CN112439015A - 冬虫夏草抗病毒组合物 - Google Patents
冬虫夏草抗病毒组合物 Download PDFInfo
- Publication number
- CN112439015A CN112439015A CN202011308763.4A CN202011308763A CN112439015A CN 112439015 A CN112439015 A CN 112439015A CN 202011308763 A CN202011308763 A CN 202011308763A CN 112439015 A CN112439015 A CN 112439015A
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- cordyceps sinensis
- composition
- cordyceps
- honey
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Classifications
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Abstract
本发明涉及冬虫夏草组合物，由包括冬虫夏草、大枣、三叶青、大叶藜、扁枝石松、炙甘草的配方制成。本发明冬虫夏草组合物对黄热病毒感染具有效果显著、副作用小等优点。
Description
技术领域
本发明属于医药领域，涉及冬虫夏草的组合物，及组合物用于抗黄热病病毒的用途。
背景技术
寨卡病毒(Zika virus，ZIKV)、登革病毒(dengue virus，DENV)及黄热病病毒(yellow fevervirus，YFV)属于黄病毒科黄病毒属，具有包膜蛋白的二十面体病毒。它的基因组为一条正向单链RNA，共有约10800个核苷酸，编码3个结构蛋白(PrM、Env、Capsid)和七个非结构蛋白(NS1、NS2A、NS2B、NS3、NS4A、NS4B、NS5)。黄热病毒可与黄病毒科其他成员如登革热病毒、西尼罗病毒、圣路易脑炎病毒、寨卡病毒等产生交叉血清学反应。
黄热病是由YFV引起的由蚊子传播的急性病毒病，症状通常包括发烧、冷颤、食欲下降、恶心、肌肉痛(特别是背部)与头痛。黄热病病毒感染期，中医理论认为属于湿热郁阻症，一般的治疗采用清热化湿，透表解肌；中毒早期，毒扰气营症，以清气凉营，泻火解毒为主；中毒期，为瘀毒入血症，采取凉血止血，解毒化瘀。有些病人会在症状改善1到2天后，发烧再度发生，出现腹痛，肝脏损伤导致黄疸，这类病患并发出血和肾衰竭的风险较高。
冬虫夏草是麦角菌科(Clavicipitaceae)真菌冬虫夏草菌Cordyceps sinensis(BerK.)Sacc.寄生在蝙蝠蛾科昆虫幼虫上的子座及幼虫尸体的干燥复合体，一种天然存在的稀有中药材，富含了虫草多糖、虫草素、虫草酸、虫草多肽、麦角甾醇等多种单体活性成分。现代研究证明冬虫夏草以其调节阴阳、肺肾双补之特性，补而不峻、温而不火之优势，广泛应用于免疫、心血管、呼吸、泌尿生殖、消化、神经系统疾病。
发明内容
本发明提供了一种冬虫夏草在制备预防和/或治疗黄热病毒感染的药物中的用途。该用途具有效果显著、副作用小、见效快等优点。
具体的，
一方面，本发明提供了一种冬虫夏草组合物，其特征在于，由包括冬虫夏草、大枣、三叶青、大叶藜、扁枝石松、炙甘草的配方制成。
在一些实施例中，本发明所述的组合物由包括冬虫夏草50-70份、大枣20-40份、三叶青10-30份、大叶藜5-20份、扁枝石松5-20份、炙甘草5-20份的配方制成。
在一些实施例中，本发明所述的组合物的所述配方还包括：疬子草5-20份、黄芩5-15份、白云花根5-20份。
在一些实施例中，本发明所述的组合物由包括冬虫夏草60份、大枣20-40份、三叶青15-20份、疬子草8-15份、大叶藜8-15份、黄芩8-10份、扁枝石松8-15份、白云花根8-15份、炙甘草8-15份的配方制成。
在一些实施例中，本发明所述的组合物按下述重量份的药材配方制成：冬虫夏草60份、大枣30份、三叶青20份、疬子草10份、大叶藜10份、黄芩8份、扁枝石松10份、白云花根10份、炙甘草10份。
在一些实施例中，本发明所述的组合物按下述重量份的药材配方制成：冬虫夏草60份、大枣40份、三叶青15份、疬子草8份、大叶藜8份、黄芩8份、扁枝石松8份、白云花根8份、炙甘草8份。
在一些实施例中，本发明所述的组合物按下述重量份的药材配方制成：冬虫夏草60份、大枣20份、三叶青20份、疬子草15份、大叶藜15份、黄芩10份、扁枝石松15份、白云花根15份、炙甘草15份。
在一些实施例中，本发明所述的组合物中所述冬虫夏草为冬虫夏草子实体。
另一方面，本发明提供了在制备治疗黄病毒属病毒感染的疾病药物中的用途。
在一些实施例中，本发明所述用途中所述的黄病毒属病毒包括寨卡病毒、登革病毒及黄热病病毒。
术语“包括”是一种开放式描述，还可以任意的包括除配方中显示的主要成分外的其他次要成分，如中药中“君臣佐使”的“佐”“使”，以及其他一些常规的要用辅料。在一些实施例中，“包括”描述的技术方案不包括其他成分。
单位“份”是重量份数。
冬虫夏草，甘，平。归肺、肾经。补肾益肺，止血化痰。用于肾虚精亏，阳痿遗精，腰膝酸痛，久咳虚喘，劳嗽咯血。
大枣：味甘，性温。能补中益气，养血生津。主治脾胃虚弱，食少便溏，气血亏虚，身体虚弱，神经衰弱，脾胃不和，劳伤咳嗽，贫血消瘦，有养肝防癌功效。
三叶青，为葡萄科崖爬藤属多年生常绿草质蔓生藤本植物。具有清热解毒、活血散结、消炎止痛、祛风化痰、理气健脾等功效,常用于小儿高热惊厥、痢疾、支气管炎、肺炎、咽喉炎、肝炎、病毒性脑膜炎、毒蛇咬伤、扁桃体炎、蜂窝织炎、跌打损伤等疾病的治疗。
疬子草，为报春花科植物延叶珍珠菜Lysimachia decurrens Forst.f.的全草，味苦、辛，性平，清热解毒，活血散结，常用于瘰疬，喉痹，疔疮肿毒，月经不调，跌打损伤。
大叶藜，为藜科植物杂配藜Chenopodium hybridum L.的全草。味甘，性平。具有调经止血，解毒消肿之功效。常用于月经不调，崩漏，吐血，衄血，尿血，血痢，便血，疮疡肿毒
黄芩：味苦，性寒，归心、肺、胆、大肠经。泻实火，除湿热，止血，壮热烦渴，肺热咳嗽，湿热泻痢，黄疸，热淋，吐衄，目赤肿痛，痈肿疔疮。
扁枝石松，石松科扁枝石松属植物，味辛、味甘温；入肺、肝二经。它全草用药具有祛风活络、散寒止咳等作用，主治风湿关节炎、跌打损伤、手足麻木、外感风寒之咳嗽等功效。
白云花根，为伞形科植物白云花的根。苦、辛，温。祛风除湿，活络止痛。风湿筋骨疼痛，跌打损伤，腰痛，胃痛，感冒，咳嗽，慢性支气管炎，哮喘，闭经，白带。
炙甘草：味甘，性温，归心、肺、胃、脾经。和中缓急，润肺解毒，调和诸药。主治脾胃虚弱，倦怠乏力，心动悸，可调节机体免疫功能，抗肿瘤和止痛作用。
具体实施方式
为描述本发明，以下列出了实施例。但需要理解，本发明不限于这些实施例，只是提供实践本发明的方法。
实施例1：冬虫夏草组合物1
将药材冬虫夏草、大枣、三叶青、大叶藜、扁枝石松、炙甘草分别粉碎，取冬虫夏草60g、大枣30g、三叶青20g、大叶藜10g、扁枝石松10g、炙甘草10g混合均匀，加水煎煮2次：第一次按药材与水质量体积比为g/10mL加入水，煎煮2小时；第二次按药材与水质量体积比为g/8mL加入水，煎煮1小时，合并第一次和第二次的煎液，冷冻干燥后，得冬虫夏草组合物1。
实施例2：冬虫夏草组合物2
将药材冬虫夏草、大枣、三叶青、疬子草、大叶藜、黄芩、扁枝石松、白云花根、炙甘草分别粉碎，取冬虫夏草60g、大枣30g、三叶青20g、疬子草10g、大叶藜10g、黄芩8g、扁枝石松10g、白云花根10g、炙甘草10g混合均匀，加水煎煮2次：第一次按药材与水质量体积比为g/10mL加入水，煎煮2小时；第二次按药材与水质量体积比为g/8mL加入水，煎煮1小时，合并第一次和第二次的煎液，冷冻干燥后，得冬虫夏草组合物2。
实施例3：冬虫夏草组合物3
将药材冬虫夏草、大枣、三叶青、疬子草、大叶藜、黄芩、扁枝石松、白云花根、炙甘草分别粉碎，取冬虫夏草60g、大枣40g、三叶青15g、疬子草8g、大叶藜8g、黄芩8g、扁枝石松8g、白云花根8g、炙甘草8g混合均匀，加水煎煮2次：第一次按药材与水质量体积比为g/10mL加入水，煎煮2小时；第二次按药材与水质量体积比为g/8mL加入水，煎煮1小时，合并第一次和第二次的煎液，冷冻干燥后，得冬虫夏草组合物3。
实施例4：冬虫夏草组合物4
将药材冬虫夏草、大枣、三叶青、疬子草、大叶藜、黄芩、扁枝石松、白云花根、炙甘草分别粉碎，取冬虫夏草60g、大枣20g、三叶青20g、疬子草15g、大叶藜15g、黄芩10g、扁枝石松15g、白云花根15g、炙甘草15g混合均匀，加水煎煮2次：第一次按药材与水质量体积比为g/10mL加入水，煎煮2小时；第二次按药材与水质量体积比为g/8mL加入水，煎煮1小时，合并第一次和第二次的煎液，冷冻干燥后，得冬虫夏草组合物4。
实施例5：冬虫夏草组合物对Vero细胞的毒性实验
试验药品：不同浓度的冬虫夏草组合物1和冬虫夏草组合物3(用含10％(v/v)胎牛血清的DMEM培养基稀释冬虫夏草组合物1和冬虫夏草组合物3成不同浓度作为试验药品)。
试验步骤：
1、接种Vero细胞(Vero细胞(非洲绿猴肾细胞)是YFV的易感细胞)：用含10％(v/v)胎牛血清的DMEM((Dulbecco's Modified Eagle Medium))培养基配成单个细胞悬液，以每孔1000-10000个细胞接种到96孔细胞培养板，每孔接种体积100μL；
2、培养Vero细胞：在37℃，5％(v/v)CO2培养条件下，培养48小时；
3、加入试验药品：吸弃每个孔中的DMEM培养基，试验组：向各个孔中加入100μL试验药品(不同浓度(6.25、12.5、25、50、100mg/L)的冬虫夏草组合物1和冬虫夏草组合物3)；对照组，向孔中加入100μL不含试验药品、含10％(v/v)胎牛血清的DMEM(可以视试验药品浓度为0)。
4、呈色：培养48小时后，每孔加MTT溶液10μL，在37℃，5％(v/v)CO2培养条件下继续孵育4小时；离心(1000转，10min)，小心吸去孔内培养液，每孔加入150μL二甲基亚砜，置摇床上低速振荡10min，直至在普通光学显微镜下观察发现Formazan全部溶解；
5、测量：在酶联免疫检测仪OD 490nm处测量各孔的吸光值，根据公式“细胞的存活率(％)＝各实验组OD值/对照组OD值×100％”计算细胞存活率(见表1)。
表1
实施例6：冬虫夏草组合物对YFV的抑制实验
试验药品：不同浓度的冬虫夏草组合物1和冬虫夏草组合物3(用含10％(v/v)胎牛血清的DMEM培养基稀释冬虫夏草组合物1和冬虫夏草组合物3成不同浓度作为试验药品)。
试验步骤：以Vero细胞为培养黄热病毒(YFV)的细胞，MOI为0.1，在24孔细胞培养板中接入Vero细胞，20小时后，细胞长至单层、覆盖孔底的面积约为80-90％；吸出培养基，接入病毒样品200μL，37℃吸附2小时；吸附完成后，吸弃各孔中的病毒液，用DMEM培养基洗去未吸附的病毒；加入试验药品(不同浓度(0、6.25、12.5、25、50、100mg/L)的冬虫夏草组合物1和冬虫夏草组合物3)，于37℃、5％(v/v)CO2的培养箱中培养48小时，收集各个孔中的上清用于病毒噬斑实验。病毒噬斑实验：在24孔板中接入Vero细胞，20小时后细胞长至单层(细胞覆盖孔底的面积约为90％)；吸弃各孔中的培养基，接入前述用含10％(v/v)胎牛血清的DMEM培养基10×系列稀释的病毒样品200μL，37℃吸附2小时；吸附完成后，弃去各孔的上清液，用10％(v/v)FBS的DMEM培养基洗去未吸附的病毒；加入新鲜的37℃预热的半固定培养基(A成分：3％(v/v)FBS，2×MEM培养基，青霉素和链霉素的终浓度分别为100U/mL、0.1mg/mL；B成分：1％(w/v)；A成分与B成分体积比为1:1))的琼脂糖，于37℃、5％(v/v)CO2的培养箱中培养48小时。待噬斑形成后，用结晶紫染色(2％(w/v)结晶紫溶于10％(v/v)甲醛)，2小时后用流水冲去结晶紫和琼脂糖；在显微镜下对每孔产生的细胞噬斑，根据噬斑数和稀释倍数计算病毒的滴度(PFU/ml，PFU：噬斑形成单位)。PFU＝病毒稀释度×P/V，(P：空蚀斑数目；V：接种量)。结果见表2。
表2
试验结果表明，本发明所述的冬虫夏草组合物够有效地抑制黄热病毒的增殖，可进一步开发为预防和/或治疗黄热病毒感染引起的疾病的药物，具有广泛的应用前景。
以上所述，仅为本发明较佳的具体实施方式，但本发明的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本发明披露的技术范围内，根据本发明的技术方案及其发明构思加以等同替换或改变，都应涵盖在本发明的保护范围之。
Claims (10)
1.一种冬虫夏草组合物，其特征在于，由包括冬虫夏草、大枣、三叶青、大叶藜、扁枝石松、炙甘草的配方制成。
2.根据权利要求1所述的组合物，其特征在于，由包括冬虫夏草50-70份、大枣20-40份、三叶青10-30份、大叶藜5-20份、扁枝石松5-20份、炙甘草5-20份的配方制成。
3.根据权利要求2所述的组合物，其特征在于，所述配方还包括：疬子草5-20份、黄芩5-15份、白云花根5-20份。
4.根据权利要求1、2或3所述的组合物，其特征在于，由包括冬虫夏草60份、大枣20-40份、三叶青15-20份、疬子草8-15份、大叶藜8-15份、黄芩8-10份、扁枝石松8-15份、白云花根8-15份、炙甘草8-15份的配方制成。
5.根据权利要求4所述的组合物，其特征在于，按下述重量份的药材配方制成：冬虫夏草60份、大枣30份、三叶青20份、疬子草10份、大叶藜10份、黄芩8份、扁枝石松10份、白云花根10份、炙甘草10份。
6.根据权利要求4所述的组合物，其特征在于，按下述重量份的药材配方制成：冬虫夏草60份、大枣40份、三叶青15份、疬子草8份、大叶藜8份、黄芩8份、扁枝石松8份、白云花根8份、炙甘草8份。
7.根据权利要求4所述的组合物，其特征在于，按下述重量份的药材配方制成：冬虫夏草60份、大枣20份、三叶青20份、疬子草15份、大叶藜15份、黄芩10份、扁枝石松15份、白云花根15份、炙甘草15份。
8.根据权利要求1所述的组合物，其特征在于，所述冬虫夏草为冬虫夏草子实体。
9.权利要求1所述的组合物在制备治疗黄病毒属病毒感染的疾病药物中的用途。
10.根据权利要求9所述的用途，其中所述的黄病毒属病毒包括寨卡病毒、登革病毒及黄热病病毒。
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| CN106074627A (zh) * | 2016-07-05 | 2016-11-09 | 青海珠峰冬虫夏草工程技术研究有限公司 | 冬虫夏草在制备预防和/或治疗黄热病毒感染的药物中的用途 |
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| CN106074627A (zh) * | 2016-07-05 | 2016-11-09 | 青海珠峰冬虫夏草工程技术研究有限公司 | 冬虫夏草在制备预防和/或治疗黄热病毒感染的药物中的用途 |
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