CN113876818A - Compound ganoderma lucidum spore oil extract for preventing stroke and preparation method and application thereof - Google Patents
Compound ganoderma lucidum spore oil extract for preventing stroke and preparation method and application thereof Download PDFInfo
- Publication number
- CN113876818A CN113876818A CN202111204054.6A CN202111204054A CN113876818A CN 113876818 A CN113876818 A CN 113876818A CN 202111204054 A CN202111204054 A CN 202111204054A CN 113876818 A CN113876818 A CN 113876818A
- Authority
- CN
- China
- Prior art keywords
- ganoderma
- ganoderma lucidum
- extract
- lucidum spore
- extraction
- 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.)
- Pending
Links
- 239000000284 extract Substances 0.000 title claims abstract description 81
- 240000008397 Ganoderma lucidum Species 0.000 title claims abstract description 67
- 235000001637 Ganoderma lucidum Nutrition 0.000 title claims abstract description 67
- 150000001875 compounds Chemical class 0.000 title claims abstract description 13
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 50
- 238000000605 extraction Methods 0.000 claims abstract description 49
- 241000222336 Ganoderma Species 0.000 claims abstract description 39
- 239000002245 particle Substances 0.000 claims abstract description 32
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 26
- 208000006011 Stroke Diseases 0.000 claims abstract description 23
- 238000002156 mixing Methods 0.000 claims abstract description 18
- 239000008187 granular material Substances 0.000 claims abstract description 17
- 239000000203 mixture Substances 0.000 claims abstract description 17
- 238000002791 soaking Methods 0.000 claims abstract description 13
- 208000032382 Ischaemic stroke Diseases 0.000 claims abstract description 12
- 230000002490 cerebral effect Effects 0.000 claims abstract description 9
- 206010008190 Cerebrovascular accident Diseases 0.000 claims abstract description 7
- 241000700159 Rattus Species 0.000 claims description 40
- 238000000034 method Methods 0.000 claims description 23
- 239000003814 drug Substances 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 230000000740 bleeding effect Effects 0.000 claims description 13
- 230000023555 blood coagulation Effects 0.000 claims description 7
- 238000000643 oven drying Methods 0.000 claims description 6
- 238000010298 pulverizing process Methods 0.000 claims description 3
- 238000007873 sieving Methods 0.000 claims description 3
- 208000037906 ischaemic injury Diseases 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 19
- 206010008118 cerebral infarction Diseases 0.000 description 27
- 206010008089 Cerebral artery occlusion Diseases 0.000 description 26
- 201000007309 middle cerebral artery infarction Diseases 0.000 description 26
- 239000003921 oil Substances 0.000 description 20
- 210000004556 brain Anatomy 0.000 description 19
- 241000699670 Mus sp. Species 0.000 description 16
- 201000006474 Brain Ischemia Diseases 0.000 description 15
- 206010008120 Cerebral ischaemia Diseases 0.000 description 15
- 241001465754 Metazoa Species 0.000 description 14
- 210000000269 carotid artery external Anatomy 0.000 description 14
- 208000026106 cerebrovascular disease Diseases 0.000 description 14
- 239000008280 blood Substances 0.000 description 13
- 210000004369 blood Anatomy 0.000 description 13
- 210000004004 carotid artery internal Anatomy 0.000 description 12
- 235000019198 oils Nutrition 0.000 description 12
- 239000005552 B01AC04 - Clopidogrel Substances 0.000 description 10
- GKTWGGQPFAXNFI-HNNXBMFYSA-N clopidogrel Chemical compound C1([C@H](N2CC=3C=CSC=3CC2)C(=O)OC)=CC=CC=C1Cl GKTWGGQPFAXNFI-HNNXBMFYSA-N 0.000 description 10
- 229960003009 clopidogrel Drugs 0.000 description 10
- 229940079593 drug Drugs 0.000 description 10
- 230000010410 reperfusion Effects 0.000 description 9
- 210000001168 carotid artery common Anatomy 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 210000005013 brain tissue Anatomy 0.000 description 7
- 238000011835 investigation Methods 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- PKDBCJSWQUOKDO-UHFFFAOYSA-M 2,3,5-triphenyltetrazolium chloride Chemical compound [Cl-].C1=CC=CC=C1C(N=[N+]1C=2C=CC=CC=2)=NN1C1=CC=CC=C1 PKDBCJSWQUOKDO-UHFFFAOYSA-M 0.000 description 6
- 206010061216 Infarction Diseases 0.000 description 6
- 206010033546 Pallor Diseases 0.000 description 6
- 210000001367 artery Anatomy 0.000 description 6
- 210000001715 carotid artery Anatomy 0.000 description 6
- 238000005520 cutting process Methods 0.000 description 6
- 238000001647 drug administration Methods 0.000 description 6
- 230000007574 infarction Effects 0.000 description 6
- 206010053567 Coagulopathies Diseases 0.000 description 5
- 230000003542 behavioural effect Effects 0.000 description 5
- 230000035602 clotting Effects 0.000 description 5
- 230000007547 defect Effects 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 208000028867 ischemia Diseases 0.000 description 5
- 238000000465 moulding Methods 0.000 description 5
- 239000002504 physiological saline solution Substances 0.000 description 5
- 239000011734 sodium Substances 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- 238000000692 Student's t-test Methods 0.000 description 4
- 238000011278 co-treatment Methods 0.000 description 4
- 230000007812 deficiency Effects 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000003550 marker Substances 0.000 description 4
- 230000003188 neurobehavioral effect Effects 0.000 description 4
- 230000007658 neurological function Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000012353 t test Methods 0.000 description 4
- 206010008092 Cerebral artery thrombosis Diseases 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- 230000005856 abnormality Effects 0.000 description 3
- 230000036770 blood supply Effects 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 3
- 229960002327 chloral hydrate Drugs 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 238000003304 gavage Methods 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 208000014674 injury Diseases 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 210000005036 nerve Anatomy 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 206010002091 Anaesthesia Diseases 0.000 description 2
- 206010062717 Increased upper airway secretion Diseases 0.000 description 2
- 206010060860 Neurological symptom Diseases 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 208000033952 Paralysis flaccid Diseases 0.000 description 2
- 206010063837 Reperfusion injury Diseases 0.000 description 2
- 230000003187 abdominal effect Effects 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- 230000037005 anaesthesia Effects 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 210000002551 anterior cerebral artery Anatomy 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 230000010102 embolization Effects 0.000 description 2
- 210000003414 extremity Anatomy 0.000 description 2
- 208000028331 flaccid paralysis Diseases 0.000 description 2
- 210000003194 forelimb Anatomy 0.000 description 2
- 238000007917 intracranial administration Methods 0.000 description 2
- 239000007928 intraperitoneal injection Substances 0.000 description 2
- 230000000302 ischemic effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 230000007971 neurological deficit Effects 0.000 description 2
- 230000000926 neurological effect Effects 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 208000026435 phlegm Diseases 0.000 description 2
- 239000000902 placebo Substances 0.000 description 2
- 229940068196 placebo Drugs 0.000 description 2
- 239000008354 sodium chloride injection Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000002269 spontaneous effect Effects 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 210000000779 thoracic wall Anatomy 0.000 description 2
- 210000001685 thyroid gland Anatomy 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 229940126680 traditional chinese medicines Drugs 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 210000000707 wrist Anatomy 0.000 description 2
- 241001552669 Adonis annua Species 0.000 description 1
- 206010048962 Brain oedema Diseases 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 208000016988 Hemorrhagic Stroke Diseases 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 210000004958 brain cell Anatomy 0.000 description 1
- 230000005978 brain dysfunction Effects 0.000 description 1
- 208000006752 brain edema Diseases 0.000 description 1
- 230000003925 brain function Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000001914 calming effect Effects 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000000658 coextraction Methods 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 230000002008 hemorrhagic effect Effects 0.000 description 1
- 230000007954 hypoxia Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 208000020658 intracerebral hemorrhage Diseases 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 208000020470 nervous system symptom Diseases 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 235000011837 pasties Nutrition 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000008289 pathophysiological mechanism Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 208000019553 vascular disease Diseases 0.000 description 1
Images
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
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/074—Ganoderma
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- 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
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Heart & Thoracic Surgery (AREA)
- Alternative & Traditional Medicine (AREA)
- Vascular Medicine (AREA)
- Cardiology (AREA)
- Neurology (AREA)
- Biomedical Technology (AREA)
- Neurosurgery (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a compound ganoderma lucidum spore oil extract for preventing cerebral apoplexy and a preparation method and application thereof, wherein the extract is prepared by the following steps: mixing Ganoderma fruiting body granule and Ganoderma spore powder granule, soaking in 75% ethanol for 2-4 hr, and subjecting the soaked mixture to supercritical CO2Extraction, wherein the extraction conditions are as follows: extracting under 28-32MPa at 40-45 deg.C for 3-4 hr, collecting extractive solution, distilling under reduced pressure, and collecting extract; the mixing ratio range of the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles is 1-2: 2-1. The invention utilizes supercritical CO2The extraction technology mixes and extracts the ganoderma lucidum fruiting body and the ganoderma lucidum spore powder in proportion to obtain the super-oil extract which has the obvious effect of preventing ischemic stroke.
Description
Technical Field
The invention belongs to the technical field of pharmacy, and particularly relates to a compound ganoderma lucidum spore oil extract for preventing stroke and a preparation method and application thereof.
Background
Cerebrovascular diseases refer to brain dysfunction caused by various vascular diseases, are common diseases and frequently encountered diseases of the nervous system, and are divided into two categories, namely ischemic stroke and hemorrhagic stroke, wherein stroke is the main clinical type of cerebrovascular diseases and is characterized by sudden onset of diseases, rapid occurrence of localized or diffuse brain function deficiency symptoms. The ischemic stroke is a common stroke type in clinic and accounts for about 70-80% of the total stroke. Ischemic stroke refers to a disease in which the blood supply to the brain is insufficient due to various reasons, and local brain tissue is necrotic or softened due to ischemia and hypoxia, and corresponding nervous system symptoms appear.
Ischemic stroke belongs to the category of stroke in the traditional Chinese medicine, has pathogenesis of deficiency in origin and excess in superficiality, and is mainly caused by deficiency of qi and blood or deficiency of liver-yin and kidney-yin, phlegm fire, qi and blood retrograde movement and obstruction of collaterals and orifice, so the treatment principles of tonifying qi and blood, nourishing liver and kidney, suppressing yang and calming endogenous wind, eliminating phlegm and removing blood stasis can be adopted. With the continuous and deep research of related experiments, the research related to the treatment of ischemic stroke by traditional Chinese medicines is increasing.
At present, the mechanism research of the traditional Chinese medicine for preventing and treating the cerebral apoplexy mostly focuses on animal experiments, model selection also has a certain gap with clinical practical conditions, but the multi-target and multi-purpose action advantages are very obvious. In recent years, the prevention and treatment action mechanisms of various traditional Chinese medicines on ischemic stroke in related literatures comprise: inhibiting inflammatory reaction, and relieving nerve function defect after apoplexy; scavenging oxygen free radicals, resisting oxidative stress, and protecting brain cells of patients; reducing blood brain barrier permeability and alleviating cerebral edema; resisting apoptosis, and reducing cerebral infarction area; promoting the regeneration of blood vessels, improving the cerebral blood reperfusion of patients and the like.
The method has become one of important ways for preventing and treating ischemic stroke by taking the pathophysiological mechanism of the stroke as a starting point. The cerebral arterial thrombosis is a disease with extremely high disability rate and mortality rate, and the traditional Chinese medicine has the characteristic of wide curative effect, and has the incomparable advantages compared with the existing western medicine when being applied to treating cerebral arterial thrombosis by combining with complex factors of cerebral arterial thrombosis. The invention utilizes supercritical CO2The extraction technology is to mix and extract the ganoderma lucidum fruiting body and the ganoderma lucidum spore powder in proportion to obtain the super-oil extract, which has the effect of preventing ischemic stroke.
Disclosure of Invention
The invention aims to overcome the defects and provide the ganoderma lucidum spore oil-containing extract for effectively preventing stroke.
Another object of the present invention is to provide a method for preparing the above extract containing ganoderma lucidum spore oil.
The aim of the invention is realized by the following method:
an extract containing ganoderma lucidum spores for preventing stroke is prepared by the following method: mixing Ganoderma fruiting body granule and Ganoderma spore powder granule, soaking in 75% ethanol for 2-4 hr, and subjecting the soaked mixture to supercritical CO2Extraction, wherein the extraction conditions are as follows: extracting under 28-32MPa at 40-45 deg.C for 2-3 hr, collecting extractive solution, distilling under reduced pressure, and collecting extract; the mixing ratio range of the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles is 1-2: 2-1.
Preferably, the mixing ratio range of the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles is 1: 2.
the ganoderma lucidum fruit body particles are prepared by the following method: pulverizing Ganoderma, and sieving with 40-50 mesh sieve to obtain Ganoderma fine powder; adding water 20-30% of the Ganoderma fine powder into the Ganoderma fine powder, granulating, and oven drying at 55-60 deg.C.
The ganoderma lucidum spore powder particles are prepared by the following method: adding Ganoderma spore powder with wall breaking rate of above 80% into water 20-30% of Ganoderma spore powder, granulating, and oven drying at 55-60 deg.C to obtain Ganoderma spore powder granule.
A method for extracting compound ganoderma lucidum spore extract for preventing cerebral apoplexy comprises the following steps: mixing Ganoderma fruiting body granule and Ganoderma spore powder granule, soaking in 75% ethanol for 2-4 hr, and subjecting the soaked mixture to supercritical CO2Extraction, wherein the extraction conditions are as follows: extracting under 28-32MPa at 40-45 deg.C for 3-4 hr, collecting extractive solution, distilling under reduced pressure, and collecting extract; the mixing ratio range of the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles is 1-2: 2-1.
The ganoderma lucidum spore extract has obvious effect on the preparation of the medicine for preventing cerebral apoplexy.
The stroke can be ischemic stroke.
The ganoderma lucidum spore extract is applied to preparing the medicine for protecting the cerebral ischemia injury of rats.
The extract containing Ganoderma spore can be used for preparing medicine for prolonging bleeding time and blood coagulation time.
Supercritical CO used in the invention2The extraction conditions are based on a large number of preliminary experiments, and the supercritical CO is influenced by 4 factors of the extraction pressure, the extraction temperature, the extraction time and the mass concentration of the entrainer2And (3) carrying out single-factor investigation on the factor of the extraction effect, optimizing the extraction process conditions by taking the yield of the extract as an index, and collecting the extract after carrying out reduced pressure distillation on the obtained extract.
1. Investigation of extraction pressure
Mixing the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles according to the proportion of 1: 2, soaking the mixture for 3 hours by using 75% ethanol with mass concentration, and performing supercritical CO treatment on the soaked mixture2And (4) extracting. Under the condition of extraction time of 3h, 10 extraction pressures (20MPa, 22MPa, 24MPa, 26MPa, 28MPa, 30MPa, 32MPa, 34MPa, 36MPa and 38MPa) with different gradients are selected for investigation, and the optimal extraction pressure is 28-32MPa by taking the extraction rate as an index.
2. Investigation of extraction temperature
Mixing the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles according to the proportion of 1: 2, soaking the mixture for 3 hours by using 75% ethanol with mass concentration, and performing supercritical CO treatment on the soaked mixture2And (4) extracting. Under the conditions of extraction time of 3h and extraction pressure of 32MPa, 8 extraction temperatures (25 ℃, 30 ℃, 35 ℃, 40 ℃, 45 ℃, 50 ℃, 55 ℃ and 60 ℃) with different gradients are selected for examination, and the optimal extraction temperature is 40-45 ℃ by taking the extraction rate as an index.
3. Investigation of extraction time
Mixing the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles according to the proportion of 1: 2, soaking the mixture for 3 hours by using 75% ethanol with mass concentration, and performing supercritical CO treatment on the soaked mixture2And (4) extracting. Under the conditions of extraction pressure of 32MPa and extraction temperature of 45 ℃, 6 extraction temperatures (1h, 1.5h, 2h, 2.5h, 3h, 3.5h and 4h) with different gradients are selected for investigation, and the optimal extraction time is obtained by taking the extraction rate as an index for 3-4 h.
4. Inspection of entrainer mass concentration
Mixing the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles according to the proportion of 1: 2, selecting 3 ethanol (50%, 75% and 95%) with different mass concentrations, soaking for 3h for investigation, and performing supercritical CO extraction on the soaked mixture under the conditions of extraction pressure of 32MPa, extraction temperature of 45 ℃ and extraction time of 3h2And (4) extracting. The best entrainer mass concentration is obtained by taking the extraction rate as an index.
5. Test results
The test shows that the supercritical CO2The optimum process conditions for extracting the super-oil extract are that the pressure of an extraction kettle is 28-32MPa, the temperature of the extraction kettle is 40-45 ℃, the extraction time is 3-4h, the extract liquid is collected, and the extract is collected after reduced pressure distillation. Wherein the Ganoderma encarpium granule and Ganoderma spore powder granule are soakedThe mass concentration of the entrainer is optimally 75 percent.
The optimal material-liquid ratio of the 75% ethanol soaking is 1: 2.
compared with the prior art, the invention has the beneficial effects that:
the invention utilizes supercritical CO2The extraction technology is to mix and extract the ganoderma lucidum fruiting body and the ganoderma lucidum spore powder in proportion to obtain the super-oil extract, which has the effect of preventing ischemic stroke.
Drawings
FIG. 12 h representative TTC staining of the brain of mice dosed with focal cerebral ischemia (MCAO)/reperfusion (n ═ 8)
FIG. 23 h representative TTC staining of the brain of mice dosed with focal cerebral ischemia (MCAO)/reperfusion (n ═ 8)
Detailed Description
The invention is further illustrated by the following specific examples.
Example 1
And (3) ganoderma lucidum fruit body particles: pulverizing Ganoderma with pulverizer, and sieving with 40-50 mesh sieve to obtain Ganoderma fine powder; adding water 20-30% of the Ganoderma fine powder into the Ganoderma fine powder, granulating, and oven drying at 55-60 deg.C to obtain Ganoderma fruiting body granule.
And (3) ganoderma lucidum spore powder particles: adding Ganoderma spore powder with wall breaking rate of above 80% into water 20-30% of Ganoderma spore powder, granulating, and oven drying at 55-60 deg.C to obtain Ganoderma spore powder granule.
Mixing the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles according to the proportion of 1: 2, soaking the mixture in 75 percent ethanol for 3 hours, and performing supercritical CO treatment on the soaked mixture2Extraction, wherein the extraction conditions are as follows: extracting at the pressure of 28-32MPa and temperature of 40-45 deg.C for 3 hr, collecting extractive solution, distilling under reduced pressure, and collecting extract to obtain Ganoderma spore extract for preventing apoplexy.
Example 2
Mixing the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles according to the proportion of 1: 3, and the other conditions were the same as in example 1.
Test example 1
1. Experimental Material
1.1 test drugs
Example 1 super oil extract (prepared according to the method of example 1), ganoderma lucidum fruiting body extract group, ganoderma lucidum spore powder extract group, example 2 super oil extract (prepared according to the method of example 2).
Ganoderma lucidum fruiting body extract group: soaking Ganoderma fruiting body granule in 75% ethanol for 3 hr, and subjecting the soaked mixture to supercritical CO2Extraction, wherein the extraction conditions are as follows: extracting at the pressure of 28-32MPa and the temperature of 40-45 deg.C for 3h, collecting extractive solution, distilling under reduced pressure, and collecting extract.
Ganoderma lucidum spore powder extract group: soaking Ganoderma spore powder in 75% ethanol for 3 hr, and subjecting the soaked mixture to supercritical CO2Extraction, wherein the extraction conditions are as follows: extracting at the pressure of 28-32MPa and the temperature of 40-45 deg.C for 3h, collecting extractive solution, distilling under reduced pressure, and collecting extract.
Preparing the medicine: the superoil extract of example 1, the ganoderma lucidum spore powder extract and the superoil extract of example 2 are yellow oily liquids, and are prepared into uniform suspensions with required concentration by 0.5 percent of CMC-Na before use; ganoderma encarpium extract, brown pasty solid, and making into uniform suspension with desired concentration with oleum Maydis before use.
1.2 Positive drugs
Clopidogrel: the characteristics are as follows: a white solid; batch number: 20191203. the preparation of the medicine comprises the following steps: 0.5 percent of CMC-Na is prepared into uniform suspension with required concentration before use.
1.3 animals
SD rats, SPF grade, male, provided by Ministry of family planning science institute of Shanghai city, production license number SCXK (Zhe) 2018-: 20180006021818, respectively;
ICR mice, SPF grade, female and male half, provided by Hangzhou medical college in Zhejiang province, production license number SCXK (Zhe) 2019-: 20200922Abzz0100000176, 20200922Abzz 0100000485;
feed: granulated feed supplied from the animal house of Chinese pharmaceutical university; feeding conditions are as follows: air-conditioned room, temperature 18-24 deg.C, relative humidity 70%.
1.4 reagents
0.9% sodium chloride injection: anhui double crane pharmaceutical industry, product of Limited liability company, batch number: 1808061V.
Chloral hydrate: shanghai Linfeng Chemicals Co., Ltd product, lot number: 20180514, pharmaceutical preparation: the mixture is dissolved by 0.9 percent sodium chloride injection to prepare clear solution with the concentration of 3 percent before use.
2,3, 5-Triphenyltetrazolium chloride (TTC): BioLINK company, lot number: 1210Y 19. The preparation method comprises the following steps: immediately before use, a 1% solution was prepared in PBS buffer.
Sodium carboxymethylcellulose (CMC-Na): national pharmaceutical group chemical reagents limited, batch number: 20180412, preparation of 0.5% CMC-Na: 2.5g of CMC-Na was uniformly scattered on the surface of 500mL of ultrapure water, left overnight at room temperature, and shaken up for use.
Medicinal corn oil: aladdin, lot number: H1912097.
1.5 instruments
GZX-9140MBE digital display drying cabinet: shanghai Bocheng Industrial Co., Ltd.
BS224s model electronic balance: product of Beijing Sidolis Instrument systems, Inc.
HH-4 digital display constant temperature water bath: products of Guohua electric appliances Limited.
BCD-254CKK25V61TI-20 ℃ refrigerator: product of SIEMENS ag, germany.
2. Research methods and results
2.1. Protection effect of administration of super oil extract 2h after MCAO reperfusion on rat cerebral ischemia injury
2.1.1 establishment of cerebral ischemia-reperfusion model
42 male SD rats (84 rats are actually needed because the molding rate is about 50%), the weight is 250-300g, and the rats are randomly divided into 7 groups according to the weight, 6 rats in each group, four groups of the super oil extract (0.25ml/kg) in example 1, the ganoderma lucidum fruiting body extract (0.25ml/kg), the ganoderma lucidum spore powder extract (10mg/kg) and the super oil extract (0.25ml/kg) in example 2, 7.5mg/kg in the positive control clopidogrel group, and 7.5mg/kg in the sham operation group and the ischemia model control group (which are all given physiological saline with equal volume). The drug administration is started 2h after MCAO re-perfusion, and the drug administration is performed by intragastric administration once a day for 7 days continuously, and the drug administration volume is 0.2ml/100g body weight.
Rat focal cerebral ischemia (MCAO) reperfusion injury model: and fasting is not forbidden for 24 hours before the molding operation. Referring to Longa et al, a rat Middle Cerebral Artery Occlusion (MCAO) model was prepared using an internal carotid artery embolization method: rats were anesthetized with 3% chloral hydrate (300mg/kg) by intraperitoneal injection (i.p), placed on a supine operating table, with a median incision in the neck, exposing the right common carotid artery, pulling the muscles of the two abdominals and sternocleidomastoid outward, sequentially freeing from the bifurcation of the common carotid artery toward the head, ligating and cutting off the branches of the external carotid artery: the inferior occipital artery and the superior thyroid artery. Ligation is carried out at the far end of the external carotid artery, and the external carotid artery is cut off to free the main trunk for standby. Then the internal carotid artery is separated, and a loose button is formed at the root of the external carotid artery by silk threads to clamp the common carotid artery and the internal carotid artery. The fishing line (40 mm long and 0.26mm diameter) is slowly pushed towards the intracranial carotid artery by the incision of the external carotid artery main trunk, and the artery clamp on the internal carotid artery is loosened after the fishing line enters the internal carotid artery. The bifurcation of the common carotid artery is used as a mark, resistance is felt when the carotid artery is pushed for about 18mm, namely, the carotid artery reaches the inside of a thinner anterior cerebral artery, all blood supply sources of MCA are blocked, and the root of the external carotid artery is fastened tightly. After 2h of ischemia, the nylon wire was pulled out and the arterial stump was tied. The skin was sutured and the MCAO was completed resulting in a focal cerebral ischemia-reperfusion model. After anesthesia, rats in the sham operation group exposed only the bifurcation of the internal and external carotid arteries without occlusion of the MCA. Preparing a fishing line: one end of a fishing line 40mm long and 0.26mm in diameter was marked with a marker, and a further marker was marked 18mm from the marked end and placed in physiological saline for later use.
2.1.2, detection index
(1) And (3) nerve function scoring:
animals were graded for neurological deficits on the second, fourth and 0.5h after the last dose of MCAO/R according to the method of Bederson, as follows:
0 minute: no neurological symptoms were observed;
1 minute: when the tail is lifted and suspended, the operation of the animal shows that the contralateral forelimb shows that the wrist and elbow are bent, the shoulder is rotated inwards, the elbow is expanded outwards and is tightly attached to the chest wall;
and 2, dividing: the animal is placed on a smooth plane, and when the side shoulder of the operation is pushed to move towards the opposite side, the resistance is reduced;
and 3, dividing: when the animal walks freely, the animal rotates towards the opposite side of the operation or rotates around;
4, dividing; flaccid paralysis, no spontaneous movement of limbs;
(2) determination of cerebral infarction rate and water content:
after the above indexes are measured, the cervical vertebrae are removed to kill the rat, and the whole brain is taken out and weighed. Making four coronary cutting knives at the positions 2mm before and after the visual intersection, cutting into five slices, quickly placing the brain slices into 5ml of phosphoric acid buffer solution containing 1% TTC, incubating in a dark place for 30 minutes, turning over every 7-8 minutes in the incubating process, taking out the brain slices after incubating for 30 minutes, taking a picture by using a digital camera, separating a pale area (infarct area) and a non-pale area (normal area) by using ophthalmic forceps, and calculating the infarct percentage as follows:
percent infarct (%) < weight in pallor area/(weight in pallor area + weight in non-pallor area) × 100%
Placing the dyed brain tissue in a 110 ℃ oven for drying, and calculating the water content of the brain by contrasting the wet weight of the brain as follows:
brain tissue water content (%) - (1-brain tissue dry weight/brain tissue wet weight) × 100%
Software used for data statistics is SPSS 22.0, single-factor variance analysis is adopted for data statistics when the variances among multiple groups are compared to be uniform, pairwise comparison is carried out, and ANOVA test is adopted; the two groups were compared using the t-test. P <0.05 shows statistical significance.
The results show that the rats in the model group have higher neurobehavioral scores at days 2, 4 and 7 after MCAO/R, which indicates that the rats in the model group have serious neurological function defects. The superoil extract (0.25ml/kg) (P <0.01) of example 1 significantly reduced the neurological score in cerebral ischemic rats compared to the model group; compared with the blank group, the cerebral infarction area and the brain water content of the rats in the model group are both obviously increased (P <0.01), compared with the model group, the super-oil extract (0.25ml/kg) in the example 1 can obviously reduce the cerebral infarction area and the water content (P <0.01), and the overall effect is similar to that of the positive drug clopidogrel. The results are shown in FIG. 1, tables 1-2.
Table 1 effect of dosing 2h after MCAO refill on neuro-behavioral in rats (data expressed as Mean ± SD, n ═ 6)
P < 0.05; p <0.01, compared to model control group
Table 2 effect of administration 2h after MCAO reperfusion on rat cerebral infarction rate and brain water content (mean ± s.d., n ═ 6)
# P <0.01, compared to placebo; p <0.05, P <0.01, compared to model control group
2.2. Protection effect of administration of super oil extract 3h after MCAO reperfusion on rat cerebral ischemia injury
2.2.1 establishment of cerebral ischemia-reperfusion model
42 male SD rats (84 rats are actually needed because the molding rate is about 50%), the weight is 250-300g, and the rats are randomly divided into 7 groups according to the weight, wherein 6 rats in each group are respectively four groups, namely the super-oil extract (0.25ml/kg) in example 1, the ganoderma lucidum fruiting body extract (0.25ml/kg), the ganoderma lucidum spore powder extract (10mg/kg) and the super-oil extract (0.25ml/kg) in example 2, the positive control clopidogrel group is 7.5mg/kg, and the sham operation group and the ischemia model control group (are respectively given physiological saline with equal volume). The drug administration is started 2h after MCAO re-perfusion, and the drug administration is performed by intragastric administration once a day for 7 days continuously, and the drug administration volume is 0.2ml/100g body weight.
Rat focal cerebral ischemia (MCAO) reperfusion injury model: and fasting is not forbidden for 24 hours before the molding operation. Referring to Longa et al, a rat Middle Cerebral Artery Occlusion (MCAO) model was prepared using an internal carotid artery embolization method: rats were anesthetized with 3% chloral hydrate (300mg/kg) by intraperitoneal injection (i.p), placed on a supine operating table, with a median incision in the neck, exposing the right common carotid artery, pulling the muscles of the two abdominals and sternocleidomastoid outward, sequentially freeing from the bifurcation of the common carotid artery toward the head, ligating and cutting off the branches of the external carotid artery: the inferior occipital artery and the superior thyroid artery. Ligation is carried out at the far end of the external carotid artery, and the external carotid artery is cut off to free the main trunk for standby. Then the internal carotid artery is separated, and a loose button is formed at the root of the external carotid artery by silk threads to clamp the common carotid artery and the internal carotid artery. The fishing line (40 mm long and 0.26mm diameter) is slowly pushed towards the intracranial carotid artery by the incision of the external carotid artery main trunk, and the artery clamp on the internal carotid artery is loosened after the fishing line enters the internal carotid artery. The bifurcation of the common carotid artery is used as a mark, resistance is felt when the carotid artery is pushed for about 18mm, namely, the carotid artery reaches the inside of a thinner anterior cerebral artery, all blood supply sources of MCA are blocked, and the root of the external carotid artery is fastened tightly. After 2h of ischemia, the nylon wire was pulled out and the arterial stump was tied. The skin was sutured and the MCAO was completed resulting in a focal cerebral ischemia-reperfusion model. After anesthesia, rats in the sham operation group exposed only the bifurcation of the internal and external carotid arteries without occlusion of the MCA. Preparing a fishing line: one end of a fishing line 40mm long and 0.26mm in diameter was marked with a marker, and a further marker was marked 18mm from the marked end and placed in physiological saline for later use.
2.2.2, detection index
(1) And (3) nerve function scoring:
animals were graded for neurological deficits on the second, fourth and 0.5h after the last dose of MCAO/R according to the method of Bederson, as follows:
0 minute: no neurological symptoms were observed;
1 minute: when the tail is lifted and suspended, the operation of the animal shows that the contralateral forelimb shows that the wrist and elbow are bent, the shoulder is rotated inwards, the elbow is expanded outwards and is tightly attached to the chest wall;
and 2, dividing: the animal is placed on a smooth plane, and when the side shoulder of the operation is pushed to move towards the opposite side, the resistance is reduced;
and 3, dividing: when the animal walks freely, the animal rotates towards the opposite side of the operation or rotates around;
4, dividing; flaccid paralysis, no spontaneous movement of limbs;
(2) determination of cerebral infarction rate and water content:
after the above indexes are measured, the cervical vertebrae are removed to kill the rat, and the whole brain is taken out and weighed. Making four coronary cutting knives at the positions 2mm before and after the visual intersection, cutting into five slices, quickly placing the brain slices into 5ml of phosphoric acid buffer solution containing 1% TTC, incubating in a dark place for 30 minutes, turning over every 7-8 minutes in the incubating process, taking out the brain slices after incubating for 30 minutes, taking a picture by using a digital camera, separating a pale area (infarct area) and a non-pale area (normal area) by using ophthalmic forceps, and calculating the infarct percentage as follows:
percent infarct (%) < weight in pallor area/(weight in pallor area + weight in non-pallor area) × 100%
Placing the dyed brain tissue in a 110 ℃ oven for drying, and calculating the water content of the brain by contrasting the wet weight of the brain as follows:
brain tissue water content (%) - (1-brain tissue dry weight/brain tissue wet weight) × 100%
Software used for data statistics is SPSS 22.0, single-factor variance analysis is adopted for data statistics when the variances among multiple groups are compared to be uniform, pairwise comparison is carried out, and ANOVA test is adopted; the two groups were compared using the t-test. P <0.05 shows statistical significance.
The results show that the rats in the model group have higher neurobehavioral scores at days 2, 4 and 7 after MCAO/R, which indicates that the rats in the model group have serious neurological function defects. Compared with the model group, the superoil extract group (0.25ml/kg) (P <0.01) in example 1 can significantly reduce the neurological score of cerebral ischemic rats; compared with the blank group, the cerebral infarction area and the brain water content of the model group rat are both obviously increased (P is less than 0.01), compared with the model group, the super-oil extract group (0.25ml/kg) can obviously reduce the cerebral infarction area and the water content (P is less than 0.01), and the overall effect is similar to that of the positive drug clopidogrel. The results are shown in FIG. 2, tables 3-4.
Table 3 effect of administration 3h after MCAO recharge on neuro-behavioural behaviour in rats (data expressed as Mean ± SD, n ═ 6)
P < 0.05; p <0.01, compared to model control group
Table 4 effect of administration 3h after MCAO reperfusion on rat cerebral infarction rate and brain water content (mean ± s.d., n ═ 6)
# P <0.01, compared to placebo; p <0.05, P <0.01, compared to model control group
2.3 Effect of the superoil extract on bleeding time and clotting time in mice
The influence of the test drug on the bleeding time of the mice is detected by a tail breaking method: 50 healthy and clean ICR mice with the weight of 18-22 g are taken and randomly divided into 6 groups according to the weight, and each group comprises 10 mice, namely four groups of the super oil extract (0.25ml/kg), the ganoderma lucidum fruiting body extract (20mg/kg), the ganoderma lucidum spore powder extract (0.25ml/kg), the super oil extract (0.25ml/kg) in example 1, 15mg/kg and a blank group (given with physiological saline with equal volume) of the clopidogrel positive control. The administration is carried out once daily by gavage for 7 days, and the administration volume is 0.2ml/10g body weight. 30min after the last administration, the tail part of the patient is vertical, the measurement is carried out by a millimeter ruler, the position 5mm away from the tail tip is marked, then the patient is vertically cut off at the marked position of the tail of the patient by an operation blade, when the blood automatically overflows, the timing is started, blood drops are sucked by filter paper for 1 time every 30s until the blood naturally stops (when the filter paper sucks the blood), namely the bleeding time, and the bleeding time is recorded. The results were compared by t-test between groups.
The influence of the test drug on the blood coagulation time of the mouse is detected by adopting a glass slide method: animal grouping, administration dosage and administration mode and bleeding time determination experiment. 30min after the last administration, inserting a glass capillary tube with the inner diameter of 1mm into the orbit of a mouse, enabling blood to automatically flow out, discarding the first drop of blood, then respectively dropping blood at the two ends of a clean glass slide, enabling the diameter of the blood drop to be 5-10 mm, immediately starting timing, then using a dry needle to pick up the blood for 1 time every 10s until the needle can pick up fibrin threads, namely blood coagulation time, and recording the blood coagulation time by using a stopwatch. The results were compared by t-test between groups.
Note: mice were observed for the presence of abnormalities in the behavioural characteristics during the dosing period.
The results show that within 7 days of administration of the superoil extract of example 1, the mice did not show abnormal behavioral changes, compared to the blank group. Example 1 the group of superoleic extracts (0.25ml/kg) had a significant prolongation of the bleeding time (P <0.01) and clotting time (P < 0.05) in mice; the positive drug clopidogrel also has obvious prolongation effect on the bleeding time and the blood coagulation time of a mouse (P <0.01), and the results are shown in a table 5.
Table 5 effect on bleeding time and clotting time in mice (mean ± s.d., n ═ 10)
*P<0.05，**P<0.01, comparison with blank group
3 small knot
1) The rats in the model group have higher neurobehavioral scores at 2, 4 and 7 days after MCAO/R, which indicates that the rats in the model group have serious neurological function defects; compared with a model group, the superoil extract in example 1 is administrated by intragastric administration 2h or 3h after MCAO/R modeling, and the superoil extract (0.5ml/kg) (P <0.01) in example 1 can obviously reduce the neurological function score of a cerebral ischemia rat;
2) compared with the blank group, the cerebral infarction area and the brain water content of the rats in the model group are obviously increased (P is less than 0.01), which indicates that the cerebral ischemia-reperfusion model is successfully established; compared with a model group, the superoil extract (0.5ml/kg) in the example 1 is administrated by intragastric administration for 2h or 3h after MCAO/R molding, so that the cerebral infarction area and the water content (P is less than 0.01) can be obviously reduced, and the overall effect is similar to that of a positive drug clopidogrel;
3) example 1 administration of the superoil extract for 7 days, mice were free of hemorrhagic behavioral abnormalities; example 1 the superoleaginous extract (0.5ml/kg) had a significant prolongation of the bleeding time (P <0.01) and clotting time (P < 0.05) in mice compared to mice in the blank control group; the positive drug clopidogrel also has obvious prolonging effect on the bleeding time and the blood coagulation time of a mouse (P < 0.01);
the experimental result shows that the super-oil extract after MCAO/R is administered by gastric gavage for 7 days, so that the cerebral infarction area and the brain water content of a rat subjected to cerebral ischemia reperfusion are obviously reduced, the behavioral change is improved, and the therapeutic effect similar to that of the positive drug clopidogrel is shown after administration for 2h or 3h after cerebral ischemia reperfusion; example 1 the superoil extract was administered by gavage for 7 days, there was no abnormality in the behavioural characteristics of the mice, and the superoil extract of example 1 was able to prolong the bleeding time and clotting time of the mice.
Claims (9)
1. A compound ganoderma lucidum spore extract for preventing cerebral apoplexy is characterized in that the extract is prepared by the following method:
mixing Ganoderma fruiting body granule and Ganoderma spore powder granule, soaking in 75% ethanol for 2-4 hr, and subjecting the soaked mixture to supercritical CO2Extraction, wherein the extraction conditions are as follows: extracting under 28-32MPa at 40-45 deg.C for 3-4 hr, collecting extractive solution, distilling under reduced pressure, and collecting extract; the mixing ratio range of the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles is 1-2: 2-1.
2. The compound ganoderma lucidum spore extract as claimed in claim 1, wherein the mixing ratio of the ganoderma lucidum fruiting body particles to the ganoderma lucidum spore powder particles is in the range of 1: 2.
3. the compound ganoderma lucidum spore extract according to claim 1, wherein the ganoderma lucidum spore particles are prepared by the following method: pulverizing Ganoderma, and sieving with 40-50 mesh sieve to obtain Ganoderma fine powder; adding water 20-30% of the Ganoderma fine powder into the Ganoderma fine powder, granulating, and oven drying at 55-60 deg.C.
4. The compound ganoderma lucidum spore extract as claimed in claim 1, wherein the ganoderma lucidum spore powder particles are prepared by the following method: adding Ganoderma spore powder with wall breaking rate of above 80% into water 20-30% of Ganoderma spore powder, granulating, and oven drying at 55-60 deg.C to obtain Ganoderma spore powder granule.
5. The method for extracting the compound ganoderma lucidum spore extract for preventing the stroke as claimed in claim 1, which is characterized by comprising the following steps: mixing Ganoderma fruiting body granule and Ganoderma spore powder granule, soaking in 75% ethanol for 2-4 hr, and subjecting the soaked mixture to supercritical CO2Extraction, wherein the extraction conditions are as follows: extracting under 28-32MPa at 40-45 deg.C for 3-4 hr, collecting extractive solution, distilling under reduced pressure, and collecting extract; the mixing ratio range of the ganoderma lucidum fruiting body particles and the ganoderma lucidum spore powder particles is 1-2: 2-1.
6. The use of the compound ganoderma lucidum spore extract for preventing stroke as claimed in claim 1 in the preparation of a medicament for preventing stroke.
7. The use according to claim 6, wherein the stroke is ischemic stroke.
8. The use of the compound ganoderma lucidum spore extract for preventing stroke as claimed in claim 1 in the preparation of a medicament for protecting rats from cerebral ischemic injury.
9. The use of the compound ganoderma lucidum spore extract for preventing stroke as claimed in claim 1 in the preparation of a medicament for prolonging bleeding time and blood coagulation time.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111204054.6A CN113876818A (en) | 2021-10-15 | 2021-10-15 | Compound ganoderma lucidum spore oil extract for preventing stroke and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111204054.6A CN113876818A (en) | 2021-10-15 | 2021-10-15 | Compound ganoderma lucidum spore oil extract for preventing stroke and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113876818A true CN113876818A (en) | 2022-01-04 |
Family
ID=79003012
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111204054.6A Pending CN113876818A (en) | 2021-10-15 | 2021-10-15 | Compound ganoderma lucidum spore oil extract for preventing stroke and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113876818A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115006874A (en) * | 2022-06-14 | 2022-09-06 | 中科健康产业集团股份有限公司 | Extraction process of ganoderma lucidum and ganoderma lucidum spores rich in ganoderic acid components |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104073344A (en) * | 2014-07-16 | 2014-10-01 | 江苏斯威森生物医药工程研究中心有限公司 | Extracting method for ganoderma lucidum spore oil |
| CN106138117A (en) * | 2016-08-03 | 2016-11-23 | 广东粤微食用菌技术有限公司 | Ganoderma spore oil application in preparing prevention and cure of cardiovascular disease medicine |
| CN110840921A (en) * | 2019-12-13 | 2020-02-28 | 福建省幸福生物科技有限公司 | High-pressure supercritical CO2Ganoderma extract and its preparation method |
| CN113150867A (en) * | 2020-12-01 | 2021-07-23 | 中科健康产业集团股份有限公司 | Preparation method of ganoderma lucidum extract oil rich in ganoderma lucidum triterpenes |
-
2021
- 2021-10-15 CN CN202111204054.6A patent/CN113876818A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104073344A (en) * | 2014-07-16 | 2014-10-01 | 江苏斯威森生物医药工程研究中心有限公司 | Extracting method for ganoderma lucidum spore oil |
| CN106138117A (en) * | 2016-08-03 | 2016-11-23 | 广东粤微食用菌技术有限公司 | Ganoderma spore oil application in preparing prevention and cure of cardiovascular disease medicine |
| CN110840921A (en) * | 2019-12-13 | 2020-02-28 | 福建省幸福生物科技有限公司 | High-pressure supercritical CO2Ganoderma extract and its preparation method |
| CN113150867A (en) * | 2020-12-01 | 2021-07-23 | 中科健康产业集团股份有限公司 | Preparation method of ganoderma lucidum extract oil rich in ganoderma lucidum triterpenes |
Non-Patent Citations (1)
| Title |
|---|
| 陈静等: "灵芝有效成分生物活性作用的研究进展", 《云南中医中药杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115006874A (en) * | 2022-06-14 | 2022-09-06 | 中科健康产业集团股份有限公司 | Extraction process of ganoderma lucidum and ganoderma lucidum spores rich in ganoderic acid components |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111040006B (en) | Extraction method of bilberry glycoside and application of bilberry glycoside | |
| CN107898782B (en) | A kind of ginkgo diterpenoid-lactone composition | |
| CN103355655A (en) | Composition with alimentary anemia improving function and preparation method of composition | |
| CN113876818A (en) | Compound ganoderma lucidum spore oil extract for preventing stroke and preparation method and application thereof | |
| CN102631525B (en) | Health care product with function of blood pressure reduction and preparation method thereof | |
| KR100949482B1 (en) | Phamaceutical composition of diabetes mellitus or hypertension using extract of taxus cuspidata and mulberry tree, and mamufacturing method thereof | |
| CN104055909A (en) | Application of compound donkey-hide gelatin pulp in preparation of medicament for menstrual period health care | |
| CN106389535A (en) | Application of industrial hemp cannaboid in preparing gout treating medicines | |
| KR100740326B1 (en) | A pharmaceutical composition for treating headache disease and process and uses thereof | |
| CN102138967A (en) | Application of dracocephalum moldavica L. general flavone in preparing medicament for preventing and treating cerebrovascular disease | |
| CN102068520B (en) | Chinese medicinal composition for treating cardiovascular and cerebrovascular diseases and preparation method thereof | |
| KR100964603B1 (en) | Phamaceutical composition of diabetes mellitus using extract of taxus cuspidata and mulberry tree, and mamufacturing method thereof | |
| CN102266386B (en) | Scutellaria baicalensis extract and preparation method and uses thereof | |
| CN113855714A (en) | Ganoderma lucidum fruiting body extract for preventing cerebral apoplexy and preparation method and application thereof | |
| CN1199979C (en) | Water-soluble safflower extract and its oral preparation for treating and preventing angiocardiopathy andcerebrova scular disease | |
| CN105287921A (en) | Medicinal composition for treating gastric cancer and preparation method thereof | |
| CN101744955A (en) | Drug for treatment of ischemic cerebrovascular disease | |
| CN111000886B (en) | Pharmaceutical composition for treating cerebral arterial thrombosis and application thereof | |
| CN108938718A (en) | A kind of preparation method of Chinese medicine composition and its preparation for treating headstroke | |
| CN105477191A (en) | Medicine composition for treating aged coronary heart disease | |
| CN109248201B (en) | Traditional Chinese medicine composition and application thereof in preparation of medicine for treating arrhythmia | |
| CN106540003A (en) | A kind of pharmaceutical composition for treating hyperosteogeny | |
| CN105832803A (en) | Medicine with function of treating cerebrovascular diseases, method for preparing medicine and application thereof | |
| CN104383247A (en) | Traditional Chinese medicine for treating phthisis | |
| CN111494457A (en) | Pharmaceutical composition containing artemisinin and herba Polygoni orientalis, preparation and preparation method thereof |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220104 |