CN102552335A - Traditional Chinese medicine health care product, its preparation method and its application - Google Patents
Traditional Chinese medicine health care product, its preparation method and its application Download PDFInfo
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Abstract
The present invention provides a traditional Chinese medicine health care product, its preparation method and its application. The traditional Chinese medicine health care product is prepared by taking Cordyceps cicadae from artificial cultivation as a main raw material. The traditional Chinese medicine health care product has the functions of improving anemia and sleep disorders. Animal experiments show that the traditional Chinese medicine health care product has the advantages of safety, effectiveness and no obvious side effect.
Description
Technical field
The invention belongs to the Chinese medicine health care product technical field, be specifically related to a kind of Chinese medicine health care product and preparation method thereof.
Background technology
Periostracum cicadae (Paecilomyces cicadae (Miquel.) Samson) belongs to worm natural disposition medicinal fungi; It is China's tradition rare Chinese medicine; Have another name called Cordyceps sobolifera, Cicadae young pilose antler, Hu Cicadae, cicada etc.; Be the nymph of Cicadae in soil by the parasitic lethal corpse that carries disease germs of the conidium of Clavicipitaceae fungus, the Stroma that it forms the head length mycelia, so the likeness in form alabastrum is the name Periostracum cicadae.Its medicinal beginning is stated from the Lei Gong's Treatise on Preparation and Broiling of Materia Medica in Liu Song of Northern and Southern Dynasties epoch.Song dynasty's Classic Classified Materia Medica for Emergencies that TANG Shen-Wei is shown speech " the Periostracum cicadae sweet in the mouth is cold, and nontoxic, main children's hangs in the sky, infantile convulsion ， clonic convulsion, night cry, cardiopalmus ".Chinese medicine the scholar think, Periostracum cicadae has the effect of dispelling wind and heat pathogens, rash, relieving spasm by subduing liver-wind, improving acuity of vision and removing nebula.Modern medicine study shows, effect such as it has immunomodulating, improves renal function, regulates lipid metabolism, promotes hemopoietic, improves the body nutriture, antipyretic-antalgic, tranquilizing soporific, strengthening by means of tonics, resisting fatigue anti-stress, nervous system regulation.
Medicinal existing more than the 1000 year history of Periostracum cicadae, but growth needs specific ecological conditions and the host insect of natural Periostracum cicadae, resource scarcity; Output descends year by year; Can only gather once in 1 year, product quantity and quality are affected by environment uncontrollable, can not satisfy people's demand.Isolating Paecilomyces cicadidae(Miquel)Samson bacterial strain can be in some basic naturally things or the top fermentation of combination culture medium from natural Periostracum cicadae, and solid culture grows similar vermivorous coremium, thereby thinks and might pass through artificial culture's approach, solves the problem of natural Periostracum cicadae inadequate resource.
Summary of the invention
The objective of the invention is to overcome defective of the prior art a kind of Chinese medicine health care product is provided.
For realizing above-mentioned purpose, the present invention takes following technical scheme:
A kind of Chinese medicine health care product, the main pharmacodynamics composition in this Chinese medicine health care product are the effective component extracts of Periostracum cicadae artificial culture product or Periostracum cicadae artificial culture thing.
Said Periostracum cicadae artificial culture product is made by following method:
1) slant strains preparation: the Paecilomyces cicadidae(Miquel)Samson inoculation to slant medium, under 20～25 ℃, was cultivated 3～10 days;
2) preparation of seed liquor: scrape from the inclined-plane and to get mycelium inoculation to fluid medium, under 20～25 ℃, 140～160rpm cultivates (being generally 3～8 days) and can make the Paecilomyces cicadidae(Miquel)Samson seed liquor;
3) with step 2) in the fermentation liquid that makes be in 5%～10% said solid phase culture medium that inserts after the sterilization by weight percentage; In temperature is 18～23 ℃; Humidity is under 60～80% the condition; Leave standstill closed cultivation 35～40 days, the irradiation of the uninterrupted full gloss spectrum of artificial light source 100-150lux artificial light source.
4) cultured products of gathering.
The said slant medium of step 1) can be PSA (potato sucrose agar culture medium), PDA (potato dextrose agar) or YMB solid medium.
Step 2) said fluid medium can be PSB (potato sucrose fluid medium) or PDB (potato glucose fluid medium).
The said solid medium of step 3) comprises the raw material components of following weight portion:
The present invention has added Semen setariae and ferrous porphyrin in solid medium, research shows, adopts this solid medium to obtain to cultivate iron content increase in the sporophore that obtains.
The said cultured products of step 4) is for cultivating the Paecilomyces cicadidae(Miquel)Samson sporophore that obtains.
Further preferred; The used Paecilomyces cicadidae(Miquel)Samson bacterial strain of step 1) is Paecilomyces cicadidae(Miquel)Samson bacterial strain Paecilomyces cicadae (Miq.) Samson CGMCC No.3453, and this bacterial strain is registered preservation in (being called for short CGMCC) on November 18th, 2009 at China Committee for Culture Collection of Microorganisms common micro-organisms center.
Paecilomyces cicadidae(Miquel)Samson bacterial strain CGMCC No.3453 adopts following method to separate:
The sources of three rivers head is gathered pure pollution-free wild Periostracum cicadae BIAO and BEN from Yunnan; The transplanting tweezer of crossing with flame sterilization on the clean work station take a morsel from sample conidium in the PSA flat board that adds rose-bengal put 22 ℃-25 ℃ cultivate 120h down after again picking list bacterium colony insert slant medium, 22 ℃-25 ℃ cultivate down and through purification repeatedly get final product Paecilomyces cicadidae(Miquel)Samson bacterial strain of the present invention [Paecilomyces cicadae (Miq.) Samson].
Paecilomyces cicadidae(Miquel)Samson bacterial strain CGMCC No.3453 belongs to Deuteromycotina Deuteromycotina, Hyphomycetes Hyphomycetes, bundle stalk spore order Stilbellales, Stilbaceae Stilbellaceae, paecilomyces Paecilomyces.
The result who identifies through Chinese Academy of Sciences's morphological characteristic that biological study is carried out is:
Paecilomyces cicadidae(Miquel)Samson [Paecilomyces cicadae (Miq.) Samson], its main morphological characteristic is:
On the PDA culture medium, cultivated 10 days for 25 ℃, bacterium colony is circular, and is open and flat, first cotton-shaped, back powdery, and white is to light yellow, and the back side is light yellow, 5～6 centimetres of diameters.Hyphae colorless, branch, tool barrier film, wide 1.2～2.5 μ m.The conidiophore multi-branched, wide 3.0～5.5 μ m are taken turns 2～5 conidiogenous cells and are formed the colyliform branch by every.The conidiogenous cell doleiform, base portion is spherical to be expanded, and it is narrow upwards to attenuate, 4.5～6.5 * 2.5～3.5 μ m.Conidium is cylindrical, and is unicellular, colourless, level and smooth, and chain is given birth to, and is straight or crooked, 6.5～8.8 (～11.0) * 2.5～3.5 μ m.
Paecilomyces cicadidae(Miquel)Samson bacterial strain CGMCC No.3453 has following microbial characteristic:
1, cultivate the characteristic of learning:
(1) puts conidium in 24 ℃, 1% C
6H
12O
6Solution is made slide glass and is cultivated, and 4h is sprouted, and the 8h long mycelia that sprouts begins to need 36h to inferior for generation of conidium from conidia germination;
(2) bacterium colony is on Rhizoma Solani tuber osi-sucrose-agar, grow prolifically, and 24 ℃ of incubation 14d, diameter reaches 60～72mm.Mycelium is greyish white to pale yellow, the fine hair shape.Late stage of culture because of producing a large amount of conidium, causes the bacterium colony outward appearance to show the opaque shape.Bacterium colony is grown good on peptone-sucrose-agar, 14d diameter 54～70mm, and local crowning or subside, yellowish pink, the later stage is covered with conidium, back side dark brown;
(3) the liquid submerged fermentation synthetic medium is with Richard culture medium (KNO
310g, KH
2PO
45g, MgSO
4.7H
2O 2.5g, sucrose 50g, FeCl
30.03g, water 1000ml) and be good;
(4) humiture:
1. temperature: 6 ℃ of this bacteria growing origin temps, 24～26 ℃ of optimum growth temps, but coremium trophophase temperature is on the low side, pure culture is handled 30h through 40 ℃, handles inactivation behind the 2.5h for 50 ℃;
2. humidity: under saturated humidity, conidia germination rate 24h is 86%, and 96h is 99%; During RH 90%, the 24h germination rate is 0%, and 96h is 78.3%; During RH 76%, the 72h germination rate is 0%, and 96h is 18%;
(5) PH: this bacterium is at the equal ability of PH 4～12 scopes normal growth, but PH 5～6 mycelium productions are high, and the slant culture bacterium colony is difficult for aging;
(6) C, N source utilize: this bacterium can utilize carbon sources such as glucose, fructose, maltose, mannose, glycerol, but does not utilize inulin, sorbose, rhamnose and lactose.As carbon source, conidium output is significantly higher than above-mentioned several kinds of carbon sources with glucose, and as carbon source, mycelium production is higher than other carbon sources with fructose.To 9 kinds of inorganic N cultivation situation such as nitro, nitroso-group, amino, this bacterium is to KNO
3Make good use of, and utilize nitro N better, but can not utilize NaNO than amino N
2And thiourea.This bacterium is quite responsive to the carbon source required amount.For example, carbon source maintained on 2% the normal growth developmental level, improve the N source or reduce the N source not quite coremium growth and sporulation quantity influence; But the N source is fixed on the normal growth level, carbon source is reduced to 0.5% from 2%, find not produce coremium, or little coremium produces.Otherwise, carbon source is improved 10 times (20%), then coremium is many and difficult aging, but generation of conidium is later;
(7) light: this bacterium has obvious phototaxis, and coremium growth is tilted towards light source direction, and it is prosperous secretly to cultivate mycelial growth, only produces sclerotium and does not form coremium.Natural light and light all can stimulate coremium growth and conidium to form;
(8) this bacterium has strong radioprotective phenomenon, and flat-plate bacterial colony shines 281h from 30Wt ultraviolet source 0.5m place, moves and does not see inactivation after growing, and does not also see growth failure;
(9) solid fermentation: do culture medium with corn such as Semen Tritici aestivis, inoculum concentration 3%～5% (percentage by weight), inoculation back 3～5 days, visible canescence fine hair shape mycelium covers stromal surface.Formed sclerotium in 20～25 days and grow with pin main body on similar coremium.Coremium is yolk yellow, upright, column or palmate branch, and 20～30 (～80) * 3～5mm produce a large amount of conidium after common 30 days, the afterwards withered gradually lodging of coremium.
Except that the effective component extracts that contains said Periostracum cicadae artificial culture product or Periostracum cicadae artificial culture thing, also can contain pharmaceutically acceptable carrier in the Chinese medicine health care product of the present invention, the weight percent content of carrier can be 1-50%, is preferably 20%-38%.This area known carrier of branch art personnel includes, but is not limited to lactose, corn starch or derivatives thereof, Talcum, vegetable oil, wax, fat, polyol for example Polyethylene Glycol, water, sucrose, ethanol, glycerol; Like that, various antiseptic, lubricant, dispersant, correctives.Preserve moisture cut to pieces, antioxidant, sweeting agent, coloring agent, stabilizing agent, salt, buffer is like that also can add wherein, these materials are used to help the stability of filling a prescription as required or help to improve activity or its biological effectiveness or under oral situation, produce acceptable mouthfeel or abnormal smells from the patient.
Chinese medicine health care product of the present invention can be processed the dosage form of any routine according to the universal method on the pharmaceutics, comprises oral formulations.Preferably capsule, tablet, granule, pill, powder, syrup most preferably are capsule.
Chinese medicine health care product of the present invention can be used for improving anemia or improves sleep.
Preferable, said anemia is an iron deficiency anemia.
The present invention also further provides the method for preparing of said Chinese medicine health care product, and said method for preparing is selected from any in the following method:
1) pulverize: with directly pulverizing after the said Periostracum cicadae artificial culture product drying, sieving makes said Chinese medicine health care product;
2) said Periostracum cicadae artificial culture product effective component extracting after drying is obtained said Chinese medicine health care product.
The method of said Periostracum cicadae artificial culture product effective component extracting can adopt conventional Chinese medicine extraction process of effective component to extract its effective ingredient, as adopting solvent extraction.
Preferable, said employing solvent extraction is extracted for the mixed liquor that adopts organic solvent, water or organic solvent and water; Said organic solvent can be selected from one or more the mixture in ethanol, ethyl acetate, chloroform, acetone, the petroleum ether; The mixed solution of said organic solvent and water is preferably ethanol water.Further being preferably water carries.
Preferably, said water extraction is: with the decocte with water of said Periostracum cicadae artificial culture thing (institute's water consumption is 6-10 a times of Periostracum cicadae artificial culture thing weight), filter, be concentrated into an amount of.Said decoction number of times is preferably 2～3 times, and each decocting time is 15～60 minutes.
Chinese medicine health care product of the present invention can have improves multiple health care such as sleeping, improve anemia.
Fermentation process of the present invention and the Paecilomyces cicadidae(Miquel)Samson of the artificial culture that obtains, growth cycle is short, output is higher, and bioactive ingredients also is much higher than wild Periostracum cicadae, has more value than wild Periostracum cicadae.Existing a large amount of health promoting product listings such as Cordyceps class such as Cordyceps, and the health promoting product of Periostracum cicadae does not almost have, the prospect of visible artificial culture Paecilomyces cicadidae(Miquel)Samson health promoting product is quite wide, can give full play to the value of its medical, edible.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction scope of the present invention.
Embodiment 1 Paecilomyces cicadidae(Miquel)Samson artificial culture
1) Paecilomyces cicadidae(Miquel)Samson bacterial strain CGMCC No.3453 is inoculated on the PSA slant medium, under 20 ℃, cultivated 10 days;
2) scrape from the inclined-plane and get mycelium inoculation to the triangular flask fluid medium that is the basis with PDB liquid, under 20 ℃, 140rpm cultivates and can make the Paecilomyces cicadidae(Miquel)Samson shake-flask seed liquid in 3 days; With the Paecilomyces cicadidae(Miquel)Samson shake-flask seed liquid by volume percentage ratio be that the 10% PSB liquid that inserts after the sterilization is in the fermentation tank of seed culture medium, under 20 ℃, stir speed (S.S.) 140rpm cultivates and can make Paecilomyces cicadidae(Miquel)Samson fermentation tank seed liquor in 5 days;
3) be in the solid phase culture medium after 5% access is sterilized by weight percentage with the fermentation tank seed liquor that makes; In temperature is 18 ℃; Humidity is under 60% the condition, during solid fermentation, to increase artificial scattering light source (increasing continuously intensity of illumination 150Lux); And regularly vented exhaust (carrying out vented exhaust at interval in 8 hours, each air-supply, aerofluxus 30 minutes) is left standstill the closed cultivation sporophore of gathering after 40 days and is obtained culture.
The used solid medium proportioning of step 3) is Semen Tritici aestivi 35 weight portions, Semen setariae 4 weight portions, and ferrous porphyrin 1 weight portion, water are 60 weight portions; Its preparation method is: Semen Tritici aestivi, Semen setariae are boiled, with ferrous porphyrin with boil proportionally mix homogeneously of Semen Tritici aestivi, Semen setariae.
Embodiment 2 Paecilomyces cicadidae(Miquel)Samson artificial cultures
1) Paecilomyces cicadidae(Miquel)Samson bacterial strain CGMCC No.3453 is inoculated on the PSA slant medium, under 22 ℃, cultivated 9 days;
2) scrape from the inclined-plane and get mycelium inoculation to the triangular flask fluid medium that is the basis with PDB liquid, under 22 ℃, 140rpm cultivates and can make the Paecilomyces cicadidae(Miquel)Samson shake-flask seed liquid in 3 days; With the Paecilomyces cicadidae(Miquel)Samson shake-flask seed liquid by volume percentage ratio be that the 10% PSB liquid that inserts after the sterilization is in the fermentation tank of seed culture medium, under 22 ℃, stir speed (S.S.) 150rpm cultivates and can make Paecilomyces cicadidae(Miquel)Samson fermentation tank seed liquor in 4 days;
3) be in the solid phase culture medium after 5% access is sterilized by weight percentage with the fermentation tank seed liquor that makes; In temperature is 21 ℃; Humidity is under 70% the condition, during solid fermentation, to increase artificial scattering light source (increasing continuously intensity of illumination 150Lux); And regularly vented exhaust (carrying out vented exhaust at interval in 8 hours, each air-supply, aerofluxus 40 minutes) is left standstill the closed cultivation sporophore of gathering after 37 days and is obtained culture.
The used solid medium of step 3) is with embodiment 1.
Embodiment 3 Paecilomyces cicadidae(Miquel)Samson artificial cultures
1) Paecilomyces cicadidae(Miquel)Samson bacterial strain CGMCC No.3453 is inoculated on the PSA slant medium, under 25 ℃, cultivated 8 days;
2) scrape from the inclined-plane and get mycelium inoculation to the triangular flask fluid medium that is the basis with PDB liquid, under 25 ℃, 140rpm cultivates and can make the Paecilomyces cicadidae(Miquel)Samson shake-flask seed liquid in 3 days; With the Paecilomyces cicadidae(Miquel)Samson shake-flask seed liquid by volume percentage ratio be that the 10% PSB liquid that inserts after the sterilization is in the fermentation tank of seed culture medium, under 25 ℃, stir speed (S.S.) 140rpm cultivates and can make Paecilomyces cicadidae(Miquel)Samson fermentation tank seed liquor in 2 days;
3) be in the solid phase culture medium after 5% access is sterilized by weight percentage with the fermentation tank seed liquor that makes; In temperature is 23 ℃; Humidity is under 80% the condition, during solid fermentation, to increase artificial scattering light source (increasing continuously intensity of illumination 150Lux); And regularly vented exhaust (carrying out vented exhaust at interval in 8 hours, each air-supply, aerofluxus 40 minutes) is left standstill the closed cultivation sporophore of gathering after 35 days and is obtained culture.
The used solid medium of step 3) is with embodiment 1.
The effective ingredient of embodiment 4 Paecilomyces cicadidae(Miquel)Samson artificial culture products detects
The artificial culture Paecilomyces cicadidae(Miquel)Samson, except the used solid culture based formulas of step 3) adopts respectively the following prescription, all the other steps are all with embodiment 3.
Comparative example 1: Semen Tritici aestivi 35 weight portions, Semen setariae 4 weight portions, ferrous porphyrin 1 weight portion, water are 60 weight portions
Comparative example 2: Semen Tritici aestivi 32 weight portions, Semen setariae 6 weight portions, ferrous porphyrin 2 weight portions, water are 60 weight portions
Comparative example 3: Semen Tritici aestivi 28 weight portions, Semen setariae 8 weight portions, ferrous porphyrin 4 weight portions, water are 66 weight portions
Comparative Examples: Semen Tritici aestivi 35 is heavy two parts, water 65 weight portions
The assay method of accordinging to ferrum, magnesium, manganese in the GB/T 5009.90-2003 food after the sporophore drying of results is respectively detected its iron content.
The result is:
Comparative example 1: iron content 190mg/100g
Comparative example 2: iron content 227mg/100g
Comparative example 3: iron content 318mg/100g
Comparative Examples: iron content 3.3mg/100g
The result shows that the Paecilomyces cicadidae(Miquel)Samson culture iron content that method for preparing of the present invention obtains is high, is beneficial to and enriches blood.
One of preparation of embodiment 5 Chinese medicine health care product
With the fresh Paecilomyces cicadidae(Miquel)Samson culture of gathering, 70 ℃ of dry 10h pulverize through pulverizer, cross 100 eye mesh screens, and irradiation sterilization is got powder 500g, incapsulates, and processes 1000, promptly gets.
Two of the preparation of embodiment 6 Chinese medicine health care product
The fresh Paecilomyces cicadidae(Miquel)Samson culture of gathering is added 8 times of distilled water decoct altogether 2 times, each 0.5 hour, decocting liquid adopted vacuum filtration, removes impurity, must filtrate, and the decompression thin film concentration is to an amount of, and spray drying promptly gets xeraphium.
Embodiment 7 Chinese medicine health care product acute oral toxicity tests
1. sample: adopt the Paecilomyces cicadidae(Miquel)Samson culture of embodiment 3 preparations to utilize the method for embodiment 5 to obtain the health care sample
2. sample character: bulk solid
3. given the test agent preparation: take by weighing sample 10000mg, adding distil water is to 40mL, fully behind the mixing as given the test agent.
4. laboratory animal:
20 of Kunming mouses, male and female half and half, body weight 19-22 gram is provided by Shanghai Slac Experimental Animal Co., Ltd..Production licence number: SCXK (Shanghai) 2007-0005.20-25 ℃ of receptacle temperature, relative humidity: 40-70%.
Laboratory animal occupancy permit number: SYXK (Shanghai) 2007-0008.Mouse feed has the two lion laboratory animal feed branches in Suzhou
Technical service affair company limited provides registration card number: the E of Soviet Union raises new word (2002) 006.
5. instrument and equipment: electronic scale ACS-3A 90401582, balance BP310S-91006909
6. laboratory method method:
6.1 animal fasting (can't help water) is after 16 hours, selects each 10 of female, male mices for use by the body weight requirement, divides to be put in two mouse cages, is no more than 3g with the difference of body weight between the sex mice.
6.2 adopt the maximum tolerated dose method that laboratory animal is contaminated given the test agent, mice is irritated gastric capacity by each 0.4mL/20g weighing machine by only weighing, secondary is to mouse stomach in 24 hours, and irritating stomach blanking time is 6 hours.
6.3 after the contamination, observe relief state, body weight change, poisoning sign and the death condition etc. of animal, the observation period is a week.
6.4 weigh to animal once more in the experiment end.The dead animal and the execution animal that expires are carried out obduction, and the perusal general pathology changes situation.
6.5 experiment overall process and observed content are all done itemized record, try to achieve chmice acute per os MTD by maximum tolerated dose method experimental result.
7. result:
Table male and female chmice acute per os toxicity test result
1) main physical signs performance: each treated animal of duration of test is movable normal, and the hair color glossiness is good, does not see any poisoning sign and death; Expire and put to death animal, each internal organs situation of gross anatomy perusal, no abnormality seen.
2) female mice: MTD＞10000mg/Kg
Female mice: MTD＞10000mg/Kg
Five, conclusion:
Sample to the maximum tolerated dose MTD of male and female chmice acute per os toxicity test all greater than 10000mg/Kg.The nontoxic level in true border.
The test of embodiment 6 health cares
One, Periostracum cicadae artificial culture thing is to the improvement effect of iron deficiency anemia
1, experiment material
1.1 laboratory animal: healthy ablactation rat, 10 every group.
1.2 sample: adopt the Paecilomyces cicadidae(Miquel)Samson culture of embodiment 3 preparations to utilize the method for embodiment 5 to prepare sample 1 respectively.Be configured to 200mg/ml solution with deionized water, subsequent use.
1.3 instrument
Instrument: spectrophotometer, spectrofluorophotometer or 930 type fluorophotometers, centrifuge, DL device;
1.4 reagent: the heparin of one 12500 unit of anticoagulant heparin agent is diluted to 25mL (1mL=500 unit) with 0.9% normal saline; 5% (W/V) kieselguhr normal saline suspension takes by weighing 5g kieselguhr and adds 0.9% normal saline to 100mL; Ethyl acetate and acetic acid mixed liquor; 0.5N HCl; The protoporphyrin titer; Protoporphyrin standard stock solution (50 μ g/mL): take by weighing the 5mg protoporphyrin, add the 4mL dehydrated alcohol and make it dissolving, be diluted to 100mL with 1.5N HCl; Protoporphyrin standard intermediate liquid (1.0 μ g/mL): get 2mL protoporphyrin stock solution, be diluted to 100mL with ethyl acetate and acetic acid (4: 1) mixed liquor; Protoporphyrin standard application liquid (0.1 μ g/mL): get 0.1mL protoporphyrin intermediate liquid, be diluted to 10mL with ethyl acetate and acetic acid (4: 1) mixed liquor.
2, experimental technique:
40 of rats are divided four groups, receive high, medium and low three dose groups of reagent and matched group; Adapt to after 5 days, feed with low iron base feedstuff and 3 weeks caused the iron deficiency model, per os filling every day stomach gives given the test agent; Dosage deionized waters such as matched group administration, 4 weeks of successive administration.Experiment is used the etherization rat after finishing, and behind abdominal aortic blood, wins liver and spleen, puts-30 ℃ of refrigerators and places subsequent use.
Low dose group dosage: irritate gastric capacity 250mg/kg body weight every day
Middle dose groups dosage: irritate gastric capacity 500mg/kg body weight every day
High dose group dosage: irritate gastric capacity 1000mg/kg body weight every day
3, test rating: body weight; Hemoglobin; Free protoporphyrin in the erythrocyte.
3.1 hemoglobinometry (the high iron processes of cyaniding)
Take by weighing sodium bicarbonate (NaHCO
3, AR) 140mg, potassium ferricyanide 200mg, potassium cyanide 50mg with dissolved in distilled water and be diluted to 1000mL. and be stored in the brown reagent bottle, are stored in 4 ℃ of refrigerators.
Draw 2.5mL reagent in 5mL test tube with cover, get rat tail blood or venous blood 10 μ L are positioned over the test tube of putting into reagent with 10 μ L hematochrome suction pipes (or quantitative capillary tube); Mixing was placed 15 minutes.
Select 0.5cm optical path cuvette for use, under the 540nm wavelength, regulate instrument zero, measure the absorbance of each sample cell, measure the absorbance of hemoglobin standard and reference standard material simultaneously, draw the standard curve of hemoglobin with reagent.Look into that standard curve can be tried to achieve testing sample and the reference standard material gets content of hemoglobin g/dl or g/l, calculate the response rate of reference material.
3.2 free protoporphyrin is measured in the erythrocyte
On special filter paper, deposit droplets of whole blood in 4 ℃ of refrigerators subsequent use (time is no more than 2 months) after to be dried.During mensuration, the filter paper that will have drop of blood with card punch is squeezed in the teat glass, adds 2 tripolite solutions; Put into 4 ℃ of refrigerator overnight eluting; Add ethyl acetate: the extraction of glacial acetic acid (4: 1) mixed liquor, centrifugal 10min gets supernatant and adds 0.5mol/L hydrochloric acid; It is centrifugal to vibrate, and takes off layer liquid and uses fluorescent spectrophotometer assay.
Calculate:
3.3 the result judges: the given the test agent group is compared with matched group; Hemoglobin difference has significance; And average rising amplitude reaches more than the 10g/L before and after it; Free protoporphyrin is compared difference with matched group in the given the test agent erythrocyte has significance, and it is positive that this given the test agent of decidable improves anemia function zoopery result.
3.4 date processing: adopt the t check that data are handled.
4, result
4.1 receive the influence of reagent: the content of hemoglobin difference not statistically significant (P＞0.05) of respectively organizing rat before the experiment to hemoglobin; Content of hemoglobin and the value added of experiment back each dose groups rat of Periostracum cicadae culture all are significantly higher than matched group (P＜0.05), see table 1.
Table 1 Periostracum cicadae culture is to the influence (g/L) of rat hemoglobin
*Compare P＜0.05 with matched group
4.2 respectively organize the comparison of rat EP content: the protoporphyrin content difference not statistically significant (P＞0.05) of respectively organizing rat before the experiment; Difference all significantly is lower than matched group (P＜0.05) before and after protoporphyrin content of experiment back each dose groups rat of Periostracum cicadae culture and the experiment, sees table 2.
Table 2 is respectively organized the comparison of rat EP content
*Compare P＜0.05 with matched group
Can find out that from table 1 and table 2 the Periostracum cicadae culture has the improvement effect to nutritional anemia.
Two, Periostracum cicadae artificial culture thing improves sleep function
1, prolongs the pentobarbital sodium experiment length of one's sleep
1.1, experiment material
Laboratory animal: adult mice, single sex, the 18-22 gram, every group of 10-15 is only.
Sample: the same.
Reagent: pentobarbital sodium (using preceding fresh)
1.2 experimental technique
Carry out preliminary experiment earlier before doing formal experiment, confirm to make animal 100% sleeping, but do not make long pentobarbital sodium dosage (30-60mg/kg) length of one's sleep, formally test with this dosage.Three dose groups and a blank group are established in experiment; After the animal last gives solvent control and variable concentrations given the test agent; The peak effect occurred preceding 10-15 minute, and gave each treated animal lumbar injection pentobarbital sodium, injection volume is 0.2mL/20g; Can with the righting reflex loss be index, observe given the test agent and prolong pentobarbital sodium length of one's sleep.
Medication: per os is irritated stomach and is given given the test agent, dosage deionized waters such as matched group administration, every day 1 time, successive administration 30 days.Basic, normal, high dose groups dosage is the same.
1.3 date processing
2, pentobarbital sodium (or barbital sodium) sub-threshold dose hypnosis experiment
2.1 experiment material
Laboratory animal and sample are the same
Reagent: pentobarbital sodium (or barbital sodium), use preceding fresh.
2.2 experimental technique
Carry out preliminary experiment earlier before the formal experiment; Confirm pentobarbital sodium (or barbital sodium) subliminal hypnosis dosage (pentobarbital sodium 16-30mg/kg.bw or barbital sodium 100-150mg/kgbw), i.e. the maximum sub-threshold dose of the pentobarbital sodium that 80-90% mice righting reflex does not disappear.Animal divides into groups the same.After the animal last gives blank and variable concentrations given the test agent, the peak effect occurred preceding 10-15 minute, the maximum subliminal hypnosis dosage of each treated animal lumbar injection pentobarbital sodium writes down sleeping number of animals (righting reflex loss reaches person more than 1 minute) in 30 minutes.Experiment should be carried out under 24-25 ℃ of quiet environment.
3, result
3.1 to the influence of the length of one's sleep of pentobarbital sodium inducing mouse: middle and high dose groups is longer than matched group (P＜0.05) length of one's sleep, sees table 3.
Table 3 Periostracum cicadae culture is to the pentobarbital sodium inducing mouse influence of the length of one's sleep (n=12)
*Compare P＜0.05 with matched group
The result shows that Periostracum cicadae is cultivated object height, middle dose groups can prolong the inductive mouse sleep time of pentobarbital sodium.
3.2 the influence to pentobarbital sodium sub-threshold dose syngignoscism: the sleeping incidence rate of the mice of three dose groups mices of Periostracum cicadae culture under the pentobarbital sodium HD is induced under threshold all is higher than the blank group, sees table 4
Table 4 Periostracum cicadae culture is to the influence (n=12) of pentobarbital sodium sub-threshold dose syngignoscism
*Compare P＜0.05 with matched group
The result shows that three dose groups of Periostracum cicadae culture all can increase the inductive mice sleep incidence rate of sub-threshold dose pentobarbital sodium.
Three, conclusion
Above experimentation shows, the Periostracum cicadae culture has and obviously improves nutritional anemia and improve the sleep effect.
Claims (10)
1. Chinese medicine health care product, the main pharmacodynamics composition in the said Chinese medicine health care product is the effective component extracts of Periostracum cicadae artificial culture product or Periostracum cicadae artificial culture product.
2. Chinese medicine health care product according to claim 1 is characterized in that said Periostracum cicadae artificial culture product is made by following method:
1) slant strains preparation: the Paecilomyces cicadidae(Miquel)Samson inoculation to slant medium, under 20-25 ℃, was cultivated 3-10 days;
2) preparation of seed liquor: scrape from the inclined-plane and to get mycelium inoculation to fluid medium, under 20-25 ℃, 140-160rpm cultivates and makes the Paecilomyces cicadidae(Miquel)Samson seed liquor;
3) with step 2) in the Paecilomyces cicadidae(Miquel)Samson seed liquor that makes insert in the solid phase culture medium after the sterilization for 5%-10% by weight percentage; In temperature is 18-23 ℃; Humidity is under the condition of 60-80%; Leave standstill closed cultivation 35-40 days, adopting intensity of illumination between culture period is the uninterrupted full gloss spectrum artificial light source irradiation of 100-150lux;
4) cultured products of gathering.
3. like the said Chinese medicine health care product of claim 2, it is characterized in that the said slant medium of step 1) is PSA, PDA or YMB solid medium; Step 2) said fluid medium is PSB or PDB fluid medium.
4. like the said Chinese medicine health care product of claim 2, it is characterized in that the used Paecilomyces cicadidae(Miquel)Samson bacterial strain of step 1) is Paecilomyces cicadidae(Miquel)Samson bacterial strain Paecilomyces cicadae (Miq.) Samson CGMCC No.3453.
5. like the said Chinese medicine health care product of claim 2, it is characterized in that the said solid medium of step 3) comprises the raw material components of following weight portion:
6. like the said Chinese medicine health care product of claim 2, it is characterized in that the said cultured products of step 4) is the Paecilomyces cicadidae(Miquel)Samson sporophore.
7. like the arbitrary said Chinese medicine health care product of claim 1-6, it is characterized in that also containing weight percent content in the said Chinese medicine health care product is the pharmaceutically acceptable carrier of 1-50%.
8. like the method for preparing of the said Chinese medicine health care product of the arbitrary claim of claim 1-7, said method for preparing is selected from any in the following method:
1) with directly pulverizing after the said Periostracum cicadae artificial culture product drying, sieving makes said Chinese medicine health care product;
2) said Periostracum cicadae artificial culture product effective component extracting after drying is obtained said Chinese medicine health care product.
9. like the method for preparing of the said Chinese medicine health care product of claim 8, it is characterized in that said method 2) in the method for Periostracum cicadae artificial culture product effective component extracting for adopting the mixed liquor extraction of organic solvent, water or organic solvent and water.
10. has the purposes in the health product that improve anemia or improve the sleep effect like the said Chinese medicine health care product of the arbitrary claim of claim 1-7 in preparation.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104872758A (en) * | 2015-05-25 | 2015-09-02 | 亳州千草药业有限公司 | Preparation method of cordyceps sobolifera and burdock oral liquid |
| CN105981584A (en) * | 2015-02-15 | 2016-10-05 | 浙江泛亚生物医药股份有限公司 | Method for cultivating calcium-rich cordyceps sobolifera |
| CN105993584A (en) * | 2015-03-26 | 2016-10-12 | 浙江泛亚生物医药股份有限公司 | Artificial culturing method for magnesium-rich cordyceps sobolifera |
| CN106318875A (en) * | 2015-06-18 | 2017-01-11 | 浙江泛亚生物医药股份有限公司 | Cordyceps sobolifera two-way artificial culture method |
| CN106797801A (en) * | 2015-11-26 | 2017-06-06 | 浙江泛亚生物医药股份有限公司 | A kind of artificial culture method of cicada fungus |
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| CN108371282A (en) * | 2016-10-12 | 2018-08-07 | 浙江泛亚生物医药股份有限公司 | A kind of cicada fungus rice and preparation method thereof |
| CN111434342A (en) * | 2018-12-25 | 2020-07-21 | 葡萄王生技股份有限公司 | Cordyceps cicadae mycelium active substance for preventing and/or improving acute lung injury |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1227869A (en) * | 1998-03-02 | 1999-09-08 | 中国农业科学院饲料研究所 | Process for producing Fe-Zn enriched trace element thallus |
| CN1563348A (en) * | 2004-04-16 | 2005-01-12 | 浙江省农业科学院 | New strain APC-20 of Paecilomyces cicadae and fementation process for artificial culture |
| CN102240301A (en) * | 2010-05-14 | 2011-11-16 | 浙江泛亚生物医药股份有限公司 | Single-composition traditional Chinese medicine preparation, and preparation method and application thereof |
-
2011
- 2011-03-11 HK HK11102500A patent/HK1146565A2/en not_active IP Right Cessation
-
2012
- 2012-02-02 CN CN201210023412.8A patent/CN102552335B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1227869A (en) * | 1998-03-02 | 1999-09-08 | 中国农业科学院饲料研究所 | Process for producing Fe-Zn enriched trace element thallus |
| CN1563348A (en) * | 2004-04-16 | 2005-01-12 | 浙江省农业科学院 | New strain APC-20 of Paecilomyces cicadae and fementation process for artificial culture |
| CN102240301A (en) * | 2010-05-14 | 2011-11-16 | 浙江泛亚生物医药股份有限公司 | Single-composition traditional Chinese medicine preparation, and preparation method and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| 《中国农技推广》 20090531 任文彦等 名贵药(食)用菌__蛹虫草速生高产栽培技术要点 28-29 1-10 第25卷, 第05期 * |
| 任文彦等: "名贵药(食)用菌――蛹虫草速生高产栽培技术要点", 《中国农技推广》, vol. 25, no. 05, 31 May 2009 (2009-05-31), pages 28 - 29 * |
| 陈玲等: "蝉花的研究进展", 《中国中医药科技》, vol. 16, no. 02, 31 March 2009 (2009-03-31), pages 159 - 160 * |
| 陈祝安等: "蝉花的人工培养及其药理作用研究", 《真菌学报》, vol. 12, no. 2, 30 June 1993 (1993-06-30), pages 138 - 144 * |
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| HK1146565A2 (en) | 2011-06-17 |
| CN102552335B (en) | 2014-12-17 |
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