CN109806283A - A kind of phellinus igniarius mycelium flavones and its preparation method and application - Google Patents
A kind of phellinus igniarius mycelium flavones and its preparation method and application Download PDFInfo
- Publication number
- CN109806283A CN109806283A CN201910219490.7A CN201910219490A CN109806283A CN 109806283 A CN109806283 A CN 109806283A CN 201910219490 A CN201910219490 A CN 201910219490A CN 109806283 A CN109806283 A CN 109806283A
- Authority
- CN
- China
- Prior art keywords
- phellinus igniarius
- igniarius mycelium
- flavones
- extracting solution
- mycelium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of phellinus igniarius mycelium flavones, preparation method includes the following steps: to carry out phellinus igniarius mycelium inoculation fermentation culture, and freeze-drying obtains phellinus igniarius mycelium powder；Phellinus igniarius mycelium powder is dissolved in distilled water, cellulase is added, carries out hot water extraction, is centrifuged, it is spare to obtain cellulase degradation extracting solution for filtering；Gained precipitating is dissolved in ethyl alcohol and carries out ultrasonic extraction, is centrifuged, and it is spare to obtain ultrasonic extraction extracting solution for filtering；Merge cellulase degradation extracting solution and ultrasound extraction extracting solution, total extract rotary evaporation is concentrated, is filtered, sterile cryo is dry, obtains phellinus igniarius mycelium flavones dry powder.The problems such as phellinus igniarius mycelium flavones of the present invention can significantly inhibit the proliferation of human liver cancer cell, can be to avoid the biggish cytotoxicity such as chemotherapy and chemical synthetic drug, side effect and high relapse rate.
Description
Technical field
The invention belongs to fungi technical field, has a kind of phellinus igniarius mycelium flavones and its preparation method and application.
Background technique
Liver cancer is worldwide one of most common, the big malignant tumour of morbidity and mortality highest three, is averaged
There is nearly 1,000,000 liver cancer patient dead every year.The most effective means of liver cancer treatment be operation excision and liver transfer operation, however due to
Onset of liver cancer concealment, tempo is fast, grade malignancy is high, and most of liver cancer patients are found and when making a definite diagnosis to be clinical evening
Phase loses the chance of surgical eradication substantially, so that 5 years survival rates of patient are down to 25%.It is removed currently, liver cancer still lacks
Other effective treatment methods of operation and liver transfer operation, trace it to its cause has high complexity and uncertainty tight with onset of liver cancer
Close correlation.And postoperative high recurrence and metastatic rate and high mortality become the important shadow of patient's long-dated survival situation after operative treatment
The factor of sound.It is lower for medicines resistant to liver cancer reactivity, it is also easy to produce drug resistance, and drug toxicity is big, therefore develop more effective liver
Cancer therapeutic agent has urgent meaning.
The antitumor research of Chinese medicine in recent years is the new direction for treating malignant tumour, finds effective anticancer drug and method,
It is the important topic that world medicine faces, Chinese medicine carrys out the powerful mean of disease preventing and treating as motherland's medical use, with chemical synthetic drug
Object is compared, other than having many advantages, such as that materials are convenient, cheap and toxic side effect is small, it is often more important that, the advantage of Chinese medicine exists
In the biological behaviour for changing tumour cell to inhibit its growth development, and the normal function of body is protected, extends patient
Service life palliates the agonizing sufferings, and keeps good quality of life.In recent years, more and more researchs are dedicated to from natural drug such as Chinese medicine
Extract low, the powerful antitumoral compounds of toxicity.Chinese medicine can protect liver, and liver cell is promoted to repair.Mitigate various chemicotherapies
Toxic side effect.Chinese medicine has multi-level, multiple target and mutually coordinated intervention effect to liver cancer, is expected to become treatment liver
The ideal type drug of cancer.
Phellinus, also referred to as Sang Chen, Phellinus mushroom etc. belong to Basidiomycotina, Hymenomycetes, Aphyllophorales, perforated Cordycepps.It is wild
Phellinus is mainly grown on the trunks such as mulberry tree, pine tree, silver birch.Contain polysaccharide, flavones, phenols, triterpenes, amino in Phellinus
Sour isoreactivity substance.Wild Phellinus has the effects that antibacterial, anti-inflammatory, anti-oxidant.In Traditional Chinese Medicine, Phellinus is for treating blood
It collapses, blood strangury, the gynecological diseases such as amenorrhoea.Ketone compounds are a kind of compounds that nature is widely present.Result of study shows Huang
Ketone compounds have the function of preferable anti-oxidant, removing free radical, antitumor, antiviral, raising immunity of organisms etc., but
Flavone compound is that body cannot synthesize, and can only be obtained from food.Therefore, flavone compound is widely used in eating
In product and medicine.As that studies flavone compound gradually gos deep into, the demand in market is continuously increased, economic benefit
Also it gets a promotion.Therefore, the research and development of flavone compound have been to be concerned by more and more people, and are expected to a variety of as treating
The potential drug of disease.
Summary of the invention
It is an object of the present invention to provide a kind of phellinus igniarius mycelium flavones and its preparation method and application.
The present invention is achieved through the following technical solutions: a kind of phellinus igniarius mycelium flavones, and preparation method includes following step
It is rapid:
1) it ferments: inoculation fermentation culture being carried out to phellinus igniarius mycelium, freeze-drying obtains phellinus igniarius mycelium powder；
2) cellulase degradation extracts: phellinus igniarius mycelium powder being dissolved in distilled water, cellulase is added, carries out hot water leaching
It mentions, is centrifuged, it is spare to obtain cellulase degradation extracting solution for filtering；
3) ultrasound assisted extraction: being dissolved in ethyl alcohol for precipitating obtained by step 2) and carry out ultrasonic extraction, be centrifuged, and filtering obtains ultrasonic
It is spare to extract extracting solution；
4) merge cellulase degradation extracting solution and ultrasound extraction extracting solution, total extract rotary evaporation be concentrated, is filtered,
Sterile cryo is dry, obtains phellinus igniarius mycelium flavones dry powder.
Preferably, above-mentioned a kind of phellinus igniarius mycelium flavones, in step 1), the specific steps of the fermented and cultured are as follows:
200g, glucose 20g, peptone 2g, MgSO are weighed after potato decortication4·7H2O1.5g, KH2PO42g, water 1000 ml, 121
DEG C sterilizing 30min, prepare PDA liquid medium.28 DEG C after inoculation, 15 d of 150r/min constant temperature oscillator culture.
Preferably, above-mentioned a kind of phellinus igniarius mycelium flavones, in step 2), by solid-liquid ratio, phellinus igniarius mycelium powder: distillation
Water=1:10-1:30 (g/ml).
Preferably, above-mentioned a kind of phellinus igniarius mycelium flavones, in step 2), using phellinus igniarius mycelium powder as substrate, fiber
The additional amount of plain enzyme is 120-180 U/g.
Preferably, above-mentioned a kind of phellinus igniarius mycelium flavones, in step 2), 50-60 DEG C of the hot water extraction temperature, enzyme
Solve time 1-2h, pH value range 4.0-6.0.
Preferably, above-mentioned a kind of phellinus igniarius mycelium flavones, in step 3), by solid-liquid ratio, the sediment: ethyl alcohol
=1:10-1:30 (g/ml).
Preferably, above-mentioned a kind of phellinus igniarius mycelium flavones, in step 3), the volume fraction of ethanol is 50%-
60%.
Preferably, above-mentioned a kind of phellinus igniarius mycelium flavones, in step 3), the condition of ultrasonic extraction is that power is 80-150
W, ultrasonic time 20-30min.
A kind of application of the above-mentioned phellinus igniarius mycelium flavones in anti-liver cancer and anti-drug or health care product.
The beneficial effects of the present invention are:
1, the initiative technology of the present invention is using phellinus igniarius mycelium flavones as the functional raw material of anti-liver cancer and anti-, and formation has
The flavones health care product or drug of anti-liver cancer and anti-.Research and inquirement phellinus igniarius mycelium flavones of the present invention to the influence of human liver cancer cell and this
Influence the effect played in cell Proliferation.The results show that phellinus igniarius mycelium flavones can significantly inhibit the increasing of human liver cancer cell
It grows, can be to avoid the biggish cytotoxicity such as chemotherapy and chemical synthetic drug, side effect and high relapse rate the problems such as.
2, the flavones that the present invention is extracted using phellinus igniarius mycelium powder prepares anti-liver cancer and anti-health care product or medicine as active material
Product, extracting method is simple, and there is no the toxic side effects of synthesis medicament, and the requirement to equipment is low, intermediate product and the equal nothing of final product
Poison, highly-safe, production cost is low, improves the rehabilitation situation and quality of life of patient, and from a wealth of sources, is easy to get,
Therefore possess very broad mass market prospect, be suitble to industrial production.Certain ginseng is provided with clinical application further to further investigate
It examines and foundation.
Detailed description of the invention
Fig. 1 is that phellinus igniarius mycelium flavones inhibits HepG2 cell Proliferation.
Specific embodiment
A kind of preparation of phellinus igniarius mycelium flavones (one) the phellinus igniarius mycelium flavones of embodiment 1
Phellinus igniarius mycelium flavones step is extracted using cellulase degradation-ultrasound assisted extraction method of fractional steps process combination
It is as follows:
1) 200g, glucose 20g, peptone 2g, MgSO are weighed after potato decortication4·7H2O1.5g, KH2PO42g, water
1000ml, 121 DEG C of sterilizing 30min prepare PDA liquid medium.Phellinus igniarius mycelium is inoculated in PDA liquid medium, 28
DEG C, about 15d is cultivated under conditions of 150r/min, is collected mycelium, is saved backup after freeze-dried.
2) cellulase degradation extracts: pressing solid-liquid ratio 1g:30ml, phellinus igniarius mycelium powder is dissolved in distilled water, cellulose
Enzyme additional amount is 150U/g, carries out hot water extraction, controls 50 DEG C of bath temperature, enzymolysis time 2h, pH value 5.0,4000r/min
Centrifugation 15min is carried out, is filtered, it is spare to obtain cellulase degradation extracting solution, and precipitating continues ultrasound assisted extraction；
3) ultrasound assisted extraction: pressing solid-liquid ratio 1g:30ml, and step 2) gained sediment is dissolved in ethyl alcohol, ethyl alcohol volume point
Number is 60%, and ultrasonic power 95W, ultrasonic time 20min, 4000r/min carry out centrifugation 15min, and filtering obtains ultrasonic
It is spare to extract extracting solution；
4) merge cellulase degradation extracting solution and ultrasound extraction extracting solution, total extract rotary evaporation be concentrated, is filtered,
Sterile cryo is dry, obtains phellinus igniarius mycelium flavones dry powder.
(2) MTT experiment
The present embodiment uses HepG 2 cell, using anticancer outside mtt assay detection phellinus igniarius mycelium flavonoids
Activity, experimental procedure are as follows:
1) liver cancer cell lines (HepG2) cell inoculation of logarithmic growth phase is placed on 37 DEG C, 5% in 96 well culture plates
CO2Being incubated overnight in cell incubator keeps its adherent；
2) it is separately added into the phellinus igniarius mycelium flavones (0.625,2.5,10 and 40mg/ml) of various concentration, is placed on culture
It is cultivated for 24 hours in case；
3) after culture for 24 hours, 20 hole μ l/ MTT (5mg/ml) is added, 96 orifice plates are then placed on 37 DEG C of constant incubators and are continued
Cultivate 4h；
4) it after cultivating, discards supernatant, the DMSO of 150 μ l is added, is completely dissolved first a ceremonial jade-ladle, used in libation crystal；
5) absorbance value (OD value) is surveyed under 490nm wavelength, growth and proliferation of cell rate is calculated according to absorbance, as a result as schemed
Shown in 1.
As shown in Figure 1, various concentration phellinus igniarius mycelium flavones (0.625,2.5,10 and 40mg/ml) is incubated for HepG2 respectively
Cell for 24 hours, detects cell proliferative conditions.The results show that concentration is the Phellinus bacterium of 10 and 40mg/ml compared with Normal group
Filament flavones significantly inhibits the proliferation of HepG2, and when concentration reaches 40mg/ml, proliferation rate is 52.31 %, as a result table
Bright, phellinus igniarius mycelium flavones can inhibit human hepatoma cell proliferation in vitro.
Claims (9)
1. a kind of phellinus igniarius mycelium flavones, which is characterized in that preparation method includes the following steps:
1) it ferments: inoculation fermentation culture being carried out to phellinus igniarius mycelium, freeze-drying obtains phellinus igniarius mycelium powder；
2) cellulase degradation extracts: phellinus igniarius mycelium powder is dissolved in distilled water, cellulase is added, carries out hot water extraction,
Centrifugation, filtering, it is spare to obtain cellulase degradation extracting solution；
3) ultrasound assisted extraction: being dissolved in ethyl alcohol for precipitating obtained by step 2) and carry out ultrasonic extraction, be centrifuged, and filtering obtains ultrasonic extraction
Extracting solution is spare；
4) merge cellulase degradation extracting solution and ultrasound extraction extracting solution, total extract rotary evaporation is concentrated, is filtered, it is sterile
Freeze-drying, obtains phellinus igniarius mycelium flavones dry powder.
2. a kind of phellinus igniarius mycelium flavones according to claim 1, which is characterized in that in step 1), the fermentation training
Feeding specific steps are as follows: 200g, glucose 20g, peptone 2g, MgSO are weighed after potato decortication4·7H2O 1.5g, KH2PO4
2g, water 1000ml, 121 DEG C of sterilizing 30min prepare PDA liquid medium.28 DEG C after inoculation, the training of 150r/min constant temperature oscillator
Support 15d.
3. a kind of phellinus igniarius mycelium flavones according to claim 1, which is characterized in that in step 2), by solid-liquid ratio, Phellinus
Erinaceus mycelium powder: distilled water=1:10-1:30 (g/ml).
4. a kind of phellinus igniarius mycelium flavones according to claim 3, which is characterized in that in step 2), with phellinus igniarius mycelium
Powder is substrate, and the additional amount of cellulase is 120-180U/g.
5. a kind of phellinus igniarius mycelium flavones according to claim 1, which is characterized in that in step 2), the hot water leaching
50-60 DEG C of temperature raising degree, enzymolysis time 1-2h, pH value range 4.0-6.0.
6. a kind of phellinus igniarius mycelium flavones according to claim 1, which is characterized in that described by solid-liquid ratio in step 3)
Sediment: ethyl alcohol=1:10-1:30 (g/ml).
7. a kind of phellinus igniarius mycelium flavones according to claim 6, which is characterized in that in step 3), the ethyl alcohol body
Fraction is 50%-60%.
8. a kind of phellinus igniarius mycelium flavones according to claim 1, which is characterized in that in step 3), the item of ultrasonic extraction
Part is that power is 80-150W, ultrasonic time 20-30min.
9. a kind of application of the phellinus igniarius mycelium flavones described in claim 1 in anti-liver cancer and anti-drug or health care product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910219490.7A CN109806283A (en) | 2019-03-21 | 2019-03-21 | A kind of phellinus igniarius mycelium flavones and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910219490.7A CN109806283A (en) | 2019-03-21 | 2019-03-21 | A kind of phellinus igniarius mycelium flavones and its preparation method and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109806283A true CN109806283A (en) | 2019-05-28 |
Family
ID=66609884
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910219490.7A Pending CN109806283A (en) | 2019-03-21 | 2019-03-21 | A kind of phellinus igniarius mycelium flavones and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109806283A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182550A (en) * | 2007-11-16 | 2008-05-21 | 上海市农业科学院 | Flavonoids from phellinus, method of producing the same and use |
| CN102363749A (en) * | 2011-10-09 | 2012-02-29 | 东北林业大学 | Preparation method of Phellinus linteus mycelium |
| CN107156842A (en) * | 2017-05-22 | 2017-09-15 | 辽宁大学 | Anti-breast cancer health products containing rice husk flavones |
-
2019
- 2019-03-21 CN CN201910219490.7A patent/CN109806283A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182550A (en) * | 2007-11-16 | 2008-05-21 | 上海市农业科学院 | Flavonoids from phellinus, method of producing the same and use |
| CN102363749A (en) * | 2011-10-09 | 2012-02-29 | 东北林业大学 | Preparation method of Phellinus linteus mycelium |
| CN107156842A (en) * | 2017-05-22 | 2017-09-15 | 辽宁大学 | Anti-breast cancer health products containing rice husk flavones |
Non-Patent Citations (2)
| Title |
|---|
| 张晶等: "《中药化学》", 30 April 2015, 中国农业大学出版社 * |
| 程俊文等: ""超声波辅助提取桑黄黄酮工艺优化"", 《浙江林业科技》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102047814B (en) | Micro ventilation lime wood antrodia camphorate cultivation method | |
| CN102731365B (en) | Hericium erinaceum biological micro-molecules for inhibiting helicobacter pylori and use of hericium erinaceum biological micro-molecules in treatment of digestive tract diseases | |
| CN101805763B (en) | Inonotus obliquus extracellular and intracellular mixing crude polysaccharide with function of strengthening immunity | |
| CN101418325A (en) | Inonotus obliquus extracellular and incellular mixed exopolysaccharide and preparation method thereof and pharmaceutical use | |
| CN107155639A (en) | A kind of cultural method for the Cordceps militaris for being rich in 1 DNJ | |
| CN1687429A (en) | Chinese traditional medicine containing glycosides transformed by ferment of microbe, and method for producing relevant aglycone preparation | |
| CN105767385A (en) | Preparation method of green-leaf gold-hypha health-care tea | |
| CN102172171B (en) | Method for culturing germanium-enriched cordyceps mycelium | |
| KR100405990B1 (en) | Processs for Preparing Organic Germanium from Cordyceps militaris | |
| CN105535035B (en) | A kind of Inonotus obliquus fermented and cultured composition and preparation method thereof | |
| CN101803531B (en) | China cryptoporus sinensis and solid cultural method and application thereof | |
| CN110477368A (en) | A kind of high activity cordyceps sinensis powder and the preparation method and application thereof | |
| CN109806283A (en) | A kind of phellinus igniarius mycelium flavones and its preparation method and application | |
| KR20120053564A (en) | Fermented ginseng with ganoderma lucidum beverage manufacturing method and that beverage | |
| KR20140129519A (en) | Cultivation method of phellinus linteus including vanadium and its phellinus linteus | |
| CN113875989A (en) | Functionalized water-soluble nano-selenium, preparation method and application | |
| CN102188485B (en) | Microbial fermentation method for improving active ingredients of schisandra chinensis | |
| KR20010110316A (en) | Artificial method to culture Cordyceps Kyushuensis Y.Kobayas and the application for its sporocarp | |
| CN101611854B (en) | Transformated mulberry leaf enriching coprinus comatus, cultivating method and application thereof | |
| KR101797663B1 (en) | Method for manufacturing of solid phase fermented herbal composition by cordyceps | |
| CN104212723B (en) | One strain Herba Artemisiae Japonicae Nei Shengyouwei can intend dish stey and application thereof | |
| CN1969956A (en) | Extract of Kadsura longepedunculata Finet et Gagnep, preparation process and use thereof | |
| KR20060022577A (en) | Composition for the culturing of phellinus linteus mycelium | |
| CN109055239A (en) | Chinese caterpillar fungus bacterial filament fast culture process | |
| CN109168954A (en) | A kind of growth promotion culture medium and its application in cordyceps sinensis culture |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190528 |
|
| RJ01 | Rejection of invention patent application after publication |