CN110522771A - Antrodia camphorata mycelium active material is used to improve the purposes of chronic obstructive pulmonary disease - Google Patents
Antrodia camphorata mycelium active material is used to improve the purposes of chronic obstructive pulmonary disease Download PDFInfo
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- antrodia camphorata
- camphorata mycelium
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- 241001486992 Taiwanofungus camphoratus Species 0.000 title claims abstract description 87
- 239000011149 active material Substances 0.000 title claims abstract description 34
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 title claims abstract description 24
- 239000007788 liquid Substances 0.000 claims abstract description 17
- 238000000855 fermentation Methods 0.000 claims abstract description 15
- 230000004151 fermentation Effects 0.000 claims abstract description 15
- 239000000203 mixture Substances 0.000 claims abstract description 14
- 238000007747 plating Methods 0.000 claims abstract description 4
- 239000000284 extract Substances 0.000 claims description 17
- 241000123370 Antrodia Species 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 239000000843 powder Substances 0.000 claims description 8
- 239000002904 solvent Substances 0.000 claims description 8
- 230000000694 effects Effects 0.000 claims description 6
- 150000001298 alcohols Chemical class 0.000 claims description 4
- 239000007789 gas Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 239000002671 adjuvant Substances 0.000 claims description 3
- 239000003085 diluting agent Substances 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 3
- 235000013373 food additive Nutrition 0.000 claims description 3
- 239000002778 food additive Substances 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 3
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 3
- 238000009423 ventilation Methods 0.000 claims description 3
- 239000001307 helium Substances 0.000 claims description 2
- 229910052734 helium Inorganic materials 0.000 claims description 2
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims description 2
- 238000000638 solvent extraction Methods 0.000 claims description 2
- 239000003570 air Substances 0.000 claims 1
- UBAZGMLMVVQSCD-UHFFFAOYSA-N carbon dioxide;molecular oxygen Chemical compound O=O.O=C=O UBAZGMLMVVQSCD-UHFFFAOYSA-N 0.000 claims 1
- 238000003756 stirring Methods 0.000 abstract description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 21
- 235000019441 ethanol Nutrition 0.000 description 21
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 14
- 238000000605 extraction Methods 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 9
- 238000003809 water extraction Methods 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 8
- 239000012887 cigarette smoke extract Substances 0.000 description 7
- 230000004083 survival effect Effects 0.000 description 7
- 241000723346 Cinnamomum camphora Species 0.000 description 5
- 210000002821 alveolar epithelial cell Anatomy 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 241000723347 Cinnamomum Species 0.000 description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 4
- 230000006872 improvement Effects 0.000 description 4
- 210000004072 lung Anatomy 0.000 description 4
- 201000005202 lung cancer Diseases 0.000 description 4
- 208000020816 lung neoplasm Diseases 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 3
- 235000010469 Glycine max Nutrition 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000005779 cell damage Effects 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 210000002919 epithelial cell Anatomy 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 231100000002 MTT assay Toxicity 0.000 description 2
- 238000000134 MTT assay Methods 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- MQTOSJVFKKJCRP-BICOPXKESA-N azithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)N(C)C[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 MQTOSJVFKKJCRP-BICOPXKESA-N 0.000 description 2
- 229960004099 azithromycin Drugs 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 208000037887 cell injury Diseases 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 235000017803 cinnamon Nutrition 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000002547 new drug Substances 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000013618 particulate matter Substances 0.000 description 2
- 201000008827 tuberculosis Diseases 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- ZCWPHDXKEDBCER-UHFFFAOYSA-N 2,5-diphenyl-2h-tetrazol-2-ium;bromide Chemical compound [Br-].C1=CC=CC=C1C1=[NH+]N(C=2C=CC=CC=2)N=N1 ZCWPHDXKEDBCER-UHFFFAOYSA-N 0.000 description 1
- 125000000972 4,5-dimethylthiazol-2-yl group Chemical group [H]C([H])([H])C1=C(N=C(*)S1)C([H])([H])[H] 0.000 description 1
- 208000014085 Chronic respiratory disease Diseases 0.000 description 1
- 241000386927 Cinnamomum micranthum f. kanehirae Species 0.000 description 1
- 241000790917 Dioxys <bee> Species 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 240000004922 Vigna radiata Species 0.000 description 1
- 235000010721 Vigna radiata var radiata Nutrition 0.000 description 1
- 235000011469 Vigna radiata var sublobata Nutrition 0.000 description 1
- 241000746966 Zizania Species 0.000 description 1
- 235000002636 Zizania aquatica Nutrition 0.000 description 1
- CKUAXEQHGKSLHN-UHFFFAOYSA-N [C].[N] Chemical compound [C].[N] CKUAXEQHGKSLHN-UHFFFAOYSA-N 0.000 description 1
- 238000003915 air pollution Methods 0.000 description 1
- 239000002249 anxiolytic agent Substances 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 229940124630 bronchodilator Drugs 0.000 description 1
- 239000000168 bronchodilator agent Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000002485 combustion reaction Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- RUTXIHLAWFEWGM-UHFFFAOYSA-H iron(3+) sulfate Chemical compound [Fe+3].[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O RUTXIHLAWFEWGM-UHFFFAOYSA-H 0.000 description 1
- 229910000360 iron(III) sulfate Inorganic materials 0.000 description 1
- 231100000225 lethality Toxicity 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 230000003137 locomotive effect Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000000414 obstructive effect Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 239000001965 potato dextrose agar Substances 0.000 description 1
- 238000010248 power generation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 230000005586 smoking cessation Effects 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000009092 tissue dysfunction Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Mycology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Alternative & Traditional Medicine (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Epidemiology (AREA)
- Pulmonology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The present invention provides a kind of Antrodia camphorata mycelium active material and is used to prepare the purposes for improving the composition of chronic obstructive pulmonary disease.The Antrodia camphorata mycelium active material is prepared with the following steps: (a) being taken an Antrodia camphorata mycelium (Antrodia cinnamomea) on plating medium, is cultivated 4-10 days at a temperature of 15-30 DEG C;(b) Antrodia camphorata mycelium after step (a) culture is seeded in a flask, in 15-30 DEG C, environment culture 3-14 days of pH 2-6;(c) Antrodia camphorata mycelium after step (b) culture is inoculated in a fermentation tank, stir culture 3-21 days in the environment of 15-30 DEG C, pH2-6 forms the Antrodia camphorata mycelium fermented liquid containing the Antrodia camphorata mycelium active material.
Description
Technical field
A kind of preparation method and its usage of the present invention about Antrodia camphorata mycelium active material.Specifically, about preparation
A kind of Antrodia camphorata mycelium active material and its purposes in food or pharmaceuticals for improving chronic obstructive pulmonary disease.
Background technique
Chronic Obstructive Pulmonary Disease (Chronic Obstructive Pulmonary Disease, abbreviation COPD) is current
The 4th of the global cause of death is occupied, is a kind of common, multiple, high disability rate and high lethality rate chronic respiratory disease.Entirely
The patient of world's chronic obstructive pulmonary disease has more than 300,000,000 people.
Smoking is the main reason for leading to COPD, so in recent years because of locomotive, the waste air of automobile and industrial power generation and combustion
Oily coal-fired use increases, and causes the air pollution of Particulate Matter (PM) 2.5 to rise, more aggravates this disease
Occur.It is chronically exposed in the environment of pollution, lung can be made to generate inflammatory response, airflow channel is caused to narrow and generate lung tissue
Dysfunction.
The therapeutic modality of COPD includes smoking cessation at present, lung rehabilitation, imbedibility bronchodilators, solid using cortex class
Alcohol, long-term oxygen supply or progress lung transplantation etc..2015 it is newest by new drug be to merge two kinds of muscle for being used in COPD in the past
Relaxant does not have breakthrough new drug development yet so far.Therefore, research effectively treats the method for chronic obstructive pulmonary disease still
It is important developing direction.
Antrodia (Antrodia cinnamomea), also known as camphor tree mushroom, camphor tree wild rice, mushroom in camphor tree, cinnamomum kanahirai hay mushroom, Antrodia camphorata, red camphor tree,
Red antrodia, the rotten heartwood inner wall of the cinnamomum kanehirai dry rot being only grown between height above sea level 450-1500 meters of TaiWan, China mountain area or it is withered fall
The dark moist surface of the Cinnamomum kanahirai hay timber of volt, belongs to the precious medicinal fungi of TaiWan, China national treasure grade.Still lack antrodia pair at present
Improve the research of chronic obstructive pulmonary disease.
Summary of the invention
The present invention provides a kind of Antrodia camphorata mycelium active material and preparation method thereof, and can be used for preparing tool improves chronic resistance
The composition of plug property tuberculosis.Compared to general Western medicine and treatment method, the invention discloses liquid state fermentation Antrodia camphorata mycelium activity
The preparation method of substance is safer, easy, and manufactured Antrodia camphorata mycelium active material is more natural, safe, and can be effectively improved
COPD。
An embodiment according to the present invention provides a kind of Antrodia camphorata mycelium active material and is used to prepare improvement chronic obstructive
The purposes of the composition of tuberculosis, wherein Antrodia camphorata mycelium active material is prepared with the following steps:
(a) Antrodia camphorata mycelium (Antrodia cinnamomea) is taken on plating medium, at a temperature of 15-30 DEG C
Culture 4-10 days;
(b) Antrodia camphorata mycelium after step (a) culture is seeded in flask, in 15-30 DEG C, the environment culture of pH 2-6
3-14 days;
(c) Antrodia camphorata mycelium after step (b) culture is inoculated in fermentation tank, in the environment of 15-30 DEG C, pH2-6
Stir culture 3-21 days, form the Antrodia camphorata mycelium fermented liquid containing Antrodia camphorata mycelium active material.
In one embodiment, the step of preparing Antrodia camphorata mycelium active material further includes step (d): will be Antrodia camphorata mycelium fermented
It is milled after liquid freeze-drying, forms the Antrodia camphorata mycelium freeze-dried powder containing Antrodia camphorata mycelium active material.
In one embodiment, the step of preparing Antrodia camphorata mycelium active material further includes step (e): Antrodia camphorata mycelium is lyophilized
Powder forms the Antrodia camphorata mycelium extract liquor containing Antrodia camphorata mycelium active material with an at least solvent extraction.
In one embodiment, the step of preparing Antrodia camphorata mycelium active material further includes step (f): Antrodia camphorata mycelium is extracted
Liquid is dry, to obtain Antrodia camphorata mycelium active material.
In one embodiment, the solvent in step (e) is selected from water or alcohols.
In one embodiment, when the solvent of step (e) is water, the effective concentration of Antrodia camphorata mycelium active material between
0.25-1mg/ml。
In one embodiment, when the solvent of step (e) is alcohols, the effective concentration of Antrodia camphorata mycelium active material between
25-100μg/ml。
In one embodiment, the flask culture of above-mentioned steps (b) is shake culture, and revolving speed is 110-130rpm.
In one embodiment, the fermentation tank in step (c) is further passed through a gas, this gas includes air, oxygen, dioxy
Change carbon, helium or combinations thereof, the slot pressure of fermentation tank is 0.5-1.0kg/cm2And Ventilation Rate is 0.01-1.5VVM.
In one embodiment, the composition is medical composition, this medical composition further includes pharmaceutically acceptable
Carrier, excipient, diluent or adjuvant.
In one embodiment, the composition is food additives.
Detailed description of the invention
Antrodia camphorata mycelium water extraction object, which is shown, in Fig. 1 improves cigarette smoke extract induction alveolar epithelium A549 cell damage
Ability as a result, wherein AcH (L), AcH (M) and AcH (H) respectively refer to Antrodia camphorata mycelium water extraction object concentration be 0.25mg/ml,
0.5mg/ml and 1.0mg/ml.
Antrodia camphorata mycelium alcohol extraction object, which is shown, in Fig. 2 improves cigarette smoke extract induction alveolar epithelium A549 cell damage
Ability as a result, wherein AcMe (L), AcMe (M) and AcMe (H) respectively refer to Antrodia camphorata mycelium alcohol extraction object concentration be 25 μ g/ml,
50 μ g/ml and 100 μ g/ml.
Specific embodiment
To keep above and other aspect of the invention apparent understandable, special embodiment below, and cooperate institute's accompanying drawings
It is described in detail.
One Antrodia camphorata mycelium culture of embodiment
The Antrodia camphorata mycelium (Antrodia cinnamomea) that the present embodiment uses is purchased from juridical person's development of food industry
Research institute's (FIRDI) living resources save and research center (BCRC, preceding title CCRC), number CCRC-35396.However, available
In antrodia kind of the invention, that it's not limited to that is individually a kind of, may also include BCRC number 35396,35398,35716,
36401,36711,36795,37848,37849,37850,37889,37890,37891,37893,37894,37941 or and its
The antrodia strain in allogene material registered data library, the present invention do not limit it simultaneously.
(1) plate culture: Antrodia camphorata mycelium is inoculated on plate, and 4-10 days (the present embodiment are cultivated at 15-30 DEG C
In, it is to be cultivated 7 days at 25 DEG C).The ingredient of plating medium may include detrine culture medium (PotatoDextrose
Agar, PDA), carbon source and nitrogen source, there is no particular restriction.
(2) flask culture: scraping (1) plate on mycelium be inoculated in flask, and with 15-30 DEG C, pH 2-6 and turn
Fast 110-130rpm condition shake culture 3-14 days (in the present embodiment, the shake culture under 25 DEG C, pH5, revolving speed 120rpm
10 days).This shake culture is that culture medium shown in following table 1 is cultivated.
1 culture medium prescription of table
In above-mentioned culture medium prescription, soybean powder and ground cinnamon are comprehensive carbon nitrogen source, can be selected from cereal (such as: flour) or
Beans (such as: soybean powder, mung bean flour, soy meal, ground cinnamon);Carbohydrate can be glucose, fructose, maltose, sucrose etc.;It is inorganic
Salt can be magnesium sulfate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, ferric sulfate etc..It illustrates, 1 culture medium prescription of table is only
One of example, ingredient can adjust on demand when use, or the commercially available culture medium of collocation uses, and there is no particular restriction.
(3) fermentation tank culture: the mycelium of flask culture in (2) is inoculated in fermentation tank, with 15-30 DEG C, slot pressure
0.5-1.0kg/cm2, under the conditions of pH 2-6 and mixing speed 50-150rpm, with the Ventilation Rate culture 3-21 of 0.1-1.5VVM
It, forming an Antrodia camphorata mycelium fermented liquid (is in 25 DEG C, slot pressure 0.5kg/cm in the present embodiment2, pH4, mixing speed 80rpm
And under conditions of 1.0VVM (air), cultivate 14 days) to get Antrodia camphorata mycelium fermented liquid.The culture medium that fermentation tank culture uses
It can be identical as the culture medium that step (2) flask culture uses.I.e. containing antrodia camphorata mycelium of the invention in this Antrodia camphorata mycelium fermented liquid
Body active material.Antrodia camphorata mycelium fermented liquid further can be prepared as Antrodia camphorata mycelium fermented liquid freeze-drying by being freeze-dried step
Powder;
(4) prepared by extract: Antrodia camphorata mycelium fermented liquid freeze-dried powder being divided into two parts, is separately added into the distillation of 20 times of weight
Water and ethyl alcohol are extracted.The extract taken water as a solvent is heated 30 minutes with 100 DEG C of temperature, centrifuging and taking supernatant after cooling
Antrodia fermentation mycelium water extraction object is dried to obtain in liquid, freeze-dried method.Using ethyl alcohol as the extract of solvent, shaken with ultrasonic
It swings 1 hour, supernatant is taken after centrifugation, antrodia fermentation mycelium alcohol extraction object is concentrated under reduced pressure to obtain in supernatant;
Via extraction step, the water extraction object and alcohol extraction object of the Antrodia camphorata mycelium active material containing higher concentration can be obtained.Camphor tree
The aspect of camphorata mycelium active material includes Antrodia camphorata mycelium fermented liquid (mycelium and clarified solution), fermentation liquid freeze-dried powder, water extraction
Object, alcohol extraction object, water extraction object and alcohol extract the mixture or other dosage forms of object.It is to extract object and alcohol extraction object with water in following embodiment two
As Antrodia camphorata mycelium active material aspect.
Two Antrodia camphorata mycelium water of embodiment, which extracts object and alcohol extraction object, improves the analysis of COPD
Have experiment at present and utilizes cigarette smoke extract (Cigarette smoke extract, CSE) and Human Lung Cancer
Epithelial cell A549 is successfully established COPD mode.It can refer to Miaomiao Chenet al. (2015) .Azithromycin
attenuates cigarette smoke extract-induced oxidative stress injury in human
This paper of alveolar epithelial cells.MOLECULAR MEDICINE REPORTS, 11:3414-3422.,
Cell survival rate analysis (MTT assay) is carried out to CSE with Human Lung Cancer epithelial cell A549, to assess antibiotic
Improvement result of the Azithromycin to COPD.CSE is important in chronic obstructive pulmonary disease mechanism to the damage of alveolar epithelium
Process.Therefore, if can inhibit CSE that can achieve the effect that improve chronic obstructive pulmonary disease to the injury of alveolar epithelium.This experiment
In cell survival rate analysis (MTT assay) is also carried out to CSE with Human Lung Cancer epithelial cell A549, assessment Antrodia camphorata mycelium is living
Improvement result of the property substance to COPD.
(1) Human Lung Cancer epithelium A549 cell is collected, cell suspending liquid concentration is adjusted, it is close with 50,000-100,000/mL
Degree is planted in 96 porose discs;
(2) 96 porose discs of (1) are placed in 37 DEG C, 5%CO2After incubator culture 16-24 hours, obtained with embodiment one
, at Antrodia camphorata mycelium water extraction object (0,0.25,0.5,1mg/ml) and the alcohol extraction object (0,25,50,100 μ g/ml) of various dose
Reason.Antrodia camphorata mycelium extract is dissolved in dimethyl Asia Shi Feng (Dimethyl sulfoxide, DMSO), DMSO when detection
Concentration is no more than 0.1% to avoid the growth of its toxic effect cell;Control group handles 0.1%DMSO, and each concentration carries out triple
It is multiple, as a result it is placed in 37 DEG C, 5%CO2Incubator culture 24 hours;
(3) result of above-mentioned (2) is first centrifuged 10min with 400 × g, removes supernatant, adds cigarette smoke extract
CSE12.5% induces the cytotoxicity of A549, and control group is not handled.Then 37 DEG C, 5%CO are placed in2Incubator culture 6 hours;
(4) supernatant of (3) is removed, 100 μ l MTT (3- (4,5-Dimethylthiazol-2-yl)-are added in every hole
2,5-diphenyltetrazolium bromide) solution, after 37 DEG C are reacted 2 hours, termination culture is carefully removed in hole
MTT solution;
(5) 100 μ l DMSO are added in every hole in (4), shake 2min, measure light absorption value in 570nm.This inhales brightness and represents
Ability (the first of cell reduction MTTForming amount), i.e. the activity (number of viable cells) of grain wire body.It is got in the suction brightness of 570nm
Height indicates that cell survival rate is higher.
Statistical analysis: experimental result data is indicated with average value (Mean) ± standard deviation (SD).Test data with
Student's t-test statisticallys analyze its group difference.It is considered as if p < 0.05 with statistically significant difference.
Water extracts the experimental result of object as shown in Fig. 1 and the following table 2.Cigarette smoke extract CSE can induce A549 cytotoxicity,
Cell survival rate is caused to decline.However, be added antrodia fermentation mycelium water extraction object can conspicuousness inhibit cellular damage, promoted survival
Rate, and the concentration being added is higher, survival rate is higher.Being had using the Antrodia camphorata mycelium water extraction object in the embodiment of the present invention one is improved
The effect of COPD, effective concentration is between 0.5-1mg/ml.
2 Antrodia camphorata mycelium water of table extracts object test result
(n=3)
# indicates statistically to have significant difference (p < 0.05) with control group (control)
* it indicates statistically to have significant difference (p < 0.05) with negative control group (CSE)
Alcohol extracts the experimental result of object as shown in Fig. 2 and the following table 3.As a result it again shows that, it is real that the antrodia zymophyte present invention is added
The Antrodia camphorata mycelium alcohol extraction object applied in example one has the function of improvement COPD, and effective concentration is between 50-100 μ g/ml.
3 Antrodia camphorata mycelium alcohol of table extracts object test result
Group | Survival rate (%) |
Control group (Control) | 100±2.93 |
CSE | 84.51±0.70# |
CSE+ Antrodia camphorata mycelium alcohol extracts 25 μ g/ml of object | 84.63±1.91 |
CSE+ Antrodia camphorata mycelium alcohol extracts 50 μ g/ml of object | 86.90±2.20 |
CSE+ Antrodia camphorata mycelium alcohol extracts 100 μ g/ml of object | 89.92±2.89* |
Antrodia camphorata mycelium alcohol extracts 100 μ g/ml of object | 103.66±2.47 |
(n=3)
# indicates statistically to have significant difference (p < 0.05) with control group (control)
* it indicates statistically to have significant difference (p < 0.05) with negative control group (CSE)
Antrodia active material of the invention has proven to have the function of improving COPD through above-mentioned experiment.It can with pharmaceutically
Acceptable carrier, excipient, diluent or adjuvant combine, and prepare medical composition, can also be used as food additives and be used.
Although the present invention has used embodiment disclosed above, so it is not intended to limiting the invention.This field general technology people
Member when can be appropriately modified to the content of above-described embodiment, and still can reach the present invention and lead after considering teachings above in light of actual conditions
The effect of opening.Therefore, protection scope of the present invention should be subject to claims.
Claims (11)
1. a kind of Antrodia camphorata mycelium active material is used to prepare the purposes for improving the composition of chronic obstructive pulmonary disease, wherein should
Antrodia camphorata mycelium active material is prepared with the following steps:
(a) it takes an Antrodia camphorata mycelium on plating medium, is cultivated 4-10 days at a temperature of 15-30 DEG C;
(b) Antrodia camphorata mycelium after step (a) culture is seeded in a flask, in 15-30 DEG C, the environment culture 3- of pH 2-6
14 days;
(c) Antrodia camphorata mycelium after step (b) culture is inoculated in a fermentation tank, is stirred in the environment of 15-30 DEG C, pH2-6
Culture 3-21 days is mixed, the Antrodia camphorata mycelium fermented liquid for containing the Antrodia camphorata mycelium active material is formed.
2. purposes as described in claim 1, wherein the step of preparing the Antrodia camphorata mycelium active material further includes step
(d): will be milled after the Antrodia camphorata mycelium fermented liquid freeze-drying, form the antrodia for containing the Antrodia camphorata mycelium active material
Mycelium freeze-dried powder.
3. purposes as claimed in claim 2, wherein the step of preparing the Antrodia camphorata mycelium active material further includes step
(e): by the Antrodia camphorata mycelium freeze-dried powder with an at least solvent extraction, being formed containing the Antrodia camphorata mycelium active material extremely
A few Antrodia camphorata mycelium extract liquor.
4. purposes as claimed in claim 3, wherein the step of preparing the Antrodia camphorata mycelium active material further includes step
(f): the Antrodia camphorata mycelium extract liquor is dry, to obtain the Antrodia camphorata mycelium active material.
5. purposes as described in claim 3 or 4, wherein in step (e), the solvent is selected from water or alcohols.
6. purposes as claimed in claim 5, wherein when the solvent of step (e) is water, the Antrodia camphorata mycelium activity
The effective concentration of substance is between 0.5-1mg/ml.
7. purposes as claimed in claim 5, wherein when the solvent of step (e) is alcohols, the Antrodia camphorata mycelium is living
The effective concentration of property substance is between 50-100 μ g/ml.
8. purposes as described in claim 1, wherein the flask culture of step (b) is shake culture, and revolving speed is 110-
130rpm。
9. purposes as described in claim 1, wherein fermentation tank described in step (c) is further passed through a gas, the gas packet
Air, oxygen, carbon dioxide, helium or combinations thereof are included, the slot pressure of the fermentation tank is 0.5-1.0kg/cm2And Ventilation Rate is
0.01-1.5VVM。
10. purposes as described in claim 1, wherein the composition is medical composition, and the medical composition is further
Include pharmaceutically acceptable carrier, excipient, diluent or adjuvant.
11. purposes as described in claim 1, wherein the composition is food additives.
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