CN106138146B - The intervention effect that Radix Rhapontici seu Radix Echinopsis ethyl acetate extract grows carcinoma of mouth - Google Patents
The intervention effect that Radix Rhapontici seu Radix Echinopsis ethyl acetate extract grows carcinoma of mouth Download PDFInfo
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Abstract
A kind of Chinese medical extract that can significantly inhibit cancer cell of oral cavity belongs to novel mouth cancer development of Chinese herb products field.Radix Rhapontici seu Radix Echinopsis ethyl acetate extract, extracting method is simple, quick, can be mass.Select the inhibited proliferation of CCK8 method detection Radix Rhapontici seu Radix Echinopsis EA oral cancer cell SCC15；Influence of the FITC/PI Flow cytometry Radix Rhapontici seu Radix Echinopsis EA to SCC15 Apoptosis, western blot detect the expression of apoptosis-related protein in MAPK signal path；The influence of invasion and scratch experiment detection Radix Rhapontici seu Radix Echinopsis EA to SCC15 cell invasion and migration, Q-PCR and western blot detect the expression of Epithelial and stromal Transformed E MT related gene and albumen.The present invention provides foundation for the research and development of clinical treatment carcinoma of mouth new drug.
Description
Technical field
The present invention relates to a kind of drugs of intervening oral epidermoid carcinoma cell growth, specifically with Chinese herbal medicine Radix Rhapontici seu Radix Echinopsis
It for raw material, is extracted through ethyl acetate, i.e. Radix Rhapontici seu Radix Echinopsis ethyl acetate extract (ethyl acetate extract, EA).The present invention is also
It is related to the preparation method of the drug.
Background technique
Rhaponticum uniflorum (Rhaponticum uniflorum, DC.) perennial herb, root are directly stretched, and stem is upright, not branch, cluster
Raw or Dan Sheng, canescence, by cotton wool.It is born in upland meadow, meadow steppe, stone matter hillside.It is distributed Heilungkiang, Jilin, Liaoning, river
The ground such as north, the Inner Mongol, Shaanxi, Gansu, Qinghai, Shanxi, Henan, Sichuan, Shandong.
Its effect is described: " Radix Rhapontici seu Radix Echinopsis, lower milk, the heat toxin that disappears, apocenosis, hemostasis, myogenic, desinsection, Gu Dongyuan in Compendium of Material Medica
Think hand, the bright medicine of foot sun, and ancient prescription controls ulcer carbuncle on the back, is claimed headed by Radix Rhapontici seu Radix Echinopsis soup.Pang Anchang treatise on Febrile Diseases controls ulcer and pre- solution
When row acne rash heat, with Radix Rhapontici seu Radix Echinopsis leaf, cloud without then with fructus gardeniae instead of, also take its it is cold can be antipyretic, lid do not know its can enter sun it is bright therefore
".Radix Rhapontici seu Radix Echinopsis is more common Chinese medicine, early there is the record of " long term usage make light of one's life by commiting suicide QI invigorating, normal hearing and eyesight, barren macrobiosis ".Radix Rhapontici seu Radix Echinopsis is both with its hardship
The property of cold tonneau, lets out clearly the heat of Yangming Channel, and because of the function of its qi and activate blood circulation, strong taste, therefore long term usage can promote longevity.But it is more
Over year it is clinical in the effect of taking its clearing heat and detoxicating, subduing inflammation, lower milk more, its antineoplastic action is studied less.
Carcinoma of mouth is the common malignant tumour of incidence, accounts for the 40% of head-neck malignant tumor.Have every year in world wide
Close to 540,000 new cases, although the technology of carcinoma of mouth treatment in recent years is continuously improved, survival rate is less than 50% within 5 years
(1).In developing country, carcinoma of mouth becomes one of main fatal disease.Currently, mainly being controlled using operation for carcinoma of mouth
It treats, radiotherapy, the Compositive sequence in treatment based on chemotherapy, but therapeutic effect is often bad.Ideal anticancer drug
Should have low toxicity, safely, effectively, the conditions such as inexpensive.Chinese medicine is China's medicine rarity, modern pharmacological studies have shown that in many tradition
Medical instrument has antitumor action, and has the advantages that excellent effect, low toxicity, low side effect.Currently, Chinese medicine has become tumor prevention and treatment
New research hotspot.The efficient therapeutic agent of novel low-toxicity is found to be of great significance to carcinoma of mouth complex treatment.
Radix Rhapontici seu Radix Echinopsis is classified as top grade first recorded in Shennong's Herbal.It is bitter, salty, it is cold in nature, enter stomach, large intestine channel has heat-clearing solution
Poison, promotes lactation effect at the carbuncle that disappears apocenosis.Studies have shown that Radix Rhapontici seu Radix Echinopsis volatile oil has alcohols, ketone, alkanes, alkenes, terpene, the change such as phenols
Closing object is wherein mainly terpenoid, terpenoid be present in plant kingdom there is the intentional of many-sided bioactivity
A kind of compound of justice, is the carypohyllene all contained in certain middle the effective elements of the medicine such as above-mentioned volatile oil, elemene isoreactivity
Effective component is respectively provided with the effects of antispastic, antiviral, expelling parasite (2).In recent years, some studies have shown that Radix Rhapontici seu Radix Echinopsis have it is anti-oxidant,
Improve immune, anti-inflammatory and anti-aging, it is antitumor the effects of (3-6).Its Anticancer Effect and Mechanism may by inducing cell apoptosis,
Reversing drug resistance cell strain to the sensibility of chemotherapeutic, inhibit the effects of tumor cell proliferation.Correlative study shows Radix Rhapontici extract
Direct tumor-inhibiting anticancer effect can be played when large dosage of.Reversing drug resistance effect outstanding can be played in low dose, help to allow
Drug resistant cell strain recaptures sensitivity to chemotherapeutic；The drug resistance to human breast carcinoma mdr cell MCF-7/ADR can be effectively reversed, because
And clinically have broad application prospects (7-8).Radix Rhapontici seu Radix Echinopsis compound preparation can effectively treat primary carcinoma of liver (9).It is dynamic in lotus knurl
In object test, Radix Rhapontici seu Radix Echinopsis and its compound preparation have apparent function of tumor inhibition, and effect may increase body immunization machine with it
Energy (10-11) related with oxidation resistance.But there is not been reported for research of the current Radix Rhapontici seu Radix Echinopsis to carcinoma of mouth effect etc..
Studies have shown that promoting EMT related after inhibiting gene participating in apoptosis Bcl-2 to be overexpressed plasmid transfection epithelial cell
Factor expression (12).When EMT occurs for rat embryo liver cell, the activation of Akt and Bcl-xL can be promoted, and then inhibit
The apoptosis of TGF-β induction, these cells have the property of liver precursor and some hepatomas.Speculate accordingly, EMT's is thin
Born of the same parents have activated the signal path (AKT and PI3K) of anti-apoptotic or have secreted the factor of anti-apoptotic.A variety of different mammals
Epithelial cell (EpH-4, NMuMG and KIM-2) has the apoptosis of ultraviolet light radiation-induced bright after EMT occurs through induction
Aobvious tolerance (13).
The study find that Radix Rhapontici seu Radix Echinopsis EA inhibits SCC15 cell Proliferation, and inhibiting effect enhances increase with time；It can significantly induce
SCC15 Apoptosis, and there is time-and concentration-dependent；Radix Rhapontici seu Radix Echinopsis EA inhibits SCC15 cell invasion and migration；And have inverse
Turn the effect of cancer cell of oral cavity SCC15 Epithelial and stromal conversion.Radix Rhapontici seu Radix Echinopsis EA causes the relevant p-p38 and p-JNK expression of rush apoptosis aobvious
Work increases, and makes to promote to be proliferated relevant p-ERK expression reduction.Radix Rhapontici seu Radix Echinopsis EA influences EMT process, to inhibit carcinoma of mouth growth, invasion
And migration.
Bibliography
(1) Fu Jinye, Gao Jing, Zheng Jiawei wait the epidemiological survey and analysis China mouth of carcinoma of mouth Related Risk Factors
Chamber decorative sursery magazine, 2011,9 (4): 316-322.
(2) Gao Yuguo, Xu Yaoshun Radix Rhapontici seu Radix Echinopsis Analysis of Essential Oil Components Anshan Normal University journal, 2013,15 (2): 38-
40.
(3) Zuo Wei, Zuo Wei, Guan Xin wait effect for reducing fat China of the rhaponticum uniflorum in adult with primary nephrotic syndrome
Natural medicine magazine, 2001.3 (1): 27-28.
(4) Quan Jihua, Jin Yanhua, Yin Xuezhe wait the Antioxidation in vitro Chinese experimental prescription of Radix Rhapontici seu Radix Echinopsis alcohol extracting thing
Magazine, 2011,17 (6): 197-199.
(5) Piao Long, Zhang Xuewu, Jin Xiangzi, treasure's traditional Chinese medical science traditional Chinese medicines when waiting the research of Radix Rhapontici extract anti-aging effects,
2006,17(10):1918-1919.
(6) Bao little Mei, Luo Suqin wait rhaponticum uniflorum chemical component and the Asia-Pacific Pharmacological Advancement traditional medicine,
2011,7(9):176-177.
(7) Jiao Zhonghua, Liu Peimin, Li Xiurong wait Radix Rhapontici seu Radix Echinopsis extractant (RHU) Contained Serum to human breast carcinoma persister
The research light Chinese medicine of MCF-7/ADR cell P170 protein expression, 2008,18 (108): 17-18.
(8) Cao Fang, Li Xiurong's Radix Rhapontici seu Radix Echinopsis extractant grind the drug resistance inversion effect of human breast carcinoma mdr cell MCF-7/ADR
Study carefully Shandong journal of traditional Chinese medicine, 2009,28 (6): 415-417.
(9) Wang Huicang, Wang Xiuming compound Radix Rhapontici seu Radix Echinopsis soup treat 30 Hunan clinical observation Journals of Traditional Chinese Medicine of primary carcinoma of liver,
2008,24 (1): 26-27.
(10) Gao Ying, Wu Mei, Tang's Bi core wait Wu Shi lumor eliminating powder to the antitumor action Chinese patent drug of tumor-bearing mice, and 2014,
26 (3): 446-450.
(11) Jin Aihua, Xu Huixian, Liu Wenjing wait rhaponticum uniflorum to the tumor-inhibiting action of H22 rat liver cancer subcutaneous transplantation tumor
And its Initial Study of Mechanism .2011,17 (5): 165-167.
(12)An J,Lv J,Li A,Qiao J,et al.Constitutive expression of Bcl-2
induces epithelial-Mesenchymal transition in mammary epithelial cells.BMC
Cancer.2015 Jun 20；15:476.
(13) RobsonH, Khaled WT, AbellKW, et a1.Epithelial-to-ruesenchymal
transition confers resistance to apoptosis in three murine mammary epithelial
Cell lines.Diferentiation, 2006,74 (5): 254-64.
Prior art background defect: carcinoma of mouth mainly uses operative treatment, radiotherapy, comprehensive based on chemotherapy
Sequence treatment is closed, but therapeutic effect is often bad.Modern pharmacological studies have shown that many traditional Chinese medicines have antitumor action, and have
There is the advantages of excellent effect, low toxicity, low side effect.People start gradually to pay attention to the exploitation in traditional Chinese medicine, and it is efficient to find novel low-toxicity
Therapeutic agent be of great significance to carcinoma of mouth complex treatment.Extraction solvent used in this experiment is fat-soluble solvent acetic acid second
Ester, more conventional water extract more can more adequately dissolve the active principle in Radix Rhapontici seu Radix Echinopsis.The present invention is that clinical treatment carcinoma of mouth is new
The research and development of medicine provide foundation.
Summary of the invention
Object of the present invention is to anti-oxidant according to Radix Rhapontici seu Radix Echinopsis, height-regulating is immune, the direct or indirect antitumor action such as antitumor, comes
Whether detection Radix Rhapontici seu Radix Echinopsis ethyl acetate extract has anti-carcinoma of mouth effect, provides to the clinical drug therapy for carcinoma of mouth new
It supports.
To achieve the above object, the technical solution adopted by the present invention is as follows.
Specifically includes the following steps:
(1) preparation of Radix Rhapontici seu Radix Echinopsis ethyl acetate extract
Radix Rhapontici seu Radix Echinopsis sample comminution takes 500 grams of coarse powder at coarse powder, with 80% ethyl alcohol it is wet after, then with 3 times of 80% alcohol refluxs of amount
It extracts twice, combined extract, ethyl alcohol is recovered under reduced pressure to no alcohol taste, extracts 2 times (supernatant liquid) with isometric ethyl acetate, closes
And acetic acid ethyl acetate extract, recycling ethyl acetate obtain acetic acid ethyl ester extract (EA) to doing.
(2) influence of the Radix Rhapontici seu Radix Echinopsis EA to SCC15 cell Proliferation
The Radix Rhapontici seu Radix Echinopsis EA of 12.5 μ g/ml, 25 μ g/ml, 50 μ g/ml, 100 μ g/ml various concentrations is acted on into SCC15 cell
Afterwards, SCC15 cell Proliferation is inhibited by different degrees of.With the raising of drug concentration, SCC15 cell proliferation inhibition rate also with
Raising.Radix Rhapontici seu Radix Echinopsis EA acts on SCC15 cell for 24 hours, and the IC50 of 48h, 72h are respectively 112.62 ± 0.21 μ g/ml, 87.71 ±
0.49 μ g/ml and 65.19 ± 0.46 μ g/ml prompts Radix Rhapontici seu Radix Echinopsis EA inhibited to SCC15 cell Proliferation, and increases at any time
Inhibiting effect is added to enhance.
(3) influence of the Radix Rhapontici seu Radix Echinopsis EA to the apoptosis of SCC15 cell
By the Radix Rhapontici seu Radix Echinopsis EA of 12.5 μ g/ml, 25 μ g/ml, 50 μ g/ml, 100 μ g/ml various concentrations, to be respectively acting on SCC15 thin
Born of the same parents for 24 hours, 48h, using FITC/PI double-staining, flow cytomery apoptosis rate.The results show that in the time, with
Drug concentration increases, and apoptosis rate dramatically increases；Under same concentrations, apoptosis rate increases obviously 48h ratio for 24 hours.Illustrate Radix Rhapontici seu Radix Echinopsis EA
SCC15 Apoptosis can be significantly induced, and there is time-and concentration-dependent.
(4) Radix Rhapontici seu Radix Echinopsis EA acts on apoptosis correlation phosphorylated protein p38, JNK and ERK1/2
Using apoptosis index of correlation in MAPK signal path in SCC15 cell after Western blot detection Radix Rhapontici seu Radix Echinopsis EA processing
Phosphorylated p38 (p-p38), phosphorylation JNK (p-JNK), the variation of phosphorylated CREB 1/2 (p-ERK).50 μ g/ml Radix Rhapontici seu Radix Echinopsis EA processing
After cancer cell of oral cavity SCC15 48h, p-p38 relevant to apoptosis is promoted and p-JNK expression are significantly increased, and relevant to proliferation is promoted
P-ERK expression reduces.The result shows that Radix Rhapontici seu Radix Echinopsis EA has the function of promoting Apoptosis.
(5) influence of the Radix Rhapontici seu Radix Echinopsis EA to SCC15 cell invasion and migration
Radix Rhapontici seu Radix Echinopsis EA is detected to the shadow of invasion and the migration of SCC15 cell using Transwell Matrigel and scratch experiment
It rings.In Matrigel, 12.5 μ g/ml, 25 μ g/ml, 50 μ g/ml, tri- kinds of various concentrations Radix Rhapontici seu Radix Echinopsis EA can inhibit in various degree
The invasive ability of SCC15 cell, compared with the control group, under Radix Rhapontici seu Radix Echinopsis EA group cell increases invasive ability gradually with drug concentration
Drop.Scratch experiment is as the result is shown: 12.5 μ g/ml, 25 μ g/ml, the Radix Rhapontici seu Radix Echinopsis EA processing SCC15 of 50 μ g/ml, tri- kinds of various concentrations are thin
For 24 hours, 48h can reduce the external healing ability of SCC15 cell to born of the same parents in various degree, as the increase of drug concentration inhibits migration to make
With being more obvious.
(6) Radix Rhapontici seu Radix Echinopsis EA is to the influence of SCC15 cell epithelia-mesenchymal transformation
50 μ g/ml Radix Rhapontici seu Radix Echinopsis EA handle cancer cell of oral cavity SCC15 48h, Q-PCR and Western blot to EMT index of correlation
The expression of the mRNA and albumen of E-cadherin, Snail, Vimentin are detected.The results show that E-cadherin expression increases
Height, Snail, Vimentin expression reduce.Illustrate that Radix Rhapontici seu Radix Echinopsis EA has the work of reversing oral cancer cell SCC15 Epithelial and stromal conversion
With.
Detailed description of the invention
Influence of Fig. 1 Radix Rhapontici seu Radix Echinopsis EA to SCC15 cell Proliferation；
Influence of Fig. 2 Radix Rhapontici seu Radix Echinopsis EA to SCC15 cell migration；
Influence of Fig. 3 Radix Rhapontici seu Radix Echinopsis EA to SCC15 cell invasion；
Fig. 4 Radix Rhapontici seu Radix Echinopsis EA, which expresses SCC15 cell EMT relevant molecule, to be influenced
Influence of Fig. 5 Radix Rhapontici seu Radix Echinopsis EA to SCC15 Apoptosis；
Fig. 6 Radix Rhapontici seu Radix Echinopsis EA influences the expression of SCC15 cell MAPK apoptosis-related protein；
Specific embodiment
(1) CCK-8 method:
Cell Proliferation is detected using CCK-8 method.Experimental setup experimental group, Radix Rhapontici seu Radix Echinopsis EA concentration are respectively 12.5 μ g/ml, 25 μ
g/ml,50μg/ml；Control group, with experimental group with the DMSO of concentration.Logarithmic growth phase cell, with pancreatin containing 0.25%EDTA
Supernatant is abandoned in digestion, centrifugation.It is 2.5 × 10 with the culture medium adjustment cell concentration containing 10% serum4A/ml.Every hole inoculation
200 μ l cell suspensions enter in 96 well culture plates, every group of 4 multiple holes.It is adherent overnight after, inhale abandon supernatant, by group be added containing difference
The culture solution (serum content 10%) of concentration Radix Rhapontici seu Radix Echinopsis EA, and set solvent control group continue cultivate different time (for 24 hours, 48h,
72h).Every hole adds 190 μ l culture mediums and 10 μ l CCK8 reagents, is placed in 37 DEG C, 5%CO2In saturated humidity incubator after culture 2.5h
Terminate culture.It is measured in microplate reader each hole absorbance (A value), selects 450nm wavelength, recorded as a result, simultaneously counting as follows
Calculate proliferation inhibition rate: proliferation inhibition rate (%)=(AExperimental group-ABlank group)/(AControl group-ABlank group) × 100%.Experiment is repeated 3 times.
(2) cell scratch experiment:
Logarithmic growth phase SCC15 cell is pressed into every hole 5 × 105It is a to be inoculated in 6 well culture plates, fusion is grown to cell
Afterwards, with the vertical scratch of 200ul yellow pipette tips.Experimental group adds respectively containing different Radix Rhapontici seu Radix Echinopsis EA concentration (12.5 μ g/ml, 25 μ g/ml, 50 μ
G/ml serum free medium), control group are added comparable sodium DMSO, are put into 37 DEG C, 5%CO2Culture in saturated humidity incubator,
By sampling for 24 hours, under 48h microscope, take pictures.Every group takes three positions to take pictures and measures relative distance between scratch hecatomeral cells, away from
From variation divided by 2 be cell relative migration distance, calculate cell relative mobility: relative mobility (%)=relative migration away from
From/0h scratching edge away from scratch middle line with a distance from, in triplicate and carry out statistical analysis.Experiment is repeated 3 times.
(3) Transwell Matrigel:
It is carried out using 24 cells orifice plate Transwell, the matrigel filter sizes that cell is coated with collagen stroma are 8 μ
m.It will freeze in 4 DEG C of BD matrigel of -80 DEG C of refrigerators overnight.Serum free medium 1:5 dilution is drawn 100 μ l and is uniformly spread
To the cell Transwell upper chamber, it is put into 37 DEG C, 5%CO2In saturated humidity incubator overnight.Be added in upper chamber 100 μ l pre-temperatures without
Serum DMEM, incubation at room temperature.The culture medium containing 12.5 μ g/ml, 25 μ g/ml, 50 μ g/ml Radix Rhapontici seu Radix Echinopsis EA is handled into training respectively respectively
Support for 24 hours, the SCC15 cell of 48h.Concentration is 5 × 10 by 0.25% trypsin digestion5The cell inoculation in a/hole is small in upper layer
Room, each 100 μ l cell suspension of cell.10% fetal calf serum (FBS) culture solution, 500 μ l, 37 DEG C of incubations are added in bottom chamber
For 24 hours, cell is taken out, the cell on filter membrane upper layer is dabbed off with cotton swab, filter membrane is washed 2 times with PBS, 10% formalin is fixed
15min, Gimsa dye 10min, and 5 visuals field are randomly selected under high power lens, and counting is migrated to the cell of filter membrane lower layer, experiment weight
It is 3 times multiple.Invasion inhibiting rate (%)=(quantity that quantity-experimental group that control group wears theca cell wears theca cell)/control group wears film
Quantity × 100% of cell.
(4) FITC/PI flow cytometry
By logarithmic growth phase SCC15 cell inoculation in middle ware, experimental group Radix Rhapontici seu Radix Echinopsis EA final concentration is respectively 12.5 μ g/ml, 25 μ
g/ml,50μg/ml,100μg/ml；Control group, with experimental group with the DMSO of concentration.Handle SCC15 cell for 24 hours, 48h.By reagent
Box specification is collected cell and is dyed, flow cytomery apoptosis rate.
(5)Q-PCR
50 μ g/ml Radix Rhapontici seu Radix Echinopsis EA extracts handle SCC15 cell 48h, it is total to extract group of cells respectively using Trizol reagent
RNA.With spectrophotometer measurement RNA concentration and quality.With ABI reverse transcription reagent box by total serum IgE reverse transcription at cDNA.Using one
Footwork RT-PCR reaction kit amplification carries out the amplification of PCR with PCR kit for fluorescence quantitative.It is carried out using relative quantitation method
Interpretation of result calculates 2-△△CtValue, GAPDH is as internal reference.Experiment is in triplicate.Gene primer sequence are as follows: GAPDH-F, 5 '-
aggtcggtgtgaacggatttg-3'；GAPDH-R,5'-tgtagaccatgtagttgaggtca-3'.Snail-F,5'-
ttaccttccagcagccctac-3'；Snail-R,5'-gacagagtcccagatgagca-3'.E-cadherin-F,5'-
ttgctactggaacagggaca-3'；E-cadherin-R,5'-gtattgggaggaaggtctgc-3'.Vimetin-F,5'-
gaagagaactttgccgttga-3'；Vimetin-R,5'-cgaaggtgacgagccatt-3'.Amplification reaction condition are as follows: pre-
95 DEG C of 10min are denaturalized, 95 DEG C of 15S are denaturalized, anneal 60 DEG C of 1min, totally 40 circulations.Melt curve analysis analysis, 95 DEG C of 15s, 60 DEG C
1min, 95 DEG C of 15s, 60 DEG C of 15s.Interpretation of result is carried out using relative quantitation method, calculates 2-△△CtValue.
(6)Western Blot
SCC15 cell processing grouping such as Q-PCR, collects group of cells respectively, adds protein lysate, mix, crack on ice
15min, 4,12000r/min are centrifuged 10min, take supernatant, as total protein of cell, dense using Bradford method measurement albumen
Degree, 1 × sds gel sample loading buffer heregulin concentration keep each group consistent, take each 50 μ g of protein sample containing same concentrations respectively, add
4 × loading buffer (3:1), 100 DEG C of 5min, 4 DEG C of 5min, through 10%SDS-PAGE electrophoresis transferring film, Ponceaux dyeing is seen
Transfer effect is examined, and determines the position of Protein Marker.2h is closed containing 5% skimmed milk power TBST fluid-tight, primary antibody rabbit-anti is added
People E-cadherin (1:1000), Snail (1:500), Vimentin (1:200), p-p38 (1:1000), p-JNK (1:
1000), p-ERK1/2 (1:1000) polyclonal antibody (1:2000) or rabbit-anti people GAPDH (1:2000), 4 DEG C of overnight incubations,
TBST is washed film 3 times, and secondary antibody horseradish peroxidase label goat anti-rabbit igg (1:3000) is added, and 37 DEG C of incubation 1h, TBST are washed film 3 times,
With the expression of ECL developer solution detection substrate, Labworks image analysis system carries out gray scale scanning to result, analyzes result.
Claims (3)
1. the application of the ethyl acetate extract of Radix Rhapontici seu Radix Echinopsis or its preparation in the drug of preparation prevention and treatment carcinoma of mouth；
The ethyl acetate extract of the Radix Rhapontici seu Radix Echinopsis is prepared by following methods:
Radix Rhapontici seu Radix Echinopsis sample comminution takes 500 grams of coarse powder at coarse powder, with 80% ethyl alcohol it is wet after, then with 3 times of 80% alcohol refluxs of amount extractions
Twice, combined extract is recovered under reduced pressure ethyl alcohol to no alcohol taste, is extracted 2 times with isometric ethyl acetate, supernatant liquid is merged
To acetic acid ethyl acetate extract, ethyl acetate is recycled to doing, obtains ethyl acetate extract.
2. application according to claim 1, which is characterized in that the preparation is by Radix Rhapontici seu Radix Echinopsis ethyl acetate extract and pharmaceutically
Acceptable carrier composition.
3. application according to claim 2, which is characterized in that the preparation is tablet, dragee, capsule, injection, molten
Liquid, lotion, ointment, emulsifiable paste, inhalant, aerosol or suppository.
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| CN103463158A (en) * | 2013-08-20 | 2013-12-25 | 天津药业集团新郑股份有限公司 | Technology for extracting effective constituents of radix rhapontici |
| CN104099180A (en) * | 2014-06-16 | 2014-10-15 | 南京泽朗医药科技有限公司 | Extraction method for Stemmacantha uniflora volatile oil |
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| CN103463158A (en) * | 2013-08-20 | 2013-12-25 | 天津药业集团新郑股份有限公司 | Technology for extracting effective constituents of radix rhapontici |
| CN104099180A (en) * | 2014-06-16 | 2014-10-15 | 南京泽朗医药科技有限公司 | Extraction method for Stemmacantha uniflora volatile oil |
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| 禹州漏芦抗炎和抑菌有效部位的筛选;李喜凤等;《中国实验方剂学杂志》;20150228;第21卷(第3期);第138-141页 * |
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