CN109200066A - Cicada fungus active material and its purposes for preventing, delaying or treating cataract - Google Patents
Cicada fungus active material and its purposes for preventing, delaying or treating cataract Download PDFInfo
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- CN109200066A CN109200066A CN201710549179.XA CN201710549179A CN109200066A CN 109200066 A CN109200066 A CN 109200066A CN 201710549179 A CN201710549179 A CN 201710549179A CN 109200066 A CN109200066 A CN 109200066A
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- cicada fungus
- active material
- cataract
- cicada
- crystalline
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/062—Ascomycota
- A61K36/066—Clavicipitaceae
- A61K36/068—Cordyceps
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Alternative & Traditional Medicine (AREA)
- Nutrition Science (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The present invention discloses a kind of CSM treated body for preventing, delaying or treating cataract, its active material, its mycelium liquid fermentation material and its medical composition, and the method for preparation before-mentioned products.CSM treated body of the invention, its active material, its mycelium liquid fermentation material simultaneously can be prepared as health food for preventing or delaying cataract.
Description
Technical field
The present invention is about a kind of active material.In particular, the present invention is about a kind of cicada fungus (Cordyceps cicadae) liquid
Fermentation mycelium active material, its medical composition, to prevent, delay or treat cataract.
Background technique
Crystalline in human eye is made of water and azelon.Azelon is arranged according to ad hoc fashion, makes water
Crystal is clear and light is allowed to penetrate.Increase with the age or the factor that faces a danger, azelon degenerate and bunch up, make
The situation that crystalline hardens, muddy forms cataract.At this point, light can not or be difficult to focus from cornea across crystalline
It projects on the retina, so that human body sensory eye-blurred or even invisible object.
Therefore, cataract refers to that the soluble protein in crystalline gradually becomes insolubility protein and formed mixed
It is turbid, cause eye-blurred, myopia degree to deepen (presbyopia's mitigation), image overlapping, the reduction of aperture halation, contrast vision etc.
Vision disorder phenomenon.Its occurrence cause includes congenital, traumatic, concurrency and senile etc., such as diabetes, wound, hair
Scorching, heredity etc..Clinically the most common cataract is cataract of old people.If muddy position occurs in crystalline with cataract
Setting can distinguish are as follows: core type cataract, cortical type cataract, rear bladder type cataract and mixed type cataract.
As the screen on hand-hold mobile device is watched in the aging increasingly of advanced country's population and human eye long-time, short distance attentively
Curtain content, stimulation of the human eye by high-intensitive light or the Nature radiation, make it easier to accelerate the generation process of cataract with
And the incidence with raising cataract.
Cataract is one of most common disease of ophthalmology.Initial symptoms include unstable eyesight, fuzzy, diplopia, photophobia, night
Between glare, be not easy to differentiate that object comparison of light and shade, tone reversal, object color be dim or deformation.Cataract advanced stage symptom is then view
Power obstacle severe exacerbation, vision are presented atomization, are only capable of distinguishing finger at the moment and only surplus light sensation eyesight, finally seriously cause to lose
It is bright.There has been no drugs or nutritional supplement confirmation can effectively prevent Cataractogenesis at present.
Cataract can not be diagnosed by directly observing outside eye.When diagnosing cataract, eye doctor is aobvious by such as slit-lamp
The instruments such as micro mirror, tonometer, ultrasonic instrument, automatic computing engine refractor, color computer cornea map are mixed to determine crystalline
Turbid shape, size and location.
The mode of existing prevention is including taking vitamin and antioxidant, with Wearing sunglasses to reduce ultraviolet light
It exposure, balanced diet and avoids smoking.But receiving cataract operation is currently the only effective treatment modus operandi.
Clinically the treatment method of cataract is to perform the operation and remove muddy opaque crystalline, including cataract at present
Cataract surgery and extracapsular cataract extraction operation, and it is implanted into aspherical or multifocal intraocular lens.Wherein, in cataract
Corneal endothelial layer is protected in resection operation using sodium hyaluronate, however has no document at present and explicitly points out sodium hyaluronate to cataract tool
There is prevention, delay or therapeutic effect.Since the expense of intraocular lens is not that the global all regions common people can bear, and still have
The risk of surgery side effect and complication.Therefore, cataract is still the causes of blindness of current human maximum.Accordingly, exploitation is pre-
Prevent, delay or treat medical composition, drug or the prevention of cataract generation or delays the health food of cataract generation undoubtedly
With large market.
Applicant in this case by carefully test and studies in view of the deficiency in known techniques, and an essence work with perseverance
Mind visualizes this case eventually, can overcome the shortcomings of prior art, and the following are the brief descriptions of this case.
Summary of the invention
In order to develop the medical composition to prevent, delay or treat cataract, the present invention prepares cicada fungus
(Cordyceps cicadae) mycelium, CSM treated body fluid state fermentation material, CSM treated body fluid state fermentation material active material
And medical composition, aforementioned CSM treated body fluid state fermentation material, CSM treated body fluid state fermentation material are confirmed with zoopery
Active material and medical composition can effectively prevent, delay or treat cataract.Therefore, aforementioned CSM treated body, cicada fungus bacterium
The active material of filament liquid state fermentation object and CSM treated body fluid state fermentation material can also be prepared as preventing or delaying the guarantor of cataract
Health food.
The present invention discloses a kind of method for preparing the medical composition for preventing, delaying or treat cataract, and medical group
Closing object includes the active material from cicada fungus, which comprises the mycelium of cicada fungus is inoculated on plating medium by (a)
Culture；(b) the CSM treated body through cultivating of step (a) is inoculated in the first culture medium with the first scale to cultivate；With
And the CSM treated body through cultivating of step (b) (c) is inoculated in the second culture medium with the second scale to cultivate, to obtain
Fermentation material comprising active material, wherein the second scale is greater than the first scale.
In one particular embodiment of the present invention, the culture of step (a) is that 5 days~14 are carried out between 15 DEG C~35 DEG C
It, and plating medium can be detrine agar (PDA) culture medium.In one particular embodiment of the present invention, step
(b) culture is to carry out between 15 DEG C~35 DEG C, 2~pH of pH 8 and 10rpm~250rpm more days.At of the invention one
In specific embodiment, the culture of step (c) be 15 DEG C~35 DEG C, the slot pressure of 0.5~1.0 kilogram/square centimeter, pH 2~8,
It is carried out 3 days~5 days under the gas of first mixing speed or the second mixing speed and the Ventilation Rate with 0.01~1.5VVM,
First mixing speed be 10rpm~250rpm, the second mixing speed be 0rpm (not stirring), and gas select free air, oxygen,
One of group composed by carbon dioxide, nitrogen and combinations thereof.
In one particular embodiment of the present invention, each of first and second culture medium includes being selected from comprehensive carbon
Nitrogen source, protein for animal, protein for animal hydrolysate, plant origin albumen, plant origin protolysate, yeast extraction
One of formed group of object, malt extract, inorganic salts, carbohydrate and combinations thereof, comprehensive carbon nitrogen source be cereal and/
Or beans, inorganic salts include selected from magnesium sulfate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate and ferric sulfate and combinations thereof formed group
One of, and carbohydrate include selected from the formed group of glucose, fructose, maltose and sucrose and combinations thereof wherein it
One.
In one particular embodiment of the present invention, the preparation method further includes: (d) is selected from by spray drying, heat
The dry fermentation material of dry technology of one of group composed by dry, roller drying and freeze-drying is air-dried, to obtain drying
Object.When step (d) is by being freeze-dried to carry out, the dried object is freeze-dried powder.
In one particular embodiment of the present invention, the preparation method further includes: (e) is described dry with water or alcohols extraction
Dry object, accordingly to obtain water extract or alcohol extract, wherein alcohols is 1%~100% methanol or 1%~100% ethyl alcohol.
In one particular embodiment of the present invention, the medical composition for preventing, delaying or treating cataract includes
Active material and pharmaceutically acceptable carrier, excipient, diluent or adjuvant from cicada fungus.In a tool of the invention
In body embodiment, the medical composition for preventing, delaying or treating cataract includes the cicada fungus bacterium comprising cicada fungus active material
Filament liquid state fermentation object and pharmaceutically acceptable carrier, excipient, diluent or adjuvant.It is specific real at of the invention one
It applies in example, the medical composition for preventing, delaying or treating cataract includes that the cicada fungus liquid comprising cicada fungus active material is sent out
The freeze-dried powder of ferment object and pharmaceutically acceptable carrier, excipient, diluent or adjuvant.In a specific implementation of the invention
Example in, the medical composition for preventing, delaying or treat cataract include comprising cicada fungus active material cicada fungus water extract object or
Cicada fungus alcohol extracts object and pharmaceutically acceptable carrier, excipient, diluent or adjuvant.
The present invention separately discloses a kind of active material preparation by cicada fungus and is used to prevent, delays or treat medical group of cataract
The purposes of object is closed, wherein active material is obtained with aforementioned preparation process.
The present invention separately discloses a kind of active material of cicada fungus for preventing, delaying or treat cataract, wherein active matter
Matter is obtained with aforementioned preparation process.
Detailed description of the invention
Fig. 1 is the schematic diagram that crystalline of the invention imitates that slit-lamp scans imaging.A: water-soluble control group, B: water-soluble injury
Group, C: cicada fungus water extracts object, D: the molten control group of oil, E: the molten injury group of oil and F: cicada fungus alcohol extracts object.
Fig. 2 is the schematic diagram of crystalline grid figure imaging of the invention.A: water-soluble control group, B: water-soluble injury group, C: cicada
Hua Shui extracts object, D: the molten control group of oil, E: the molten injury group of oil and F: cicada fungus alcohol extracts object.
Fig. 3 is crystalline scattergram imaging schematic diagram of the invention.A: water-soluble control group, B: water-soluble injury group, C: cicada fungus
Water extracts object, D: the molten control group of oil, E: the molten injury group of oil and F: cicada fungus alcohol extracts object.
Fig. 4 is the schematic diagram for the crystalline light transmittance that cicada fungus water of the present invention extracts object.
Fig. 5 is the schematic diagram for the crystalline light transmittance that cicada fungus alcohol of the present invention extracts object.
Fig. 6 is the schematic diagram for the crystalline lumens difference that cicada fungus water of the present invention extracts object.
Fig. 7 is the schematic diagram for the crystalline lumens difference that cicada fungus alcohol of the present invention extracts object.
Specific embodiment
This case proposed invention can be illustrated by embodiment below and is fully understood, so that technical field
Middle tool usually intellectual can complete accordingly, however the implementation of this case not can be limited its implementation type by the following example
State, those of ordinary skill in the art can still deduce out other implementations according to the spirit for removing both disclosed embodiments
Example, the embodiments such as described are all when belonging to the scope of the present invention.
Cicada fungus (Cordyceps cicadae) is mycota (Fungi), Ascomycota (Ascomycota), excrement shell Gammaproteobacteria
(Sordariomycetes), Hypocreales (Hypocreales), Clavicipitaceae (Clavicipitaceae), Cordyceps
(Cordyceps) fungi.It infects cicada pupa or Cicadidae mountain cicada (Cicada flammate), a kind of cicadahun (Platypleura
Kaempferi), the larvas such as black grasshopper (Crytotympana pustulata) and bamboo cicada (Platylomia pieli) keep it dead
It dies, forms bud shape stroma, therefore named cicada fungus then at cicada pupa front end or polypide head.Therefore, cicada fungus is a kind of bacterium worm complex.Cicada
Flower can be classified as big cicada fungus or golden cicada grass (C.cicadae), small cicada fungus according to different hosts and infection bacteria species
(C.sobolifera) and three kinds of cicada grass or cicada infested careless (C.cicadicola).Cicada fungus fecund in the drought torrid zone and
The ground such as subtropical zone, such as Chinese Fujian, Zhejiang, Sichuan, Yunnan and Jiangsu.Also there is wild cicada fungus in Taiwan part mountain area
Entity trace.
Cicada fungus is rare traditional Chinese medicine, cold in nature, sweet in flavor, nontoxic, can be used as medicine after drying, there is dissipating wind-heat, relieving convulsion, promoting eruption
The effect of, cure mainly children day hang, frightened epilepsy, palpitaition, the morbid night crying of babies.Modern pharmacology experiment also indicates that cicada fungus and its artificial culture have
There is apparent adjust be immunized, nervous system adjusting, antifatigue, calm, Antipyretic, improve renal function, is hypoglycemic, reducing blood
Pressure, reducing heart rate, inhibition atherogenesis, antitumor, anti-radiation and nourishing and fit keeping function and other effects.
Cicada fungus and cordyceps sinensis belong to entomogenous fungi complex, the functionality of cicada fungus and application no less than cordyceps sinensis and
Cordyceps militaris has similar medicinal health care function, and contains similar chemical component, so frequently as the substitute of cordyceps sinensis.
However, the yield of natural cordyceps sinensis and natural cicada fungus is few and need to rely on host, and host and the climate factor or people
For the influence of factor.Therefore, the yield and its economic value for how promoting cicada fungus become the industry and strongly solve the problems, such as.
The present invention discloses a kind of method for preparing cicada fungus liquid state fermentation mycelium active substance (abbreviation active material), a kind of
The method of medical composition of the preparation comprising the active material and a kind of the active material or medical composition are used for
Prevent, delay or treat the purposes of the induced cataract of physical injury.Compared to surgical removal or western medicine, the present invention
Preparation method it is safer, easy, the active material of acquisition is more natural, safety, and reaches prevention, delays or treat eyes and is sick
The effect of change.The active material can be also prepared as with general health food for preventing or delaying the health care of cataract to eat
Product.
(1) experimental procedure:
1. cicada fungus material:
CSM treated body used in specific embodiments of the present invention is acquisition from the wild cicada fungus fructification in Taiwan, through separating
And its mycelium, and subculture is on plating medium, and identifies its base through Taiwan Foodstuff Industrial Development Inst. of Financial Group Legal Persons
Cicada fungus is confirmed as because of sequence.However, can be applied to cicada fungus active material of the invention is not limited to thus strain (C.cicadae)
Gained.
2. the liquid state fermentation culture of CSM treated body:
Firstly, CSM treated body aseptic inoculation is trained under 15 DEG C~35 DEG C (preferably 25 DEG C) on plating medium
It supports 5 days to 2 weeks (14 days), then CSM treated body is scraped with aseptic inoculation technology and is inoculated in the culture medium with the first scale
(such as culture medium in flask).Then 15 DEG C~35 DEG C (preferably 25 DEG C), pH 2~8 (preferably pH 4~7, preferably
Person about pH 4.5), cultivate a couple of days (such as 3 days or 3 days or more under concussion rate 10rpm~250rpm.It then, will be in flask
Culture aseptic inoculation is to the culture medium (such as culture medium in fermentation tank) with the second scale, (preferably at 15 DEG C~35 DEG C
Be 25 DEG C), 0.5-1.0kg/cm2Slot pressure, pH 2~8, the mixing speed of 10rpm~150rpm or do not stir (air lift)
The case where, gas (such as the mixing of air, air and oxygen, carbon dioxide or nitrogen is passed through with 0.01-1.5VVM Ventilation Rate
Object), it cultivates 3 days~5 days, obtains CSM treated body fluid state cultivation and fermentation object comprising CSM treated body and clarified solution.The cicada
Flower mycelium liquid cultivation and fermentation object contains the active material from cicada fungus.In order to expand CSM treated body fluid state cultivation and fermentation object
Yield, the second scale is typically larger than the first scale.Gas, which is passed through, with 0.01-1.5VVM Ventilation Rate is preferably passed through sky
Gas.
It is aforementioned that have the culture medium of the first scale and the second scale may include identical or different ingredient and formula.Have
Culture medium prescription as shown in Table 1 can be used in the culture medium of first scale and the second scale, but not limited to this formula.
Table 1, culture medium prescription
Wherein comprehensive carbon nitrogen source can be cereal (such as flour class) or beans (such as soybean powder, mung bean flour, soy meal
Deng), inorganic salts can for magnesium sulfate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, ferric sulfate etc., and carbohydrate can for glucose, fructose,
Maltose, sucrose etc..
3. the drying of CSM treated body fluid state cultivation and fermentation object:
The freeze-dried powder for fermentation material can be further dried in CSM treated body fluid state cultivation and fermentation object.Used dry technology
Including but not limited to spray drying, heated-air drying, roller drying, freeze-drying or other be applicable in and of the invention commonly use dry skill
Art.
4. the extraction of CSM treated body fluid state cultivation and fermentation object:
Distilled water is added in the freeze-dried powder of CSM treated body fluid state cultivation and fermentation object, back dissolving is allowed to and is suspended in distilled water
In, it is heated several minutes with 90 DEG C~121 DEG C of temperature.It is concentrated under reduced pressure after cooling or obtains cicada through aforementioned dry technology again
Flower mycelium water extraction object (abbreviation cicada fungus water extracts object).
Alternatively, alcohols (1% (v/v)~100% (v/ is added in the freeze-dried powder of CSM treated body fluid body cultivation and fermentation object
V) methanol or ethyl alcohol), it is allowed to back dissolving and is suspended in alcohols, with several minutes of immersion, stirring, concussion or ultrasonic extraction, then pass through
Vacuum concentration method or aforementioned dry technology and obtain CSM treated body alcohol extraction object (abbreviation cicada fungus alcohol extract object).
5. the preparation of the medical composition containing cicada fungus active material:
Those of ordinary skill in the art can be active by cicada fungus of the invention according to its pharmacy knowledge and technology
Substance, the cicada fungus liquid cultivation and fermentation object comprising cicada fungus active material, the cicada fungus lyophilized products comprising cicada fungus active material or comprising
The cicada fungus water extraction object or cicada fungus alcohol of cicada fungus active material extract object, with pharmaceutically acceptable carrier, excipient, diluent or adjuvant
It is prepared as the medical composition for preventing, delaying or treating cataract.The dosage form of the medical composition may include but unlimited
In capsule, pastille, pill, lotion, suspension dispersive agent, solvent.By taking pastille as an example, pharmaceutically acceptable carrier includes but unlimited
In lactose, cornstarch, lubricant and magnesium stearate.And the diluent that capsule uses includes but is not limited to lactose, drying
Cornstarch.
6. the preparation of the health food containing cicada fungus active material:
Those of ordinary skill in the art can be according to its food industry knowledge and technology by cicada fungus of the invention
Active material, the cicada fungus liquid cultivation and fermentation object comprising cicada fungus active material, the cicada fungus lyophilized products comprising cicada fungus active material or
Cicada fungus water extraction object or cicada fungus alcohol comprising cicada fungus active material extract object, with the additive of food-grade (such as incremental agent, colorant,
Sweetener, fragrance, preservative etc.) it is prepared as health food for preventing or delaying cataract.
(2) zoopery:
Ultraviolet light divides into UVA (315~380nm), UVB (280~315nm) and UVC (100 according to its wavelength section
~280nm).The excessive radiation of ultraviolet light will cause photochemistry to injure, especially most obvious with injury of the UVB to eyeball.
Nearly whole UVC can be absorbed in cornea, but for longer wavelengths of ultraviolet light, permeability can rapidly increase
Add.Such as the ultraviolet-transparent 60% of 320nm.Crystalline then further absorbs most into intraocular ultraviolet light.Year
Wavelength 370nm ultraviolet light below can be absorbed in the crystalline of light people, but as the age is elongated, and the crystalline of the elderly becomes yellow
Brown and the blue light for absorbing more UVA and shorter wavelength.
Zoopery, which has proven to ultraviolet radiation, can cause cataract, especially be easy to produce in crystalline cortex and rear capsule position
Raw muddy phenomenon.Undifferentiated crystalline epithelial cell can be observed under the microscope rearward to migrate, cause post gelating time
Cataract.The formation of research on epidemiology and observation clinically, cataract is related with sun exposure amount.It stays in more
There is higher incidence of cataract close to the bigger region of region of the equator, sunshine amount and the more frequent people of outdoor activities.Further
Studies have shown that cataract is related with UVB ultraviolet radiation, then generates muddiness most with crystalline cortex and rear capsule position in kenel
It is more.
In the present invention, with the crystal bulk damage of ultraviolet induction mouse, assess CSM treated body active material in prevention,
Delay or treat the effect of cataract caused by ultraviolet light, and its in the related application of eye health.
Embodiment one, cicada fungus liquid state fermentation culture and the preparation of active material
1. plate culture: by CSM treated body aseptic inoculation in detrine agar (potato dextrose agar,
PDA it) on plating medium, is cultivated at 25 DEG C about 5 days.
2. flask culture: with the CSM treated body on aseptic technique scraping plating medium and being inoculated in flask
Culture medium (contains 2.0 weight % of sucrose, 0.5 weight % of yeast extract, 1.0 weight % of soybean powder, add water to 100 weight %),
The shake culture three days under 25 DEG C, pH 4.5,120rpm.
3. tank culture of fermenting: by the CSM treated body aseptic inoculation in flask in the culture medium of fermentation tank.In fermentation tank
Medium component it is identical as the medium component in flask.In 25 DEG C, 0.5-1.0kg/cm2Slot pressure, pH 4.5,10rpm
The mixing speed of~150rpm or do not stir under (air lift) and the gas of the Ventilation Rate with 0.5-1.0VVM culture and
Fermentation 3 days obtains CSM treated body fluid state cultivation and fermentation object comprising CSM treated body and clarified solution.The CSM treated body
Liquid cultivation and fermentation object contains the active material from cicada fungus.It is dry that CSM treated body fluid state cultivation and fermentation object is subjected to freezing again
It is dry, the freeze-dried powder of fermentation material is obtained, in favor of saving.CSM treated body fluid state culture after 20 tonnes of fermentation tank cultivation and fermentations
After fermentation material is freeze-dried, about 110 kilograms of fermentation material freeze-dried powder can get.
4. the preparation of cicada fungus water extraction object: CSM treated body fermentation material freeze-dried powder is dissolved in the distilled water of 20 times of volumes, with
100 DEG C are heated 30 minutes, are freeze-dried after cooling, are obtained cicada fungus water and are extracted object.
5. cicada fungus alcohol extraction object preparation: by CSM treated body fermentation material freeze-dried powder back dissolving in 20 times of volumes ethyl alcohol (1% to
100% ethyl alcohol), with ultrasonic vibrating extraction 1 hour.Extraction suspension takes supernatant after being centrifuged, and obtains cicada through being concentrated under reduced pressure
Flower alcohol extracts object.Aforementioned 1% ethyl alcohol or mean lower than 100% ethyl alcohol mixes ethyl alcohol with solvent (such as water) with volume ratio, aforementioned
100% ethyl alcohol means the straight alcohol for being not added with solvent (such as water).Those of ordinary skill in the art can be by methanol
Or ethyl alcohol is replaced by the compound for having similar physical chemical property with methanol or ethyl alcohol.
The analysis of embodiment two, cataract zootype and associated pointers
1. the foundation of cataract mouse species mode:
The present embodiment (is purchased from Taiwan Le Sike biotechnology stock using ICR strain, 6 week old, 25~33 grams of female mice
Part Co., Ltd), raising middle mountain medical university Experimental Animal Center in Taiwan provides normal clean feed and drinking-water,
Feeding environment is that illumination in 12 hours and 12 hours dark circulation lights shine, and temperature is controlled in 20 ± 2 DEG C, humid control 50 ±
5%.
2. feeding dosage and experimental procedure:
The ultraviolet (UV) B induction projected with ultraviolet lamp (French Vilber Lourmat company, fluorescent tube model VL-6MCUVB)
Mouse generates cataract.This test carries out 29 days altogether, and mouse is randomly divided into 6 groups, every group 3 before test:
(1) water-soluble control group: daily pipe feeds 0.9%NaCl solution；
(2) water-soluble injury group: daily pipe fed 0.9%NaCl solution, in progress ultraviolet (UV) B injury in the 6th day to the 28th day；
(3) cicada fungus water extracts object: daily pipe feeds the cicada fungus water extraction object for the 100mg/kg weight for being dissolved in 0.9%NaCl solution, in
Progress ultraviolet (UV) B injury in 6th day to the 28th day；
(4) the molten control group of oil: daily pipe feeds commercially available soybean oil；
(5) the molten injury group of oil: daily pipe fed commercially available soybean oil, in progress ultraviolet (UV) B injury in the 6th day to the 28th day；With
And
(6) cicada fungus alcohol extracts object: daily pipe feeds the cicada fungus alcohol extraction object for the 100mg/kg weight for being dissolved in commercially available soybean oil, in the 6th
It was to progress ultraviolet (UV) B injury in the 28th day.
In the processing of ultraviolet (UV) B (i.e. water-soluble injury group, cicada fungus water extraction object, the molten injury group of oil and cicada fungus alcohol extract object),
The mouse anaesthetized through 2.5%Avertin is placed in camera bellows, eyeball irradiates 0.72J/cm upward2The ultraviolet light of intensity 90 seconds, makes
Mouse crystalline is damaged.All groups of other mouse were sacrificed on 29th.
3. the detection of crystalline degree of injury and cataract assessment index of correlation analysis:
This divides into eyeball imaging (imitative slit-lamp) and crystalline imaging (grid figure and scattergram) and detects.
The purpose that imitative slit-lamp scanning imaging compares is to judge crystalline in eyeball via the mode of lateral projection
The degree of light transmission in portion, so compare cataract development degree be in image sharpness.After mouse is sacrificed, right eye eyeball juxtaposition is taken out
In on red line white background backboard, and wheel portion is aligned red line, and shoot the side imaging result of right eye eyeball.
Crystalline imaging compares divides into grid figure imaging and scattergram imaging again.The main purpose that grid figure imaging compares
It is detect whether crystalline center imaging effect is normal, the main purpose that scattergram imaging compares is to detect crystalline periphery
Whether imaging effect is excellent.After mouse is sacrificed, the crystalline of left eye eyeball is taken out, is placed on grid figure and scattergram, and shoot
The imaging result of left eye eyeball crystalline.
4. luminance meter analyzes (light transmittance and light transmittance):
The purpose that light transmittance is tested with light transmittance is detection crystalline after ultraviolet (UV) B injury and via preparatory
The difference of resistance degree of injury after feeding cicada fungus active material.
Light transmittance is that light passes through the percentage after crystalline at different wavelengths with luminance meter measurement.By a mouse left side
Crystallization is placed on slide, and is set in luminance meter inducing probes, is irradiated crystalline with fixed light source and is captured its light transmission
Degree and light transmittance.
Light transmittance measurement be calculate how many lumen value light pass through crystalline.When the light passed through is fewer, table
Show that the saturating clear degree of crystalline is lower, also represent that cataract degree is higher, also just makes under the lumen value by the light of crystalline
Drop.Total lumen value (coloured light of comprehensive all wavelengths) of original ray subtracts the lumen value measured after light source irradiation crystalline i.e.
Indicate the translucency of crystalline.Crystalline light transmittance is higher, and lumens difference is smaller.Therefore, the severity of cataract can be with
Lumens difference indicates.
Embodiment three, cicada fungus active material are for preventing, delaying or treating the recruitment evaluation of cataract and index of correlation
1. eyeball imaging (imitative slit-lamp) and crystalline imaging (grid figure and scattergram) detect
Referring to Fig. 1, it imitates the schematic diagram that slit-lamp scans imaging for crystalline of the invention.In Fig. 1, water-soluble control
Both group (A) and the molten control group (D) of oil are all without the crystal body characteristics for exception occur.Opposite, water-soluble injury group (B) and the molten wound of oil
All there is crystalline research of chaotic phenomenon in both evils group (E), indicate successfully to induce mouse cataract with ultraviolet (UV) B.In edible cicada
Hua Shui extracts in the mouse group (C, F) of object and cicada fungus alcohol extraction object, and cicada fungus water extraction object and cicada fungus alcohol extraction object all can effectively maintain mouse
The degree of light transmission of eyeball side.Therefore, it the cicada fungus water extraction object containing cicada fungus active material and cicada fungus alcohol extraction object and thus prepares
Medical composition can effectively prevent, delay or treat the induced cataract of physical injury.
Referring to Fig. 2, it is the schematic diagram of crystalline grid figure imaging of the invention.In Fig. 2, in addition to can be observed with
It is similar as a result, working as the clarity of crystalline central area imaging compared with grid figure distortion level that eyeball imaging compares (Fig. 1)
When, it is possible to find water-soluble control group (A) and the molten control group (D) of oil and with clearly grid figure imaging and there are few distortion distort, water
Both molten injury group (B) and the molten injury group (E) of oil are unsharp grid figure imaging and distortion and distortion.However, cicada fungus water extracts
Object (C) and cicada fungus alcohol extraction object (F) then have clearly grid figure imaging and there are few distortion and distortions.Therefore, containing cicada fungus activity
The cicada fungus water extraction object and cicada fungus alcohol extraction object of substance and the medical composition thus prepared can effectively prevent, delay or treat physical
Injure induced cataract.
Referring to Fig. 3, it is the schematic diagram of crystalline scattergram imaging of the invention.The crystalline scattergram periphery of Fig. 3
Imaging also shows that consistent with the crystalline grid figure imaging of Fig. 2.Therefore, preparatory feeding cicada fungus water extraction object or cicada fungus alcohol extract object
Group all can effectively prevent, delay or treat injury of the ultraviolet (UV) B induction early stage cataract to crystalline.
2. luminance meter analyzes (light transmittance and light transmittance):
Fig. 4 and Fig. 5 is please referred to, is respectively the crystalline light transmittance of cicada fungus water extraction object and cicada fungus alcohol extraction object of the present invention
Schematic diagram.In Fig. 4 and Fig. 5, crystalline is irradiated to fix the light source (400nm to 725nm wavelength) of warm colour light, is put locally
Compare the light transmittance between each group in big 550nm~600nm wavelength region, shows and scan imaging ratio with imitative slit-lamp
(Fig. 2, Fig. 3) consistent result is compared to (Fig. 1) and crystalline imaging.Water-soluble control group and the molten control group of oil and cicada fungus water extract
Object and cicada fungus alcohol extraction object have consistent light transmittance result.Therefore, preparatory feeding cicada fungus water extraction object or cicada fungus alcohol extract object
Group can make the mouse crystalline after ultraviolet (UV) B is injured maintain due translucency, keep its imaging relatively sharp.Therefore,
Ultraviolet (UV) B induction early stage property can effectively be prevented, delay or be treated to the group of preparatory feeding cicada fungus water extraction object or cicada fungus alcohol extraction object
Injury of the cataract to crystalline.
Fig. 6 and Fig. 7 is please referred to, is respectively the crystalline lumens difference of cicada fungus water extraction object and cicada fungus alcohol extraction object of the present invention
Schematic diagram.It is more each in 425nm~550nm and 600nm~650nm range of wavelengths of partial enlargement in Fig. 6 and Fig. 7
Lumens difference between group shows result corresponding with above-mentioned data.This means, extract object and cicada in preparatory feeding cicada fungus water
Even if injury of the mouse of flower alcohol extraction object by ultraviolet (UV) B, can still maintain the translucency of its crystalline, therefore include that cicada fungus is living
Property substance cicada fungus water extraction object or cicada fungus alcohol extraction object and the medical composition that thus prepares can effectively prevent, delay or treat it is ultraviolet
Injury of the cataract of line induction to crystalline.
In summary, CSM treated body disclosed in the specific embodiment of the invention, CSM treated body fluid state fermentation material (and freeze-drying
Object), the active material of CSM treated body fluid state fermentation material and can effectively be prevented by medical composition prepared by preceding product, be prolonged
Slow or treatment cataract and the cataract induced by physical uv damage.Accordingly, aforementioned CSM treated body, cicada fungus
The active material of mycelium liquid fermentation material (and lyophilized products) and CSM treated body fluid state fermentation material can also be according to the guarantor of this field
Health food manufacturing technology is prepared as preventing or delaying the health food of cataract.
The present invention belongs to the innovation of difficult energy in fact, deep to have industrial value, helps and files an application in accordance with the law.In addition, the present invention can be with
Any modification is made by those of ordinary skill in the art, but is not departed from such as the claimed model of appended claims
It encloses.
Claims (10)
1. a kind of method for preparing the medical composition for preventing, delaying or treat cataract, the medical composition include
Active material from cicada fungus (Cordyceps cicadae), which comprises
(a) mycelium of the cicada fungus is inoculated on plating medium to cultivate；
(b) the CSM treated body through cultivating of step (a) is inoculated in the first culture medium with the first scale to cultivate；And
(c) the CSM treated body through cultivating of step (b) is inoculated in the second culture medium with the second scale to cultivate, to obtain
The fermentation material comprising the active material is obtained, wherein second scale is greater than first scale.
2. according to the method described in claim 1, wherein the culture of step (a) is to carry out 5 days~14 between 15 DEG C~35 DEG C
It, and the plating medium is detrine agar (PDA) culture medium.
3. according to the method described in claim 1, wherein the culture of step (b) be 15 DEG C~35 DEG C, 2~pH of pH 8 and
It is carried out between 10rpm~250rpm more days.
4. according to the method described in claim 1, wherein the culture of step (c) is at 15 DEG C~35 DEG C, 0.5~1.0 kilogram/flat
One of slot pressure, pH 2~8, the first mixing speed and second mixing speed of Fang Gongfen and with 0.01~1.5VVM's
It is carried out 3 days~5 days under the gas of Ventilation Rate, first mixing speed is 10rpm~250rpm, second mixing speed
For 0rpm, and the gas selects one of group composed by free air, oxygen, carbon dioxide, nitrogen and combinations thereof.
5. according to the method described in claim 1, wherein each of described first and second culture medium include being selected from
Comprehensive carbon nitrogen source, protein for animal, protein for animal hydrolysate, plant origin albumen, plant origin protolysate,
One of formed group of yeast extract, malt extract, inorganic salts, carbohydrate and combinations thereof, the comprehensive carbon
Nitrogen source be cereal and beans at least one, the inorganic salts include being selected from magnesium sulfate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate
And one of formed group of ferric sulfate and combinations thereof, and the carbohydrate includes being selected from glucose, fructose, maltose and sugarcane
One of formed group of sugar and combinations thereof.
6. according to the method described in claim 1, further including:
(d) with the drying selected from one of the group composed by spray drying, heated-air drying, roller drying and freeze-drying
The dry fermentation material of technology, to obtain dried object.
7. according to the method described in claim 6, the dried object is to freeze wherein when step (d) is by being freeze-dried to carry out
Dry powder.
8. according to the method described in claim 6, further including:
(e) dried object is extracted with one of water and alcohols, with accordingly obtain water extract and alcohol extract wherein it
One, wherein the alcohols is one of 1%~100% methanol and 1%~100% ethyl alcohol.
9. a kind of active material by cicada fungus (Cordyceps cicadae) is prepared for preventing, delaying or treating cataract
The purposes of medical composition, wherein the active material is that the method according to any one of claims 1 to 8 obtains.
10. a kind of active material of the cicada fungus (Cordyceps cicadae) for preventing, delaying or treating cataract, wherein
The active material is obtained with the method according to any one of claims 1 to 8.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710549179.XA CN109200066A (en) | 2017-07-07 | 2017-07-07 | Cicada fungus active material and its purposes for preventing, delaying or treating cataract |
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| CN201710549179.XA CN109200066A (en) | 2017-07-07 | 2017-07-07 | Cicada fungus active material and its purposes for preventing, delaying or treating cataract |
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| CN201710549179.XA Pending CN109200066A (en) | 2017-07-07 | 2017-07-07 | Cicada fungus active material and its purposes for preventing, delaying or treating cataract |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110279724A (en) * | 2019-05-31 | 2019-09-27 | 兰溪市立顺生物有限公司 | Cicada fungus active material and its purposes for preventing, delaying or treating cataract |
| EP3946414A4 (en) * | 2019-04-03 | 2022-12-07 | Zih Yuan Tang Biotechnology Co., Ltd. | Method for treating ocular diseases |
-
2017
- 2017-07-07 CN CN201710549179.XA patent/CN109200066A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3946414A4 (en) * | 2019-04-03 | 2022-12-07 | Zih Yuan Tang Biotechnology Co., Ltd. | Method for treating ocular diseases |
| CN110279724A (en) * | 2019-05-31 | 2019-09-27 | 兰溪市立顺生物有限公司 | Cicada fungus active material and its purposes for preventing, delaying or treating cataract |
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