CN106974943A - A kind of purposes and preparation method of trametes robinioplila alcohol extract - Google Patents
A kind of purposes and preparation method of trametes robinioplila alcohol extract Download PDFInfo
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- CN106974943A CN106974943A CN201710054449.XA CN201710054449A CN106974943A CN 106974943 A CN106974943 A CN 106974943A CN 201710054449 A CN201710054449 A CN 201710054449A CN 106974943 A CN106974943 A CN 106974943A
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- 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
- A61K36/07—Basidiomycota, e.g. Cryptococcus
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- 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
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- 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
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
Abstract
The invention discloses a kind of purposes and preparation method of trametes robinioplila alcohol extract, trametes robinioplila alcohol extract is used as the application for preparing antineoplastic.Present invention discover that trametes robinioplila alcohol extract has significant suppression stomach cancer BGC823 cells, the effect of colon cancer cell HCT116 propagation, illustrate that trametes robinioplila alcohol extract is likewise supplied with antitumor action, further exploitation have been carried out to the medicinal ingredient of trametes robinioplila.
Description
Technical field
The present invention relates to a kind of active ingredient of Chinese herbs, the more particularly to a kind of purposes and preparation method of trametes robinioplila alcohol extract.
Background technology
Trametes robinioplila is China's important medicinal fungi among the people, and trametes robinioplila is On Polyporaceae Chinese scholartree bolt bacterium Trametes
Robiniophila Murr. drying fructification, alias Chinese scholartree Xiao, Chinese scholartree bacterium, Chinese scholartree chicken, Chinese scholartree moth, red chicken etc., be distributed mainly on Shaanxi,
The ground such as Shandong, Hebei.《Tang materia medica》Record：Bacterium on trametes robinioplila, this Chinese scholartree, it is hard as mulberry ear person is good when taking.Mulberry, Chinese scholartree, paper mulberry, elm,
Willow, this is five ears, and soft person may simultaneously eat, and paper mulberry ear people's often to eat, trametes robinioplila controls hemorrhoid.It is visible according to this, it is hard enter medicinal trametes robinioplila should be simultaneously
It is non-to be used as edible agaric.The cap of Chinese scholartree bolt bacterium is semicircle, like ear sample, is grown in Gu on Chinese scholar tree, so ancient claim " trametes robinioplila ",
Because whole fructification is similar to moth, so being called " Chinese scholartree bacterium ", " Chinese scholartree moth ".It is to sum up known, " the Chinese scholartree of history tree institute record description
Ear ", should be the Chinese scholartree bolt bacterium being grown on ancient middle Chinese scholar tree.
Trametes robinioplila is mild-natured, bitter, pungent, with the effect such as hemostasis, stop dysentery, anticancer.Effect of trametes robinioplila early has in successive dynasties medical book
Record,《Haigoushen》Record：" wind, blood-breaking, beneficial power can be controlled.”《Bencao Tujing》Cloud：" control big have blood in stool and five hemorrhoid, prolapse of the anus etc.." make
To have applied the traditional Chinese medicine trametes robinioplila in more than 1600 years in China, either single medicine and its extract still have been used for cancer
The huaier granules of auxiliary treatment, trade name " Jin Ke " all receives the extensive concern of educational circles, especially antitumor side in recent years
Face[2]。
So far, separation is carried out to trametes robinioplila chemical composition to be rarely reported with careful research, it is most of to study sight
Focus on its crude extract trametes robiniophila, its good anticancer therapeutic has obtained clinical extensive affirmative with extremely low side effect.
The preparation of huaier granules is that trametes robinioplila hot water, ethanol extract clear cream, is concentrated to relative density more than 1.33, plus the appropriate dextrin of people,
The auxiliary materials such as Icing Sugar, are made particle, the brown yellow granule after drying, every bag of 20g is huaier granules agent, chief active after being well mixed
Composition is GL-PP.Through S180 sarcoma inhibition test, inhibitory rate 40%~47% illustrates that GL-PP has anticancer
Activity, is the useful effect position of locust tree microellobosporia and trametes robiniophila.It is a kind of chocolate brown powder, without obvious fusing point, easily
Water is dissolved in, the ethanol of low concentration is slightly soluble in, it is organic molten insoluble in high concentration ethanol, acetone, ether, ethyl acetate, n-butanol etc.
Agent.Its transparent aqueous solution is in sticky shape, and pH value is 5.5, no optical activity.Through high performance liquid chromatography and electrophoretic analysis show its it is main into
Divide essentially homogeneous polysaccharide, and prove that Huai Er is a protein binding heteroglycan by spectrum and gas chromatography method, wherein many
Sugared mass fraction is 41.53%, and amino acid masses fraction is 12.93%, and moisture 8.72%, its hydrolysate contains L-arabinose
(arabinose), D- xyloses (xylose), D-MANNOSE (mannose), D- galactolipins (galactose), D-Glucose
(glucose) 6 kinds of monose such as, and proline (proline), glycine (glycine), alanine (alanine), cystine
(cystine), isoleucine (isoleucine), aspartic acid (aspartic acid), threonine (threonine), silk ammonia
Sour (serine), 18 kinds of amino acid such as glutamic acid (glutamicacid).
The existing extracting method of trametes robinioplila has common water extraction, HTHP water extraction, cold caustic soda method, thermokalite method, alkaline extraction water
The yield of soluble extract is higher than water extraction, and the yield of wherein thermokalite method extract is up to 29.65%, but its Thick many candies content
Minimum, the net yield of cold caustic soda method Thick many candies is up to 6.93%.
At present, for the research of trametes robinioplila, huaier granules or medicinal extract are concentrated mainly in recent years to tumor cell resistance aspect,
Trametes robinioplila extracting method and chemical composition carry out separation and are rarely reported with careful research, and it is thick that sight is focused on its by most of research
On extract trametes robiniophila, trametes robiniophila is the hot water extract after locust tree microellobosporia ferments, and it contains polysaccharide, protein, alkaloid
Mineral matter element is planted etc. a variety of organic principles and more than 10, main active is GL-PP.
The content of the invention
It is an object of the invention to provide a kind of purposes of trametes robinioplila alcohol extract, trametes robinioplila alcohol extract has significant suppression stomach cancer
The effect of BGC823 cells, colon cancer cell HCT116 propagation, illustrates that trametes robinioplila alcohol extract is likewise supplied with antitumor action, to trametes robinioplila
Medicinal ingredient carried out further exploitation, the exploitation for anti-gastric cancer, colon cancer drug provides new approach.
Present invention also offers the preparation method of trametes robinioplila alcohol extract.
The technical solution adopted for the present invention to solve the technical problems is：
A kind of purposes of trametes robinioplila alcohol extract, trametes robinioplila alcohol extract is used as the application for preparing antineoplastic.
Prior art is studied using trametes robinioplila water extract or polysaccharide, and for antitumor, does not disclose trametes robinioplila alcohol extracting
Whether thing has medical value, present invention firstly discovers that trametes robinioplila alcohol extract has preferable antitumor action, and recognizes better than existing
For trametes robinioplila water extract or polysaccharide antitumor action.Further exploitation have been carried out to the medicinal ingredient of trametes robinioplila, have been anti-
Stomach cancer, the exploitation of colon cancer drug provide new approach.
Preferably, the antineoplastic is anti-gastric cancer medicament or drugs against colon cancer.
Preferably, the trametes robinioplila alcohol extract is prepared by the following method and obtained：Trametes robinioplila fructification is crushed, Chinese scholartree of sieving to obtain
Ear crushed material, according to 1g trametes robinioplila crushed materials：After the solid-liquid ratio of 30mL ethanol mixes trametes robinioplila crushed material with ethanol, it is heated to reflux carrying
Take 2-3 times, 2 hours every time, filter, merge extract solution, the extract solution revolving of merging reclaims ethanol, is concentrated into paste, freezes
Dry.
Preferably, the mass concentration of the ethanol is 70%.
A kind of preparation method of the trametes robinioplila alcohol extract with antitumaous effect, trametes robinioplila fructification is crushed, and sieve to obtain trametes robinioplila powder
Mince, according to 1g trametes robinioplila crushed materials：After the solid-liquid ratio of 20-40mL ethanol mixes trametes robinioplila crushed material with ethanol, it is heated to reflux carrying
Take 2-3 times, 2 hours every time, filter, merge extract solution, the extract solution revolving of merging reclaims ethanol, is concentrated into paste, freezes
Dry.
Preferably, the mass concentration of the ethanol is 60-90%.
Preferably, trametes robinioplila fructification is crushed, sieve to obtain trametes robinioplila crushed material, according to 1g trametes robinioplila crushed materials：30mL mass
After concentration mixes trametes robinioplila crushed material with ethanol for the solid-liquid ratio of 70% ethanol, 80 DEG C of refluxing extractions are heated to 3 times, every time 2
Hour, filtering merges extract solution, and the extract solution revolving of merging reclaims ethanol, is concentrated into paste, is freeze-dried.
A kind of preparation method of the trametes robinioplila extract with antitumaous effect, step is as follows：
(1) alcohol extracting：Trametes robinioplila fructification is crushed, sieve to obtain trametes robinioplila crushed material, according to 1g trametes robinioplila crushed materials：30mL mass is dense
Spend the ethanol for 70% solid-liquid ratio trametes robinioplila crushed material is mixed with ethanol after, be heated to 80 DEG C of refluxing extractions 3 times, 2 is small every time
When, filtering merges extract solution and obtains alcohol extract；
(2) water extraction：After the remaining dregs of a decoction are dried after step (1) alcohol extracting, according to the 1g dregs of a decoction：The solid-liquid ratio of 30mL distilled water
Plus distilled water, heating and refluxing extraction 3 times, each 2.5h, filtering, merge extract solution and obtain Aqueous extracts；
(3) alcohol extract and Aqueous extracts are mixed, vacuum-concentrcted to paste, freeze-drying.
Acted on preferably, antitumaous effect is anti-gastric cancer or inhibitor against colon carcinoma cells.
The beneficial effects of the invention are as follows：Trametes robinioplila alcohol extract has significant suppression stomach cancer BGC823 cells, colon cancer cell
The effect of HCT116 propagation, illustrates that trametes robinioplila alcohol extract is likewise supplied with antitumor action, the medicinal ingredient of trametes robinioplila has been carried out entering one
The exploitation of step, the exploitation for anti-gastric cancer, colon cancer drug provides new approach.
Brief description of the drawings
Fig. 1 is influence of the concentration of alcohol to general flavone recovery rate.
Fig. 2 is that ethanol measures influence to general flavone recovery rate again.
Fig. 3 is influence of the extraction time to general flavone recovery rate.
Fig. 4 is influence of the extraction time to general flavone recovery rate.
Embodiment
Below by specific embodiment, technical scheme is described in further detail.
In the present invention, if not refering in particular to, raw material and equipment used etc. is commercially available or commonly used in the art.
Method in following embodiments, is the conventional method of this area unless otherwise instructed.
Embodiment：
Using general flavone recovery rate as inspection target, by single factor exploration and orthogonal experiment, trametes robinioplila alcohol extract is determined most
Good extraction process.
1. single factor exploration
Concentration of alcohol is selected, ethanol is measured again, and extraction time, extraction time is investigated, each level of each factor is put down
3 experiments of row.It is each Factor Experiment result below.
1.1. concentration of alcohol is investigated
Trametes robinioplila fructification is crushed, 20 mesh sieves is crossed, weighs 8 parts of trametes robinioplila fructification powder, it is about 1g/ parts, accurately weighed, investigate not
Influence with concentration of alcohol to general flavone recovery rate, respectively select 60%, 65%, 70%, 75%, 80%, 85%, 90%,
95% ethanol, solid-liquid ratio 1g:30mL, 80 DEG C of water-bath refluxing extractions 1 time, 1h/ times, lets cool, and mends weight, filtering.Precision pipettes continuous
Appropriate filtrate, the content of determined by ultraviolet spectrophotometry general flavone, parallel three experiments.Calculate general flavone recovery rate, general flavone
Recovery rate (mg/g)=general flavone/crude drug, is as a result shown in Fig. 1.
1.2. ethanol measures investigation again
Trametes robinioplila fructification is crushed, 20 mesh sieves is crossed, weighs 5 parts of trametes robinioplila fructification powder, it is about 1g/ parts, accurately weighed, investigate second
Alcohol measures the influence to general flavone recovery rate again, is separately added into 10,20,30,40,50 times of amount (g:ML 70% ethanol), 80 DEG C of water
Bathe refluxing extraction 1 time, 1h/ times, let cool, mend weight, filtering.Precision pipettes subsequent filtrate in right amount, and determined by ultraviolet spectrophotometry is always yellow
The content of ketone, parallel three experiments.Calculate general flavone recovery rate, general flavone recovery rate (mg/g)=general flavone/crude drug.As a result see
Fig. 2.
1.3. the investigation of extraction time
Trametes robinioplila fructification is crushed, 20 mesh sieves is crossed, weighs 5 parts of trametes robinioplila fructification powder, about 1g/ parts, accurately weighed, investigation is carried
Influence of the time to general flavone recovery rate is taken, 5 horizontal 0.5h, 1h, 1.5h, 2h, 2.5h are selected, 80 DEG C of water-bath backflows, liquid is solid
Than 30:1, concentration of alcohol 70%.Precision pipettes subsequent filtrate in right amount, the content of determined by ultraviolet spectrophotometry general flavone, parallel three
Secondary experiment.Calculate general flavone recovery rate, general flavone recovery rate (mg/g)=general flavone/crude drug.As a result Fig. 3 is seen.
1.4. extraction time is investigated
Trametes robinioplila fructification is crushed, 20 mesh sieves is crossed, weighs 3 parts of trametes robinioplila fructification powder, it is about 1g/ parts, accurately weighed, add 30
70% ethanol measured again, 80 DEG C of water-baths backflow 2h, it is respectively 1 time, 2 times, 3 times to investigate extraction time, 2h/ times, calculates general flavone
Recovery rate, general flavone recovery rate (mg/g)=general flavone/crude drug.As a result Fig. 4 is seen.
2. orthogonal
Tested using the horizontal quadrature of four factor three, parallel test 3 times, using trametes robinioplila general flavone content as index, in single factor test
On the basis of investigation, the investigation of optimised process is carried out with heating water bath reflow method, determines that concentration of alcohol (A), solvent are measured again
(B), extraction time (C), extraction time (D) are investigation factor, from L9(34) orthogonal arrage tested, factor level table is shown in Table
1。
The factor of table 1 and water-glass
2.1. orthogonal experiments are analyzed
Select L9(34) orthogonal arrage can obtain 9 groups of process conditions.Analysis in table 2 directly perceived, variance analysis is shown in Table 3.
The orthogonal design result table of table 2
The results of analysis of variance table of table 3
Note：*P<0.1, * * P<0.05, * * * P<0.01.
General flavone counts content as inspection target using rutin in trametes robinioplila, and from analysis directly perceived, each factor is to the total Huang of alcohol extract
The influence size that ketone is extracted is followed successively by extraction time (D)>Extraction time (C)>Concentration of alcohol (A)>Solvent is measured (B) again.By variance
Analysis is understood, is shown through F assays, and extraction time, extraction time and concentration of alcohol are principal element, and solvent measures no shadow again
Ring, therefore the extraction optimize technique of trametes robinioplila general flavone is A1B3C2D3。
Orthogonal test significance analysis understands that solvent is measured again not to make significant difference to the content of general flavone, it is considered to which the energy is timely
Between save, consider, it is thus determined that trametes robinioplila alcohol extracting optimal extract process be A1B1C2D3, i.e., 30 times are measured, and 70% ethanol is molten
Liquid, heating and refluxing extraction 3 times, each 2h.
2.2. optimum extraction process verification experimental verification
According to result of the test, it is A to determine trametes robinioplila alcohol extracting optimum extraction process1B1C2D3.Three batches of trametes robinioplila medicinal materials are taken, according to above-mentioned
Optimum extraction process carries out checking test.It the results are shown in Table 4.
The checking test result table of table 4
General flavone average content is 25.76mg/g in parallel 3 measure, trametes robinioplila, and RSD is 1.70%, shows extraction process
Reappearance is good, stablizes feasible.
Test portion：
Using colon cancer cell HCT116 and stomach cancer BGC823 cells as experiment cell line, trametes robinioplila different component extract is thin
Sample is administered in born of the same parents, using MTT colorimetric methods, and research trametes robinioplila different component extract is made to HCT116 cells and BGC823 cells propagation
With filtering out trametes robinioplila active component, the Study on extraction for trametes robinioplila active component provides scientific basis.
First, experiment material
(1) experimental drug and reagent
Trametes robinioplila medicinal material, originates in Shandong, lot number 20131001
MTT (Sigma Co., USA, model M2123)
Trypsase (Amresco companies of the U.S., 1:250)
Hyclone (Fetal Bovine Serun, FBS) (Chinese holly, lot number 20151112)
Dual anti-(Penicillin-Streptomycin Solution) (Hyclone companies, lot number J130043)
DMSO (Dimethyl Sulfoxide) (ɑ MRESCO companies, lot number 3544B028)
Phosphate buffer salt (PBS) (silent winged generation that biochemistry product (Beijing) Co., Ltd, lot number NZA1064 of match)
96 orifice plates (the silent winged generation that Thermo Fisher of match).
(2) experiment equipment
High-speed multifunctional pulverizer (Shanghai Li Jian Machinery Co., Ltd.s, model YB-1000A)
Ultrasonic cleaner (Kunshan ultrasonic instrument Co., Ltd, model KQ5200DE).
2nd, method and result
1 cell culture
Colon cancer HCT116 cells and stomach cancer BGC823 cell lines, derive from life science institute of the Chinese Academy of Sciences thin
Born of the same parents storehouse.HCT116 cells (BGC823 cells) are accessed into 25cm2Tissue Culture Plate, adds the complete culture containing 10% hyclone
Liquid 6mL, puts 37 DEG C, 5%CO2In incubator, partly outward winding the bottle cap of blake bottle and keep humidity in incubator, 1-2d changes one
Secondary nutrient solution.Observation cell growth condition daily, after carrying out passage or freezing after cell fusion 80%.
2 passages
Cell growth to 80% fusion need to be passed on.When cell is in exponential phase, nutrient solution in blake bottle is abandoned,
Washed with PBS 2~3 times, 2~3mL, is careful not to directly rinse cell every time, add 0.25% trypsin solution 2mL,
2-5min or so (depending on specific cell) is acted on, isometric nutrient solution is added and is neutralized.Cell suspension is moved into 15mL
Centrifuge tube, setting rotating speed is 800rmin-1, centrifuges 5min, and abandoning supernatant is gently washed 1~2 time with nutrient solution.Cell sinks
Form sediment and slowly dispelled with a little new complete culture solution, sub-bottle Secondary Culture or biography plate grouping experiment after being counted with cell counting count board.
It is prepared by 3 cell samples
Trametes robinioplila fructification powder about 10g is weighed, is extracted by optimum extraction process, be i.e. 30 times of 70% ethanol of amount of alcohol extracting addition are molten
Liquid, heating and refluxing extraction 3 times, each 2h, revolving reclaims ethanol, is concentrated into after paste, freeze-drying and obtains sample A (trametes robinioplilas
Alcohol-extracted extract powder)；
Add 30 times of amount distilled water after the medicine dregs of a decoction are dried after alcohol extracting to flow back 3 times, each 2.5h, revolving reclaims ethanol, concentration
To paste, sample B (trametes robinioplila the water extracted immersing paste powder) is obtained after freeze-drying；
It is another to prepare a alcohol extract and Aqueous extracts with same procedure, alcohol extract and Aqueous extracts are merged, vacuum-concentrcted
To paste, freeze-drying obtains sample A+B (the double extracted extract powder of trametes robinioplila alcohol water), and using commercially available huaier granules C as control, will
The cell sample of preparation is used for HCT-116 cells (BGC-823 cells) growth inhibition test.
4 cell administration concentration gradients
Accurately weighed trametes robinioplila difference turnover position sample extract powder 0.064g (huaier granules are converted in proportion), is dissolved in 10ml
Not in serum-containing media, the μ g/mL mother liquors of medicinal extract 6400, gradient dilution to each experimental concentration are configured to.Cell administration concentration ladder
Degree is shown in Table 5.
The trametes robinioplila of table 5 difference leaches position cell administration concentration gradient
Using above pharmaceutical samples, compare each extract part to HCT116 cells (BGC823 cells) proliferation inhibition test
(MTT experiment), the IC50 values of the inhibiting rate by comparing each group filter out the leaching position of drug effect optimal (IC50 values are minimum),
The research of next step trametes robinioplila optimum extraction process is carried out with this active component.
5 screening techniques
Inhibiting tumour cells proliferation experiment is determined with mtt assay, the tumor control rate IC50 values to each extract are compared,
And then filter out the leaching position of drug effect most preferably.
6 cell growth inhibition tests
HCT116 cells (BGC823 cells) in exponential phase are used into 0.25% Trypsin Induced, centrifuges, adopts
After being counted with cell counting count board, concentration of cell suspension is adjusted, every 5 × l04 of hole/mL cell suspension inoculation is formulated as in 96 holes
Plate, to prevent edge effect, 96 orifice plate outer rims one circle adds 200 μ L PBS phosphate buffers, and it is thin that remaining adds 100 μ L per hole
Born of the same parents' suspension, is positioned over CO2 concentration for 5%, temperature for culture 24-48h in 37 DEG C of CO2 incubators (visual cell's upgrowth situation and
It is fixed), nutrient solution is abandoned in suction, adds the different quality concentration samples without serum, respectively the μ g/mL of medicinal extract 640,320 μ g/mL, 160
μ g/mL, 80 μ g/mL, 40 μ g/mL, 20 μ g/mL are parallel to set 6 multiple holes per the μ L of hole 100, at the same set blank control group without
Cell suspension, only adds 100 μ L fresh cultures, parallel determination 3 times.Continue to cultivate 24h in the incubator of similarity condition, in
Cellular morphology is observed under inverted microscope.20 μ L MTT solution (5mg/mL, i.e. 5% concentration) are added per hole, continue to be incubated 4h.Eventually
Only cultivate, carefully suck nutrient solution in culture hole.150 μ L DMSO are added per hole, is placed on low speed shaking table and vibrates 10min, with molten
The MTT- formazans crystallization of solution residual, in optical density (OD) value for determining each hole on multi-function microplate reader at 490nm, with normal control
The OD values that group is measured are control, and each extract group of trametes robinioplila is calculated for HCT116 cells (BGC823 cells) according to inhibiting rate formula
The inhibition of propagation.The action time of trametes robinioplila different component is respectively set as 24h and 48h, calculates IC50 values.
Inhibiting rate=[1- (experimental group MTT detection OD values-blank group OD)/(control group MTT detection OD values-blank group OD)]
× 100%.
Improve Kou Shifa：LgIC50=Xm-I (P- (3-Pm-Pn)/4)
Xm:Lg maximum doses
I:Lg (maximum dose/adjacent dosage)
P:Positive reaction rate sum
Pm:Maximum positive reaction rate
Pn:Minimum positive reaction rate.
Colon cancer HCT116 cell results are shown in Table 6；Stomach cancer BGC823 cell results are shown in Table 7.
The trametes robinioplila of table 6 difference extract is to HCT116 cell inhibitory effect IC50 values
The trametes robinioplila of table 7 difference extract is to BGC823 cell inhibitory effect IC50 values
3rd, discuss and analysis
This part experimental studies results show that trametes robinioplila respectively leaches position under 640~20 μ gmL-1 concentration to colon cancer
HCT116 cells and stomach cancer BGC823 cells have an obvious inhibited proliferation, and with the increase of drug concentration, in dosage according to
The relation of relying.
Trametes robinioplila difference extract is respectively provided with different inhibited proliferations to HCT116 cells and BGC823 cells, and HCT116 is thin
Two kinds of cells of born of the same parents and BGC823 cells show that action time 48h is good compared with 24h effects, and in dose-dependence, trametes robinioplila alcohol water
It is double to carry extract I C50Value is minimum, next to that alcohol extract, difference leaches between positions that there were significant differences (trametes robinioplila water extract and trametes robinioplila
There is no significant difference between particle).Therefore, IC is selected50The double extract conducts of alcohol water of value minimum (antitumous effect is optimal)
Later stage tests active component.
This result shows that trametes robinioplila alcohol extract has significant suppression stomach cancer BGC823 cells, colon cancer cell HCT116
The effect of propagation, illustrates that trametes robinioplila alcohol extract is likewise supplied with antitumor action.And research is only limitted to trametes robinioplila water extraction both at home and abroad before this
Other medicinal parts of trametes robinioplila are not carried out constituent analysis, cause related medicinal part to be abandoned by thing and its polysaccharide component.This
Research prompting trametes robinioplila alcohol extract possesses the effect for suppressing tumour, and this undoubtedly points out the chemical composition of trametes robinioplila alcohol extract to separate with being
System research is valuable, possesses DEVELOPMENT PROSPECT.
Embodiment described above is a kind of preferably scheme of the present invention, not makees any formal to the present invention
Limitation, also has other variants and remodeling on the premise of without departing from the technical scheme described in claim.
Claims (9)
1. a kind of purposes of trametes robinioplila alcohol extract, it is characterised in that trametes robinioplila alcohol extract is used as the application for preparing antineoplastic.
2. purposes according to claim 1, it is characterised in that the antineoplastic is anti-gastric cancer medicament or inhibitor against colon carcinoma cells
Medicine.
3. purposes according to claim 1 or 2, it is characterised in that the trametes robinioplila alcohol extract be prepared by the following method and
：Trametes robinioplila fructification is crushed, sieve to obtain trametes robinioplila crushed material, according to 1g trametes robinioplila crushed materials：The solid-liquid ratio of 30mL ethanol is by trametes robinioplila
After crushed material is mixed with ethanol, heating and refluxing extraction 2-3 times, 2 hours every time, filtering merged extract solution, the extract solution rotation of merging
Steam and reclaim ethanol, be concentrated into paste, be freeze-dried.
4. purposes according to claim 3, it is characterised in that the mass concentration of the ethanol is 70%.
5. a kind of preparation method of the trametes robinioplila alcohol extract with antitumaous effect, it is characterised in that crush trametes robinioplila fructification, sieving
Trametes robinioplila crushed material is obtained, according to 1g trametes robinioplila crushed materials：After the solid-liquid ratio of 20-40mL ethanol mixes trametes robinioplila crushed material with ethanol, plus
Circumfluence distillation 2-3 times, 2 hours every time, filtering merged extract solution, and the extract solution revolving of merging reclaims ethanol, is concentrated into thick paste
Shape, freeze-drying.
6. preparation method according to claim 5, it is characterised in that the mass concentration of the ethanol is 60-90%.
7. the preparation method according to claim 5 or 6, it is characterised in that crush trametes robinioplila fructification, sieve to obtain trametes robinioplila powder
Mince, according to 1g trametes robinioplila crushed materials：30mL mass concentrations mix trametes robinioplila crushed material with ethanol for the solid-liquid ratio of 70% ethanol
Afterwards, 80 DEG C of refluxing extractions are heated to 3 times, 2 hours every time, filtering merged extract solution, and the extract solution revolving of merging reclaims ethanol,
Paste is concentrated into, is freeze-dried.
8. a kind of preparation method of the trametes robinioplila extract with antitumaous effect, it is characterised in that step is as follows：
(1) alcohol extracting：Trametes robinioplila fructification is crushed, sieve to obtain trametes robinioplila crushed material, according to 1g trametes robinioplila crushed materials：30mL mass concentrations are
After the solid-liquid ratio of 70% ethanol mixes trametes robinioplila crushed material with ethanol, 80 DEG C of refluxing extractions are heated to 3 times, every time 2 hours, mistake
Filter, merges extract solution and obtains alcohol extract；
(2) water extraction：After the remaining dregs of a decoction are dried after step (1) alcohol extracting, according to the 1g dregs of a decoction：The solid-liquid ratio of 30mL distilled water adds steaming
Distilled water, heating and refluxing extraction 3 times, each 2.5h, filtering merges extract solution and obtains Aqueous extracts；
(3) alcohol extract and Aqueous extracts are mixed, vacuum-concentrcted to paste, freeze-drying.
9. the preparation method according to claim 5 or 8, it is characterised in that antitumaous effect is that anti-gastric cancer or inhibitor against colon carcinoma cells are made
With.
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| CN115531427B (en) * | 2022-11-04 | 2023-11-24 | 山东中医药大学 | Application of pagodatree ear extract in preparation of anti-influenza virus drugs |
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