CN110338398B - 一种灵芝新菌种 - Google Patents
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Abstract
本发明公开了一种灵芝新菌种——药赤芝1号,它是保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)、保藏登记入册编号为CGMCC No.16972的菌种。本发明还提供了所述灵芝子实体的水提物。本发明的灵芝(水提物)具有十分优秀的免疫调节能力,还能有效治愈咳嗽变异性哮喘,应用前景良好。
Description
技术领域
本发明涉及中药领域,尤其涉及一种灵芝新菌种。
背景技术
灵芝(Ganoderma sp.)隶属于担子菌纲(Basidiomycetes)多孔菌目(Polyporales)多孔菌科(Polyporaceae)灵芝属(Ganoderma),是一种重要的药用真菌,在传统中医中被广泛应用,其药理作用多有论述。《神农本草经》中记载灵芝具有扶正固本、滋补强壮、延年益寿等功效,现代药理学与临床试验进一步证明灵芝具有抗衰老、抗病毒、抗氧化、抗肿瘤、增强免疫力、保肝等作用。其中赤芝是仅有的两个被《中华人民共和国药典》收录的灵芝属之一。
灵芝水提物中富含灵芝多糖,是主要的免疫调节物质,因此,灵芝水提物的免疫调节作用是评价灵芝质量好坏的重要标准之一。而灵芝的产地、品种使得我国灵芝品质良莠不齐。
发明内容
本发明的目的在于提供一种新的优质的灵芝菌种,其具有良好的免疫调节能力。
本发明提供了一种灵芝菌种——药赤芝1号,它于2018年12月04日保 藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),地址为北 京市朝阳区北辰西路1号院3号,分类命名为灵芝(Ganoderma lucidum), 保藏登记入册编号为CGMCC No.16972。
本发明还提供了前述灵芝在制备免疫调节药物或保健品中的用途。
本发明还提供了前述灵芝在制备治疗咳嗽变异性哮喘的药物中的用途。
本发明还提供了前述灵芝制备得到的水提物。
本发明还提供了前述灵芝的水提物的制备方法,其特征在于:包括:
1)取子实体粉末,加热回流3~5小时,优选4小时;
2)趁热过滤;
3)滤液水浴蒸干,即得。
前述的制备方法,其特征在于:所述方法还包括:
步骤2)过滤后,对滤渣再次加热回流,再重复步骤2)和步骤3),合并滤液。
本发明还提供了一种免疫调节药物,它是以前述灵芝、所述灵芝的粉末和/或其提取物为活性成分,加上药学上可接受的辅料制备而成的制剂。
本发明还提供了一种免疫调节保健品,它是以前述灵芝、所述灵芝的粉末和/或其提取物为活性成分,加上药学上可接受的辅料制备而成的制剂。
本发明还提供了一种治疗咳嗽变异性哮喘的药物,它是以前述灵芝、所述灵芝的粉末和/或其提取物为活性成分,加上药学上可接受的辅料制备而成的制剂。
本发明的水提物具有十分良好的免疫调节活性。经验证,其对大鼠淋巴细胞增殖促进率均能大于20%,最高促进率可达到39.78%。
本发明还能有效治愈咳嗽变异性哮喘,有效率可达86.7%。
本发明在药物或保健品制备中具有很好的应用价值。
显然,根据本发明的上述内容,按照本领域的普通技术知识和惯用手段,在不脱离本发明上述基本技术思想前提下,还可以做出其它多种形式的修改、替换或变更。
以下通过具体实施方式对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
具体实施方式
灵芝样品:驯化栽培后符合灵芝生物学形态标准的24个灵芝子实体,其编号为:代GL1、代GL2、代GL4、代GL5、代GL6、代GL13、代GL14、代GL23、代GL28、代GL70、代YCZ1、代SXLZ;段GL1、段GL2、段GL4、段GL5、段GL6、段GL13、段GL14、段GL23、段GL28、段GL70、段 YCZ1、段SXLZ。
(编号中,“代”表示代料(栽培的)灵芝;“段”表示段木(栽培的)灵芝)
表1菌株编号与来源
实验例1灵芝水提物对脾淋巴细胞增殖的影响
1.方法
(1)灵芝水提物制备
取灵芝粉末2g,精密称定,置圆底烧瓶中,加水60mL,静置1小时,加热回流4小时,趁热滤过,用少量热水洗涤滤器和滤揸,将滤渣及滤纸置烧瓶中,加水60mL,加热回流3小时,趁热滤过滤,合并滤液,置水浴上蒸干,得水提物。
水提物按每10mg加入1ml量分别加入无菌水,过滤除菌,-20℃保存。
(2)细胞悬液制备
取正常大鼠,断颈处死,无菌取脾。以10%胎牛血清RPMI-1640培养液洗涤取出血污,剪碎。在70μm尼龙细胞分离筛上用专用细胞杵,轻轻磨碎脾脏,并用0.83%Tris-NH4C1溶液洗下细胞。静置5min后,加入等量10%胎牛血清RPMI-1640培养液,1500rpm离心10min。弃去上清,细胞以10%胎牛血清RPMI-1640培养液重悬后,加入细胞培养瓶中,培养30-60min。收集悬浮细胞,1500rpm离心10min。弃去上清,加入10%胎牛血清RPMI-1640 培养液制备单细胞悬液。
(3)细胞培养实验
盼蓝染色计数(活细胞数应>95%),调整细胞浓度,在96孔板中加入 5×105个细胞(体积为180μl)。同时,加入20μl灵芝水提样品,使终浓度分别为500,158,50,15.8,5μg/ml,每孔总体积为200μl,每个剂量设3个复孔。另设空白孔,加入20μl培养液。
细胞培养板置37℃5%CO2饱和湿度空气条件下培养2d。于培养结束前6h取出,1500rpm离心10min。小心吸取上清液,加入100μl Earle’s缓冲液,再加入10μl5mg/ml MTT,继续培养6h。培养结束后,每孔加10%SDS(用 0.01M的HCl配制)100μl,37℃下放置过夜。在570nm波长处用酶标仪测定每孔OD值。
2.结果
2.1段木和代料灵芝样品对脾淋巴细胞增殖的影响
试验结果如表2。试验结果表明,段木灵芝样品段GL2、段GL13、段 GL14、段GL23、段YCZ1、段SXLZ和代料灵芝样品代YCZ1、代SXLZ对大鼠淋巴细胞增殖促进率均能大于20%,具有较强的促进作用。其中仅YCZ1 和SXLZ两个灵芝菌株在段木和代料两种栽培模式下,均具有较为显著的脾淋巴细胞增殖促进作用,段YCZ1更是在15.8μg/ml终浓度下达到39.78%的促进率。
表2段木和代料灵芝水提样品对大鼠脾淋巴细胞增殖的作用
实验例2本发明药物组合物治疗咳嗽变异性哮喘的临床疗效
1实验药物和对象
1.1实验药物实验例1中药赤芝1号的水提物。
1.2患者资料观察病例共30例,年龄平均(35.8±13.4)岁。
2实验方法
2.1纳入病例标准:①符合咳嗽变异性哮喘诊断:慢性咳嗽时间大于8 周,常伴夜间刺激性咳嗽,支气管舒张试验阳性,支气管舒张剂治疗有效。②患者年龄在18-65岁之间。③患者知情并同意参加。
2.2排出病例标准:①合并肺部感染、肺结核等肺部疾病者。②合并有严重心、肝、肾和造血系统等严重原发性疾病及精神病患者。③妊娠期妇女。
2.3疗效判断
按照症状积分(《中医病证诊断疗效标准》,南京大学出版社,1994:7,48.) 及疗效判定方法,将咳嗽变异性哮喘的治疗效果分为临床控制、显效、有效、无效四个等级,分别按照患者临床症状改善程度进行评定。临床控制:I≥90%。显效:I为60%-89%。有效:I为30%-59%。无效:I<30%。总有效率=临床控制率+显效率+有效率。其中,I是中医证候疗效指数,I=[(治疗前积分-治疗后积分)/治疗前积分]×100%。
3实验结果
实验结果如表3所示,结果表明,服用3个月可见明显效果,本发明药物组合物,有效26例,总有效率86.7%,本发明药物组合物能有效治疗咳嗽变异性哮喘。
表3治疗前与治疗后疗效比较
综上,本发明的灵芝子实体的水提物,对脾淋巴细胞的促增殖作用显著, 15.8μg/ml终浓度下最高达到39.78%的促进率,且高于已审定灵芝新品种“三祥灵芝”和野生分离的赤芝菌株。本发明的灵芝还能有效治疗咳嗽变异性哮喘。表明本发明的灵芝品种具有优秀的药用性能,应用前景良好。
Claims (3)
1.一种灵芝菌种,它是保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)、保藏登记入册编号为CGMCC No.16972的灵芝菌种,分类命名为灵芝(Ganodermalucidum)。
2.权利要求1所述灵芝菌种在制备免疫调节药物或保健品中的用途。
3.权利要求1所述灵芝菌种在制备治疗咳嗽变异性哮喘的药物中的用途。
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