CN110522771A - 樟芝菌丝体活性物质用于改善慢性阻塞性肺病的用途 - Google Patents
樟芝菌丝体活性物质用于改善慢性阻塞性肺病的用途 Download PDFInfo
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Abstract
本发明提供一种樟芝菌丝体活性物质用于制备改善慢性阻塞性肺病的组合物的用途。所述樟芝菌丝体活性物质是以下列步骤制备而成:(a)取一樟芝菌丝体(Antrodia cinnamomea)于平板培养基上,于15‑30℃的温度下培养4‑10天;(b)将步骤(a)培养后的樟芝菌丝体接种至一烧瓶内,于15‑30℃、pH 2‑6的环境培养3‑14天;(c)将步骤(b)培养后的樟芝菌丝体接种于一发酵槽内,于15‑30℃、pH2‑6的环境下搅拌培养3‑21天,形成含有该樟芝菌丝体活性物质的樟芝菌丝体发酵液。
Description
技术领域
本发明关于一种樟芝菌丝体活性物质的制备方法及其用途。具体来说,关于制备一种用于改善慢性阻塞性肺病的樟芝菌丝体活性物质,以及其在食品或医药品中的用途。
背景技术
慢性阻塞性肺疾病(Chronic Obstructive Pulmonary Disease,简称COPD)目前居全球死亡原因的第4位,是一种常见、多发、高致残率和高致死率的慢性呼吸***疾病。全世界慢性阻塞性肺病的患者有超过三亿人。
吸烟是导致COPD的主要原因,然近年因机车、汽车的排废气以及工业上发电与燃油燃煤的使用增加,导致Particulate Matter(PM)2.5的空气污染上升,更加重此一疾病的发生。长期暴露在污染的环境下,会使肺部产生发炎反应,导致气流通道变窄而产生肺组织功能障碍。
COPD的治疗方式目前包含戒烟、肺康复治疗、吸入性支气管扩张药、使用皮质类固醇、长期供氧或进行肺移植等。2015年最新通过的新药是合并过去两种使用于COPD的肌肉松弛剂,截至目前仍未有突破性的新药发展。因此,研究有效治疗慢性阻塞性肺病的方法仍是重要的发展方向。
樟芝(Antrodia cinnamomea),又称为樟菇、樟菰、樟内菇、牛樟菇、牛樟芝、红樟、红樟芝,只生长于中国台湾山区海拔450-1500公尺间的牛樟树干腐朽的心材内壁或枯死倒伏的牛樟木材阴暗潮湿的表面,属于中国***宝级的珍贵药用真菌。目前仍缺乏樟芝对改善慢性阻塞性肺病的研究。
发明内容
本发明提供一种樟芝菌丝体活性物质及其制备方法,其可用于制备具改善慢性阻塞性肺病的组合物。相较于一般西药及治疗方法,本发明揭露的液态发酵樟芝菌丝体活性物质的制备方法更为安全、简便,制成的樟芝菌丝体活性物质更天然、安全,并可有效改善COPD。
根据本发明的一实施例,提供一种樟芝菌丝体活性物质用于制备改善慢性阻塞性肺病的组合物的用途,其中樟芝菌丝体活性物质是以下列步骤制备而成:
(a)取樟芝菌丝体(Antrodia cinnamomea)于平板培养基上,于15-30℃的温度下培养4-10天;
(b)将步骤(a)培养后的樟芝菌丝体接种至烧瓶内,于15-30℃、pH 2-6的环境培养3-14天;
(c)将步骤(b)培养后的樟芝菌丝体接种于发酵槽内,于15-30℃、pH2-6的环境下搅拌培养3-21天,形成含有樟芝菌丝体活性物质的樟芝菌丝体发酵液。
一实施例中,制备樟芝菌丝体活性物质的步骤还包括步骤(d):将樟芝菌丝体发酵液冷冻干燥后磨粉,形成含有樟芝菌丝体活性物质的樟芝菌丝体冻干粉。
一实施例中,制备樟芝菌丝体活性物质的步骤还包括步骤(e):将樟芝菌丝体冻干粉以至少一溶剂萃取,形成含有樟芝菌丝体活性物质的樟芝菌丝体萃取液。
一实施例中,制备樟芝菌丝体活性物质的步骤还包括步骤(f):将樟芝菌丝体萃取液干燥,以获得樟芝菌丝体活性物质。
一实施例中,步骤(e)中的溶剂选自水或醇类。
一实施例中,当步骤(e)的溶剂为水时,樟芝菌丝体活性物质的有效浓度介于0.25-1mg/ml。
一实施例中,当步骤(e)的溶剂为醇类时,樟芝菌丝体活性物质的有效浓度介于25-100μg/ml。
一实施例中,上述步骤(b)的烧瓶培养为震荡培养,且转速为110-130rpm。
一实施例中,步骤(c)中的发酵槽进一步通入一气体,此气体包括空气、氧气、二氧化碳、氦气或其组合,发酵槽的槽压为0.5-1.0kg/cm2且通气速率为0.01-1.5VVM。
一实施例中,所述组合物为医药组合物,此医药组合物进一步包含药学上可接受的载剂、赋形剂、稀释剂或辅剂。
一实施例中,所述组合物为食品添加剂。
附图说明
图1显示的是樟芝菌丝体水萃物改善香烟烟雾萃取物诱导肺泡上皮A549细胞伤害的能力的结果,其中AcH(L)、AcH(M)及AcH(H)分别指樟芝菌丝体水萃物浓度为0.25mg/ml、0.5mg/ml及1.0mg/ml。
图2显示的是樟芝菌丝体醇萃物改善香烟烟雾萃取物诱导肺泡上皮A549细胞伤害的能力的结果,其中AcMe(L)、AcMe(M)及AcMe(H)分别指樟芝菌丝体醇萃物浓度为25μg/ml、50μg/ml及100μg/ml。
具体实施方式
为使本发明的上述及其它方面更为清楚易懂,下文特举实施例,并配合所附图式详细说明。
实施例一樟芝菌丝体培养
本实施例采用的樟芝菌丝体(Antrodia cinnamomea)购自财团法人食品工业发展研究所(FIRDI)生物资源保存及研究中心(BCRC,前称CCRC),编号为CCRC-35396。然而,可用于本发明的樟芝品种并不限定于此单独一种,其还可包括BCRC编号35396、35398、35716、36401、36711、36795、37848、37849、37850、37889、37890、37891、37893、37894、37941或及其他生物材料寄存数据库的樟芝菌种,本发明并不对其限制。
(1)平板培养:将樟芝菌丝体接种于平板上,于15-30℃下培养4-10天(本实施例中,是于25℃下培养7天)。平板培养基的成分可包含马铃薯糊精培养基(PotatoDextroseAgar,PDA)、碳源及氮源,并无特别限制。
(2)烧瓶培养:刮取(1)平板上的菌丝体接种于烧瓶中,并以15-30℃、pH 2-6及转速110-130rpm的条件震荡培养3-14天(本实施例中,于25℃、pH5、转速120rpm之下震荡培养10天)。此震荡培养是以下表1所示的培养基进行培养。
表1培养基配方
上述培养基配方中,黄豆粉及肉桂粉为综合性碳氮源,其可选自谷类(如:麦粉)或豆类(如:黄豆粉、绿豆粉、大豆粉、肉桂粉等);糖类可为葡萄糖、果糖、麦芽糖、蔗糖等;无机盐类可为硫酸镁、磷酸氢二钾、磷酸二氢钾、硫酸铁等。特别说明的是,表1培养基配方仅为其中一种范例,使用时成分可依需求调整,或搭配市售培养基使用,并无特别限制。
(3)发酵槽培养:将(2)中烧瓶培养的菌丝体接种于发酵槽内,以15-30℃、槽压0.5-1.0kg/cm2、pH 2-6及搅拌速度50-150rpm条件下,以0.1-1.5VVM的通气速率培养3-21天,形成一樟芝菌丝体发酵液(本实施例中,是在25℃、槽压0.5kg/cm2、pH4,搅拌速度80rpm及1.0VVM(空气)的条件下,培养14天),即得樟芝菌丝体发酵液。发酵槽培养使用的培养基可与步骤(2)烧瓶培养使用的培养基相同。此樟芝菌丝体发酵液内即含本发明的樟芝菌丝体活性物质。樟芝菌丝体发酵液可进一步通过冷冻干燥步骤制备为樟芝菌丝体发酵液冻干粉;
(4)萃取物制备:将樟芝菌丝体发酵液冻干粉分为两份,分别加入20倍重量的蒸馏水及乙醇进行萃取。以水为溶剂的萃取物以温度100℃加热三十分钟,待冷却后离心取上清液,经冷冻干燥法进行干燥得樟芝发酵菌丝体水萃物。以乙醇为溶剂的萃取物,以超音波震荡1小时,离心后取上清液,将上清液经减压浓缩得樟芝发酵菌丝体醇萃物;
经由萃取步骤,可取得含较高浓度的樟芝菌丝体活性物质的水萃物及醇萃物。樟芝菌丝体活性物质的态样包含樟芝菌丝体发酵液(菌丝体与澄清液)、发酵液冻干粉、水萃物、醇萃物、水萃物及醇萃物的混合物或其他剂型。以下实施例二中,是以水萃物及醇萃物作为樟芝菌丝体活性物质态样。
实施例二樟芝菌丝体水萃物及醇萃物改善COPD的分析
目前已有实验利用香烟烟雾萃取物(Cigarette smoke extract,CSE)及人类肺癌上皮细胞A549成功建立COPD模式。可参照Miaomiao Chenet al.(2015).Azithromycinattenuates cigarette smoke extract-induced oxidative stress injury in humanalveolar epithelial cells.MOLECULAR MEDICINE REPORTS,11:3414-3422.此篇论文,其以人类肺癌上皮细胞A549对CSE进行细胞存活率分析(MTT assay),用以评估抗生素Azithromycin对COPD的改善结果。CSE对肺泡上皮的损伤是慢性阻塞性肺病机制中的重要过程。因此,若能抑制CSE对肺泡上皮的伤害即能达到改善慢性阻塞性肺病的效果。本实验中亦以人类肺癌上皮细胞A549对CSE进行细胞存活率分析(MTT assay),评估樟芝菌丝体活性物质对COPD的改善结果。
(1)收集人类肺癌上皮A549细胞,调整细胞悬浮液浓度,以50,000-100,000/mL密度植种于96孔盘内;
(2)将(1)的96孔盘置于37℃、5%CO2培养箱培养16-24小时后,以实施例一所获得,不同剂量的樟芝菌丝体水萃物(0、0.25、0.5、1mg/ml)及醇萃物(0、25、50、100μg/ml)处理。樟芝菌丝体萃取物溶解于二甲基亚石枫(Dimethyl sulfoxide,DMSO)中,检测时DMSO的浓度不超过0.1%以避免其毒性影响细胞生长;对照组处理0.1%DMSO,每个浓度进行三重复,结果置于37℃、5%CO2培养箱培养24小时;
(3)上述(2)的结果先以400×g离心10min,移除上清液,再加入香烟烟雾萃取物CSE12.5%诱导A549的细胞毒性,对照组不处理。接着置于37℃、5%CO2培养箱培养6小时;
(4)将(3)的上清液移除,每孔加入100μl MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)溶液,于37℃反应2小时后,终止培养,小心移除孔内MTT溶液;
(5)于(4)中每孔加入100μl DMSO,震荡2min,于570nm测定吸光值。此吸亮度代表细胞还原MTT的能力(甲的形成量),即粒线体的活性(活细胞数目)。于570nm的吸亮度越高,表示细胞存活率越高。
统计分析:实验结果数据以平均值(Mean)±标准偏差(SD)表示。试验数据以Student's t-test统计分析其组间差异。若p<0.05则视为具有统计上的显著差异。
水萃物的实验结果如图1及下表2所示。香烟烟雾萃取物CSE会诱导A549细胞毒性,造成细胞存活率下降。然而,加入樟芝发酵菌丝体水萃物可显著性抑制细胞损伤,提升存活率,且加入的浓度越高,存活率越高。采用本发明实施例一中的樟芝菌丝体水萃物具有改善COPD的作用,有效浓度介于0.5-1mg/ml。
表2樟芝菌丝体水萃物测试结果
(n=3)
#表示与对照组(control)统计上具显著差异(p<0.05)
*表示与负对照组(CSE)统计上具显著差异(p<0.05)
醇萃物的实验结果如图2及下表3所示。结果同样表明,加入樟芝发酵菌本发明实施例一中的樟芝菌丝体醇萃物具有改善COPD的作用,有效浓度介于50-100μg/ml。
表3樟芝菌丝体醇萃物测试结果
组别 | 存活率(%) |
对照组(Control) | 100±2.93 |
CSE | 84.51±0.70# |
CSE+樟芝菌丝体醇萃物25μg/ml | 84.63±1.91 |
CSE+樟芝菌丝体醇萃物50μg/ml | 86.90±2.20 |
CSE+樟芝菌丝体醇萃物100μg/ml | 89.92±2.89* |
樟芝菌丝体醇萃物100μg/ml | 103.66±2.47 |
(n=3)
#表示与对照组(control)统计上具显著差异(p<0.05)
*表示与负对照组(CSE)统计上具显著差异(p<0.05)
本发明的樟芝活性物质经上述实验,已证实具有改善COPD的作用。其可与药学上可接受的载剂、赋形剂、稀释剂或辅剂结合,制备医药组合物,亦可作为食品添加剂之用。
虽然本发明已用实施例揭露如上,然其并非用以限制本发明。本领域一般技术人员,于参酌以上教示后,当能对上述实施例的内容进行适当修改,而仍然能达到本发明所主张的功效。因此,本发明的保护范围应以权利要求书为准。
Claims (11)
1.一种樟芝菌丝体活性物质用于制备改善慢性阻塞性肺病的组合物的用途,其中,该樟芝菌丝体活性物质是以下列步骤制备而成:
(a)取一樟芝菌丝体于平板培养基上,于15-30℃的温度下培养4-10天;
(b)将步骤(a)培养后的樟芝菌丝体接种至一烧瓶内,于15-30℃、pH 2-6的环境培养3-14天;
(c)将步骤(b)培养后的樟芝菌丝体接种于一发酵槽内,于15-30℃、pH2-6的环境下搅拌培养3-21天,形成含有所述樟芝菌丝体活性物质的樟芝菌丝体发酵液。
2.如权利要求1所述的用途,其中,制备所述樟芝菌丝体活性物质的步骤还包括步骤(d):将所述樟芝菌丝体发酵液冷冻干燥后磨粉,形成含有所述樟芝菌丝体活性物质的樟芝菌丝体冻干粉。
3.如权利要求2所述的用途,其中,制备所述樟芝菌丝体活性物质的步骤还包括步骤(e):将所述樟芝菌丝体冻干粉以至少一溶剂萃取,形成含有所述樟芝菌丝体活性物质的至少一樟芝菌丝体萃取液。
4.如权利要求3所述的用途,其中,制备所述樟芝菌丝体活性物质的步骤还包括步骤(f):将所述樟芝菌丝体萃取液干燥,以获得所述樟芝菌丝体活性物质。
5.如权利要求3或4所述的用途,其中,在步骤(e)中,所述溶剂选自水或醇类。
6.如权利要求5所述的用途,其中,当步骤(e)的所述溶剂为水时,所述樟芝菌丝体活性物质的有效浓度介于0.5-1mg/ml。
7.如权利要求5所述的用途,其中,当步骤(e)的所述溶剂为醇类时,所述樟芝菌丝体活性物质的有效浓度介于50-100μg/ml。
8.如权利要求1所述的用途,其中,步骤(b)的烧瓶培养为震荡培养,且转速为110-130rpm。
9.如权利要求1所述的用途,其中,步骤(c)中所述发酵槽进一步通入一气体,该气体包括空气、氧气、二氧化碳、氦气或其组合,所述发酵槽的槽压为0.5-1.0kg/cm2且通气速率为0.01-1.5VVM。
10.如权利要求1所述的用途,其中,所述组合物为医药组合物,所述医药组合物进一步包含药学上可接受的载剂、赋形剂、稀释剂或辅剂。
11.如权利要求1所述的用途,其中,所述组合物为食品添加剂。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101803528A (zh) * | 2009-02-16 | 2010-08-18 | 葡萄王生技股份有限公司 | 一种樟芝菌丝体的新颖培养方法 |
US20150050315A1 (en) * | 2013-08-13 | 2015-02-19 | Industrial Technology Research Institute | Method for modulating th17 cells and method for treating a disease related to modulation of th17 cells |
CN104367600A (zh) * | 2013-08-13 | 2015-02-25 | 财团法人工业技术研究院 | 牛樟芝萃取物用于制备调节Th17细胞或治疗与Th17细胞调节相关之疾病的药物的用途 |
CN106309508A (zh) * | 2015-07-09 | 2017-01-11 | 葡萄王生技股份有限公司 | 樟芝菌丝体活性物质及其制法、含其的医药组合物及用途 |
-
2018
- 2018-05-25 TW TW107117897A patent/TWI698243B/zh active
- 2018-07-12 CN CN201810761894.4A patent/CN110522771A/zh active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101803528A (zh) * | 2009-02-16 | 2010-08-18 | 葡萄王生技股份有限公司 | 一种樟芝菌丝体的新颖培养方法 |
US20150050315A1 (en) * | 2013-08-13 | 2015-02-19 | Industrial Technology Research Institute | Method for modulating th17 cells and method for treating a disease related to modulation of th17 cells |
CN104367600A (zh) * | 2013-08-13 | 2015-02-25 | 财团法人工业技术研究院 | 牛樟芝萃取物用于制备调节Th17细胞或治疗与Th17细胞调节相关之疾病的药物的用途 |
CN106309508A (zh) * | 2015-07-09 | 2017-01-11 | 葡萄王生技股份有限公司 | 樟芝菌丝体活性物质及其制法、含其的医药组合物及用途 |
Non-Patent Citations (1)
Title |
---|
黄志伟等: "樟芝菌丝体的固体培养特性研究 ", 《中国食用菌》 * |
Cited By (1)
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---|---|---|---|---|
CN113082015A (zh) * | 2020-01-08 | 2021-07-09 | 深圳市罗湖区人民医院 | 金雀异黄素在预防慢性阻塞性肺病中的应用及验证方法 |
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