CN108785332A - 包含羊肚菌提取物作为有效成份且具有抗癌功能的组成物 - Google Patents
包含羊肚菌提取物作为有效成份且具有抗癌功能的组成物 Download PDFInfo
- Publication number
- CN108785332A CN108785332A CN201810421421.XA CN201810421421A CN108785332A CN 108785332 A CN108785332 A CN 108785332A CN 201810421421 A CN201810421421 A CN 201810421421A CN 108785332 A CN108785332 A CN 108785332A
- Authority
- CN
- China
- Prior art keywords
- extract
- morchella esculenta
- constituent
- anticancer
- esculenta extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 239000000284 extract Substances 0.000 title claims abstract description 62
- 240000002769 Morchella esculenta Species 0.000 title claims abstract description 46
- 235000002779 Morchella esculenta Nutrition 0.000 title claims abstract description 46
- 239000000470 constituent Substances 0.000 title claims abstract description 33
- 230000001093 anti-cancer Effects 0.000 title claims description 24
- 239000004615 ingredient Substances 0.000 title claims description 6
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 15
- 201000011510 cancer Diseases 0.000 claims abstract description 15
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 36
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 18
- 241000723418 Carya Species 0.000 claims description 16
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- 239000000126 substance Substances 0.000 claims description 11
- 235000019441 ethanol Nutrition 0.000 claims description 7
- 235000020710 ginseng extract Nutrition 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 abstract description 21
- 230000006907 apoptotic process Effects 0.000 abstract description 14
- 102000004169 proteins and genes Human genes 0.000 abstract description 10
- 239000004480 active ingredient Substances 0.000 abstract description 6
- 230000002401 inhibitory effect Effects 0.000 abstract description 4
- 210000004027 cell Anatomy 0.000 description 52
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 description 12
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 11
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 11
- 101000721661 Homo sapiens Cellular tumor antigen p53 Proteins 0.000 description 11
- 108700000707 bcl-2-Associated X Proteins 0.000 description 10
- 230000004083 survival effect Effects 0.000 description 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- 102000055102 bcl-2-Associated X Human genes 0.000 description 9
- 238000012545 processing Methods 0.000 description 9
- 108050006400 Cyclin Proteins 0.000 description 8
- 102100036691 Proliferating cell nuclear antigen Human genes 0.000 description 8
- 239000000463 material Substances 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 238000007792 addition Methods 0.000 description 6
- 235000013376 functional food Nutrition 0.000 description 6
- 230000036541 health Effects 0.000 description 6
- 241001494479 Pecora Species 0.000 description 5
- 229940093499 ethyl acetate Drugs 0.000 description 5
- 235000019439 ethyl acetate Nutrition 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 210000003734 kidney Anatomy 0.000 description 5
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 4
- 241000283707 Capra Species 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000001737 promoting effect Effects 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 241000208340 Araliaceae Species 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- 241000233866 Fungi Species 0.000 description 3
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 3
- 235000003140 Panax quinquefolius Nutrition 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 230000000996 additive effect Effects 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000001640 apoptogenic effect Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000002552 dosage form Substances 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 239000010408 film Substances 0.000 description 3
- 235000008434 ginseng Nutrition 0.000 description 3
- 230000002438 mitochondrial effect Effects 0.000 description 3
- 229920000136 polysorbate Polymers 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 238000001262 western blot Methods 0.000 description 3
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 2
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 2
- 102000011727 Caspases Human genes 0.000 description 2
- 108010076667 Caspases Proteins 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- 108700041737 bcl-2 Genes Proteins 0.000 description 2
- 230000022131 cell cycle Effects 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 231100000433 cytotoxic Toxicity 0.000 description 2
- 230000001472 cytotoxic effect Effects 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 239000000539 dimer Substances 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 230000036737 immune function Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 208000032839 leukemia Diseases 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- -1 nitrite anions Chemical class 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- QAPSNMNOIOSXSQ-YNEHKIRRSA-N 1-[(2r,4s,5r)-4-[tert-butyl(dimethyl)silyl]oxy-5-(hydroxymethyl)oxolan-2-yl]-5-methylpyrimidine-2,4-dione Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O[Si](C)(C)C(C)(C)C)C1 QAPSNMNOIOSXSQ-YNEHKIRRSA-N 0.000 description 1
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 102000018832 Cytochromes Human genes 0.000 description 1
- 108010052832 Cytochromes Proteins 0.000 description 1
- 208000003098 Ganglion Cysts Diseases 0.000 description 1
- 101000911513 Homo sapiens Uncharacterized protein FAM215A Proteins 0.000 description 1
- 241000221637 Morchellaceae Species 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 101150008755 PCNA gene Proteins 0.000 description 1
- 238000009004 PCR Kit Methods 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 241000123107 Phellinus Species 0.000 description 1
- 230000018199 S phase Effects 0.000 description 1
- 208000001203 Smallpox Diseases 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 208000005400 Synovial Cyst Diseases 0.000 description 1
- ATJFFYVFTNAWJD-UHFFFAOYSA-N Tin Chemical compound [Sn] ATJFFYVFTNAWJD-UHFFFAOYSA-N 0.000 description 1
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 1
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 1
- 102100026728 Uncharacterized protein FAM215A Human genes 0.000 description 1
- 241000870995 Variola Species 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 229940124650 anti-cancer therapies Drugs 0.000 description 1
- 238000011319 anticancer therapy Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 101150024147 bax gene Proteins 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000007975 buffered saline Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000002026 chloroform extract Substances 0.000 description 1
- NEHMKBQYUWJMIP-UHFFFAOYSA-N chloromethane Chemical class ClC NEHMKBQYUWJMIP-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000002035 hexane extract Substances 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 229950008882 polysorbate Drugs 0.000 description 1
- 229940068965 polysorbates Drugs 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 230000001686 pro-survival effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 150000003536 tetrazoles Chemical class 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- A61K36/062—Ascomycota
-
- 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
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- 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
- 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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
-
- 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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/80—Scrophulariaceae (Figwort family)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- 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
- 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
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/308—Foods, ingredients or supplements having a functional effect on health having an effect on cancer prevention
-
- 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
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/208—Fungi extracts
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Mycology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Epidemiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Alternative & Traditional Medicine (AREA)
- Immunology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
本发明涉及包含羊肚菌提取物作为有效成分的组成物。尤其是,根据本发明的组成物具有抑制癌细胞增殖的功能,并且具有可以调节与癌细胞的细胞凋亡有关的基因及蛋白质的功能。
Description
技术领域
本发明涉及作为有效成份包含羊肚菌(Morchella esculenta)提取物的组成物。
尤其是,根据本发明的组成物具有抑制癌细胞增殖的功能,并且具有可调节与癌细胞的细胞凋亡有关的基因及蛋白质的功能。
背景技术
羊肚菌在分类学上是属于真菌门中的子囊菌亚纲羊肚菌科羊肚菌的野生食用菌,是全世界性分布的菌类。羊肚菌在形成子囊果的子实体的表皮形成子囊和8个子囊孢子,子实体的形态为菌盖周边形成如地垄的深沟,其特征在于像地垄那样形成的周围形成为如火山喷发的喷火口或痘疮印,学名为羊肚菌,俗称为羊肚蘑。
子实体的直径为4~5cm,高为8~15cm,头部是宽阔的卵形,具有看起来像刻成网眼的样子的多个凹陷的地方,在其下方子实体层发达呈浅黄褐色或灰黄色(图1)。菌柄的长度为2.5~5cm且为白色,下部膨出,表面为浊黄色,有褶皱,并附着有如米糠的鳞片,内部为中空到头部。孢子的大小为20~25×12~15μm,是无色的椭圆形,表面光滑,是美味的食用菌。羊肚菌子实体的生长经常是在早春4月初至中旬之间生长在自然状态的土壤或地垄、或者发生过山火的第二年的山区,但是生长期间短,单生型且分散性生长,因此收集子实体非常困难。
本发明人研究羊肚菌的抗癌活性,从而提出作为有效成分包含羊肚菌的抗癌活性组成物。若观察在先技术文献,在大韩民国公开专利公报第10-2014-0081137号记载有羊肚菌子实体提取物或羊肚菌菌丝体提取物的皮肤外用剂组成物。另外,在大韩民国注册专利公报第10-1629793号记载有本申请人申请并注册的用于生产羊肚蘑的培养基组成物。但是未检索到关于羊肚菌或羊肚蘑的抗癌活性的专利文献。
在先技术文献
专利文献1:大韩民国公开专利公报第10-2014-0081137号(2014.07.01公开
专利文献2:大韩民国注册专利公报第10-1629793号(2016.06.15公告)
发明内容
所要解决的技术课题
本申请所要解决的技术课题是提供一种包含羊肚菌提取物作为有效成份的抗癌组成物。
课题解决方案
本发明通过提供一种包含羊肚菌提取物作为有效成份的抗癌组成物,解决技术课题。
在本发明中,上述羊肚菌提取物为将羊肚菌通过乙醇、乙酸乙酯、三氯甲烷或己烷提取出的提取物。
在本发明中,上述羊肚菌提取物的含量为按照癌细胞5×104~3×106细胞/孔基准观察时,包含1~100μg/mL。
在本发明中,在上述组成物中还包含羊肚菌发酵物。
在本发明中,在上述组成物还包含免疫增进物质。
本发明具有如下的有益效果:
根据本发明的组成物,具有抑制癌细胞增殖的功能。
根据本发明的组成物,具有可以调节与癌细胞的细胞凋亡有关的基因及蛋白质的功能。
附图说明
图1是在自然状态下的羊肚菌照片。
图2是羊肚菌提取物的癌细胞增殖抑制实验的结果图表。
图3是羊肚菌提取物对正常细胞生存率的影响的实验结果图表。
图4及图5是羊肚菌提取物对细胞凋亡相关基因表达的影响的实验结果照片。
图6是羊肚菌提取物对细胞凋亡相关基因表达的影响的实验结果照片。
具体实施方式
本说明书及权利要求书所使用的用语或单词不应限定性地解释为一般或词典上的意义,应立足于发明人可以为了用最优选的方法说明自己的发明适当定义用语的概念的原则解释为符合本发明的技术思想的意义和概念。
因此,本说明书记载的实施例、参照例及附图中记载的事项不过是本发明的最优选的一例,不代表本发明的全部技术思想,因此,应当理解在本申请提交时可以存在可代替它们的多样的等同物和变形例。
[实施例]
1.包含羊肚菌提取物作为有效成分的抗癌组成物
根据本发明的实施例1,提供一种包含羊肚菌提取物作为有效成分的组成物。上述羊肚菌提取物可以反映下述的实验结果来提供。例如,羊肚菌提取物采用通过乙醇、乙酸乙酯、三氯甲烷或己烷提取出的提取物。另外,羊肚菌提取物的含量为按照癌细胞5×104~3×106细胞/孔基准观察时,可包含1~100μg/mL。
根据本发明的组成物可以用于药剂材料或保健功能食品,这时可以分别添加药学上或食品学上可允许的添加剂。利用了根据本发明的组成物的药剂材料或保健功能食品能够以能被人体摄入或可以服用的多种剂型提供。
2.包含羊肚菌提取物及羊肚菌发酵物作为有效成分的抗癌组成物
根据本发明的实施例2,提供一种在羊肚菌提取物添加羊肚菌发酵物组成的抗癌组成物。羊肚菌提取物可以根据上述实施例1记载的方案提供。羊肚菌发酵物是指将羊肚菌利用可食用的微生物通过多种方法进行发酵的发酵物。随着添加羊肚菌发酵物,可以同时提供基于羊肚菌提取物的抗癌功能和基于发酵物的免疫功能。
根据本发明的组成物可以用于药剂材料或保健功能食品,这时可以分别添加药学上或食品学上可允许的添加剂。利用了根据本发明的组成物的药剂材料或保健功能食品能够以能被人体摄入或服用的多种剂型提供。
3.包含羊肚菌提取物及免疫增进物质作为有效成分的抗癌组成物。
根据本发明的实施例3,提供一种在羊肚菌提取物添加免疫增进物质组成的抗癌组成物。羊肚菌提取物可以通过上述实施例1记载的方案提供。免疫增进物质是指黑参、发酵人参、发酵黑参或发酵桑黄菌等物质的提取物。随着添加免疫增进物质,可以同时提供基于羊肚菌提取物的抗癌功能和基于免疫增进物质的免疫功能。
根据本发明的组成物可以用于药剂材料或保健功能食品,这时可以分别添加药学上或食品学上可允许的添加剂。利用了根据本发明的组成物的药剂材料或保健功能食品能够以能被人体摄入或服用的多种剂型提供。
[实验方法]
1.试料
本实验使用的试料是将从中国进口的干燥羊肚菌提取到乙醇层、乙酸乙酯层、三氯甲烷层及己烷层,并溶解在包含10%DMSO的PBS中使用。
2.细胞株
本实验所使用的细胞株是分别将从韩国细胞株银行分开出售的Molt-4(人体急性淋巴母细胞白血病:Human acute lymphoblastic leukemia)在5%的FBS-RPMI1640培养基灭菌培养,将293(鼠转化原胚肾:Mouse transformed primary embryonal kidney)细胞在5%的FBS-MEM培养基灭菌培养的同时使用。
3.试剂及仪器
本实验所使用的试剂为RPMI1640、MEM媒、胎牛血清(fetal bovine serum:FBS)、水缓冲液(phpsphate buffered saline:PBS)(GIBCO)、3-[4-5-甲基噻唑-2-基]-2,-二苯基四氮唑溴(MTT)(Amresco)、亚硝酸根(Griess ragent for nitrite)(Sigma),PCR试剂盒使用TaKaRa公司的RNA to cDNA EcoDry premix(Oligo dT)、EmeraldAmp GT PCR MasterMix,DNA标记使用了TaKaRa公司的产品。
另外,p53、Bax、Bcl-2、PCNA引物利用PRIMER3程序制作。所使用的抗体为山羊抗兔IgG HRP抗体(goat anti-rabbit IgG-HRP antibody)(Santactuz)、PCNA、Bax、p53、Bcl-2Rabbit Ab(Santactuz),设备使用ELISA检测仪(分子器件(Molecular Device),VERSAmax)、PCR设备(TP-600,Takara,日本),微型凝胶电泳装置(Mupid-21)、显色机(自动X-ray胶片处理机,JP-33,韩国)等。
4.细胞生存率(MTT分析)
将继代培养中的Molt-4(人体急性淋巴细胞白血病)、293(鼠转化原胚肾)细胞在96孔板上调节细胞数使其成为5×104细胞/孔之后,将试料处理为1~100μg/mL浓度的羊肚菌提取物(下面,称之为试料),在37℃的5%的CO2培养箱内培养24小时。在结束培养4小时之前,将以5㎎/mL的浓度稀释于PBS(pH7.4)的MTT溶液20μL处理到各孔,用溶解于0.1N盐酸的10%的SDS100μL溶解且用锡纸遮光18小时。利用ELISA检测仪在570nm测量显色的各孔的吸光度,并与对照组的吸光度比较而按百分率换算细胞生存率。
5.反转录(RT)-PCR
在Molt-4细胞(3×106细胞/孔)添加1~100μg/mL浓度的试料之后,培养24小时,分离总RNA确认了p53、Bax、Bcl-2、PCNA基因。总RNA利用了RNA异加法,遵照制造公司的方法。cDNA利用RNA to cDNA EcoDry premix(Oligo dT)合成,利用EmeraldAmp GT PCRMaster Mix在Dice(TaKaRa)中放大30周期。各周期是在98℃使其变性10秒之后,在55℃退火30秒,在72℃延长30秒。PCR产品在0.5%琼脂糖凝胶中电泳,用Loading STAR(戴恩生物公司:DYNE Bio Co.)染色。使用于PCR的引物如下(表1)。
【表1】
6.蛋白质印迹(western blotting)法
在Molt-4细胞(5×106细胞/孔)添加1~100μg/mL浓度的试料之后,培养24小时,使用PRO-PRAP遵照制造公司的方法提取出蛋白质。将提取出的蛋白质利用布拉德福(Bradford)定律法定量之后,每孔装载20μg之后,通过10%丙烯酰胺凝胶执行SDS-PAGE。在电泳之后,转化为PVDF薄膜(0.2μm)之后,用5%脱脂乳进行封闭(blocking)1小时。然后,将各1级抗体过夜反应(1:1000,4℃),并且用TBS-T(0.05%吐温(注:聚山梨酸酯(polysorbates)制剂的商品名)20)洗涤4次,使山羊抗兔IgG HRP(goat anti-rabbit IgG-HRP)共轭抗体(1:3000)反应1小时。将洗涤4次后的薄膜利用ECL法在暗室中感光到X射线胶片上。
7.统计处理
统计处理是通过学生检验(又称为T检验(student's t-test))执行,将p<0.05以下判定为显著性。
[实验结果]
1.羊肚菌提取物的癌细胞增殖抑制功能
在Molt-4细胞添加1~100μg/mL浓度的试料之后,培养24小时后用MTT方法予以分析。用酶标分析仪(micro plate reader)在570nm测量了各孔的直径。测量结果是3次测量以平均±标准偏差(SD)表示。(与对照组的显著差异为***p<0.001,**p<0.01,*p<0.05)。
为了解羊肚菌提取物对抗癌活性的影响,在Molt-4细胞按各浓度(1~100μg/mL)处理羊肚菌提取物且处理24小时之后,测量细胞生存率的结果,确认到羊肚菌提取物在显著地减少细胞生存率。因此,可知羊肚菌提取物具有抑制白血病细胞的增殖而显示抗癌活性的功能。进而,乙醇、乙酸乙酯、三氯甲烷及己烷分级提取出的提取物均显示出功能,其中,可知三氯甲烷分级提取物及己烷分级提取物的功能比乙醇分级提取物及乙酸乙酯分级提取物更加优秀。
2.羊肚菌提取物对正常细胞生存率的影响
图3是羊肚菌提取物对正常细胞生存率的影响的实验结果图表。在培养的正常细胞施加1~100μg/mL浓度的试料并培养24小时之后用MTT法分析了细胞。用酶标分析仪在570nm测量各孔的外径,结果是测量3次以平均±SD表示。(在此,正常细胞为293鼠转化原胚肾:Mouse transformed primaryembryonal kidney,***表示p<0.001,**表示p<0.01,*表示p<0.05。)
为了解羊肚菌提取物对正常细胞生存率的影响,在293细胞中按各浓度(1~100μg/mL)处理羊肚菌提取物并处理24小时之后测量细胞生存率的结果,确认羊肚菌提取物不减少正常细胞的细胞生存率。该结果表明羊肚菌提取物作为抗癌疗法并行剂是没有问题的。
3.羊肚菌提取物对细胞凋亡相关基因的表达造成的影响
图4及图5是羊肚菌提取物对细胞凋亡相关基因的表达的影响的实验结果照片。在Molt-4细胞施加适当试料浓度并培养24小时。分离总RNA,用RT-PCR测量了PCNA、p53、Bcl-2mRNA表达。(C;细胞用10%的DMSO/PBS处理。)
PCNA(增殖细胞核:proli-ferating cell nuclear)在细胞周期中经过G1前半期和S期的同时开始合成,在整个细胞周期的进行中作为良性调节因子执行重要的功能。尤其,PCNA蛋白质被认为是可以判断有无细胞增殖的重要的靶蛋白,如Sp-1蛋白质,在引起细胞周期干扰或凋亡的细胞中作为被激活的称为半胱天冬蛋白酶(caspase)的酶的基质利用。p53是抗癌基因中最有名的因子。据报告,位于正常细胞内的p53平时不具有活性,但是如果p53引起突变或消失,则正常细胞变异为癌细胞。即使在癌细胞中p53也以非活性状态存在。但是,据报告若癌细胞受到外部刺激或凋亡信号,则p53变为活性型,会主导细胞死灭。
Bax在Bcl-2家族群中属于促凋亡(pro-apoptosis)群。Bax平时在细胞内线粒体外膜以单位体存在,被Bcl-2阻碍作用。在受到凋亡信号时引起形态上的变化而形成二聚体(dimer)而作用于线粒体外膜。这种作用是在线粒体外膜造孔,使线粒体内部的物质流出到外面。这时出来的物质为细胞色素C(cytochrome C)、凋亡蛋白酶激活因子1(Apaf-1)等,均为诱发凋亡的物质。
对抗Bax的基因即Bcl-2基因在Bcl-2家族群中属于保生存(pro-survival)群,平时与Bid或Bim结合。当收到凋亡信时,Bik或Bad与Bcl-2结合,从Bcl-2分开的Bid或Bim与Bax结合引起形态上的变化来引起凋亡。即,Bcl-2具有捕捉Bid或Bim来防止凋亡机制的作用。其中Bid或Bim是为激活Bax而结合的。
观察图4,可确认乙醇分级提取物及乙醇乙酯分级提取物中的结果。在Molt-4细胞中处理试料24小时之后,提取mRNA并确认基因的结果,在PCNA、p53、Bcl-2、Bax基因中均未显示大影响。其结果与上述的实验结果MTT分析实验对比,判断羊肚菌EtOH、E.An层提取物对癌细胞的细胞毒性效果造成了影响,但是对细胞内抗癌相关基因DNA的表达没有显示大影响。
观察图5,可以确认三氯甲烷分级提取物及己烷分级提取物中的结果。Bcl-2基因显示随着浓度减少的趋势,在Bax基因中显示随着浓度增加的结果。该结果被判断为从基因表达方面支持在上述的实验结果——MTT分析实验中显示的羊肚菌三氯甲烷、己烷分级提取物的细胞毒性效果。
4.羊肚菌提取物对细胞凋亡相关蛋白质表达的影响
图6是实验羊肚菌提取物对细胞凋亡相关蛋白质的表达的影响的结果照片。在Molt-4细胞施加适当浓度的试料并培养24小时。在PCNA、p53、Bcl-2、Bax和β-actin的条件下通过使用特异抗体的蛋白质印迹法测量了蛋白质表达图案。(将C(+);细胞用10%的DMSO/PBS处理。)
在Molt-4细胞处理试料24小时之后,执行蛋白质印迹分析了凋亡相关蛋白质。PCNA在己烷层处理组显示出比对照组稍微减少的趋势,p53蛋白质在己烷层处理组比对照组增加很多。Bcl-2在己烷层处理组显示出浓度依赖性减少的趋势,Bax蛋白质在己烷层处理组显示出浓度依赖性增加的趋势。根据该结果可知在己烷层中抗癌相关物质的活性最大,可以解释为再次确认羊肚菌提取物的抗癌活性的结果。
Claims (5)
1.一种抗癌组成物,其特征在于,包含羊肚菌(Morchella esculenta)提取物作为有效成分。
2.如权利要求1所述的抗癌组成物,其特征在于,
上述羊肚菌提取物是将羊肚菌用乙醇、乙酸乙酯、三氯甲烷或己烷提取的提取物。
3.如权利要求1所述的抗癌组成物,其特征在于,
上述羊肚菌提取物的含量为按照癌细胞5×104~3×106细胞/孔基准观察时,包含1~10μg/mL。
4.如权利要求1所述抗癌组成物,其特征在于,
在上述组成物还包含羊肚菌发酵物。
5.如权利要求1所述抗癌组成物,其特征在于,
在上述组成物还包含发酵人参提取物等免疫增进物质。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR10-2017-0056838 | 2017-05-04 | ||
KR1020170056838A KR101969506B1 (ko) | 2017-05-04 | 2017-05-04 | 곰보버섯 추출물을 유효성분으로 포함하며 항암효능을 갖는 혼합 조성물 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN108785332A true CN108785332A (zh) | 2018-11-13 |
Family
ID=64093328
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810421421.XA Withdrawn CN108785332A (zh) | 2017-05-04 | 2018-05-04 | 包含羊肚菌提取物作为有效成份且具有抗癌功能的组成物 |
Country Status (2)
Country | Link |
---|---|
KR (1) | KR101969506B1 (zh) |
CN (1) | CN108785332A (zh) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1507772A (zh) * | 2002-12-16 | 2004-06-30 | 北京市食品研究所 | 液体深层发酵制备羊肚菌的方法及其产品 |
WO2017031399A1 (en) * | 2015-08-20 | 2017-02-23 | Genomatica, Inc. | Compositions and multiplexed systems for coupled cell-free transcription-translation and protein synthesis and methods for using them |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102067283B1 (ko) | 2012-12-21 | 2020-01-16 | 주식회사 코씨드바이오팜 | 곰보버섯 자실체 추출물 또는 곰보버섯 균사체 추출물을 함유하는 피부 외용제 또는 화장료 조성물 |
KR20150080206A (ko) * | 2013-12-31 | 2015-07-09 | 웅진식품주식회사 | 기능성 사포닌이 다량 함유된 발효 인삼 |
KR101629793B1 (ko) | 2014-03-17 | 2016-06-15 | 주식회사 앤코스메슈 | 모렐버섯의 생산을 위한 배지 조성물 |
-
2017
- 2017-05-04 KR KR1020170056838A patent/KR101969506B1/ko active IP Right Grant
-
2018
- 2018-05-04 CN CN201810421421.XA patent/CN108785332A/zh not_active Withdrawn
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1507772A (zh) * | 2002-12-16 | 2004-06-30 | 北京市食品研究所 | 液体深层发酵制备羊肚菌的方法及其产品 |
WO2017031399A1 (en) * | 2015-08-20 | 2017-02-23 | Genomatica, Inc. | Compositions and multiplexed systems for coupled cell-free transcription-translation and protein synthesis and methods for using them |
Non-Patent Citations (4)
Title |
---|
NITHA B ET AL.: "Anti-inflammatory and antitumour activities of cultured mycelium of morel mushroom, Morchella esculenta", 《CURRENT SCIENCE》 * |
张力田等: "《碳水化合物化学 第2版》", 31 July 2013, 中国轻工业出版社 * |
陈彦等: "羊肚菌胞外多糖抗肿瘤作用的研究", 《食品科学》 * |
陈长勋等: "《中药药理学 第2版》", 31 January 2015, 上海科学技术出版社 * |
Also Published As
Publication number | Publication date |
---|---|
KR20180122799A (ko) | 2018-11-14 |
KR101969506B1 (ko) | 2019-08-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Lezoul et al. | Extraction processes with several solvents on total bioactive compounds in different organs of three medicinal plants | |
Buenz et al. | The traditional Chinese medicine Cordyceps sinensis and its effects on apoptotic homeostasis | |
Bains et al. | Bioactives from mushroom: health attributes and food industry applications | |
Bhat et al. | Tongkat Ali (Eurycoma longifolia Jack): a review on its ethnobotany and pharmacological importance | |
CN102099046B (zh) | 含有来自紫杉形成层或原形成层的细胞系作为活性成分的抗氧化、抗炎或抗衰老组合物 | |
Adachi et al. | The “Prunus mume Sieb. et Zucc”(Ume) is a rich natural source of novel anti-cancer substance | |
CN101842112A (zh) | 含有来自紫杉形成层或原形成层的植物干细胞系的抗癌组合物 | |
Lai et al. | Effects of cyclic AMP on development and secondary metabolites of Monascus ruber M‐7 | |
US9084798B2 (en) | Use of lanostane and Poria extract in treating diabetes | |
Suzuki et al. | Discovery and genetic analysis of non-bitter Tartary buckwheat (Fagopyrum tataricum Gaertn.) with trace-rutinosidase activity | |
CN107001413A (zh) | 环肽衍生物及其制造方法和组合物 | |
Turrini et al. | Hemidesmus indicus induces immunogenic death in human colorectal cancer cells | |
Pintathong et al. | The feasibility of utilizing cultured Cordyceps militaris residues in cosmetics: Biological activity assessment of their crude extracts | |
Kimira et al. | Citrus limonoid nomilin inhibits osteoclastogenesis in vitro by suppression of NFATc1 and MAPK signaling pathways | |
Śliwiński et al. | Orchidaceae-derived anticancer agents: A review | |
Orlando et al. | Unravelling the phytochemical composition and the pharmacological properties of an optimized extract from the fruit from Prunus mahaleb L.: From traditional liqueur market to the pharmacy shelf | |
Kaewmalakul et al. | Melanization and morphological effects on antifungal susceptibility of Penicillium marneffei | |
Lee et al. | Inhibition of melanogenesis by Aster yomena callus pellet extract in melanoma cells and patients with skin pigmentation | |
Bautista-Pérez et al. | Spinach methanolic extract attenuates the retinal degeneration in diabetic rats | |
Péter et al. | Natural compounds as target biomolecules in cellular adhesion and migration: from biomolecular stimulation to label-free discovery and bioactivity-based isolation | |
Hazekawa et al. | Neuroprotective effect of repeated treatment with Hericium erinaceum in mice subjected to middle cerebral artery occlusion | |
CN108785332A (zh) | 包含羊肚菌提取物作为有效成份且具有抗癌功能的组成物 | |
Okafor et al. | Flavonoid and phenolic acid profiles of dehulled and whole Vigna subterranea (L.) Verdc seeds commonly consumed in South Africa | |
KR101095034B1 (ko) | 구멍쇠미역(Agarum clathratum) 추출물을 포함하는 치매 예방 또는 치료 및 인지능 개선용 약학적 조성물 또는 기능성 식품 조성물 | |
Fitmawati et al. | Traditional medicinal formulation: Obat pahit from lingga Malay ethnic in riau archipelago, Indonesia |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WW01 | Invention patent application withdrawn after publication | ||
WW01 | Invention patent application withdrawn after publication |
Application publication date: 20181113 |