CN110123982A - 一种草果精油的应用 - Google Patents
一种草果精油的应用 Download PDFInfo
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- CN110123982A CN110123982A CN201910461887.7A CN201910461887A CN110123982A CN 110123982 A CN110123982 A CN 110123982A CN 201910461887 A CN201910461887 A CN 201910461887A CN 110123982 A CN110123982 A CN 110123982A
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Abstract
本发明公开了一种草果精油的应用,包括草果精油在制备治疗或预防脑功能及其障碍疾病的药物中的应用、草果精油在制备改善脑功能及其障碍疾病的健康产品中的应用;本发明首次发现草果精油在实验动物脑功能及其障碍疾病中表现出活性的医药或大健康产品新用途,首次发现草果精油或含草果精油或其任意的组合物在动物体内模型中能改善老年痴呆症、抑郁焦虑等脑功能及其障碍动物的症状。与目前治疗脑功能及其障碍的药物相比,所述草果精油或其任意组合物有抑制胆碱酯酶、调节神经递质等多分子协同作用于多靶点的机制,且具有良好的药代参数,易于透过血脑屏障,因此具有毒理学安全性、代谢稳定,较长的半衰期、较小的副作用的特点。
Description
技术领域
本发明涉及生物医药技术领域,特别涉及一种草果精油在制备治疗和预防脑功能及其障碍疾病药物和健康产品中的用途。
背景技术
姜科植物草果(Amomum tsao-ko Crevost et Lemaire)是一种重要的药食两用植物,我国80%以上草果主要用于食品添加,只有不到20%应用于中成药的配制和其他领域。草果目前主要栽培于云南、贵州、广西及越南等地。草果的药用部位为其成熟干燥的果实,历年《中华人民共和国药典》均有收载。草果有燥湿祛寒、除痰截疟之功效,主要用于治疗寒湿内阻,脘腹胀痛,痞满呕吐,疟疾寒热,瘟疫发热等症。《中国药典》2010年版把测定挥发油总量作为评价草果品质的重要指标,精油的化学组成是草果的主要有效成分之一。现代药理研究表明,草果精油具有抑制细菌和真菌、抗肿瘤、抗氧化、抗辐射诱变、调节胃肠功能、改变药物通透性等多种药理作用。主要机制涉及诱导细胞凋亡、清除DPPH和ABTS自由基、在一定程度上增加血清胃泌素水平、抑制小鼠小肠推进、萜类小分子改变表皮蛋白对药物分子的通透性等。虽有文献报道“草果知母汤”可纠正CNS失衡状态,起到抗惊厥、抗癫痛的作用,但与本发明所涉及的机制不同。文献认为其机制可能涉及抑制神经元兴奋性,从而达到阻断皮层异常放电的作用;上调GABAA受体α1亚单位蛋白的表达,增加与GABAA受体的结合率;抑制癫痫形成中大脑海马区神经元数目的减少及神经元凋亡等。
迄今,已有数篇专利涉及草果精油或复方草果精油的医药用途,主要包括促进肠蠕动、抑菌、抗菌作用。目前,尚无专利涉及草果精油对脑功能及其障碍疾病的缓解或治疗作用。
芳香药用植物在脑功能及其障碍疾病的治疗或缓解中发挥着重要作用。近期,我们应用胆碱酯酶活性测定模型和化合物诱导的老年痴呆症斑马鱼体内筛选模型研究草果精油改善CNS疾病时,首次发现,姜科豆蔻属植物草果(Amomum tsao-ko Crevost etLemaire)的精油可抑制胆碱酯酶活性,能显著改善老年痴呆症斑马鱼的认知和反应能力。草果精油能有效调节脑功能及其障碍,可以用于治疗/预防或缓解包括焦虑、抑郁、失眠、神经退行性疾病等脑功能及其障碍疾病。之前,此药理作用尚未被报道过。
发明内容
本发明的目的在于针对现有技术的不足,提供一种草果精油的应用。
本发明的目的是通过以下技术方案来实现的:一种草果精油的应用,所述应用包括草果精油在制备治疗或预防脑功能及其障碍疾病的药物中的应用、草果精油在制备改善脑功能及其障碍疾病的健康产品中的应用。
进一步地,所述草果精油通过以下方法提取得到:
(1)发酵:选取新鲜或干燥的草果果实,渥堆放置约1-2天,然后粉碎至30-60目的粗颗粒备用。
(2)浸提:将粗颗粒浸泡于7-9倍质量的pH值为5.0-6.0盐酸或硫酸水溶液中,浸提过夜,得浸提混合液。
(3)酶解:向浸提混合液中加入纤维素酶和果胶酶,纤维素酶、果胶酶和浸提混合液的质量配比为0.5-1:0.1-0.5:100,在35-37℃酶解约2-3小时。
(4)提取:将步骤3酶解后的混合液放入水蒸气蒸馏提取仪器中,100℃-120℃回流提取约5-7个小时,收集蒸馏液,冷却至室温后置于油水分离器中分离,油层用无水硫酸钠干燥、分离,即得草果精油;或将步骤3酶解后的混合液放入二氧化碳超临界提取仪器中,提取的温度45℃-48℃,压力为13.70MPa-17.80MPa,时间为50min-100min,二氧化碳的流速为25kg/h-35kg/h;收集提取物,置于分子蒸馏仪器中精制,进料温度为40℃-45℃,进样速度为480-530mL/h,一段蒸发温度为110℃-115℃,二段蒸发温度为60℃-65℃,收集二段蒸馏部分,得草果精油。
进一步地,所述草果为姜科豆蔻属植物草果。
进一步地,所述草果精油用气相色谱质谱联用仪分析检测,按质量百分比计,其组分比例为:1,8-桉叶素>35%、柠檬醛>8%、香叶醇>5%、橙花叔醇>1%。
进一步地,所述脑功能及其障碍疾病包括焦虑、抑郁、失眠、狂躁症、精神分裂症、痉挛、双相障碍、中风、亨廷顿病、脑外伤、脊柱损伤和神经退行性疾病。
进一步地,所述神经退行性疾病意包括老年痴呆、帕金森病、亨廷顿病和肌萎缩性侧索硬化。
进一步地,所述药物包括口服制剂和非口服给药制剂;所述口服制剂包括片剂、丸剂、胶囊剂、颗粒剂、微囊片剂、混悬剂、滴丸、口服液体;所述非口服给药制剂包括吸入剂、注射剂、气雾剂、栓剂、皮下给药剂型。
进一步地,所述健康产品包括化妆品、洗浴剂和香薰剂。
本发明的有益效果是:本发明提供了一种草果精油的制备方法,并首次发现草果精油在实验动物脑功能及其障碍疾病中表现出活性的医药或大健康产品新用途。首次发现草果精油或含草果精油或其任意的组合物在动物体内模型中能改善老年痴呆症、抑郁焦虑等脑功能及其障碍动物的症状。与目前治疗脑功能及其障碍的药物相比,所述草果精油或其任意组合物有抑制胆碱酯酶、调节神经递质等多分子协同作用于多靶点的机制,且具有良好的药代参数,易于透过血脑屏障,因此具有提高/改善的毒理学安全性(即毒性降低)、提高/改善的代谢稳定,较长的半衰期和/或较小的副作用的特点,同时产生类似或提高的生物活性(药效)。制备的药物或健康产品可用于预防或治疗脑功能及其障碍疾病和病症,如焦虑、抑郁、失眠、惊厥、狂躁症、精神分裂症、痉挛和双相障碍。还可用于认知功能的提高和改善。
附图说明
图1是草果精油的抑制乙酰胆碱酯酶活性示意图;
图2是草果精油的抑制丁酰胆碱酯酶活性示意图;
图3是草果精油恢复老年痴呆症斑马鱼的反应能力的示意图;
图4是草果精油恢复老年痴呆症斑马鱼的移动速度的示意图。
具体实施方式
1、概述
本申请发明人发现,动物体内施用本发明的草果精油可以显著抑制胆碱酯酶,在化合物诱导老年痴呆症斑马鱼模型中,能显著改善老年痴呆症斑马鱼的认知和反应能力。此外,也能在抑郁焦虑斑马鱼模型中,能有效改善斑马鱼的抑郁焦虑症状。也就是说,草果精油通过多分子协调,多靶点作用,治疗/缓解脑功能及其障碍疾病的相关症状,而不会完全抑制所有活性,也没有出现毒副作用。
许多脑功能及其障碍疾病的病理状态,例如抑郁、焦虑、焦虑、抑郁、失眠、狂躁症、精神分裂症、痉挛和双相障碍、老年痴呆等,发病机制复杂,单一成分或分子难与治愈或缓解。草果精油由多个活性分子组成,因此,患有脑功能及其障碍疾病的个体可接受含有本发明中涉及的草果精油,或含本发明中涉及的含草果精油的组合物。
2、示例性病理状况
情感情绪障碍:情感情绪障碍的临床表现为情感高涨或低落,伴有思维奔逸或迟缓,精神运动性兴奋或抑制。这包括从焦虑、抑郁、精神分裂等一系列病症,如躁狂症、分裂情感性障碍、精神分裂症、创双极性情绪障碍、恐慌状态和行为阵发性失控综合征等。根据本发明的草果精油以及药物配方和组合物可用于治疗这些疾病、症状及病况,并且其与这一治疗类别中的现有治疗剂相比,应当显示出对副作用的改善。
神经退行性疾病:神经退行性疾病是神经元或其髓鞘的丧失所致,以运动障碍为特征的运动异常。随着时间的推移而恶化,以导致功能障碍,比如阿尔兹海默病(AD)等。本发明中的草果及其衍生物组合缓解运动障碍的一种或多种症状。
本发明中草果精油及其组合物也可用作焦虑缓解(抗焦虑)剂。
本发明中草果精油也可用作脑功能及其障碍发病机制研究的化学工具
脑功能及其障碍疾病和病症意指神经系统的障碍或疾病,包括但不限于焦虑、抑郁、失眠、狂躁症、精神分裂症、痉挛、双相障碍和阿尔兹海默病。
神经退行性疾病意指诸如但不限于以下的疾病:老年痴呆症、帕金森症等。
本发明的有效药物制剂和组合物可用来治疗脑功能及其障碍疾病和病症。虽然这些药物制备通常用于人患者的治疗,但它们也可以用于治疗其他动物的相似或相同的疾病,所述其他动物例如灵长类动物、家禽(如鸡、鸭、鹅)、农场动物(如猪、牛)、体育用动物(如赛马)和宠物(如狗和猫)。
本发明所述可药用载体包括但不限于碳酸钙、磷酸钙、硫酸钙、蔗糖、葡萄糖、乳糖、果糖、木糖醇、山梨醇、淀粉、淀粉糊、纤维素衍生物、明胶、聚乙烯吡咯烷酮、氯化钠、糊精、硬脂酸、硬脂酸镁、硬脂酸钙、植物油、聚乙二醇、无菌磷酸盐缓冲盐水、盐水、林格氏溶液以及它们的组合。
本发明的所述口服剂型包括但不局限于固体口服剂型(如肠溶衣片、滴丸、口服片、咀嚼片、颗粒剂、粉末或胶囊)或液体口服剂型(如糖浆或酊剂)。此外,本发明中的草果精油或其组合物还可以添加到化妆品、洗浴用品或大健康产品中,用于香薰、嗅吸、按摩、涂抹、泡浴使用。此外,本发明中的草果精油或其组合物也可以配制成口香糖,以促进口服递送和吸收。
本发明所述的非口服剂型包括但不局限于通过注射或其他全身途径给药,所述其它全身途径例如经皮施用或经粘膜施用(例如经鼻、舌下、口含、经阴道或直肠,通过栓剂)。其他施用途径(例如可在兽医应用中使用的)包括肠内和胃肠外递送,包括肌肉、皮下和/或髓内注射,以及鞘内注射、直接脑室注射、静脉内注射、腹膜内注射、鼻内注射或眼球内注射。
本发明涉及的草果精油也可与其他活性成分联用,以制备其他的新药物组合物。
本发明的草果精油或其组合用以缓解上述症状的证实性疗效和治疗相关活性,是通过斑马鱼、小鼠动物老年痴呆症和抑郁模型试验和筛选方法进行证实的。
上述本发明草果精油或其组合的治疗作用、良好的药物代谢参数和通常无毒性使得本发明化合物成为用于治疗上述病症的理想药物。
下面以本发明的实施例来进一步说明本发明的实质性内容,但不以任何方式对本发明加以限制,基于本发明所作的任何变换或替换,均属于本发明的保护范围。
本发明所述的草果精油的制备方法,包括水蒸气蒸馏提取、二氧化碳超临界提取。
本发明所述的草果精油用气相色谱质谱联用仪器检测,按质量百分比计其组分比例为1,8-桉叶素(eucalyptol)(>35%)、柠檬醛(citral)(>8%)、香叶醇(geraniol)(>5%)、橙花叔醇(nerolidol)(>1%)。
实施例1
取草果(Amomum tsao-ko Crevost et Lemaire)干燥的果实100Kg,渥堆放置2天,粉碎至50目的粗颗粒,将粗颗粒浸泡于8质量倍量pH值为5.0盐酸水溶液中,浸提过夜。向浸提混合液中加入质量百分比分别为0.8%纤维素酶和0.3%果胶酶,在37℃酶解2.5小时。将上述样品及其混合液放入水蒸气蒸馏提取仪器中,110℃回流提取7个小时,收集蒸馏液,冷却至室温后置于油水分离器中分离。油层用无水硫酸钠干燥、分离,即得即得草果精油。
实施例2
取草果(Amomum tsao-ko Crevost et Lemaire)干燥的果实50Kg,渥堆放置2天,粉碎至50目的粗颗粒,将粗颗粒浸泡于8质量倍量pH值为5.0盐酸水溶液中,浸提过夜。向浸提混合液中加入质量百分比分别为0.8%纤维素酶和0.3%果胶酶,在37℃酶解2.5小时。将上述样品及其混合液放入二氧化碳超临界提取仪器中,提取的温度46℃,压力为15.70MPa,时间为85min,二氧化碳的流速为30kg/h;收集精油,置于分子蒸馏仪器中精制,进料温度为43℃,进样速度为510mL/h,一段蒸发温度为112℃,二段蒸发温度为63℃,收集二段蒸馏部分,即得草果精油。
实施例3
以实施例1-2制备得到的草果精油进行GC-MS分析。按质量百分比计其组分比例为1,8-桉叶素(eucalyptol)(>35%)、柠檬醛(citral)(>8%)、香叶醇(geraniol)(>5%)、橙花叔醇(nerolidol)(>1%)。
实施例4
以实施例1制备得到的草果精油进行抗乙酰胆碱酯酶活性测试:
取NaH2PO4·2H2O 3.121g加水溶解,定溶至100mL得到0.2moL/L的NaH2PO4;Na2HPO4·12H2O 71.64g加水溶解,定溶至1000mL得到0.2mol/L的Na2HPO4。取上述配制的26.5mL NaH2PO4和473.5mL Na2HPO4充分混合后,得到0.2moL/L pH=8.0的PBS,再将此溶液稀释1倍得到0.1moL/L pH=8.0的PBS;b.取上述配制的39mL NaH2PO4和61mL Na2HPO4充分混合后,得到0.2moL/L pH=7.0的PBS,再将此溶液稀释1倍,得到0.1moL/L pH=7.0PBS。
先用pH=8.0的PBS配制成1000U/mL乙酰胆碱酯酶(AchE)的母液,再稀释成4U/mL的储备液-80℃保存,实验时再将储备液稀释成0.4U/mL的乙酰胆碱酯酶液。准确称取8.8mgATCI(碘化硫代乙酰胆碱),溶于去离子水中,定溶至50mL,4℃放置(现配现用);0.6mmoL/LDTNB[5,5'-二硫代双(2-硝基苯甲酸)]的配制:准确称取11.9mg DTNB和0.75g NaHCO3溶于pH=7.0的PBS中,定溶至50mL,避光保存(现配现用)。阳性药石杉碱甲用色谱级甲醇配成10mg/mL的初始浓度,待测物用色谱级甲醇配成100mg/mL的初始浓度。实验时阳性对照和待测物均稀释为5个浓度(阳性对照:100,10,1,0.1,0.01mg/mL;待测物:100,50,25,12.5,6.25mg/mL),每个浓度做3组平行实验。分别分为如下几个组测试:实验组:40μL PBS(pH=8.0),10μL待测物,10μL 0.4U/mL的AchE溶液,20μL 0.6mmoL/L DTNB依次加入96孔板中,37℃预孵10min,然后再加20μL 0.6mmoL/L ATCI于37℃恒温反应30min,然后再加50μL无水乙醇终止反应。实验控制组:40μL PBS(pH=8.0),10μL待测物,10μL PBS(pH=8.0),20μL0.6mmoL/L DTNB依次加入96孔板中,37℃预孵10min,然后再加20μL 0.6mmoL/L ATCI于37℃恒温反应30min,然后再加50μL无水乙醇终止反应。空白组:40μL PBS(pH=8.0),10μL甲醇,10μL 0.4U/mL的AchE溶液,20μL 0.6mmoL/L DTNB依次加入96孔板中,37℃预孵10min,然后再加20μL 0.6mmoL/L ATCI于37℃恒温反应30min,然后再加50μL无水乙醇终止反应。空白对照组:40μL PBS(pH=8.0),10μL甲醇,PBS(pH=8.0),20μL 0.6mmoL/L DTNB依次加入96孔板中,37℃预孵10min,然后再加20μL 0.6mmoL/L ATCI于37℃恒温反应30min,然后再加50μL无水乙醇终止反应。
用酶标仪在412nm处检测波长,计算抑制率公式如下:
结果表明(表1和图1)所有测试的草果精油在1mg/mL浓度下均显示抑制乙酰胆碱酯酶的作用,与阳性对照石杉碱甲相当。
表1:乙酰胆碱酯酶抑制活性测试
化合物名称 | IC<sub>50</sub>(mg/mL) |
草果(Amomum tsao-ko Crevost et Lemaire)果实水蒸气蒸馏精油 | 0.0623±0.0110 |
阳性对照组(石杉碱甲) | 0.0032±0.0001 |
实施例5
以实施例1制备得到的草果精油进行抗丁酰胆碱酯酶活性测试:
取NaH2PO4·2H2O 3.121g加水溶解,定溶至100mL得到0.2moL/L的NaH2PO4;Na2HPO4·12H2O 71.64g加水溶解,定溶至1000mL得到0.2mol/L的Na2HPO4。取上述配制的26.5mL NaH2PO4和473.5mL Na2HPO4充分混合后,得到0.2moL/L pH=8.0的PBS,再将此溶液稀释1倍得到0.1moL/L pH=8.0的PBS;b.取上述配制的39mL NaH2PO4和61mL Na2HPO4充分混合后,得到0.2moL/L pH=7.0的PBS,再将此溶液稀释1倍,得到0.1moL/L pH=7.0PBS。
先用pH=8.0的PBS配制成1000U/mL丁酰胆碱酯酶(BchE)的母液,再稀释成4U/mL的储备液-80℃保存,实验时再将储备液稀释成0.4U/mL的丁酰胆碱酯酶液。准确称取2.5mgBTCI(碘化硫代丁酰胆碱),溶于去离子水中,定溶至50mL,4℃放置(现配现用);将丁酰胆碱酯酶(BChE)粉末用pH=8.0的PBS充分溶解,配制成500U/mL母液,再稀释成8U/ml储备液-80℃保存,实验时再将储备液稀释成0.8U/mL的丁酰胆碱酯酶液;DTNB配置方法同上。阳性药他克林用色谱级甲醇配成10μM的初始浓度,实验时稀释5个梯度,每个浓度做3组平行实验。分别分为如下几个组测试:实验组:40μL PBS(pH=8.0),10μL待测物,10μL 0.4U/mL的BchE溶液,20μL 0.6mmoL/L DTNB依次加入96孔板中,37℃预孵10min,然后再加20μL 0.6mmoL/LATCI于37℃恒温反应30min,然后再加50μL无水乙醇终止反应。实验控制组:40μL PBS(pH=8.0),10μL待测物,10μL PBS(pH=8.0),20μL 0.6mmoL/L DTNB依次加入96孔板中,37℃预孵10min,然后再加20μL 0.6mmoL/L ATCI于37℃恒温反应30min,然后再加50μL无水乙醇终止反应。空白组:40μL PBS(pH=8.0),10μL甲醇,10μL 0.4U/mL的BchE溶液,20μL 0.6mmoL/L DTNB依次加入96孔板中,37℃预孵10min,然后再加20μL 0.6mmoL/L ATCI于37℃恒温反应30min,然后再加50μL无水乙醇终止反应。空白对照组:40μL PBS(pH=8.0),10μL甲醇,PBS(pH=8.0),20μL 0.6mmoL/L DTNB依次加入96孔板中,37℃预孵10min,然后再加20μL0.6mmoL/L ATCI于37℃恒温反应30min,然后再加50μL无水乙醇终止反应。
用酶标仪在412nm处检测波长,计算抑制率公式如下:
结果表明(表2和图2)所有测试的草果精油在1mg/mL浓度下均显示抑制丁酰胆碱酯酶的作用,与阳性对照他克林相当。
表2:丁酰胆碱酯酶抑制活性测试
化合物名称 | IC<sub>50</sub>(mg/mL) |
草果(Amomum tsao-ko Crevost et Lemaire)果实水蒸气蒸馏精油 | 0.0216±0.0060 |
阳性对照组(他克林) | 0.0194±0.0030 |
实施例6
以实施例1制备得到的草果精油进行体内抗老年痴呆症活性测试:
1)实验动物
实验用的斑马鱼幼鱼为斑马鱼种鱼所产胚胎自然孵育而成。养鱼用水水质:每1L反渗透水中加入200mg速溶海盐,电导率为480~510μS/cm;pH为6.9~7.2;硬度为53.7~71.6mg/L CaCO3。实验完成后,用0.25mg/ml的三卡因甲磺酸对各个发育阶段的斑马鱼进行麻醉处死。麻醉处死的操作步骤符合美国兽医协会(AVMA)对动物麻醉处死的规范要求。
2)实验设计
本研究共设置5组实验,每组实验包括1个空白对照组、1个溶剂对照组、1个老年痴呆模型组、1个阳性对照组(多奈哌齐)、2个草果精油。草果精油的初筛浓度为6μg/mL,若出现死亡或畸形,则选取0.6μg/mL,进行实验。三氯化铝处理的斑马鱼为老年痴呆模型组;三氯化铝与DPZ共同处理的斑马鱼为阳性对照组;三氯化铝与待测化合物共同处理的斑马鱼为待测化合物组;0.1%的DMSO处理的斑马鱼为阴性对照组(溶剂对照组);未处理的斑马鱼为空白对照组,每个实验组均处理30尾斑马鱼。利用行为分析仪记录60min内斑马鱼的运动轨迹,通过光暗周期性变化,黑暗10min,光照10min,60min分为三个光暗周期,然后对斑马鱼的运动速度(运动功能障碍)、明暗周期速度变化(反应能力)进行统计学分析。
活性测试结果符合质量控制标准:①整个实验过程中所有实验组均未出现斑马鱼死亡或畸形;②模型组和溶剂对照组比较具有统计学差异(p<0.001);③所有实验中空白对照组与溶剂对照组相比无统计学差异(p>0.05);④阳性对照组(多奈哌齐,DPZ)与模型组比较具有统计学差异(p<0.05)。
3)对老年痴呆症斑马鱼运动功能障碍的恢复作用
所有实验老年痴呆模型组斑马鱼运动速度显著下降,与溶剂对照组比较均有统计学意义(p<0.001);阳性药(DPZ)组斑马鱼运动速度显著回升,与模型组比较具有统计学意义(p<0.01或p<0.001)。斑马鱼在60min明暗周期内,其运动速度具有规律性、周期性和稳定性。以实施例1制备得到的草果精油处理后的AD斑马鱼的运动速度与模型组相比显著提高(p<0.05,p<0.01或p<0.001),显著恢复老年痴呆症斑马鱼的运动速度(表3和图3)。
表3:老年痴呆症斑马鱼运动速度的治疗效率
化合物名称 | 浓度(μg/mL) | 治疗效率% | p值 |
草果(Amomum tsao-ko Crevost et Lemaire)果实水蒸气蒸馏精油 | 6.00 | 70.25 | 8.04E-05 |
阳性对照组(多奈哌齐) | 3.79 | 68.95 | 5.11E-07 |
4)对老年痴呆症斑马鱼反应能力的影响
所有实验中,老年痴呆模型组斑马鱼反应能力下降(明暗周期速度差值显著下降),与溶剂对照组比较均有统计学意义(p<0.001);阳性药(DPZ)组斑马鱼反应能力部分恢复(明暗周期速度差值显著回升),与模型组比较具有统计学意义(p<0.05或p<0.001)。以实施例1制备得到的草果精油处理后可以不同程度的恢复AD斑马鱼的反应能力(明暗周期速度差值显著回升),与模型组相比,统计学差异显著或极显著(p<0.05,p<0.01或p<0.001)(表4和图4)。
表4:老年痴呆症斑马鱼反应能力的治疗效率
化合物名称 | 浓度(μg/mL) | 治疗效率% | p值 |
草果(Amomum tsao-ko Crevost et Lemaire)果实水蒸气蒸馏精油 | 6.00 | 58.57 | 8.20E-06 |
阳性对照组(多奈哌齐) | 3.79 | 73.84 | 9.55E-11 |
实施例7
分别以实施例2制备得到的草果精油进行活性测试,测试方法与实施例4、实施例5、实施例6相同,结果均表明,本发明制备得到的草果精油能显著改善三氯化铝诱导老年痴呆症斑马鱼的运动能力和反应能力。
实施例8:草果精油片剂的制备
草果精油10mL,药用淀粉100g,混合均匀,用适量乙醇作为粘合剂制粒,干燥,经整粒机整粒,压片,每片0.30g,口服,每次1-2片,每日两次。
实施例9:草果精油胶囊剂的制备
草果精油10mL,药用淀粉100g,混合均匀,用适量乙醇作为粘合剂制粒,干燥,经整粒机整粒,装胶囊,每粒0.30g,口服,每次1-2粒,每日两次。
实施例10:草果精油颗粒剂的制备
草果精油10mL,药用淀粉100g,糖粉500g,混合均匀,用适量乙醇作为粘合剂制粒,干燥,经整粒机整粒,分装即得,口服,每次5g,每日两次
实施例11:草果精油吸入制剂的制备
草果精油10mL,基础油500mL,适当稳定剂和调香剂,混合均匀,分装即得,吸入,每次10mL,每日两次。
实施例12:草果精油按摩油的制备
草果精油10mL,基础油500mL,混合均匀,分装即得,按摩,每日2-3次。
实施例13:草果精油泡浴剂的制备
草果精油100mL,分装即得,泡浴时,滴入水中使用,每周2-3次。
实施例14:草果精油香薰油的制备
草果精油100mL,分装即得,香薰时,滴入香薰机中使用,每天2-3次。
上述实施例用来解释说明本发明,而不是对本发明进行限制,在本发明的精神和权利要求的保护范围内,对本发明做出的任何修改和改变,都落入本发明的保护范围。
Claims (8)
1.一种草果精油的应用,其特征在于,所述应用包括草果精油在制备治疗或预防脑功能及其障碍疾病的药物中的应用、草果精油在制备改善脑功能及其障碍疾病的健康产品中的应用。
2.根据权利要求1所述的应用,其特征在于,所述草果精油通过以下方法提取得到:
(1)发酵:选取新鲜或干燥的草果果实,渥堆放置约1-2天,然后粉碎至30-60目的粗颗粒备用。
(2)浸提:将粗颗粒浸泡于7-9倍质量的pH值为5.0-6.0盐酸或硫酸水溶液中,浸提过夜,得浸提混合液。
(3)酶解:向浸提混合液中加入纤维素酶和果胶酶,纤维素酶、果胶酶和浸提混合液的质量配比为0.5-1:0.1-0.5:100,在35-37℃酶解约2-3小时。
(4)提取:将步骤3酶解后的混合液放入水蒸气蒸馏提取仪器中,100℃-120℃回流提取约5-7个小时,收集蒸馏液,冷却至室温后置于油水分离器中分离,油层用无水硫酸钠干燥、分离,即得草果精油;或将步骤3酶解后的混合液放入二氧化碳超临界提取仪器中,提取的温度45℃-48℃,压力为13.70MPa-17.80MPa,时间为50min-100min,二氧化碳的流速为25kg/h-35kg/h;收集提取物,置于分子蒸馏仪器中精制,进料温度为40℃-45℃,进样速度为480-530mL/h,一段蒸发温度为110℃-115℃,二段蒸发温度为60℃-65℃,收集二段蒸馏部分,得草果精油。
3.根据权利要求1所述的应用,其特征在于,所述草果为姜科豆蔻属植物草果。
4.根据权利要求1所述的应用,其特征在于,所述草果精油用气相色谱质谱联用仪分析检测,按质量百分比计,其组分比例为:1,8-桉叶素>35%、柠檬醛>8%、香叶醇>5%、橙花叔醇>1%。
5.根据权利要求1所述的应用,其特征在于,所述脑功能及其障碍疾病包括焦虑、抑郁、失眠、狂躁症、精神分裂症、痉挛、双相障碍、中风、亨廷顿病、脑外伤、脊柱损伤和神经退行性疾病。
6.根据权利要求5所述的应用,其特征在于,所述神经退行性疾病意包括老年痴呆、帕金森病、亨廷顿病和肌萎缩性侧索硬化。
7.根据权利要求1所述的应用,其特征在于,所述药物包括口服制剂和非口服给药制剂;所述口服制剂包括片剂、丸剂、胶囊剂、颗粒剂、微囊片剂、混悬剂、滴丸、口服液体;所述非口服给药制剂包括吸入剂、注射剂、气雾剂、栓剂、皮下给药剂型。
8.根据权利要求1所述的应用,其特征在于,所述健康产品包括化妆品、洗浴剂和香薰剂。
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WO2023272798A1 (zh) * | 2021-06-29 | 2023-01-05 | 中国热带农业科学院热带作物品种资源研究所 | 一种抑制热带假丝酵母菌的草果精油的提取方法及草果精油 |
CN113913245B (zh) * | 2021-06-29 | 2023-10-10 | 中国热带农业科学院热带作物品种资源研究所 | 一种抑制热带假丝酵母菌的草果精油的提取方法及草果精油 |
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