CN105125942A - Extracting method and application of plant composite containing mycelia of cultured cordyceps sinensis - Google Patents
Extracting method and application of plant composite containing mycelia of cultured cordyceps sinensis Download PDFInfo
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- CN105125942A CN105125942A CN201510643115.7A CN201510643115A CN105125942A CN 105125942 A CN105125942 A CN 105125942A CN 201510643115 A CN201510643115 A CN 201510643115A CN 105125942 A CN105125942 A CN 105125942A
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- cordyceps mycelium
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Abstract
The invention provides an extracting method of a plant composite containing mycelia of cultured cordyceps sinensis. The plant composite is prepared from the raw material herbs of 100 g of mycelia of cultured cordyceps sinensis, 60 g of gentiana apiata, 80 g of salvia chinensis and 160 g of Chinese pennisetum. The plant composite is prepared by the adoption of the microwave extracting method, in this way, the content is greatly increased, and the usage amount is reduced. The invention further provides an application of the plant composite containing the mycelia of cultured cordyceps sinensis in preparation of medicine for restraining C57BL/6 mice fibrosarcoma S180 cell proliferation.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to a kind of vegetable composition extracting method containing Cordyceps mycelium and application.
Background technology
Containing the vegetable composition of Cordyceps mycelium by Cordyceps mycelium 100g, Herba Gentianae tsinglingensis 60g, Herba Salviae Chinensis 80g, Herba penniseti 160g make as crude drug, and current extracting method decocts with water secondary, each 2-3 hour, collecting decoction, filters, is condensed into thick paste, granulate, drying, is pressed into 1000, sugar coating, obtain, can immunity be improved.
In prior art, not yet there is the vegetable composition containing Cordyceps mycelium extracting the report adopting microwave technology in preparation, and adopt the method for soak by water, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, be inconvenient to take, had a strong impact on this product and applied clinically.
In prior art, the every sheet 0.4g of vegetable composition containing Cordyceps mycelium, each 4,3 times on the one, adopt the every sheet 0.4g of vegetable composition containing Cordyceps mycelium that the present invention is prepared into, but the medical material amount contained is original 2 times, therefore only needs 2 at every turn, within 1st, take 3 times, under the condition with more active component, greatly reduce dose.
Summary of the invention
Goal of the invention: in order to solve the problem, the object of the present invention is to provide a kind of vegetable composition extracting method containing Cordyceps mycelium.
Another object of the present invention is to provide a kind of application of vegetable composition in preparation suppression C57BL/6 mouse fibrosarcoma S180 cell proliferation containing Cordyceps mycelium.
Technical scheme: the object of the invention is by following scheme realize:
A kind of vegetable composition extracting method containing Cordyceps mycelium, vegetable composition is by Cordyceps mycelium 100g, Herba Gentianae tsinglingensis 60g, Herba Salviae Chinensis 80g, Herba penniseti 160g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
Above-mentioned a kind of vegetable composition extracting method containing Cordyceps mycelium, in preparation method, microwave extracting power is 500W, each extraction 6 minutes.
A kind of vegetable composition containing Cordyceps mycelium suppresses the application in C57BL/6 mouse fibrosarcoma S180 cell proliferation in preparation, containing the vegetable composition of Cordyceps mycelium by Cordyceps mycelium 100g, Herba Gentianae tsinglingensis 60g, Herba Salviae Chinensis 80g, Herba penniseti 160g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
Above-mentioned a kind of vegetable composition containing Cordyceps mycelium suppresses the application in C57BL/6 mouse fibrosarcoma S180 cell proliferation in preparation, the power of microwave extracting described in preparation method is 500W, each extraction 6 minutes.
Detailed description of the invention
Form by the following examples, foregoing of the present invention is described in further detail again, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Cordyceps mycelium 100g, Herba Gentianae tsinglingensis 60g, Herba Salviae Chinensis 80g, Herba penniseti 160g, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, every sheet 0.4g.
Embodiment 2
Get Cordyceps mycelium 100g, Herba Gentianae tsinglingensis 60g, Herba Salviae Chinensis 80g, Herba penniseti 160g, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into capsule.
Embodiment 3
Get Cordyceps mycelium 100g, Herba Gentianae tsinglingensis 60g, Herba Salviae Chinensis 80g, Herba penniseti 160g, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into granule.
The Herba Gentianae tsinglingensis is the herb of Gentianaceae agueweed Herba Gentianae tsinglingensis GentianaapiataN.E.Brown.Summer gathers, and dries.Regulating menstruation and activating blood, clearing away heat to improve acuity of vision, diuresis.For menoxenia, dysmenorrhea, dizzy, insomnia, dysuria, stranguria, metrorrhagia, leucorrhea, dysentery, stomachache.
Herba Salviae Chinensis, Latin is called SalviachinensisBenth., annual herb, high 20-70cm.Root multi-branched, taproot is not obvious, yellowish-brown.Herb is by the pubescence of giving birth to or long pubescence.Single or the branch of stem, upright or base portion tilts, four prismatics.Heat-clearing and toxic substances removing, invigorating blood circulation analgesia.For icterohepatitis, cancer, nephritis, leucorrhea, dysmenorrhea, tuberculous lymphadenitis, elephantiasis; Facial paralysis is controlled in external, mastitis, furuncle and phyma.The present invention is with all herbal medicine.
Herba penniseti is the herb of grass Herba penniseti Pennisetumalopecuroides (L.) Spreng..Sweet; Property is put down.Clearing away lung-heat to relieve cough; Cooling blood for improving eyesight.Cure mainly cough due to lung-heat; Conjunctival congestion and swelling pain.
Cordyceps mycelium is exactly that the Cordyceps fungus be separated in Cordyceps body obtains through cultivating.
Embodiment 4: the vegetable composition containing Cordyceps mycelium suppresses the experimentation data of C57BL/6 mouse fibrosarcoma S180 cell proliferation
1 experiment material
1.1 experiment cell strains
C57BL/6 mouse fibrosarcoma S180 cell, Nanjing Medical University's Experimental Animal Center laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: the present invention is containing the vegetable composition of Cordyceps mycelium: prepare by embodiment 1 method.
Medicinal liquid liquid storage: take 100mg containing the vegetable composition of Cordyceps mycelium, be dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ ldoff pipe subpackages ,-20 DEG C of storage, 0.2 μm of frit dehydrated alcohol prepares against the use of matched group simultaneously.
1.3 experiment reagent
DMEM (GIBCO company Cat.No.12100-061Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHCO3 (Shanghai hundred million chemical reagent company limited Cat.No.11810-033Lot.No.1088387 of a specified duration); Trypsin (AMRESCO company lot number: 2010/04); EDTA (AMRESCO company lot number: 2009/10); PenicillinGSodiumSalt (AMRESCO company lot number: 2010242); StreptomycinSulfate (AMRESCO company lot number: 2010382); Dehydrated alcohol (Nanjing Chemistry Reagent Co., Ltd.'s lot number: 080310182); MTT (Biosharp lot number: 0793); PBS (laboratory autogamy);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRAMAX190); CO2 incubator (FORMA model: 3111); Super-clean bench (safe and sound Inc. of Su Jing group moulding number: SW-CJ-ZFD); Pure water instrument (Spring company of U.S. model: S/N020579); Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi company limited model: BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air dry oven (the accurate experimental facilities company in Shanghai model: DHG9123A); Refrigerator (Siemens Company's model: KG18V21TI); Liquid nitrogen container (CBS model: 2001); Low speed centrifuge (Anting Scientific Instrument Factory, Shanghai's model: KA-1000); 0.2 μm of filter (MILLIPORE model: SLGP033RB); 10cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2 experimental techniques
1) C57BL/6 mouse fibrosarcoma S180 cell DMEM+10%FBS in 37 DEG C, 5%CO2 carries out cellar culture (10cm culture dish), when Growth of Cells is to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml0.25% trypsin-0.04%EDTA, after 37 DEG C of digestion 2min, add 5ml complete medium neutralization reaction wherein, proceeded in centrifuge tube after piping and druming cell, the centrifugal 5min of 1000rpm, adjustment concentration of cell suspension 3 × 10
4individual/ml.
2) enter in 96 well culture plates by cell kind, every hole adds cell suspension 180 μ l, and culture plate puts into cell culture incubator (37 DEG C, 5%CO2) cellar culture.
3) according to cell growth status, generally grow to 50%-70%, add the vegetable composition solution containing Cordyceps mycelium, continue to cultivate 24h.
4) add 20 μ lMTT solution (5mg/ml, i.e. 0.5%MTT) after 24h, continue to cultivate 4h.
5) after 4h, buckle method removes supernatant, pats dry gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, crystal is fully dissolved.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm place.
6) background (do not add cell, only add culture fluid) is set, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide) simultaneously, often organizes the multiple hole of setting 6.
7) result represents with the suppression ratio of medicine to cell:
Cell proliferation suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%.Experiment repetition 3 times.
3 statistical dispositions
Adopt the correlation analysis in MicrosoftExcel2003 software and Studentt inspection, data represent with mean ± S.D..
4 experimental results
Statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to C57BL/6 mouse fibrosarcoma S180 cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), has pole significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 contains the vegetable composition of Cordyceps mycelium to C57BL/6 mouse fibrosarcoma S180 cell inhibitory effect influence research
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
Vegetable composition containing Cordyceps mycelium can suppress C57BL/6 mouse fibrosarcoma S180 cell proliferation, and reduce the Growth of Cells number of C57BL/6 mouse fibrosarcoma S180 cell, this effect is dose dependent.
Claims (4)
1. the vegetable composition extracting method containing Cordyceps mycelium, it is characterized in that vegetable composition is by Cordyceps mycelium 100g, Herba Gentianae tsinglingensis 60g, Herba Salviae Chinensis 80g, Herba penniseti 160g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
2. a kind of vegetable composition extracting method containing Cordyceps mycelium according to claim 1, is characterized in that in preparation method, microwave extracting power is 500W, each extraction 6 minutes.
3. one kind is being suppressed the application in C57BL/6 mouse fibrosarcoma S180 cell proliferation containing the vegetable composition of Cordyceps mycelium, it is characterized in that the vegetable composition containing Cordyceps mycelium is by Cordyceps mycelium 100g, Herba Gentianae tsinglingensis 60g, Herba Salviae Chinensis 80g, Herba penniseti 160g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
4. a kind of vegetable composition containing Cordyceps mycelium, suppressing the application in C57BL/6 mouse fibrosarcoma S180 cell proliferation, is characterized in that the power of microwave extracting described in preparation method is 500W, each extraction 6 minutes according to claim 3.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101049336A (en) * | 2006-04-03 | 2007-10-10 | 黄振华 | New composition of medication for anti cancer |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101049336A (en) * | 2006-04-03 | 2007-10-10 | 黄振华 | New composition of medication for anti cancer |
Non-Patent Citations (1)
| Title |
|---|
| 李琴等: ""不同原料来源的虫草素的提取研究概况"", 《西北民族学报（自然科学版）》 * |
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