CN102091102A - Glossy ganoderma extract and pharmaceutical application thereof - Google Patents
Glossy ganoderma extract and pharmaceutical application thereof Download PDFInfo
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Abstract
The invention relates to new preparation technology of glossy ganoderma extract and pharmaceutical application thereof, in particular to fat soluble extract of glossy ganoderma spore powder, a preparation method thereof, a preparation method of various formulations of the extract and application of the extract in the preparation of health-care products and medicaments which are used for senile dementia, Parkinson, Huntington chorea, cerebral palsy, stroke hemiplegia sequela or cerebral trauma sequela.
Description
Technical field
The present invention relates to Ganoderma extract and product-use thereof.Be specifically related to a kind of liposoluble extract of Ganoderma spore powder, also relate to described preparation method of extract, the preparation method of the various dosage forms of extract and in preparation senile dementia, parkinson, Huntington Chorea, cerebral palsy, the apoplectic hemiplegia sequela or, the health product of cerebral trauma sequela, the application on the medicine.
Background technology
Arrival along with the senescence epoch, the sickness rate of various nervus centralis degenerative disease increases gradually, doctor trained in Western medicine thinks that these diseases comprise: Alzheimer's disease and Parkinson's disease, learning and memory decline and brain aging cause serious diseases such as senile insomnia disease, and clinical manifestation is dementia, movement disorder, hypomnesis, muscular tremor.Though doctor trained in Western medicine adopts his Kelin, galantamine, eptastigmine to treat Alzheimer's disease etc., adopt trihexyphenidyl, levodopa to wait and treat Parkinson's disease; Promote proliferation of neural stem cells with nerve growth factor, still,
Nervus centralis becomes row property disease course and is carrying out property, is generally 2-8, and long-term Western medicine has certain toxic and side effects.Therefore, become for the treatment nervus centralis row property disease Chinese medicine be developed to focus in order to study, people wish to develop the Chinese medicine that can reduce the Western medicine toxic and side effects and can take for a long time.
At present, bibliographical information has been arranged, Ganoderma extract is used to develop the medicine of treatment nervus centralis degenerative disease.But the preparation method of existing Ganoderma spore oil can cause the Ganoderma spore oil oxidation deterioration that is heated, and step is tediously long, be easy to generate dissolvent residual, the cost height is difficult to industrialization promotion and uses, therefore, these preparation methoies have limited the further development and use of Ganoderma extract.
Ganoderma is the treasure in the Chinese medicine and pharmacy treasure-house, and Ganoderma sporophore has long history in China as medicinal application.Ganoderma spore (Spores of Ganoderma lucidum) is that the Ganoderma development later stage launches the seed that discharges, and biologically is called basidiospore, puts together to be Powdered the common name Ganoderma spore powder.Ganoderma spore powder is collected very difficulty under natural environment, about 1000 kilograms of Ganodermas just can be collected 1 kilogram of spore powder, and is therefore very precious.In recent years, along with the development of Ganoderma artificial cultivation technique, the difficult problem of scale collection Ganoderma spore is solved substantially.
Ganoderma spore powder contains compositions such as abundant polysaccharide, polypeptide, triterpenes, organic germanium.Modern pharmacology and clinical proof Ganoderma spore powder have many-sided effect such as blood sugar lowering, antitumor, defying age, adjusting body's immunity; For the treatment chronic hepatitis, eliminate chemicals to the infringement of liver, prevent tissue fibering and in treatment of diabetes, all show good result.
It is filbert to yellowish-brown, avette that Ganoderma spore generally is, and (8. 5 ~ 11. 2) μ m * (5.2 ~ 6.9) μ m includes oil droplet.Ganoderma spore oil is the lipide actives that extracts from Ganoderma spore, and per 20 g Ganoderma spore powders just can extract 1 g Ganoderma spore oil usually.Ganoderma spore oil has been concentrated the triterpenes in the Ganoderma spore, ucleosides and part polysaccharide compound, is the aggregation of Ganoderma spore effective ingredient.
In recent years, the Ganoderma spore oil product has become the treasure in the high-grade health product, has crucial value and economic worth.Ganoderma spore oil has obvious effect for aspects such as human body immunity improving power, hypoxia-bearing capability, cholesterol reducing.Current research shows that the lipide actives matter in the Ganoderma spore oil also has the activity of significant inhibition tumor and anti AIDS virus.
The quantitative analysis results of Ganoderma spore powder liposoluble constituent shows, the unsaturated fatty acid total amount is 73. 6%, wherein main body composition oleic acid (C18: 1), linoleic acid (C18: 2) and Palmic acid (C16: equal size 0) is respectively 57. 5%, 13. 4% and 19. 6%; The unsaturated fatty acid hexadecenoic acid (C16: 1) and linolenic acid (C18: content 3) is respectively 2. 2% and 0.5%.Therefore, the easy oxidative deformation of Ganoderma spore powder liposoluble constituent, also destroyed in leaching process easily.
The shortcoming of the preparation method of existing Ganoderma spore oil
The Ganoderma spore that Ganoderma sporophore discharges when ripe, its content of effective exceeds 75 times of Ganodermas, and the quantity that generally can collect only is 1% of Ganoderma entity.Because it has the hard chitin shell of one deck, the utilization that is difficult to be absorbed by the body of effective ingredient wherein.The not spore of breaking cellular wall is taken in evidence, have only the effective ingredient about 12% to be absorbed by the body, and the effective ingredient absorbance of sporoderm-broken Ganoderma spore is more than 95%.
The technology of breaking trachytectum of glossy ganoderma has squeezing and pressing method, high pressure homogenization method, ultrasound wave at present, microwave heating method etc., these wall-breaking methods can cause the heat radiation difficulty, cause the Ganoderma spore oil oxidation deterioration that is heated, thereby reduced the physiologically active of Ganoderma spore oil and preparation thereof, be unfavorable for the development and use of Ganoderma spore oil.
Summary of the invention
The invention provides the preparation method of various dosage forms of liposoluble extract, preparation method, the extract of Ganoderma spore powder, the liposoluble extract of Ganoderma spore powder is in preparation senile dementia, parkinson, Huntington Chorea, cerebral palsy, apoplectic hemiplegia sequela, or the health product of cerebral trauma sequela, the application on the medicine.
Creationary characteristics of the present invention are:
The present invention is not a raw material with the breaking trachytectum of glossy ganoderma powder of market sale, but is raw material with the Ganoderma spore powder, in the process of breaking cellular wall, adds surfactant in enzymatic solution, has improved breaking cellular wall efficient.
The present invention adds surfactant in the mixed solution of alcohol-water in leaching process, improved extraction efficiency, and has avoided the oxidation destruction of heating to the Ganoderma spore powder liposoluble constituent.
The present invention adopts as above step, is in higher level with the content that guarantees liposoluble constituent in the raw material.
Surfactant toxicity of the present invention is low, is widely used in food, pharmaceuticals industry.HLB VALUE OF SURFACTANTS is at 11-22, be dissolved in the water easily, be not easy to be dissolved in the oil, therefore, the present invention reclaims organic solvents such as ethanol earlier, again by the mode of oil-water separation, makes the surfactant enrichment be dissolved in the water, surfactant can be not residual in liposoluble constituent, can not have influence on the safety of Ganoderma spore liposoluble constituent of the present invention.
The present invention adopts modes such as stirring, percolation, makes the breaking trachytectum of glossy ganoderma powder fully contact mixed solvent, and the lower temperature of control, can not make that the Ganoderma spore liposoluble constituent is subjected to thermooxidizing destruction.
The present invention adopts modes such as stirring, percolation to extract the Ganoderma spore liposoluble constituent, stirred tank or, carry out in the percolation bucket, can be used for suitability for industrialized production, and organic solvent reclaims, can not cause environmental pollution and waste.
The present invention makes soft capsule with the Ganoderma spore liposoluble constituent, and taking convenience is easy to preservation, and is not perishable.
Technical scheme of the present invention is: Ganoderma extract is prepared by following method:
With the Ganoderma spore powder is raw material, adopt the method for nonionic surfactant, organized enzyme to carry out breaking cellular wall, lyophilizing, get the breaking trachytectum of glossy ganoderma powder, in the mixed solvent of alcohol-water, add the surfactant of hydrophile-lipophile balance value at 11-22, fully contact, centrifugal, filter, obtain the Ganoderma spore liposoluble constituent.
The preparation method of the breaking cellular wall of Ganoderma spore powder
When breaking cellular wall, nonionic surfactant, organized enzyme all are dissolved in the mixed solvent of alcohol-water.
When breaking cellular wall, organized enzyme is pectase and cellulase
When breaking cellular wall, the nonionic surfactant of employing is a tween 80, and concentration is 0.1-10%, preferably 1%.Tween 80: have the characteristics of low toxicity, be widely used in medical industry, food industry and biochemical test, can not cause the toxicity of residue to human body.Tween 80 is a non-ionic surface active agent, and neutral own does not combine with pheron, can not make enzyme deactivation yet.
When breaking cellular wall, the alcohol in the mixed solvent of alcohol-water is: ethanol, propylene glycol.The concentration of alcohol is 1-20%, preferably 10%.
When breaking cellular wall, organized enzyme is pectase, cellulase,
The pectase consumption is the 1000-20000IU/g Ganoderma spore, preferably 8000 IU/g Ganoderma spores
The cellulase consumption is the 1000-20000IU/g Ganoderma spore, preferably 5000 IU/g Ganoderma spores
When breaking cellular wall, hydrolysis temperature 25-50 ℃, preferably 35 ℃,
When breaking cellular wall, enzymolysis pH value 6.0-8.0, preferably 7.5,
When breaking cellular wall, enzymolysis time 1.0-5 hour, preferably 3 hours.
After the breaking cellular wall of Ganoderma spore powder, the method for extracting liposoluble constituent is
After breaking cellular wall, extract the Ganoderma spore liposoluble constituent, after breaking cellular wall, during extraction, in the mixed solvent of alcohol-water, alcohol is ethanol,
The volume ratio of ethanol--water is: (1-10): (1-10):
The volume ratio of ethanol--water is preferably: 4:6
In the mixed solvent of alcohol-water, preferably can add petroleum ether-ethyl acetate, form the mixed solvent of petroleum ether-ethyl acetate-alcohol-water,
The volume ratio of petroleum ether-ethyl acetate-alcohol-water is:
（1-10）：（1-10）：（7-70）：（1-90）
The preferred volume ratio of petroleum ether-ethyl acetate-alcohol-water is: 5:15:25:55
During extraction,
Hydrophile-lipophile balance value is 11-22'sSurfactant is, dodecyl sodium sulfate, tween 20, Tween-40, polysorbate60 or, tween 80, Myrij 35, Myrij 49, Myrj 52, preferably tween 80
Hydrophile-lipophile balance value is 11-22'sThe consumption of surfactant is the 0.1-5% of mixed solvent, preferably 1%.
Fully be 1-6 hour time of contact, preferably 2 hours
The mode that the breaking trachytectum of glossy ganoderma powder fully contacts mixed solvent is: stirring, backflow, percolation, ultrasonic, microwave or, column chromatography.Preferably stir, percolation.Temperature is that 15-40 ℃, preferably 30 ℃, time are 6-24 hour, preferably 12 hours.
Filtrate is separated out the Ganoderma spore liposoluble constituent after reclaiming organic solvent from water, surfactant dissolves washes with water to remove and removes surfactant remaining in the Ganoderma spore liposoluble constituent in water, obtains the Ganoderma spore liposoluble constituent.
Therefore, the method for the method of the breaking cellular wall of comprehensive above Ganoderma spore powder and extraction liposoluble constituent, the extracting method of Ganoderma spore powder liposoluble constituent of the present invention specifically is following two kinds,
In the mixed solution of the alcohol-water of pectase and cellulase, the dissolving tween 80, handle Ganoderma spore powder with this mixed solution, make ganoderma lucidium spore powder wall breaking, lyophilizing, get the breaking trachytectum of glossy ganoderma powder, in the mixed solvent of alcohol-water, add tween 80, fully contact, centrifugal, filter, behind the filtrate recycling ethanol, from water, separate out the Ganoderma spore liposoluble constituent, tween 80 is dissolved in the water, washes with water except that tween 80 remaining in the Ganoderma spore liposoluble constituent, obtains the Ganoderma spore liposoluble constituent.
Tween-80 is dissolved in the mixed solution of pectase and cellulase, handle Ganoderma spore powder with this mixed solution, make ganoderma lucidium spore powder wall breaking, lyophilizing, get the breaking trachytectum of glossy ganoderma powder, in the mixed solvent of petroleum ether-ethyl acetate-alcohol-water, add tween 80, fully contact is centrifugal, filter, filtrate is separated out the Ganoderma spore liposoluble constituent after reclaiming petroleum ether-ethyl acetate-ethanol from water, tween 80 is dissolved in the water, wash with water and remove remaining tween 80 in the Ganoderma spore liposoluble constituent, obtain the Ganoderma spore liposoluble constituent.
The present invention adopts two kinds of dosage forms such as soft capsule, lyophilized powder to the Ganoderma spore powder liposoluble constituent, and their formulation method is respectively:
The preparation of soft capsule method of Ganoderma extract is
In high pressure homogenize equipment, 30～35 ℃, 5 weight portion Ganoderma spore liposoluble constituents are carried out homogenizing, get core liquid;
Get gelatin 100 weight portions and water 100 weight portions are put into container, be heated to 80～90 ℃, constantly stir and make it dissolving, add glycerol 40 weight portions, Polyethylene Glycol 10 weight portions continuation stirring then, dissolving fully stops heating, naturally cool to room temperature, obtain the soft capsule material;
Core liquid and soft capsule material are cleaned in compacting on the soft capsule press, and drying gets soft capsule, and the Co60 irradiation sterilization is handled, and becomes product.
The preparation method of the lyophilized powder of Ganoderma extract is: the Ganoderma spore liposoluble constituent is earlier at-45 ℃ of pre-freeze 2-4 hours, and dry 24-28 hour of reduced vacuum under-45 ℃～10 ℃ conditions then is at last at 30 ℃ of dry 6-8 hours.
The Ganoderma spore liposoluble constituent can be prepared into various forms of compositionss, concrete dosage form can be selected from injection, oral liquid, tablet, capsule, granule, aerosol, powder spray, spray, patch, it is long-acting or quick-acting that dosage form such as slow releasing preparation, solid dispersion reaches, and improve the effect that absorbs.
Ganoderma spore powder liposoluble constituent of the present invention is used for preparation treatment senile dementia, parkinson, Huntington Chorea, cerebral palsy, the apoplectic hemiplegia sequela or, the application on the medicine of cerebral trauma sequela.
But technical scheme of the present invention comprises the above technology that is not restricted to, and has comprised extracting method, the formulation method of this area routine, the selection of adjuvant certainly.
Specific embodiment
Embodiment 1
The preparation of breaking trachytectum of glossy ganoderma powder:
Get the 100g Ganoderma spore powder, in the mixed solvent 1000ml of 10% alcohol-water, fully stir, soaked 3 hours, and made the spore imbibition, outside organization is softening, add 8000 IU/g pectases, 5000 IU/g cellulase, 1% tween 80,35 ℃ of hydrolysis temperatures, pH7.5, enzymolysis time 3 hours, centrifugal, filter, lyophilizing obtains the breaking trachytectum of glossy ganoderma powder.
The spore powder that lyophilizing is good is agglomerating granular, brown, standby.
Embodiment 2
The detection of breaking trachytectum of glossy ganoderma powder:
At area is lmm
2, the degree of depth is the counting plate of 1.10mm.1 big grid of central authorities is divided into 25 medium squares with two-wire, and each medium square is divided into 16 lattices again, for the usefulness of counting.
Method of counting
Take by weighing the 1mg Sporoderm-broken Ganoderma Lucidum Spore powder with photoelectric analytical balance (precision 0.0lmg) and put into the 5mL test tube, in test tube, add the distilled water of 1mL, sway about 10min, Sporoderm-broken Ganoderma Lucidum Spore powder is dissolved in water evenly, obtain the suspension of Ganoderma spore.
Draw suspension 2 μ L with microsyringe (5 μ L), suspension splashed in the counting plate, cover slide, leave standstill 5min, treat that suspension is evenly distributed in counting plate after, counting.
Under optical microscope, whether the breaking cellular wall of discriminating spore, and the conidial cell wall of breaking cellular wall is smooth not complete, plentiful, and double-walled construction is clear, the individual morphology unanimity; Behind the breaking cellular wall, conidial cell wall is imperfect, content is irregular bulk.
The calculating of wall breaking rate of ganoderma lucidum spores
The computing formula of wall breaking rate of ganoderma lucidum spores: sporoderm-broken rate=[(X-Y)/X] * 100%
X is a complete spore count before the breaking cellular wall, and Y is a complete spore count behind the breaking cellular wall.
In specific embodiment 1, utilize tween 80, the compound enzyme that associating pectase and cellulase are formed is handled Ganoderma spore, through microscopy and calculate its sporoderm-broken rate.
The result shows, adopt pectase, cellulase that spore powder is carried out broken wall treatment separately, sporoderm-broken rate is respectively 82% and 87%, after the The combined, and increase the tween 80 processing, spore powder outer wall deliquescing depression, germinal furrow breaks, content is bulk and discharges, and the shell-broken effect of pollen is obvious, and sporoderm-broken rate reaches 94%.
Embodiment 3
The extracting method of the liposoluble constituent of Ganoderma spore, adopt alcohol-water for extracting solvent:
Get the 100g Sporoderm-broken Ganoderma Lucidum Spore powder, in the mixed solvent 1000ml of the alcohol-water of volume ratio 4:6, add 0.5% tween 80, fully stirred 2 hours, centrifugal, filter, behind the filtrate recycling ethanol, from water, separate out the Ganoderma spore liposoluble constituent, the tween 80 major part is dissolved in the water, have only in the considerably less tween 80 dissolving Ganoderma spore liposoluble constituent, remaining tween 80 in washing with water except that the Ganoderma spore liposoluble constituent obtains the Ganoderma spore liposoluble constituent.
Embodiment 4
The extracting method of the liposoluble constituent of Ganoderma spore, adopt petroleum ether-ethyl acetate-alcohol-water for extracting solvent:
Get the 100g Sporoderm-broken Ganoderma Lucidum Spore powder, in the mixed solvent 1000ml of petroleum ether-ethyl acetate-alcohol-water of volume ratio 5:15:25:55, add 0.5% tween 80, fully stirred 2 hours, centrifugal, filter, after filtrate is reclaimed petroleum ether-ethyl acetate-ethanol, from water, separate out the Ganoderma spore liposoluble constituent, the tween 80 major part is dissolved in the water, have only in the considerably less tween 80 dissolving Ganoderma spore liposoluble constituent, remaining tween 80 in washing with water except that the Ganoderma spore liposoluble constituent obtains the Ganoderma spore liposoluble constituent.
Explanation about the advantage of the extracting method of the liposoluble constituent of Ganoderma spore
For the Ganoderma spore powder liposoluble constituent, can be according to the extracting method of general volatile oil, adopt steam distillation, but length consuming time, extraction efficiency is low, easily cause unsaturated fatty acid in the Ganoderma spore powder generation oxidation deterioration that is heated, reduced the pharmacologically active of Ganoderma spore powder liposoluble substance.
For the Ganoderma spore powder liposoluble constituent, can adopt supercritical fluid extraction, though can save time, extract fully.But the cost height is difficult to suitability for industrialized production.
For the Ganoderma spore powder liposoluble constituent, also available ether, ethyl acetate, organic solvent extraction such as chloroform, but cost height, complex technical process causes dissolvent residual easily, the toxicity height.
Parents' structure of surfactant can reduce surface tension, strengthens wettability and the permeability of solvent to material, and the effective ingredient of natural product is had solubilization, leaches usefulness and extraction yield thereby increase.
At present; still there is not the relevant report of using surfactant assisted extraction Ganoderma spore powder liposoluble constituent; the present invention extracts the Ganoderma spore powder liposoluble constituent with surfactant; improved the extraction ratio of Ganoderma spore powder liposoluble constituent; deep processing and environmental conservation for this series products are significant; and cost is low, and suitability for industrialized production is promoted easily.
In the leaching process of breaking trachytectum of glossy ganoderma powder, add surfactant, make the abundant moistening of Ganoderma spore powder, and make enough the approaching of thickness of liquid film between medical material and the extractant, reduce the resistance the process of effective ingredient from the medical material diffusion into the surface to extractant, thereby accelerated the speed of extracting.
Embodiment 5
The preparation of soft capsule of Ganoderma spore liposoluble constituent:
In high pressure homogenize equipment, 30～35 ℃, 5 weight portion Ganoderma spore liposoluble constituents are carried out homogenizing, get core liquid; Get gelatin 100 weight portions and water 100 weight portions are put into container, be heated to 80～90 ℃, constantly stir and make it dissolving, add glycerol 40 weight portions, Polyethylene Glycol 10 weight portions continuation stirring then, dissolving fully stops heating, naturally cool to room temperature, obtain the soft capsule material; Core liquid and soft capsule material are cleaned in compacting on the soft capsule press, and drying gets soft capsule, and the Co60 irradiation sterilization is handled, and becomes product.
Embodiment 6
The preparation of the lyophilized powder of Ganoderma spore liposoluble constituent:
The Ganoderma spore liposoluble constituent is earlier at-45 ℃ of pre-freeze 2-4 hours, and dry 24-28 hour of reduced vacuum under-45 ℃～10 ℃ conditions then is at last at 30 ℃ of dry 6-8 hours.
Embodiment 7
The Ganoderma spore liposoluble constituent is to the neural effect of mice:
Confirm that through the animal pharmacodynamics Ganoderma spore liposoluble constituent of specific embodiments of the invention 3,4 becomes row property disease to the senile nervus centralis function of aged mouse remarkable effect.It improves brain estrogen beta receptor, and the neuronic quantity significance of estrogen receptor is increased; Receptor such as granulocyte colony-stimulating factor, VEGF significance increases.The Ganoderma spore liposoluble constituent can be used as treatment senile dementia, parkinson, Huntington Chorea, cerebral palsy, the apoplectic hemiplegia sequela or, the health product or the medicine of cerebral trauma sequela.
Test method: subjects is divided into 4 groups: 3 groups of specific embodiments, 4 groups of specific embodiments, the blank group of aged Mus, young Mus blank group, every group of 10 mices.
3,4 groups of specific embodiments, Ganoderma spore liposoluble constituent and normal diet mixing, every day, amount was 50mg/kg, 2 months persistent period.
3,4 groups of specific embodiments and aged blank group compare, and the hypothalamus estrogen receptor positive neuron number that specific embodiment is 3,4 groups increases, and has the significance difference opposite sex.
3,4 groups of specific embodiments and aged blank group are relatively, observe the staining power of the positive astrocyte of granulocyte colony-stimulating factor in the brain, the staining power of finding the positive astrocyte of 3,4 groups of granulocyte colony-stimulating factors of specific embodiment weakens, and illustrates that medicine has the effect that suppresses neuronal apoptosis.With aged blank group relatively, it is remarkable that diversity appears in 3,4 groups of specific embodiments.Compare with old blank group, the area and the staining power of the positive vessels of 3,4 groups of mices of specific embodiment have significant difference.
Behind 3,4 groups of the specific embodiments with aged blank group relatively, the number of VEGF positive neuron significantly increases; The relative area of VEGF positive vessels and staining power all have remarkable increase.
3,4 groups of specific embodiments and aged blank group are relatively, animal grouping, modeling, administration are the same, after administration finishes, sacrificed by decapitation, take out brain, make brain tissue homogenate, to specifications Gui Ding step time-and-motion study mice activity of monoamine oxidase, superoxide dismutase activity, catalase activity.The result is as follows:
Aged Mus blank is organized activity of monoamine oxidase apparently higher than young Mus blank group,
3,4 groups of activity of monoamine oxidase of specific embodiment are starkly lower than the blank group of aged Mus.
The blank group of aged Mus superoxide dismutase, catalase activity are starkly lower than young Mus blank group,
3,4 groups of superoxide dismutase of specific embodiment, catalase activity are apparently higher than the blank group of aged Mus.
This show specific embodiment 3,4 can be in alleviating cerebral tissue the aging of neurocyte, strengthen the vigor of neurocyte.
Embodiment 8
The Ganoderma spore liposoluble constituent is to the improvement of learning and memory of little mouse function
Pharmacological testing shows that the Ganoderma spore liposoluble constituent can improve the learning and memory function of mouse aging; Toxicological test shows that Ganoderma spore liposoluble constituent of the present invention is safe and effective.
(D-galactose, D-gal) artificial aging model and naturally-aged model (17 monthly age) as index, are observed the effect of specific embodiment 3,4 with mice coordination exercise and learning and memory to adopt mice D-galactose.
Artificial aging model: establish (6 monthly age) normal control group of the same age (normal diet), D-gal model group, positive controls (levodopa), specific embodiment 3 (40mg/kg
/D), specific embodiment 4 (40mg/kg/d) is 5 groups; Specific embodiment 3,4 is with after normal diet mixes, and administration detects following various indexs after March.
Rotating stick test: the long 80cm of transfer rod, liftoff high 40cm, 15 rev/mins of rotating speeds, the time is 2min.Write down every group of number of mice that falls down, calculate the rate of falling.
Step down test: choose 50 of Kunming kind white mice, be divided into 5 groups at random, 10 every group, ♀ ♂ half and half.Mice is put into reaction chamber (be divided into 5 with the black plastic plate, 4.5 * 4.5cm valve rubber is put as the place of safety in every right back, lay 0.5cm copper grid at interval at the bottom of the case) in, after adapting to 5min, be put on the platform, during extremity contact copper grid, the 36V electricity irritation, write down the number of animals that skips to the copper grid in 5 minutes, the wrong number of times that two forelimbs contact the copper grid simultaneously reached from the place of safety to the time (incubation period) of contact copper grid.Resurvey behind the 24h once, diving tower errors number in incubation period that the record mice jumps off and the 5min is estimated the learning and memory achievement.
The result shows, compares with normal control group of the same age, and artificial mouse aging is fallen rate and significantly raise, and obviously shorten incubation period, and errors number obviously raises.Specific embodiment 3,4 can improve significantly artificial mouse aging and reduce the rate of falling, and prolongs the incubation period of mouse aging and reduces errors number, improves the motion and the memory ability of artificial mouse aging.
The result shows, compares with normal control group of the same age, and artificial mouse aging escape latency prolongs, and crosses over the platform number of times and reduces.Specific embodiment 3,4 can significantly shorten escape latency, obviously increases and crosses over the platform number of times, can obviously improve the learning and memory function of D-gal mouse aging.
Specific embodiment 3,4 (40mg/ kg) is organized the effect of having clear improvement of the acquired obstacle of the learning and memory of artificial mouse aging as a result,
Specific embodiment 4 performances are especially obvious: specific embodiment 4 treated animals prolong 22.1% from the place of safety to the incubation period (172.7 ± 38.97 seconds) in the district of getting an electric shock than artificial mouse aging group (134.5 ± 37.2 seconds), the errors number that specific embodiment is 4 groups (0.192 ± 0.237) reduces 86.0% than artificial mouse aging group (1.374 ± 1.256), and the error rate (20.1%) that specific embodiment is strayed into the district of getting an electric shock for 4 groups reduces 66.6% than artificial mouse aging group (60.2%).
Claims (9)
1. Ganoderma extract is prepared by following method:
With the Ganoderma spore powder is raw material, adopts nonionic surfactant, organized enzyme to carry out breaking cellular wall, lyophilizing,
Get the breaking trachytectum of glossy ganoderma powder, in the mixed solvent of alcohol-water, add the surfactant of hydrophile-lipophile balance value at 11-22, fully contact, centrifugal, filter,
Filtrate is separated out the Ganoderma spore liposoluble constituent after reclaiming organic solvent from water, surfactant dissolves washes with water to remove and removes surfactant remaining in the Ganoderma spore liposoluble constituent in water, obtains the Ganoderma spore liposoluble constituent.
2. Ganoderma extract according to claim 1 is characterized in that:
Organized enzyme is pectase and cellulase,
The nonionic surfactant that adopts during breaking cellular wall is a tween 80,
In the mixed solvent of alcohol-water, add petroleum ether-ethyl acetate, form the mixed solvent of petroleum ether-ethyl acetate-alcohol-water, surfactant be dodecyl sodium sulfate, tween 20, Tween-40, Tween-60 or, tween 80.
3. Ganoderma extract according to claim 1 is characterized in that: the mode that the breaking trachytectum of glossy ganoderma powder fully contacts mixed solvent is: stirring, backflow, percolation, ultrasonic, microwave or column chromatography.
4. Ganoderma extract according to claim 1, it is characterized in that: tween 80 is dissolved in the mixed solution of pectase and cellulase, handle Ganoderma spore powder with this mixed solution, make ganoderma lucidium spore powder wall breaking, lyophilizing, get the breaking trachytectum of glossy ganoderma powder, in the mixed solvent of alcohol-water, add tween 80, fully contact, centrifugal, filter, behind the filtrate recycling ethanol, from water, separate out the Ganoderma spore liposoluble constituent, tween 80 is dissolved in the water, washes with water except that tween 80 remaining in the Ganoderma spore liposoluble constituent, obtains the Ganoderma spore liposoluble constituent.
5. Ganoderma extract according to claim 1, it is characterized in that: tween 80 is dissolved in the mixed solution of pectase and cellulase, handle Ganoderma spore powder with this mixed solution, make ganoderma lucidium spore powder wall breaking, lyophilizing, get the breaking trachytectum of glossy ganoderma powder, in the mixed solvent of petroleum ether-ethyl acetate-alcohol-water, add tween 80, fully contact, centrifugal, filter, after filtrate is reclaimed petroleum ether-ethyl acetate-ethanol, from water, separate out the Ganoderma spore liposoluble constituent, tween 80 is dissolved in the water, washes with water except that tween 80 remaining in the Ganoderma spore liposoluble constituent, obtains the Ganoderma spore liposoluble constituent.
6. Ganoderma extract according to claim 5 is characterized in that: its dosage form is soft capsule, lyophilized powder.
7. Ganoderma extract according to claim 5 is characterized in that: the preparation of soft capsule method is
In high pressure homogenize equipment, 30～35 ℃, 5 weight portion Ganoderma spore liposoluble constituents are carried out homogenizing, get core liquid;
Get gelatin 100 weight portions and water 100 weight portions are put into container, be heated to 80～90 ℃, constantly stir and make it dissolving, add glycerol 40 weight portions, Polyethylene Glycol 10 weight portions continuation stirring then, dissolving fully stops heating, naturally cool to room temperature, obtain the soft capsule material;
Core liquid and soft capsule material are cleaned in compacting on the soft capsule press, and drying gets soft capsule, and the Co60 irradiation sterilization is handled, and becomes product.
8. Ganoderma extract according to claim 5 is characterized in that: its dosage form is
Injection, oral liquid, tablet, capsule, granule, aerosol, powder spray, spray, patch, slow releasing preparation, solid dispersion.
9. Ganoderma extract according to claim 1 is characterized in that: be used for preparation treatment senile dementia, parkinson, Huntington Chorea, cerebral palsy, apoplectic hemiplegia sequela, or the health product of cerebral trauma sequela, the application on the medicine.
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| CN104920687A (en) * | 2015-05-29 | 2015-09-23 | 陈文创 | Health-maintenance jasmine black tea and processing method thereof |
| CN104957298A (en) * | 2015-05-29 | 2015-10-07 | 陈文创 | Health preserving jasminum sambac flower tea |
| CN104920687B (en) * | 2015-05-29 | 2018-10-19 | 陈文创 | A kind of health jasmine black tea and its processing method |
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| CN109588376A (en) * | 2018-12-27 | 2019-04-09 | 广西融安县金鼎制丝有限责任公司 | Silkworm egg preservation method |
| CN109588376B (en) * | 2018-12-27 | 2021-09-21 | 融安县德源农业科技发展有限公司 | Silkworm egg preservation method |
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