsiao或膜荚黄芪Astragalus membranaceus(Fisch.)Bge.的干燥根,味甘,微温,归肺、脾经。具有补气升阳,固表止汗,利水消肿,脱毒排脓,敛疮生肌之功效,主治肺脾气虚,中气下陷,表虚自汗,气虚水肿,疮痈难溃或久溃不敛等证。
甘草(拉丁学名:Glycyrrhiza uralensis Fisch.),性平,味甘。归心经、胃经、脾经、肺经。补脾益气、止咳祛痰、缓急定痛、调和药性,属补虚药下分类的补气药。
白术(拉丁学名:Atractylodes macrocephala Koidz.),根茎略呈圆柱状块形,下部两侧膨大,质坚实,不易折断,气清香,味甜微辛,嚼之略带黏液性。性温,味甘、苦。归脾经、胃经。健脾、益气、燥湿利水、止汗、安胎,属补虚药下属分类的补气药。
黄芩(拉丁学名:Scutellaria baicalensis Georgi),圆锥形或不规则条形,常有分枝,扭曲,质硬而脆,易折断,断面纤维性,鲜黄色或微带绿色。气微,味苦。性寒,味苦。归肺经、胆经、脾经、大肠经、小肠经。清热燥湿、泻火解毒、止血、安胎。属清热药下属分类的清热燥湿药。
五倍子(拉丁学名:Rhus chinensis Mill.),肚倍呈长圆形或纺锤形囊状,质硬而脆,易破碎,气特异,味涩。主治敛肺降火,涩肠止泻,敛汗止血,收湿敛疮。用于肺虚久咳,肺热痰嗽,久泻久痢,盗汗,消渴,便血痔血;外伤出血,痈肿疮毒,皮肤湿烂。
陈皮,为芸香科植物橘(Citrus reticulata Blanco)及其栽培变种的干燥成熟果皮,常3瓣相连,形状整齐,厚度均匀。点状油室较大,对光照视,透明清晰。质较柔软。味辛、味苦,性温。归脾经、肺经。主治理气开胃,燥湿化痰,治脾胃病。属理气药。
所有中草药均购自安徽省亳州市中药材市场。
2-甲基-4-(苯基硒基)-3-丁炔-2-醇,CAS:1719-19-3,纯度:95%,CAPOT化学。
壳聚糖,脱乙酰度>90%,Mw=50000~10000g/mol,浙江金壳药业。
稀盐酸,31wt%,河北墨星生物科技。
对照例1
一种含有金银花和黄芪的兽类药物组合物的制备方法,包括如下步骤:
称取金银花800g,黄芪600g,甘草500g,白术500g,黄岑500g,五倍子500g,陈皮500g,洗净后在40℃烘干6h,粉碎,过200目筛,混合均匀后,即得兽类药物组合物。
实施例1
一种含有金银花和黄芪的兽类药物组合物的制备方法,包括如下步骤:
称取金银花800g,黄芪600g,甘草500g,白术500g,黄岑500g,五倍子500g,陈皮500g,洗净后在40℃烘干6h,粉碎,过200目筛,再加入有机硒壳聚糖复合物150g,混合均匀后,即得兽类药物组合物。
所述有机硒壳聚糖复合物的制备方法,包括如下步骤:
S1称取壳聚糖100g分散在2.46kg水/乙酸乙酯的混合溶液中(水与乙酸乙酯质量比=1:7),加入稀盐酸100mL,混合均匀后加入到380mL浓度为4.93mol/L的NaNO2水溶液中,降温至4℃下搅拌1h,滴加浓度为4.64mol/L的NaN3水溶液400mL,在4℃下继续搅拌60min,反应结束后,过滤,滤饼加入1mol/LNaOH水溶液调节pH至中性,随后在40℃下干燥4h,得到叠氮化壳聚糖;
S2称取2-甲基-4-(苯基硒基)-3-丁炔-2-醇160g,KOH 60g,己烷3L,混合均匀后升温至50℃反应5h,反应结束后,减压浓缩至干得到炔丙基苯基硒;
S3称取CuI 50g,三乙胺1L,甲苯2L,二甲基甲酰胺1L,混合均匀后升温至50℃下搅拌60min,得到溶液A;称取步骤S2中的炔丙基苯基硒0.5g溶于1L的甲苯中,得到溶液B;将溶液B加入到溶液A中,再加入步骤S1中的叠氮化壳聚糖500g,升温至50℃下搅拌12h,反应结束后降至室温,过滤,将滤饼分散在500mL水中,并进行透析,透析袋截留分子量=12000~14000,透析48h后在10Pa,﹣55℃下冷冻干燥48h,得到有机硒壳聚糖复合物。
实施例2
一种含有金银花和黄芪的兽类药物组合物的制备方法,包括如下步骤:
称取金银花800g,黄芪600g,甘草500g,白术500g,黄岑500g,五倍子500g,陈皮500g,洗净后在40℃烘干6h,粉碎,过200目筛,再加入硒化壳聚糖复合物150g,混合均匀后,即得兽类药物组合物。
所述硒化壳聚糖复合物的制备方法,包括如下步骤:
将100g壳聚糖置于5L水中,充分溶胀后加入100mL冰乙酸,在搅拌下形成透明溶液,取133g***钠溶于100mL水中混合均匀,然后将其缓慢加入到壳聚糖溶液中,反应2h后加入无水乙醇500mL,使其完全沉淀,过滤,再用200mL无水乙醇洗涤2次,﹣55℃下冷冻干燥48h,即得硒化壳聚糖。
实施例3
一种含有金银花和黄芪的兽类药物组合物的制备方法,包括如下步骤:
按上述配方称取金银花800g,黄芪600g,甘草500g,白术500g,黄岑500g,五倍子500g,陈皮500g,洗净后在40℃烘干6h,粉碎,过200目筛,再加入有机硒壳聚糖复合物100g,混合均匀后,即得兽类药物组合物。
所述有机硒壳聚糖复合物的制备方法,包括如下步骤:
S1称取壳聚糖100g分散在2.46kg水/乙酸乙酯的混合溶液中(水与乙酸乙酯质量比=1:7),加入稀盐酸100mL,混合均匀后加入到380mL浓度为4.93mol/L的NaNO2水溶液中,降温至4℃下搅拌1h,滴加浓度为4.64mol/L的NaN3水溶液400mL,在4℃下继续搅拌60min,反应结束后,过滤,滤饼加入1mol/LNaOH水溶液调节pH至中性,随后在40℃下干燥4h,得到叠氮化壳聚糖;
S2称取2-甲基-4-(苯基硒基)-3-丁炔-2-醇160g,KOH 60g,己烷3L,混合均匀后升温至50℃搅拌,反应5h,反应结束后,减压浓缩至干得到炔丙基苯基硒;
S3称取CuI 50g,三乙胺1L,甲苯2L,二甲基甲酰胺1L,混合均匀后升温至50℃下搅拌60min,得到溶液A;称取步骤S2中的炔丙基苯基硒0.5g溶于1L的甲苯中,得到溶液B;将溶液B加入到溶液A中,再加入步骤S1中的叠氮化壳聚糖500g,升温至50℃下搅拌12h,反应结束后降至室温,过滤,将滤饼分散在500mL水中,并进行透析,透析袋截留分子量=12000~14000,透析48h后在10Pa,﹣55℃下冷冻干燥48h,得到有机硒壳聚糖复合物。
测试例1
对对照例和实施例所制备的兽药组合物进行免疫力增强的实验,选用健康1日龄AA(爱拔益加)肉鸡1000羽,饲养试验在某肉鸡养殖场进行,所有肉鸡均饲养在人工控制的环境中,采用网上平养,前3d温度控制(33±1)℃,以后每周平均降低3℃,直至24℃,并保持此温度到42日龄,环境湿度维持在65%左右,给予24h光照制度,自由采食和饮水。日常管理与免疫接种按常规程序进行。将肉鸡随机分为对照组和实验组,对照组玉米-豆粕型基础日粮,实验组则在基础日粮中添加1.0%的兽药组合物,试验期42d。分别于试验第21和42天从每个处理组随机抽取15羽(每个重复3羽),禁食12h,称重后采用分离胶真空促凝分离采血管从翅下静脉采血5mL用于分离血清。颈动脉放血致死,迅速解剖采取胸腺、脾、法氏囊,分析天平称重,固定于4%多聚甲醛磷酸缓冲液(pH=7.4)。血液样品室温静置2h后,于4℃条件下3000rpm离心15min,分离血清,﹣70℃保存待用。胸腺、脾和法氏囊等免疫器官被分离后,迅速吸干器官表面水分,剥离脂肪和无关组织,称重,记录免疫器官质量,并计算免疫器官指数,器官指数=(器官质量/屠体重)×100%。肉鸡血清经室温融化后,采用全自动血清生化分析仪测量血清IgA、IgM、IgG含量。具体测试结果见表1.
表1免疫力增强实验结果表
胸腺的主要功能是产生和培育T淋巴细胞,参与机体的细胞免疫功能。法氏囊的主要功能是产生和培育特异性的B淋巴细胞,参与机体的体液免疫功能。脾是免疫应答的场所,是机体对抗病原入侵而产生体液免疫反应和细胞免疫反应的主要部位,胸腺、法氏囊和脾的生长发育状况对禽类机体的免疫功能状态可产生极其重要的直接影响,而三种免疫器官的生长发育状况可通过质量与器官指数来反映。通过表1中的测试数据可以发现,对照例和实施例日粮中添加1.0%的兽药组合物可显著增加肉鸡各免疫器官指数,这表明日粮中适量添加该兽药组合物能明显促进肉鸡中枢和外周免疫器官生长发育。其原因主要是由于免疫器官的生长发育需要大量的营养物质,而本研究所用的兽药组合物中所含的主要成分能够促进动物小肠结构发育,提高肠道消化酶活性和消化能力,进而为免疫器官生长发育提供充足的必需营养物质。动物机体的特异性免疫反应包括体液免疫和细胞免疫,体液免疫主要由B淋巴细胞产生特异性抗体来介导,细胞免疫主要由T淋巴细胞及其分泌的细胞因子来介导。因此,通过检测血清中免疫球蛋白含量可以直接反映动物机体的体液免疫和细胞免疫功能状态。日粮中添加适量兽药组合物剂能明显增加肉鸡血清中免疫球蛋白含量,表明其可明显促进肉鸡生长后期免疫球蛋白和细胞因子的分泌,进而增强机体非特异性免疫功能和细胞免疫功能。实施例1中由于添加的有机硒壳聚糖活性更高因此更易被吸收,因此,提升效果优于其他例。
测试例2
对对照例及测试例所制备的兽药组合物进行免疫应激实验,免疫应激(immunechallenge)又叫免疫激发,在免疫学中是指向动物体内注射抗原来引起机体产生免疫反应。在动物生长过程中,许多因素都能引起免疫应激。动物机体发生过度的免疫反应的过程中会产生大量的自由基,破坏体内的自由基平衡,过量的自由基会导致蛋白质变性、脂质过氧化和细胞解体等,从而破坏了机体氧化-抗氧化平衡,使机体发生氧化应激。购买300只1日龄体重相近的健康京红一号雏鸡,适应性饲养一周后,随机分为6组,即空白对照组、阳性对照组及本发明中的4组对照例及实施例的实验组,除空白组外,其余组雏鸡于试验第8、10、12天腹腔注射细菌脂多糖(1.5mg/kg BW)建立免疫应激模型,从12日龄开始,多糖组连续肌肉注射给药7d,1mL/只,正常组给予相同体积的生理盐水。采用网上平养,前3d温度控制(33±1)℃,以后每周平均降低3℃,直至24℃,并保持此温度到46日龄,环境湿度维持在65%左右,给予24h光照制度,自由采食和饮水。日常管理与免疫接种按常规程序进行。空白对照组及阳性对照组喂食玉米-豆粕型基础日粮,实验组则在基础日粮中添加1.0%的兽药组合物,分别于试验第18、46天,每组随机选取10只鸡进行心脏采血,倾斜静置,常温下制备血清。待血清析出后,3000r/min离心10min,收集上清液,-20℃保存待测。采用生化法测定血清中T-AOC、SOD、GSH-Px和MDA含量或活性。
表2抗氧化实验测试结果表
机体发生氧化应激时氧化还原平衡紊乱,并且伴随着活性氧、活性氮和有毒自由基的过量生成,超过了机体抗氧化防御***的清除能力,机体细胞膜损伤,从而出现氧化应激。总抗氧化能力(T-AOC)是作为衡量机体抗氧化能力的综合指标。超氧化物歧化酶(SOD)是一种特异性抗氧化酶,可以将超氧自由基转化成过氧化氢,再被体内的过氧化物酶和过氧化氢酶水解,维持机体的平衡状态。谷胱甘肽过氧化物酶(GSH-Px)作为机体生物抗氧化防御体系中最重要的抗氧化酶,能够水解体内的过氧化氢,减少脂质过氧化丙二醛(MDA)的沉积,保护机体不受氧化损伤,其活性可以作为评判机体抗氧化状态的指标。丙二醛(MDA)是脂质过氧化的终产物之一,血清MDA的含量能够反映出机体内的氧化程度,间接地说明细胞的损伤程度。通过表2的实验结果可以看出,实施例1中通过在日常饲料中添加兽类组合物可以显著提高血清中T-AOC、SOD和GSH-Px的活性,显著降低MDA含量,说明本发明中的兽类组合物可以通过提高氧化应激条件下血清抗氧化酶的活性和降低MDA的含量来降低血清中过氧化物的产生,缓解由于细菌脂多糖引起的氧化应激,维持体内的氧化和抗氧化***的平衡,实施例1中的有机硒壳聚糖复合物由于其中***环状基团和硒原子的共价官能化赋予了多糖同时转移单个电子或氢原子以配对自由基中的未配对电子的能力,提升了抗氧化性,与实施例2中的硒化壳聚糖相比生物活性更高,吸收性更好,因此抗氧化能力更佳。
Claims (8)
1.一种含有金银花和黄芪的兽类药物组合物,其特征在于,包括以下组分:金银花、黄芪、甘草、白术、黄岑、五倍子、陈皮、有机硒壳聚糖复合物。
2.如权利要求1所述的含有金银花和黄芪的兽类药物组合物,其特征在于,包括以下重量份的组分:金银花5~10份,黄芪5~10份,甘草5~8份,白术5~8份,黄岑5~8份,五倍子5~8份,陈皮5~8份,有机硒壳聚糖复合物1~3份。
3.如权利要求1或2所述的含有金银花和黄芪的兽类药物组合物,其特征在于,所述有机硒壳聚糖复合物的制备方法,包括以下步骤:
S1称取壳聚糖分散在24~30重量份的水/乙酸乙酯的混合溶液中,其中水与乙酸乙酯质量比=1:7,加入稀盐酸0.5~2重量份,混合均匀后加入到3~10重量份浓度为4.5~5mol/L的NaNO2水溶液中,降温至0~5℃下搅拌1~2h,滴加浓度为4.5~5mol/L的NaN3水溶液1~5重量份,在0~5℃下继续搅拌30~60min,反应结束后,过滤,滤饼加入NaOH水溶液调节pH至中性,随后在40℃下干燥2~4h,得到叠氮化壳聚糖;
S2称取2-甲基-4-(苯基硒基)-3-丁炔-2-醇1~2重量份,KOH 0.6~1重量份,己烷15~30重量份,混合均匀后升温至50℃搅拌,反应1~5h,反应结束后,减压浓缩至干得到炔丙基苯基硒;
S3称取CuI 0.5~1重量份,三乙胺7~10重量份,甲苯8~20重量份,二甲基甲酰胺9~10重量份,混合均匀后升温至40~50℃下搅拌30~60min,得到溶液A;称取步骤S2中的炔丙基苯基硒0.005~0.05重量份溶于8~10重量份的甲苯中,得到溶液B;将溶液B加入到溶液A中,再加入步骤S1中的叠氮化壳聚糖,升温至40~50℃下搅拌10~12h,反应结束后降至室温,过滤,将滤饼分散在5~10重量份的水中,并进行透析,透析袋截留分子量=12000~14000,透析24~48h后冷冻干燥,得到有机硒壳聚糖复合物。
4.如权利要求3所述的含有金银花和黄芪的兽类药物组合物,其特征在于:所述步骤S1中壳聚糖为1~5重量份。
5.如权利要求3所述的含有金银花和黄芪的兽类药物组合物,其特征在于:所述步骤S3中叠氮化壳聚糖为0.5~1重量份。
6.一种制备如权利要求1~5任一项所述的含有金银花和黄芪的兽类药物组合物方法,其特征在于,包括如下步骤:
按上述配方称取金银花、黄芪、甘草、白术、黄岑、五倍子、陈皮,洗净后烘干,粉碎,过筛,再加入有机硒壳聚糖复合物,混合均匀后,即得兽类药物组合物。
7.如权利要求6所述的含有金银花和黄芪的兽类药物组合物的制备方法,其特征在于:所述烘干温度为30~40℃,时间为4~8h。
8.如权利要求6所述的含有金银花和黄芪的兽类药物组合物的制备方法,其特征在于:所述过筛目数为200~400目。
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