CN111903810A - 中药组合物在制备具有抗应激功效的食品、药品、保健食品或饲料中的应用 - Google Patents
中药组合物在制备具有抗应激功效的食品、药品、保健食品或饲料中的应用 Download PDFInfo
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Abstract
本发明涉及一种中药组合物在制备具有抗应激的功效的食品、药品、保健食品或饲料中的应用,所述中药组合物,由包括如下重量份的原料组分制成:仙草1000~3500份、鸡蛋花135~900份、布渣叶45~900份、菊花180~900份、金银花180~900份、夏枯草180~900份、甘草45~900份。所述中药组合物能够显著提高秀丽线虫的氧化应激或热应激能力,具有良好的抗应激的作用,能够用于制备具有抗应激功效的食品、药品、保健食品或饲料。
Description
技术领域
本发明涉及中药组合物的新应用,特别是涉及一种中药组合物在制备具有抗应激功效的食品、药品、保健食品或饲料中的应用。
背景技术
应激反应是机体在受到各种内外环境、社会及心理因素刺激时所出现的一类非特异性反应,涉及、神经、心血管、内分泌和免疫系统等多方面。它是生命为了生存和发展所必须的,是机体启动适应和保护机制的重要组成部分,主要包括生理反应和心理反应两大类。其中,生理反应则表现为交感神经兴奋、垂体和肾上腺皮质激素分泌增多、血糖升高、血压上升、心率呼吸加快、皮肤及内脏血管收缩等;心理反应可表现为焦虑、恐惧、愤怒、抑郁、逃避与回避、麻木、退化与依赖、警觉性和敏感性增高等。
应激反应根据影响程度可分为良性(积极)和恶性(消极)。适当强度的应激反应对人有积极意义,可提高警觉性、增强身体的抵抗和适应能力,以维持机体的身心平衡;而如果应激反应过于强烈或过于持久,那么不管这些反应是生理性的、还是心理性的,都是有害的,易引发心血管疾病(原发性高血压、冠心病、动脉粥样硬化、雷诺病、心源性猝死)、胃肠道疾病(功能性消化不良、肠易激综合征、应激性溃疡)、自身免疫性疾病(内风湿性关节炎、过敏性哮喘)、内分泌性疾病(肥胖、糖尿病、甲状腺功能亢进)、癌症、抑郁、癔症等多种身心疾病。同样地,除了人之外,牲畜和水产动物在养殖过程中也会因受到各种外界因素改变(包括热刺激)而引起应激反应。应激反应是影响其生产力因素之一,能够干扰养殖对象的正常代谢,降低进食量,影响体重增长,甚至导致免疫力低下而引发疾病,从而最终影响经济效益。
因此,亟待以提高人、牲畜或水产动物的抗应激能力的食品、药品、保健食品或饲料的研究和开发。抗应激性是指能够减轻应激源刺激和增强在应激负荷下的适应能力。目前,抗应激产品仍处于试验阶段或经验应用水平,常用的种类包括:糖类(葡萄糖、活性多糖、几丁聚糖等),氨基酸类(蛋氨酸、色氨酸、赖氨酸、牛磺酸等)、有机酸类(柠檬酸、果酸、延胡索酸、月桂酸等)、维生素类(Vit C、Vit E和Vit B)、皮质激素(地塞米松、肾上腺素等)、电解质和矿物质(氯化钠、氯化钾、钙、铁、锌、硒等)、抗生素(阿莫西林、恩诺沙星、氟苯尼考等)、镇静剂(氯丙嗪、安定、利血平等)、微生态制剂(乳酸菌、芽孢杆菌)等。
中国专利CN102793038A公开一种中药组合物,其原料组成包括仙草1000~3500份、鸡蛋花135~900份、布渣叶45~900份、菊花180~900份、金银花180~900份、夏枯草180~900份、甘草45~900份。各原料组分互相配伍,具有祛火除湿、清热解毒、生津止渴的功效。目前,尚无关于该中药组合物的抗应激功效报道。
发明内容
基于此,有必要提供一种中药组合物在制备具有抗应激功效的食品、药品、保健食品或饲料中的应用。
一种中药组合物在制备具有抗应激功效的食品、药品、保健食品或饲料中的应用,所述中药组合物,由包括如下重量份的原料组分制成:
仙草1000~3500份、鸡蛋花135~900份、布渣叶45~900份、菊花180~900份、金银花180~900份、夏枯草180~900份、甘草45~900份。
在其中一个实施例中,所述中药组合物,由包括如下重量份的原料组分制成:
仙草1500~2500份、鸡蛋花300~500份、布渣叶300~500份、菊花300~500份、金银花300~500份、夏枯草300~500份、甘草300~500份。
在其中一个实施例中,所述中药组合物,由包括如下重量份的原料组分制成:
仙草1900~2500份、鸡蛋花350~450份、布渣叶350~450份、菊花350~450份、金银花350~450份、夏枯草350~450份、甘草350~450份。
在其中一个实施例中,所述中药组合物的制备方法包括如下步骤:
按照配方量配制各原料组分后,加水提取;
浓缩所得提取液,得浓缩液,即可。
在其中一个实施例中,所述水的用量为所述原料组分重量的6-20倍。
在其中一个实施例中,所述提取的时间为0.5-3h。
在其中一个实施例中,加水后,先浸泡10-50分钟,再进行所述提取。
在其中一个实施例中,所述浓缩液的浓度为9-14Be。
在其中一个实施例中,所述应激为氧化应激或热应激。
在其中一个实施例中,所述中药组合物还包括食品、药品、保健食品上可接受的辅料或辅助性成分。
在其中一个实施例中,所述食品为凉茶。
与现有技术相比,本发明具有如下有益效果:
本发明提供了活性成分主要为仙草、鸡蛋花、布渣叶、菊花、金银花、夏枯草、甘草的中药组合物的新应用。通过研究发现:该中药组合物能够显著延长高温或氧化胁迫条件下线虫的存活时间,具有良好的抗应激的作用,能够用于制备具有抗应激功效的药品、食品、保健食品或饲料,提供了该中药组合物的新用途,也为抗应激提供了新的途径。
附图说明
图1A为对照组和中药组合物组(P10)秀丽线虫的抗氧化应激能力对比;
图1B为对照组和中药组合物组(P10)秀丽线虫的抗热应激能力。
具体实施方式
以下结合具体实施例对本发明的中药组合物在制备具有抗应激的功效的食品、药品、保健食品或饲料中的应用作进一步详细的说明。
实施例1
本实施例为所述中药组合物在制备具有抗应激功效的食品、药品、保健食品或饲料中的应用研究。
1、材料与方法
1.1实验试剂及设备
1.1.1实验试剂
秀丽线虫生长固体培养基(NGM)组成包括:NaCl,蛋白胨,琼脂,链霉素,胆固醇,CaCl2,MgSO4,制霉菌素。秀丽线虫裂解液组成包括:NaOH,家用漂白剂。秀丽线虫为N2野生型秀丽线虫(C.elegans,the Bristol strain N2),雌雄同体。大肠杆菌OP50(Escherchiacoli OP50;E.coli OP50)为尿嘧啶合成缺陷菌株,在NGM上生长缓慢,不影响在显微镜下观察秀丽线虫的清晰度。
中药组合物试样:由如下重量份的原料组分制成:仙草2200份、鸡蛋花400份、布渣叶400份、菊花400份、金银花400份、夏枯草400份、甘草400份。制备方法如下:
1)原料处理:选择用于制备中药组合物试样的中药原材料,洗净、切碎之后按照配方量配制;
2)水提:于所述原材料中加入6倍于原材料重量的水,浸泡15分钟后,提取1h,得水提液;
3)浓缩:将所述水提液浓缩至浓度为9Be,得中药组合物试样。该中药组合物试样经过灭菌包装后即为凉茶。也可进一步加入药品、保健食品、食品上可接受的辅料制成药品、保健食品或凉茶之外的其它食品,又可以加入其它的饲料原料制成饲料。其中,所述的食品是指一般意义上的食品,不强调特定功能和特定人群;保健食品是指具有一定保健功效的食品,具有特定的功能且一般有特定的食用人群。
1.1.2设备(表1)
表1实验主要仪器
2.2实验溶液及培养基的配制
①1M磷酸钾缓冲液:KH2PO4 108.39g,K2HPO4 35.69g,加水至1L,调pH值至6.0。
②M9缓冲液:KH2PO4 3g,Na2HPO4 6g,NaCl 5g,MgSO4 0.12g,加入蒸馏水至1L,121℃高压灭菌20min。
③秀丽线虫裂解液:NaOH 0.1g,加4mL纯水和1mL NaCl,混合摇匀。秀丽线虫裂解液现配现用。
④LB液体培养基:LB肉汤21g,加水至1L,121℃灭菌15分钟,宜现用现配。
⑤秀丽线虫生长固体培养基(Nematode Growth Medium,NGM):氯化钠3g,琼脂粉17g,胰蛋白胨2.5g,加入975mL蒸馏水,121℃,高压灭菌30min,然后在无菌条件下加入5mg/mL胆固醇1mL(用无水乙醇配制),1M CaCl2、1M MgSO4各1mL和1M pH6.0磷酸钾缓冲液25mL。摇匀后,在无菌条件下,将培养基分装到培养皿(直径55mm平板)中,10mL/板,用前在室温下放置2至3天,再涂布菌液。
2.3实验方法
2.3.1秀丽线虫的准备
①秀丽线虫的培养与传代
参照文献方法(S.Brenner,The genetics of caenorhabditis elegans,Genetics77:71-94,May 1974),在无菌条件下,切一块含有秀丽线虫较多的培养基,转移新的OP50的NGM培养基上即可,在标准培养条件(20℃,湿度40~60%)下培养,秀丽线虫会自动由食物较少的培养基边缘爬到食物较多菌苔中去。由于秀丽线虫非常容易通过培养基表面空隙钻入琼脂内,因此培养秀丽线虫时,应尽量选择表面光滑,无气泡、划痕的琼脂板。约培养48小时,可进行同期化。
②秀丽线虫的同期化
采用高氯酸钠漂白法(Brian Onken et al.,Metformin induces a dietaryrestriction-like state and the oxidative stress response to extend C.eleganshealthspan via AMPK,LKB1,and SKN-1,PLOS one:January 2010,Volume5,Issue1,8758):在无菌条件下,用已高温灭菌的M9缓冲溶液将正处于产卵期的秀丽线虫洗至2mL无菌管中,加1mL裂解液裂解秀丽线虫,振荡3~5min,置低速离心机上3000rpm离心1min,弃上清,再用M9冲洗秀丽线虫2次,置低速离心机上3000rpm离心1min,弃上清后用移液枪吸取2mL无菌管底部剩余的0.1mL秀丽线虫滴于NGM的无菌区,在标准培养条件下培养,约51h后裂解的秀丽线虫体内的受精卵基本发育成L4期幼虫,完成同期化用于试验。
2.3.2不含与含所述中药组合物试样的NGM的配制
①E.coli OP50的培养
挑取OP50的菌种接于100mL LB液体培养基中,在37℃,170rpm条件下,振荡培养12h,至OD600=0.4,用于接种NGM喂养正常组秀丽线虫。培养好的菌液若不是立即使用完,用10mL无菌管分装约每管8mL置于4℃医用冷藏箱中备用。
②不含所述中药组合物试样NGM的配制(对照组)
在每个NGM平板上加适量的OP50菌液(一般直径55mm平板加100μL),用移液器取100μL OP50菌液均匀地涂布在NGM平板中央,尽量使菌液边缘距离平板边缘0.5cm左右。涂布好细菌的NGM平板在室温(21~25℃)下过夜后即可使用,若不是立即使用则将其封膜倒置置于4℃医用冷藏箱中备用。
③含所述中药组合物试样NGM的配制(中药组合物组)
在无菌环境下用10mL容量瓶分别称取100mg所述中药组合物试样,用OP50菌液定容至10mL,配制为浓度为10mg/mL的中药组合物样品(P10)。涂布方法同不含所述中药组合物试样NGM的配制。置于4℃冷藏箱内的平板2周内尽快用完。
2.3.3所述中药组合物对秀丽线虫抗应激能力的影响
(1)在高温胁迫下检测所述中药组合物对秀丽线虫抗应激能力的影响
热应激实验:参照文献方法(Honda Y,Honda S.,The daf-2gene network forlongevity regulates oxidative stress resistance and Mn-superoxide dismutasegene expression in Caenorhabditis elegans[J].,The FASEB journal,1999,13(11):1385-1393),经所述中药组合物处理4天的野生型秀丽线虫从20℃转移到35℃。转移后,每小时记录秀丽线虫的存活率并及时移走那些因为脱水而死亡的秀丽线虫。
(2)在氧化胁迫下检测所述中药组合物对秀丽线虫抗应激能力的影响
氧化应激实验:在挑虫第10天将虫转移至加入了诱导氧化胁迫剂-百草枯(最终浓度:10mM)的固体培养基中,百草枯是一种细胞内游离的物质,能诱导多种氧化应激的自由基生成化合物(Wu J Z,Huang J H,Khanabdali R,et al.,Pyrroloquinoline quinoneenhances the resistance to oxidative stress and extends lifespan upon DAF-16and SKN-1activities in C.elegans[J].,Experimental gerontology,2016,80:43-50)。之后每12小时观察秀丽线虫并记录秀丽线虫的存活情况。
2.3.4数据统计
在秀丽线虫的生命周期内,每48h对其存活、死亡数量进行详细记录,使用GraphPad Prism 5软件、SPSS Statistics 19.0软件和Excel表格对数据进行统计分析。
3、结果与分析
3.1在氧化胁迫下所述中药组合物对秀丽线虫抗应激能力的影响
注:同一列不同的小写字母表示显著差异。
分别转移第7天的秀丽线虫到涂有样品的百草枯固体培养基中,由表2可知,对照组的秀丽线虫平均寿命时间是4.43±0.26d,而涂有所述中药组合物的培养基中秀丽线虫的平均寿命时间是4.95±0.09d,具有显著性差异(P<0.001)。由生长曲线图1A可发现所述中药组合物明显出现让生长曲线右移的情况。
3.2在高温胁迫下所述中药组合物对秀丽线虫抗应激能力的影响
注:同一列不同的小写字母表示显著差异。
从表3可知,在热应激实验中,与对照组相比(平均寿命15.36±0.28d;最大寿命21.67±0.58d),中药组合物组的平均寿命为17.28±0.84d,最大寿命为23.33±0.58d,能显著性提高秀丽线虫的死亡时间(P<0.05)。由图1B可知,与对照组相比,中药组合物组能显著使秀丽线虫生长曲线右移(p<0.001)。
综上可知,所述的活性成分为仙草、鸡蛋花、布渣叶、菊花、金银花、夏枯草和甘草的中药组合物能够显著提高对热力胁迫和百草枯诱导的氧化胁迫的应激能力,具有好的抗热应激和抗氧化应激能力,能够用于制备具有抗应激功效的食品、药品、保健食品或饲料。
以上所述实施例的各技术特征可以进行任意的组合,为使描述简洁,未对上述实施例中的各个技术特征所有可能的组合都进行描述,然而,只要这些技术特征的组合不存在矛盾,都应当认为是本说明书记载的范围。
以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 (10)
1.一种中药组合物在制备具有抗应激功效的食品、药品、保健食品或饲料中的应用,所述中药组合物,由包括如下重量份的原料组分制成:
仙草1000~3500份、鸡蛋花135~900份、布渣叶45~900份、菊花180~900份、金银花180~900份、夏枯草180~900份、甘草45~900份。
2.根据权利要求1所述的应用,其特征在于,所述中药组合物,由包括如下重量份的原料组分制成:
仙草1500~2500份、鸡蛋花300~500份、布渣叶300~500份、菊花300~500份、金银花300~500份、夏枯草300~500份、甘草300~500份。
3.根据权利要求2所述的应用,其特征在于,所述中药组合物,由包括如下重量份的原料组分制成:
仙草1900~2500份、鸡蛋花350~450份、布渣叶350~450份、菊花350~450份、金银花350~450份、夏枯草350~450份、甘草350~450份。
4.根据权利要求1所述的应用,其特征在于,所述中药组合物的制备方法包括如下步骤:
按照配方量配制各原料组分后,加水提取;
浓缩所得提取液,得浓缩液,即可。
5.根据权利要求4所述的应用,其特征在于,所述水的用量为所述原料组分重量的6-20倍;及/或,所述提取的时间为0.5-3h。
6.根据权利要求4所述的应用,其特征在于,加水后,先浸泡10-50分钟,再进行所述提取。
7.根据权利要求4所述的应用,其特征在于,所述浓缩液的浓度为9-14Be。
8.根据权利要求1-7任一项所述的应用,其特征在于,所述应激为氧化应激或热应激。
9.根据权利要求1-7任一项所述的应用,其特征在于,所述中药组合物还包括食品、药品、保健食品或饲料上可接受的辅料或辅助性成分。
10.根据权利要求9所述的应用,其特征在于,所述食品为凉茶。
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