CN109497511A - 一种真菌提取物组合物及其在制备功能性食品和保健品中的应用 - Google Patents
一种真菌提取物组合物及其在制备功能性食品和保健品中的应用 Download PDFInfo
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Abstract
本发明涉及一种真菌提取物组合物及其在制备功能性食品和保健品中的应用。该组合物中各组分含量按重量份计为蛹虫草提取物9~11份,灰树花提取物6~8份,绣球菌提取物0.5~1.5份,金耳提取物0.5~1.5份,虎奶菇提取物0.5~1.5份。本发明的组合物可用于慢性支气管炎,哮喘相关疾病的功能性食品和保健品的制备。
Description
技术领域
本发明涉及一种真菌提取物组合物,该组合物中各组分含量按重量份计为蛹虫草提取物9~11份,灰树花提取物 6~8份,绣球菌提取物0.5~1.5份,金耳提取物0.5~1.5份,虎奶菇提取物0.5~1.5份。可用于制作肺部保健的各种功能性食品和保健品制剂。本发明属于食品加工,医药工业相关领域。
背景技术
蛹虫草(Cordyceps militaris)又称北虫草,北冬虫夏草,隶属于真菌界、子囊菌门(Ascomycota)、肉座菌目(Hypocreales)、麦角菌科(Clavicipitaceae),在世界各地均有分布。蛹虫草是中国的传统中药材,早在500年前李时珍所著《本草纲目》中就有记载蝉花主治“小儿天吊惊痫,夜啼,心悸”。1986年吉林省蚕科所以家蚕和柞蚕为寄主培养获得了与天然蛹虫草相一致的子实体,随后人工栽培蛹虫草技术不断发展形成产业化,其保健功能得到了广大人民群众的认可,近些年来在超市、菜场和中药店均能买到蛹虫草子实体鲜品或干品。同时,蛹虫草的研究开发也备受关注,在药理、药效、成分研究方面都取得了相当的进展,比如戴瑛等(中国临床药理学与治疗学,2004 Apr;9(4):386- 388)通过小鼠实验证实蛹虫草提取物可以调节机体组织的免疫功能,缓解由于内毒素或类似物质造成的肺部炎症,对肺具有一定的保护作用。本发明中提到的蛹虫草提取物采用蛹虫草子实体干品为原料,经粉碎,水提,过滤,提取液浓缩,干燥等工艺流程获得。
灰树花(Grifola frondosa)又名贝叶多孔菌,舞茸,属担子菌纲,多孔菌科,其子实体味道鲜美,风味独特,具有抗肿瘤、免疫调节等药理作用,深受国内外市场青睐。目前,国内灰树花子实体栽培已经形成产业化,大大推动了灰树花子实体为原料的各类功能性食品,保健品和药品的开发应用。自20世纪80年代始,国内外学者对灰树花多糖进行了大量研究, 从灰树花子实体和菌丝体中提取了几十种活性多糖,其中包括 D-组分、MD组分、Grifolan组分、T-2组分等具有显著生理活性的组分。劳华均等(上海农业学报,1997,13,(1):25~30)从灰树花子实体中提取得到灰树花粗多糖GFP,实验证明该粗多糖对于小鼠Lewis肺癌和Colon癌均有显著的抑制作用。本发明中提到的灰树花提取物采用灰树花子实体干品为原料,经粉碎,水提,过滤,提取液浓缩,干燥等工艺流程获得。
绣球菌(Sparassis crispa))又名绣球菇,因形似巨大的绣球而得名,隶属于担子菌亚门,异隔担子菌纲,绣球菌科。绣球菌不仅味道鲜美,而且营养价值高,特别是子实体中含有的β-葡聚糖能提高人体免疫力及机体造血功能,对某些肿瘤也有一定功效(赵慧慧,中国农学通报,2013,29(21):149-153)。由于野生绣球菌稀少,价格昂贵,人工栽培形成规模以后绣球菌才逐渐走入了寻常百姓家,由于其味美,营养价值高而深受人们喜爱。本发明中提到的绣球菌提取物采用绣球菌子实体干品为原料,经粉碎,水提,过滤,提取液浓缩,干燥等工艺流程获得。
金耳((Tremella aurantialba)又称云耳、黄木耳、金木耳、金银耳,属担子菌纲,银耳目,银耳科,银耳属。主要分布在四川、西藏、云南、福建、江西等地,为珍贵稀有食用菌。上世纪80年代昆明食用菌研究所人工栽培试验成功,之后若干年成功推向市场。金耳胶质细腻、滑润可口,营养丰富,且有一定药用价值。《西藏常用中草药》记载:金耳味甘,性平,可滋阴润肺、生津、治虚劳、咳嗽、肺结核。现代中医认为具有化痰止咳、定喘调气、平肝阳的功效,适用于肺热痰多,感冒咳嗽,气喘,高血压等症。蒋剑平等(中草药,第40卷,第10期,2009年10月)发现金耳多糖对大鼠过敏性气道炎症具有抑制作用,为进一步开展金耳治疗哮喘和气管炎的研究提供了药理学依据。本发明中提到的金耳提取物采用金耳子实体干品为原料,经粉碎,水提,过滤,提取液浓缩,干燥等工艺流程获得。
虎奶菇(pleurotus tuber – regium(Fr.)Sing)又名虎乳菌、虎奶菌,南洋茯苓等,属真菌界,层菌纲,侧耳科。子实体呈漏斗状至杯状,表面多黄白色,成熟时外皮红褐色,外皮薄且多粗糙,气微,味淡,嚼之有颗粒感,微黏性。虎奶菇对咳嗽、哮喘有神奇疗效,可提高免疫力,防痴呆,消除肺部的污染物。临床可用于肺癌,哮喘,心血管病,神经衰弱等病症。目前虎奶菇人工栽培技术已经成熟,市面上可以看到各式虎奶菇菜肴及其他功能性保健品。本发明提到的虎奶菇提取物采用虎奶菇子实体为原料,经粉碎,水提,过滤,提取液浓缩,干燥等工艺流程获得。
以上是对单个真菌的介绍,将以上几种真菌提取得到提取物,并按本发明所述比例将提取物混合组成提取物组合物未见文献报道。
发明内容
本发明提取物所用原料均为子实体干品,提取溶剂均为水,提取工艺类似,可以归纳为:
1、子实体干品粉碎
粉碎的目的是为了原料中有效物质在水中能更好的溶出,粒度大小没有特殊要求,粒度只要不过大或者过细即可,即便粒度过大或过小也只是影响提取物得率,优选的,对通常生产过程中以能通过20目筛网为标准。
2、 水提
不同加水量和提取温度会影响提取物得率,但是对组合物并没有本质影响,优选的,按原料10倍重量加水,100摄氏度提取2小时。
3、固液分离
固液分离的目的是除去残渣以获得水提物,过滤的方式有多种,常见的有板框过滤,离心分离等,采用何种过滤方式对提取物并无影响,仅仅是为了去除不溶物。
4、提取液浓缩
浓缩的目的是为了最后的干燥,在干燥的过程中降低能耗,缩短时间,常见的有薄膜浓缩,纳滤膜浓缩等。
5、 干燥
将提取物浓缩液干燥,常见的有烘干,喷雾干燥,冻干等,无论那种干燥方式,目的均为脱水,只要能获得干品即可,若干燥后为块状则需另加粉碎步骤,优选的常用喷雾干燥,产物喷干即为粉末状无需另行粉碎。
以上几种物质通过水提得到粉末状水提物,在实际使用过程中相对原药材具有免煎煮使用方便的特点。实际上,使用以上物质的原药材按组合物各组分的比例配制复方也一样具有类似效果,只是需要另行煎煮,不利于商业化。本发明所述提取物采用中药常用的水煎法,故其创新点不在于对原药材的提取,而在于创新性的把几种提取物按一定比例配料得到组合物,该组合物在肺部保健具有显著作用,比该组合物中任何单个成分均强。
该组合物中各组分含量按重量份计为蛹虫草提取物9~11份,灰树花提取物6~8份,绣球菌提取物0.5~1.5份,金耳提取物0.5~1.5份,虎奶菇提取物0.5~1.5份。优选的,各组分含量按重量份计为蛹虫草提取物10份,灰树花提取物7份,绣球菌提取物1份,金耳提取物1份,虎奶菇提取物1份。
支气管炎,哮喘的病症共同点之一就是均伴随有呼吸道炎症,实施例7和实施例8的动物实验表明本发明提取物组合物能抑制致敏大鼠抗原攻击后支气管肺泡灌流液中嗜酸性粒细胞升高,能降低哮喘大鼠血浆中IL- 4水平及IL- 4/ IFN-γ值,减少体内IgE的合成及嗜酸性粒细胞的活化,进而减轻气道炎症,降低气道高反应性,减轻发病症状。
(三)说明书附图
无
(四)具体实施方式
下面结合具体实施例对本发明进行进一步描述,但本发明的保护范围并不仅限于此:
实施例1
称量蛹虫草,灰树花,绣球菌,金耳子和虎奶菇实体中任意一种按重量份计1份,加10倍重量的水90摄氏度提取2小时,固液分离得提取液,提取物浓缩,浓缩液烘干并粉碎即得相应的提取物。
实施例2
称量蛹虫草,灰树花,绣球菌,金耳子和虎奶菇实体中任意一种按重量份计1份,加10倍重量的水100摄氏度提取2小时,固液分离得提取液,提取物浓缩,浓缩液喷雾干燥即得相应的提取物。
实施例3
实施例1或实施例2中所得各种提取物按重量份计蛹虫草提取物10份,灰树花提取物7份,绣球菌提取物1份,金耳提取物1份,虎奶菇提取物1份的配比,混合均匀即得本发明所述组合物。
实施例4
实施例1或实施例2中所得各种提取物按重量份计蛹虫草提取物9份,灰树花提取物6份,绣球菌提取物0.5份,金耳提取物0.5份,虎奶菇提取物0.5份的配比,混合均匀即得本发明所述组合物。
实施例5
实施例1或实施例2中所得各种提取物按重量份计蛹虫草提取物11份,灰树花提取物8份,绣球菌提取物1.5份,金耳提取物1.5份,虎奶菇提取物1.5份的配比,混合均匀即得本发明所述组合物。
实施例6
实施例1或实施例2中所得各种提取物按重量份计蛹虫草提取物11份,灰树花提取物6份,绣球菌提取物1.5份,金耳提取物0.5份,虎奶菇提取物0.5份的配比,混合均匀即得本发明所述组合物。
实施例7 提取物组合物对支气管炎的药理作用
一、实验目的
评价提取物组合物对致敏大鼠抗原攻击后支气管肺灌流液中的炎症细胞
的作用。组合物中主要为灰树花提取物和蛹虫草提取物,故将两者作为受试材料列入对比。
二、材料:
1, 本发明中所述提取物组合物(按重量份计蛹虫草提取物10份,灰树花提取物7份,绣球菌提取物1份,金耳提取物1份,虎奶菇提取物1份)、灰树花提取物、蛹虫草提取物。
2, 卵白蛋白(V级),美国Sigma chemical Co.。
3, 地塞米松,高邮制药厂。
4, 乌拉坦(Urethane),中国医药公司上海化学试剂公司。
5, SD大鼠,雌性,清洁级。
三、实验方法
1, 分组:SD大鼠共60只,分为6组,分别为正常组,模型组,提取物组合物组,灰树花提取物组,蛹虫草提取物组和地塞米松组,每组10只。
2, 致敏:将卵白蛋白溶于10%氢氧化铝凝胶中,配成2%卵白蛋白凝胶溶液。每只大鼠四足掌皮下注射,每点0.1 ml;10天后再重复致敏一次。
3, 给药:从致敏第10天开始给药。提取物组合物、灰树花提取物、蛹虫草提取物或生理盐水溶液0.5 g / kg灌胃(ig),或地塞米松0.5 mg / kg腹腔注射 (ip),以上受试物均给药10天。
4, 抗原攻击:致敏第18天,给药后1小时将致敏大鼠置于4L玻璃钟罩内,用 PARIMASTER 压缩吸入机气雾1.0% 卵蛋白30 min,次日重复攻击一次。将大鼠放回鼠笼再养24h后供实验用。
5, 支气管肺泡灌流:乌拉坦1g / kg ip大鼠,麻醉后切开颈部皮肤,分离出气管,切开气管,插入气管插管,用含肝素的Hanks溶液10 ml,分2次,每次5 ml,从气管插管注入气道,每次来回冲洗3回,冲洗液收集于试管内,回收率约70-80%(7-8 ml)。
6, 白细胞计数和分类计数:1%冰醋酸1:1稀释灌流液,用计数板在显微镜下计总数。将灌流液涂于玻片上,干后用染色液染色,然后在油镜下分类计数。
四、结果与讨论:
模型组经1.0%卵蛋白2次攻击后,嗜酸性粒细胞明显增多、淋巴单核细胞百分率明显降低,表明致敏大鼠抗原攻击有非常明显的过敏性炎症反应过程。对各实验组致敏大鼠给药后,结果表明受试样品均能抑制致敏大鼠抗原攻击后支气管肺泡灌流液中嗜酸性粒细胞升高,但作用强度有明显区别,地塞米松最强,其次是提取物组合物组,单个提取物稍弱。具体结果见表-1。
表-1: 提取物组合物对支气管肺灌流液中炎症细胞的抑制作用( x±SD)。
组别 | 白细胞总数(个/mm<sup>3</sup>) | 嗜酸性粒细胞(%) | 淋巴单核细胞(%) |
模型组 | 444.0<u>+</u>201.1 | 11.05<u>+</u>7.11 | 71.55<u>+</u>14.25 |
正常组 | 142.3<u>+</u>63.1 | 0.00<u>+</u>0.00** | 89.01<u>+</u>14.22* |
地塞米松组 | 322.5<u>+</u>144.3 | 0.00<u>+</u>0.00** | 87.71<u>+</u>17.73* |
提取物组合物组 | 360.0<u>+</u>203.3 | 3.01<u>+</u>1.78** | 87.22<u>+</u>9.57* |
灰树花提取物组 | 402.8<u>+</u>223.3 | 3.88<u>+</u>4.01* | 74.25<u>+</u>11.04 |
蛹虫草提取物组 | 487.0<u>+</u>258.5 | 4.42<u>+</u>2.93* | 77.22<u>+</u>19.84 |
统计方法:T-test,与模型组比较*P<0.05,**P<0.01
实施例8 提取物组合物对哮喘大鼠的药理作用
一、实验目的
评价提取物组合物对大鼠Th1/Th2型细胞因子水平的影响。组合物中主要
提取物为灰树花提取物和蛹虫草提取物,故将两者作为受试材料列入对比。
二、材料:
1, 本发明中所述提取物组合物(按重量份计蛹虫草提取物10份,灰树花提取物7份,绣球菌提取物1份,金耳提取物1份,虎奶菇提取物1份)、灰树花提取物、蛹虫草提取物。
2, 地塞米松,高邮制药厂。
3, IL-4和IFN-γ 酶联免疫试剂盒 北京晶美。
4, 卵白蛋白(V级),美国Sigma chemical Co.。
5、 乌拉坦(Urethane),中国医药公司上海化学试剂公司。
6, SD大鼠,雌性,清洁级。
三、实验方法
1, 分组:SD大鼠共60只,分为6组,分别为正常组,模型组,提取物组合物组,灰树花提取物组,蛹虫草提取物组和地塞米松组,每组10只。
2, 模型制备:第0、7天给大鼠腹腔注射0.2 ml的造模液(含卵白蛋白100 mg、氢氧化铝干粉100 mg、灭活百日咳杆菌5×109个)致敏,第7~21天用超声雾化器给大鼠雾化吸入2%卵蛋白生理盐水悬液,每天3 min,进行激发。最后1次激发后24h采集标本。
3, 给药:从第7天开始给药。提取物组合物、灰树花提取物、蛹虫草提取物或生理盐水溶液0.5 g / kg灌胃(ig),或地塞米松0.5 mg / kg腹腔注射 (ip),以上受试物均给药14天。
4, 血浆采集:大鼠麻醉后仰卧位固定,剪开腹部皮肤,肝素湿润后用1次性注射器抽取腹主动脉血3 ml立即注入抗凝管中,静置2h后离心分取血浆。
5,细胞因子的测定:IL- 4和IFN-γ水平的检测均按试剂盒说明进行,基本过程类似。在酶标板上,分别加入标本及不同浓度的标准品(100μl/孔),室温(20~ 25℃)孵育2小时,洗板4次,加入辣根过氧化物酶标记的单克隆抗体(100μl/孔),室温孵育60min,洗板4次,加显色剂,避光室温10~30min,加终止液,混匀,5min内用酶标仪测光密度值,绘制标准曲线,查出标本浓度。
四、结果与讨论:
T细胞以Th为中心的免疫应答与哮喘炎症的形成密切相关(Bochner BSUndem等,Immunological aspects of allergic asthma,Annual review of immunology 1994 12:295-335.)。T细胞分为Th1和Th2两个亚群。Th1主要分泌IL- 2和IFN-γ介导细胞免疫应答,Th2主要分泌IL- 4、5、6、9、10、13主导B细胞介导的体液免疫应答。哮喘患者由于这些细胞因子分泌失衡导致Th1/ Th2失衡。因此通过评价受试物对细胞因子的影响可以评估该受试物对于哮喘的药理作用。
实验结果(详见表2)表明受试物能降低哮喘大鼠血浆中IL- 4水平及IL- 4/ IFN-γ值,这可能跟受试物抑制Th2细胞的分泌功能有关,从而降低炎性细胞因子IL- 4水平和IL- 4/ IFN-γ比值,减少体内IgE的合成及嗜酸性粒细胞的活化,进而减轻气道炎症,降低气道高反应性,减轻哮喘的症状。
表2 提取物组合物对哮喘大鼠血浆细胞因子的影响
组别 | IL-4(pg/ml) | IFN-γ(pg/ml) | IL-4/IFN-γ |
模型组 | 59.25<u>+</u>8.31 | 113.38<u>+</u>16.05 | 0.53<u>+</u>0.10 |
正常组 | 19.14<u>+</u>3.41 | 100.02<u>+</u>16.21 | 0.19<u>+</u>0.07 |
地塞米松 | 23.05<u>+</u>4.12** | 108.03<u>+</u>14.32 | 0.22<u>+</u>0.09** |
提取物组合物组 | 34.16<u>+</u>6.28** | 110.01<u>+</u>17.03 | 0.31<u>+</u>0.10* |
灰树花提取物组 | 49.14<u>+</u>6.11* | 111.04<u>+</u>15.22 | 0.45<u>+</u>0.12* |
蛹虫草提取物组 | 48.22<u>+</u>5.36* | 112.03<u>+</u>15.47 | 0.43<u>+</u>0.10* |
统计方法:T-test,与模型组比较*P<0.05,**P<0.01
实施例9
取实施例3~6中所述提取物组合物500克,加辅料400克,混匀,制粒,干燥,压制成3000片,即得提取物组合物片,可用于支气管炎和哮喘相关疾病的保健品。
实施例10
取实施例3~6中所述提取物组合物500克,加辅料400克,混匀,制粒,干燥,分装成600袋,每袋1.5克,即得提取物组合物固体饮料,可以作为辅助治疗支气管炎和哮喘相关疾病的功能性食品。
Claims (6)
1.一种真菌提取物组合物及其在制备功能性食品和保健品中的应用,该组合物中各组分含量按重量份计为蛹虫草提取物9~11份,灰树花提取物 6~8份,绣球菌提取物0.5~1.5份,金耳提取物0.5~1.5份,虎奶菇提取物0.5~1.5份。
2.如权利要求书1中所述的组合物,其特征在于,所述各组分含量按重量份计为蛹虫草提取物10份,灰树花提取物7份,绣球菌提取物1份,金耳提取物1份,虎奶菇提取物1份。
3.如权利要求书1中所述的组合物,其特征在于,蛹虫草提取物,灰树花提取物,绣球菌提取物,金耳提取物和虎奶菇提取物所用原料均为各自相应菌类的干燥子实体。
4.如权利要求书1中所述的组合物,其特征在于,蛹虫草提取物,灰树花提取物,绣球菌提取物,金耳提取物和虎奶菇提取物的生产工艺为相应干燥子实体原料经粉碎,水提,固液分离,提取液浓缩,浓缩液干燥得到粉末状物。
5.如权利要求书1~4所述的组合物在制备支气管炎相关疾病功能性食品和保健品中的应用。
6.如权利要求书1~4所述的组合物在制备哮喘相关疾病功能性食品和保健品中的应用。
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