CN108837037A - 一种具有增强免疫力功能的组合物及其制备方法 - Google Patents
一种具有增强免疫力功能的组合物及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种具有增强免疫力功能的组合物及其制备方法,该组合物主要有效成分由以下重量份的原料制备而成:北冬虫夏草1‑5份,甘草1‑5份,枸杞子1‑5份,西洋参2‑10份,淫羊藿10‑30份,黄芪1‑5份。该组合物能有效增强学习记忆力作用,同时还是抗衰老抗氧化和抗肿瘤的代表保健品,尤其适用于免疫力低下患者。
Description
技术领域
本发明涉及组合物,具体是增强免疫功能的组合物,属于保健品领域。
背景技术
随着生活压力的增大及生活节奏的加快,现代人们常常会出现精神萎靡、疲乏无力、食欲降低、睡眠障碍等现象。这些现象的深层原因是免疫力低下。免疫系统主要包括中央和外周淋巴器官、淋巴组织和全身各处的淋巴细胞、抗原呈递细胞等,还包括血液中的白细胞。淋巴细胞经血液和淋巴使各处的淋巴器官和淋巴组织连成一个功能整体。免疫是指机体接触“抗原性异物”或“异己成分”的一种特异性生理反应。免疫系统对维持机体正常生理功能具有重要意义。当人们营养素摄入不足(蛋白质、维生素、微量元素等缺乏)时会造成机体抵抗力下降,导致免疫活性降低,对免疫机制产生不良影响。如今,免疫力减退已成为临床热点问题之一,开发改善或治疗这类疾病的保健品及药物已刻不容缓。
中医中药在我国有着悠久的历史,有“药食同源”的理论作基础,来源于天然产物,毒副作用较小,可以长期服用,因此特别适合于免疫力减弱等亚健康状态的调理和治疗。因此,从天然中药及食材中寻找具有增强免疫力的活性成分,并据此开发药物和保健食品已成为研究热点,具有十分重要的现实意义。
发明内容
基于此,本发明的目的在于克服现有技术的缺陷,提供一种用于提高免疫力的组合物,该组合物具有很好的提高免疫力的作用。为实现上述目的,本发明采取以下技术方案:
一种用于提高免疫力的组合物,所述组合物的有效成分由以下重量份的原料制备而成:
北冬虫夏草1-5份,甘草1-5份,枸杞子1-5份,西洋参2-10份,淫羊藿10-30份,黄芪1-5份。
另外的,一种用于增强学习记忆力的组合物的有效成分由以下重量份的原料制备而成:
北冬虫夏草1-1.2份,甘草1-1.2份,枸杞子1-1.2份,西洋参2-3份,淫羊藿10-15份,黄芪1-1.2份。
另外的,一种用于增强学习记忆力的组合物的有效成分由以下重量份的原料制备而成:
北冬虫夏草1份,甘草1份,枸杞子1份,西洋参2份,淫羊藿10份,黄芪1份。
本发明还公开一种上述的用于增强学习记忆力的组合物的制备方法,包括以下步骤:
制备浸膏一:按重量份称取北冬虫夏草,粉碎,得粗粉,按北冬虫夏草原料重量的5-15倍加水浸泡,随后以盐酸调节溶液pH值为1.0-3.0,再按北冬虫夏草重量百分比的0.2-0.8%加入胃蛋白酶和按北冬虫夏草重量百分比的0.2-0.8%加入胰蛋白酶,37℃酶解2-8小时,然后加热使酶失活,过滤,得酶解液,将酶解液浓缩,即得浸膏一;
制备浸膏二:按重量份称取西洋参和淫羊藿,按西洋参和淫羊藿总重量的6-10倍加入体积百分含量为60%-80%的乙醇溶液,回流提取2-4次,每次回流提取时间为2-4小时,合并滤液,将滤液浓缩,即得浸膏二;
制备浸膏三:按重量份称取甘草、枸杞子和黄芪,按甘草、枸杞子和黄芪总重量的5-15倍加入水,在80-100℃下浸提2-4次,每次浸提时间为2-4小时,合并滤液,合并滤液,将滤液浓缩,即得浸膏三;
混合:将上述得到的浸膏一、浸膏二和浸膏三混合均匀,即得具有提高免疫力功能的组合物。该组合物再经真空干燥,粉碎,得到粉末状的浸膏,命名为组合物1。
本发明的一种用于提高免疫力的组合物及其制备方法,采用复合生物酶解超滤技术提取北冬虫夏草有效成分,工艺方法独特、条件温和,可在有效提取北冬虫夏草活性成分的同时保留其生物活性,使得北冬虫夏草有效成分得到最大限度的利用。且该制备方法中,工艺科学合理,质量稳定、可控、便于操作,可实现工业化连续生产,生产成本低。制备的组合物具有良好的提高免疫力作用,同时还是抗氧化抗衰老的代表中药,尤其适用于免疫力减退患者。并且,这些原料均为食材或药食两用的药材,符合中医“药食同源”理论,天然、毒副作用低,适合长期服用。
具体实施方式
下面详细描述本发明的具体实施方式,本发明不受下述实施案例限制,可根据本发明的技术方案与实际情况确定具体的实施方式。在描述中,在此所使用的某些术语仅用于参照,并不意味着进行限制。
本发明采取以下技术方案:
一种用于提高免疫力的组合物,所述组合物的有效成分由以下重量份的原料制备而成:
北冬虫夏草1-5份,甘草1-5份,枸杞子1-5份,西洋参2-10份,淫羊藿10-30份,黄芪1-5份。
方中:北冬虫夏草(Cordyceps militaris)也叫蛹虫草或蛹草,是虫与菌结合的药用真菌,为现代珍稀中草药,与冬虫夏草的化学成分、营养价值和药理功能非常相似。研究证实,北冬虫夏草具有明显的抗衰老作用,能有效清除人体自由基,抑制过氧化脂质产生,保护细胞免受过氧化物损害,同样能够调节血液黏稠度,扩张心脑血管,保护心肌细胞,增强强免疫力等,具有好的经济价值,药用和保健功能具有很大的开发潜力,目前有用其研制开发出的保健品投入市场。北冬虫夏草多肽为北冬虫夏草的主要活性成分之一,经过酶解工艺直接制得的水解物,有极强的药理作用和生理功能多样性。
甘草为豆科(Legum inosae)蝶形花亚科(Papiliantae Taub)草本植物,又名甜草根、粉草、灵通、国老等,具补脾益气,润肺止咳,缓急止痛,缓和药性,泻火解毒之功效,其有效成分主要为黄酮、三萜和多糖类化合物。研究表明一定剂量的甘草多糖能显著抑制肿瘤细胞的生长,剂量过高会影响其抑瘤效果,甘草多糖对移植小鼠体液免疫功能有增强作用,以直接诱导小鼠B淋巴细胞进行增殖,说明甘草多糖具有提高机体免疫抗肿瘤的作用机制。可见甘草多糖具有较强的抗癌活性,以及增强移植瘤小鼠免疫功能。
枸杞子系茄科枸杞属植物宁夏枸杞(Lycium barbarum L)的干燥成熟果实,为传统常用中药,具滋肾润肺、补肝明目、强筋健骨的功效。其主要成分有甜菜碱、枸杞多糖、胡萝卜素、多种氨基酸、维生素、钙、磷、铁等。枸杞子味甘,性平,归肝、肾经,为滋补、强壮之佳品,延年益寿之良药。现代药理研究证明,枸杞多糖具有免疫调节作用,为抗衰老的活性成分。其活性成分LBCP(枸杞子粗多糖)是一种有效的免疫增强剂,在细胞免疫和体液免疫调节中具有重要作用。LBCP主要使用于T细胞,激活T细胞产巨嗜细胞活化因子介导以增强巨嗜细胞的活性。枸杞多糖在所用剂量下可增强巨嗜细胞释放NO能力及胞内溶菌酶活性,说明枸杞多糖有活化巨嗜细胞作用。
西洋参(Panax quinquefolium L)又名美国参、花旗参、洋参,为五加科人参属多年生草本植物。西洋参的多种化学成分与人参相似,含有多种人参皂苷、多糖、挥发油、维生素及微量元素等生理活性物质,在安神益智、抗疲劳、抗氧化、抗体克、抗应激、调节血脂、降血压、降血糖和抗肿瘤等方面具有较好的生理功能。西洋参能抑制免疫细胞内游离钙水平,有利于防止细胞内游离钙浓度过高造成免疫功能衰退。
淫羊藿(EpimediumdavidiiFranch)又名仙灵脾,小檗科淫羊藿属的多年生草本植物,《神农本草经》即有记载,包括箭叶淫羊藿、巫山淫羊藿、朝鲜淫羊藿、柔毛淫羊藿、心叶淫羊藿等,有广泛的药用价值。《中国药典》中记载,淫羊蕾味辛、甘,性温,归肝、肾经,有助阳益精、补肝肾、强筋骨和祛风湿等很多种药理作用,可以使机体的免疫力增强,降压理气。其在临床上经常用来治疗老年腰膝酸软、肾虚阳痪、风寒湿痹及老年神经衰弱、冠心病、慢性支气管炎、高血压、外阴白斑、病毒性心肌炎、更年期综合征、中年健忘、骨质疏松等疾病,特别是在很多重大疾病如癌症、心血管疾病的预防和治疗方面有很好的应用前景。
黄芪(Leguminosae)为豆科植物蒙古黄芪或膜荚黄芪的干燥根,主要药效成分为黄芪多糖,含有皂苷类成分,近年来研究发现黄芪中这些活性成分均有促进抗体生成和免疫反应的作用。黄芪味甘性温,有益气补虚、托毒生肌、利水退肿的功效。黄芪的成分包括黄芪多糖(APS)、生物碱、黄酮、叶酸以及多种微量元素,中黄芪多糖是主要成分,有广泛的药理作用。临床研究和实验表明,黄茂多糖能够改善心、肝、肾脏的功能,调节血糖和血压,增强机体免疫力,还具有抗氧化、抗病毒、防衰老的作用。其中黄芪多糖的主要生物活性是调节机体免疫力,并且黄芪多糖的其他生物活性很大程度上都基于其对机体免疫系统的调节作用。
中医学里认为疾病的发生在于人体正气的虚衰,正气是指人体自身机能的活动以及对于外界的适应、抗病和康复能力,正气充足则人体阴阳协调,五脏六腑气血运作正常,抵抗外来异物,保持健康。正气不足则人体阴阳失衡,机体功能失常,易受外界病菌感染,产生疾病。免疫力低下患者体内阴阳失调,肺虚致感冒不断,肝肾虚致精力不济,脾胃虚致抵抗力下降,这些情况导致患者体质虚弱,种病症反复发作。其中,北冬虫夏草补肾精、益元阳,甘草补养心气、和中缓急,二者共为君药;淫羊藿补肾阳,强筋骨,祛风湿,枸杞子益精明目,滋补肝肾,共为臣药;西洋参补益气血,清热生津,强壮后天之本为佐药;黄芪补肺健脾,实卫敛汗为使药。上述几种药材或食材通过合理的药物配伍,达到了提高免疫力的作用。
在其中一个实施例中,所述组合物的有效成分由以下重量份的原料制备而成:
北冬虫夏草1-5份,甘草1-5份,枸杞子1-5份,西洋参2-10份,淫羊藿10-30份,黄芪1-5份。
在其中一个实施例中,所述组合物的有效成分由以下重量份的原料制备而成:
北冬虫夏草1份,甘草1份,枸杞子1份,西洋参2份,淫羊藿10份,黄芪1份。
本发明还公开一种上述的用于提高免疫力的组合物的制备方法,包括以下步骤:
制备浸膏一:按重量份称取北冬虫夏草,粉碎,得粗粉,按北冬虫夏草原料重量的5-15倍加水浸泡,随后以盐酸调节溶液pH值为1.0-3.0,再按北冬虫夏草重量百分比的0.2-0.8%加入胃蛋白酶和按北冬虫夏草重量百分比的0.2-0.8%加入胰蛋白酶,37℃酶解2-8小时,然后加热使酶失活,过滤,得酶解液,将酶解液浓缩,即得浸膏一;
制备浸膏二:按重量份称取西洋参和淫羊藿,按西洋参和淫羊藿总重量的6-10倍加入体积百分含量为60%-80%的乙醇溶液,回流提取2-4次,每次回流提取时间为2-4小时,合并滤液,将滤液浓缩,即得浸膏二;
制备浸膏三:按重量份称取甘草、枸杞子和黄芪,按甘草、枸杞子和黄芪总重量的5-15倍加入水,在80-100℃下浸提2-4次,每次浸提时间为2-4小时,合并滤液,合并滤液,将滤液浓缩,即得浸膏三;
混合:将上述得到的浸膏一、浸膏二和浸膏三混合均匀,即得具有提高免疫力功能的组合物。该组合物再经真空干燥,粉碎,得到粉末状的浸膏,命名为组合物1。
本发明的制备方法,采用现代制药技术提取原料中的有效成份,在保证疗效的前提下,提高了便利性和口感。并且在制备北冬虫夏草浸膏一时,以胃蛋白酶和胰蛋白酶进行酶解,使北冬虫夏草蛋白质中的部分肽键水解,获得吸收速度更快、生物活性更强的小分子多肽,使其更利于人体吸收,增加北冬虫夏草的生物利用度。此外,酶解反应条件温和、过程易控制,酶解过程不会破坏北冬虫夏草中其他有效成分的生物活性。
在其中一个实施例中,制备浸膏一步骤中,所述胃蛋白酶和胰蛋白酶的重量比为2:1-1:2。
在其中一个实施例中,制备浸膏一步骤中,将酶解液以截留分子量为100000的超滤柱进行过滤,得滤液,滤液减压浓缩,即得浸膏一。去除大分子杂质,保留北冬虫夏草酶解所得的多肽、腺苷、多糖、多种维生素等主要活性成份,对北冬虫夏草的有效成份进行纯化。
在其中一个实施例中,制备浸膏一步骤中,加热至100℃保持15-30分钟使酶失活。
在其中一个实施例中,制备浸膏一步骤中,滤液减压浓缩至55-65℃时密度为1.15-1.25g/mL的稠浸膏,再经真空干燥,粉碎,得到粉末状的浸膏一;
制备浸膏二步骤中,滤液减压浓缩至55-65℃时密度为1.15-1.25g/mL的稠浸膏,再经真空干燥,粉碎,得到粉末状的浸膏二;
制备浸膏三步骤中,滤液减压浓缩至55-65℃时密度为1.15-1.25g/mL的稠浸膏,再经真空干燥,粉碎,得到粉末状的浸膏三。
本发明还公开了一种上述的组合物在制备用于提高免疫力的药物或保健食品中的应用。
在其中一个实施例中,所述药物或保健食品的剂型为片剂、颗粒剂、硬胶囊、软胶囊、口服液、丸剂或滴丸。与现有技术相比,本发明具有以下有益效果:
本发明的一种用于提高免疫力的组合物,原料由北冬虫夏草,甘草,枸杞子,西洋参,淫羊藿,黄芪组成。这几种药材和食材相互配合,协同作用,经动物实验结果证实:经口服灌胃给小鼠本发明的组合物30天后,进行免疫器官指数测定、血液中白细胞数量血清中溶血素含有量测定和迟发型变态反应实验,实验结果均显示本发明的组合物具有提高小鼠免疫功能的作用。
另外,本发明的组合物中诸药不仅具有良好的提高免疫力作用,同时还是抗衰老抗氧化和抗肿瘤的代表中药。所以,本发明的组合物在传统的调补心血基础上,还辅以滋补肾精,诸药配伍合理,是治疗免疫力低下的上佳之选。
并且,这些原料均为食材或药食两用的药材,符合中医“药食同源”理论,天然、毒副作用低,适合长期服用。
本发明的一种用于提高免疫力的组合物的制备方法,采用复合生物酶解超滤技术提取北冬虫夏草有效成分,工艺方法独特、条件温和,可在有效提取北冬虫夏草活性成分的同时保留其生物活性,使得北冬虫夏草有效成分得到最大限度的利用。
且该制备方法中,工艺科学合理,质量稳定、可控、便于操作,可实现工业化连续生产,生产成本低。
具体实施方式
以下结合具体实施例来详细说明本发明。
本实施例中所采用的材料和仪器如下:
胃蛋白酶(长春鼎国生物技术有限责任公司,3000u/mg),胰蛋白酶(长春鼎国生物技术有限责任公司,250u/mg),超滤柱(北京赛普瑞特设备有限公司,UF2型,截留分子量100000)。
实施例1
一种用于提高免疫力的组合物,所述组合物的有效成分由以下原料制备而成:
北冬虫夏草500g,甘草500g,枸杞子500g,西洋参1000g,淫羊藿5000g,黄芪500g。
具体制备方法如下:
(1)制备浸膏一
称取北冬虫夏草500g,洗净后,于60℃烘干,时间4h;在室温下粉碎,过80目筛,得北冬虫夏草粗粉;加入北冬虫夏草原料重量10倍重量的纯水,室温浸泡2小时后,用0.1mol/L的盐酸调节pH=2.0;再分别加入北冬虫夏草原料重量百分比0.5%的胃蛋白酶和北冬虫夏草原料重量百分比0.5%的胰蛋白酶,在37℃下恒温震荡酶解5小时后,加热至100℃灭酶20分钟,过滤,得酶解液,酶解液经截留分子量100000的超滤柱过滤,得到分子量在100000以下的北冬虫夏草滤液;滤液在0.06MPa的压力下以60℃浓缩至密度为1.15-1.25g/mL(60℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的浸膏一,备用;
(2)称取西洋参1000g、淫羊藿5000g,洗净后加西洋参和淫羊藿总重量8倍量的体积百分含量为60%的乙醇溶液,回流提取(90℃下)3小时,100目筛过滤,收集滤液;滤渣加6倍重量的60%乙醇,在相同条件下再回流提取2次,每次2小时,100目筛过滤,收集滤液;合并3次的滤液,在0.06MPa的压力下以60℃浓缩至密度为1.15-1.25g/mL(60℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的浸膏二,备用;
(3)称取甘草500g、枸杞子500g、黄芪500g,洗净后按甘草、枸杞子和黄芪总重量的10倍加入水,在室温浸泡2小时,用提取罐加热至100℃,浸提2小时,100目筛过滤,收集滤液;滤渣加10倍重量的水,在相同温度下再浸提2小时,100目筛过滤,收集滤液;合并2次的滤液,在0.06MPa的压力下以60℃浓缩至密度为1.15-1.25g/mL(60℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的浸膏三,备用;
(4)将上述得到的浸膏一、浸膏二和浸膏三混合均匀,按常规制剂方法,加入糊精、蔗糖适量,并以95%乙醇制颗粒,低温干燥,过筛,分装,即得颗粒剂。
实施例2
一种用于提高免疫力的组合物,所述组合物的有效成分由以下原料制备而成:
北冬虫夏草500g,甘草100g,枸杞子500g,西洋参200g,淫羊藿3000g,黄芪100g。
具体制备方法如下:
(1)制备浸膏一
称取北冬虫夏草500g,洗净后,于50℃烘干,时间5h;在室温下粉碎,过80目筛,得北冬虫夏草粗粉;加入北冬虫夏草原料重量5倍重量的纯水,室温浸泡1小时后,用0.1mol/L的盐酸调节pH=1.0;再分别加入北冬虫夏草原料重量百分比0.2%的胃蛋白酶和北冬虫夏草原料重量百分比0.4%的胰蛋白酶,在37℃下恒温震荡酶解8小时后,加热至100℃灭酶15分钟,过滤,得酶解液,酶解液经截留分子量100000的超滤柱过滤,得到分子量在100000以下的北冬虫夏草滤液;滤液在0.06MPa的压力下以55℃浓缩至密度为1.15-1.25g/mL(55℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的浸膏一,备用;
(2)称取西洋参200g,淫羊藿3000g,洗净后加西洋参和淫羊藿总重量5倍量的体积百分含量为80%的乙醇溶液,回流提取5小时,100目筛过滤,收集滤液;滤渣加5倍重量的80%乙醇,在相同条件下再回流提取3次,每次5小时,100目筛过滤,收集滤液;合并4次的滤液,在0.06MPa的压力下以55℃浓缩至密度为1.15-1.25g/mL(55℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的浸膏二,备用;
(3)称取甘草100g,枸杞子500g,黄芪100g,洗净后按甘草、枸杞子和黄芪总重量的5倍加入水,在室温浸泡3小时,用提取罐加热至90℃,浸提4小时,100目筛过滤,收集滤液;滤渣加5倍重量的水,在相同温度下再浸提4小时,100目筛过滤,收集滤液;合并2次的滤液,在0.06MPa的压力下以55℃浓缩至密度为1.15-1.25g/mL(55℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的浸膏三,备用;
(4)将上述得到的浸膏一、浸膏二和浸膏三混合均匀,按常规制剂方法,加入糊精、蔗糖适量,并以95%乙醇制颗粒,低温干燥,过筛,装胶囊,即得硬胶囊剂。
实施例3
一种用于提高免疫力的组合物,所述组合物的有效成分由以下原料制备而成:
北冬虫夏草100g,甘草500g,枸杞子100g,西洋参1000g,淫羊藿1000g,黄芪500g。
具体制备方法如下:
(1)制备浸膏一
称取北冬虫夏草100g,洗净后,于40℃烘干,时间6h;在室温下粉碎,过80目筛,得北冬虫夏草粗粉;加入北冬虫夏草原料重量15倍重量的纯水,室温浸泡3小时后,用0.1mol/L的盐酸调节pH=3.0;再分别加入北冬虫夏草原料重量百分比0.8%的胃蛋白酶和北冬虫夏草原料重量百分比0.4%的胰蛋白酶,在37℃下恒温震荡酶解2小时后,加热至100℃灭酶30分钟,过滤,得酶解液,酶解液经截留分子量100000的超滤柱过滤,得到分子量在100000以下的北冬虫夏草滤液;滤液在0.06MPa的压力下以65℃浓缩至密度为1.15-1.25g/mL(65℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的浸膏一,备用;
(2)称取西洋参1000g,淫羊藿1000g,洗净后加西洋参和淫羊藿总重量10倍量的体积百分含量为40%的乙醇溶液,回流提取3小时,100目筛过滤,收集滤液;滤渣加6倍重量的60%乙醇,在相同条件下再回流提取1次,每次1小时,100目筛过滤,收集滤液;合并2次的滤液,在0.06MPa的压力下以65℃浓缩至密度为1.15-1.25g/mL(65℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的浸膏二,备用;
(3)称取甘草500g,枸杞子100g,黄芪500g,洗净后按甘草、枸杞子和黄芪总重量的15倍加入水,在室温浸泡3小时,用提取罐加热至80℃,浸提1小时,100目筛过滤,收集滤液;在0.06MPa的压力下以65℃浓缩至密度为1.15-1.25g/mL(65℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的浸膏三,备用;
(4)将上述得到的浸膏一、浸膏二和浸膏三混合均匀,按常规制剂方法,加入糊精、蔗糖适量,并以95%乙醇制颗粒,低温干燥,加润滑剂适量,压片,即得片剂。
对比例1
参芪枸杞组合物,通过以下原料制备而成:
西洋参120克,枸杞子180克,黄芪180枚。
具体制备方法如下:以上三味,洗净后加重量8倍量的体积百分含量为60%的乙醇溶液,回流提取(90℃下)3小时,100目筛过滤,收集滤液;滤渣加6倍重量的60%乙醇,在相同条件下再回流提取2次,每次2小时,100目筛过滤,收集滤液;合并3次的滤液,在0.06MPa的压力下以60℃浓缩至密度为1.15-1.25g/mL(60℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的参芪枸杞组合物。
对比例2
甘草提取物,通过以下方法制备:
甘草500g,按原料总重量的10倍量加入水,煎煮2小时,过滤,滤渣再以同样的条件提取2次,合并滤液,在0.06MPa的压力下以60℃浓缩至密度为1.15-1.25g/mL(60℃测)的稠浸膏;再在0.06MPa的压力下以65℃干燥成干浸膏;干浸膏经粉碎,过80目筛,得粉末状的甘草提取物。
实验例1对血液细胞数以及血清溶血素的影响
将上述实施例和对比例制备得到的组合物粉末进行血液细胞数以及血清溶血素的测定
一、材料
1.组合物,通过上述实施例和对比例制备得到的组合物1-5;
2.注射用环磷酰胺,上海新亚药业有限公司;
3.SPF级ICR小鼠,体重16-18g,雄性,购自长春市亿斯实验动物技术有限责任公司。
二、方法
1.受试物
实施例1制备得到的用于提高免疫力的组合物颗粒剂,棕色颗粒,1.0g/袋(组合物有效成分0.5g),对比例1制备得到的粉末状参芪枸杞组合物,对比例2制备得到的粉末状甘草提取物,密封,置阴凉、通风、干燥处保存。
2.动物分组和处理
将ICR雄性小鼠随机分为7组:每组18只,分别为空白对照组、免疫低下模型组、组合物1-5组(2.0g·kg-1·d-1)。对照组和模型组小鼠灌胃给予生理盐水(20mL·kg-1·d-1),组合物1-6组分别灌服2.0g·kg-1·d-1的组合物1-5,连续灌胃14d,第15d除对照组外,其余各组均腹腔注射环磷酰胺(20mg·kg-1)1次,建立免疫低下动物模型。
3.血液细胞数测定
末次给药后,以摘除小鼠眼球的方法采集全血,采用干净无菌的眼科镊子摘取小鼠一侧眼球,让血液自由滴入抗凝管中,采血过程中要不断混匀血液,每只小鼠采集抗凝全血约1mL。在24h内,用全血细胞分析仪检测白细胞总数。
4.血清溶血素的测定
给药结束后,每只小鼠腹腔注射2%SRBC 0.2mL进行免疫。5d后处死小鼠,摘除眼球取血于离心管中,放置使血清充分析出,2 000r/min离心10min,收集血清。取血清10μL,用1mL 1∶5稀释的SA(水杨酸)缓冲液稀释,从1mL中吸取100μL加入离心管,再依次加入10%SRBC 50μL、补体100μL(用SA溶液按1∶8稀释),37℃恒温水浴保温30min,1500r/min离心10min。每管各取上清液50μL,加入另一个96孔培养板中,加都氏试剂150μL。另设半数溶血孔,加10%SRBC 12.5μL,再加都氏试剂至200μL。充分混匀后,用酶标仪在540nm波长下测定光密度值,公式为半数溶血值=样品光密度值/SRBC半数溶血时的光密度值×稀释倍数。
三、结果
实验结果如下表所示。
表1各组小鼠血液细胞数以及血清溶血素影响结果(n=18)
VS对照组:*p<0.05,**p<0.01;VS模型组:#p<0.05,##p<0.01。
结果显示,与正常对照组相比,模型组小鼠血液白细胞总数以及半数溶血值均显著降低,表明小鼠免疫力低下模型建模成功;
与模型组相比,实施例1低、中、高剂量组戊巴小鼠血液白细胞总数以及半数溶血值分别提高21.4%(p﹤0.01)、26.4%(p﹤0.01)和42.0%(p﹤0.01)。不同剂量实施例组小鼠血液白细胞总数均高于模型组,而且高剂量组与模型组比较,具有显著性差异(P<0.05)。表明该实施例组合物能增加小鼠血液中白细胞的数量。
不同剂量实施例组对小鼠血清溶血素的影响数据显示,各剂量组对小鼠血清半数溶血值有不同程度的影响,其中中剂量组与模型组比较,有显著性差异(P<0.05),表明实施例组能提高小鼠血清中溶血素的含有量。
与模型组相比,对比例1组和2组可增加小鼠血液白细胞总数以及半数溶血值(P<0.05),但效果没有实施例组明显。由此说明实施例组有提高小鼠免疫力的作用。
实验例2北冬虫夏草多肽对小鼠迟发型变态反应的影响
将上述实施例和对比例制备得到的组合物粉末进行氧化应激相关酶活性及含量测定实验。
一、方法
绵羊红细胞(SRBC)诱导小鼠迟发型变态反应(足肿胀法)
给药结束后,小鼠用2%SRBC腹腔注射0.2mL。4d后在右后足趾部位皮下注射20%SRBC 20μL,24h后处死小鼠,剪掉左右后足分别称定质量,左右后足质量之差表示足肿胀程度。公式为肿胀度=右后足质量(mg)-左后足质量(mg)。
二、结果
实验结果如下表所示。
表2对小鼠迟发型变态反应的影响(x±s)
组合物与对照组对小鼠迟发型变态反应的影响见表2。数据显示,各组对小鼠足肿胀有不同程度的影响,实施例1中、高剂量组与模型组比较具有显著性差异(P<0.05,P<0.01),但对比例1组和2组效果没有实施例组明显。表明该组合物能增强SRBC诱导小鼠足肿胀程度,具有提高小鼠免疫力的功能。
上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对本发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
Claims (9)
1.一种用于增强免疫力的组合物,其特征在于:所述组合物的有效成分由以下重量份的原料制备而成:北冬虫夏草1-5份,甘草1-5份,枸杞子1-5份,西洋参2-10份,淫羊藿10-30份,黄芪1-5份。
2.一种用于增强免疫力的组合物,其特征在于:所述组合物的有效成分由以下重量份的原料制备而成:北冬虫夏草1-1.2份,甘草1-1.2份,枸杞子1-1.2份,西洋参2-3份,淫羊藿10-15份,黄芪1-1.2份。
3.一种用于增强免疫力的组合物,其特征在于:所述组合物的有效成分由以下重量份的原料制备而成:北冬虫夏草1份,甘草1份,枸杞子1份,西洋参2份,淫羊藿10份,黄芪1份。
4.一种如权利要求1-3中任一项所述用于增强免疫力的组合物的制备方法,其特征在于:包括以下步骤:
制备浸膏一:按重量份称取北冬虫夏草,粉碎,得粗粉,按北冬虫夏草原料重量的5-15倍加水浸泡,随后以盐酸调节溶液pH值为1.0-3.0,再按北冬虫夏草重量百分比的0.2-0.8%加入胃蛋白酶和按北冬虫夏草重量百分比的0.2-0.8%加入胰蛋白酶,37℃酶解2-8小时,然后加热使酶失活,过滤,得酶解液,将酶解液浓缩,即得浸膏一;
制备浸膏二:按重量份称取西洋参和淫羊藿,按西洋参和淫羊藿总重量的6-10倍加入体积百分含量为60%-80%的乙醇溶液,回流提取2-4次,每次回流提取时间为2-4小时,合并滤液,将滤液浓缩,即得浸膏二;
制备浸膏三:按重量份称取甘草、枸杞子和黄芪,按甘草、枸杞子和黄芪总重量的5-15倍加入水,在80-100℃下浸提2-4次,每次浸提时间为2-4小时,合并滤液,合并滤液,将滤液浓缩,即得浸膏三;
混合:将上述得到的浸膏一、浸膏二和浸膏三混合均匀,再经真空干燥,粉碎,得到粉末状的浸膏,即可制得为所述组合物。
5.根据权利要求4所述的制备方法,其特征在于:在制备北冬虫夏草浸膏一时,以胃蛋白酶和胰蛋白酶进行酶解,使北冬虫夏草蛋白质中的部分肽键水解;制备浸膏一步骤中,所述胃蛋白酶和胰蛋白酶的重量比为2:1-1:2;制备浸膏一步骤中,将酶解液以截留分子量为100000的超滤柱进行过滤,得滤液,滤液减压浓缩;制备浸膏一步骤中,加热至100℃保持15-30分钟使酶失活;制备浸膏一步骤中,滤液减压浓缩至55-65℃时密度为1.15-1.25g/mL的稠浸膏,再经真空干燥,粉碎,得到粉末状的浸膏一。
6.根据权利要求5所述的制备方法,其特征在于:制备浸膏二步骤中,滤液减压浓缩至55-65℃时密度为1.15-1.25g/mL的稠浸膏,再经真空干燥,粉碎,得到粉末状的浸膏二。
7.根据权利要求6所述的制备方法,其特征在于:制备浸膏三步骤中,滤液减压浓缩至55-65℃时密度为1.15-1.25g/mL的稠浸膏,再经真空干燥,粉粹,得到粉末状的浸膏三。
8.一种如权利要求1-3中任一项所述用于增强免疫力的组合物在制备用于增强记忆力的保健食品中的应用。
9.如权利要求8所述保健食品的剂型为片剂、颗粒剂、硬胶囊、软胶囊、口服液、丸剂或滴丸。
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