CN104147127B - A kind of Chinese medicine composition and its preparation method and application for treating malignant tumour - Google Patents
A kind of Chinese medicine composition and its preparation method and application for treating malignant tumour Download PDFInfo
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Abstract
The present invention provides a kind of Chinese medicine composition and its preparation method and application for treating malignant tumour.The Chinese medicine composition is prepared by following bulk pharmaceutical chemicals: rhizoma menispermi, Radix Notoginseng and Radix Astragali, auxiliary material needed for preparations shaping is added after each bulk pharmaceutical chemicals are crushed, clinically suitable various preparations are made into according to pharmaceutical preparation conventional method, Chinese medicine composition of the invention its full side is clearing heat and detoxicating, strengthens the body resistance to consolidate the constitution and gets rid of evils, the function of the powdered repellent-antalgesic medicine stasis of blood, according to clinical verification satisfactory effect.Meanwhile the present invention, by studying the Chinese medicine composition to the antitumor action and mechanism of action of internal and external tumor model, the new Chinese medicine for further research and development treatment malignant tumour provides objective basis.
Description
Technical field
The present invention relates to Chinese medicine compositions, and in particular to a kind of Chinese medicine composition for treating malignant tumour further relates to its system
Preparation Method and application.
Background technique
Malignant tumour is to seriously endanger the formidable enemy of human health, oneself warp becomes the first killer of urban and rural residents, China death,
It and is in rise year by year trend.Anti- curing oncoma is the key subjects of contemporary medical science circle, extensively the concern by domestic and international medical field.So
And the treatment of malignant tumour is very difficult.Surgical operation, radiotherapy, the chemotherapy of doctor trained in Western medicine are currently treat malignant tumour three big basic
Means.However surgical operation not only brings huge physiology and psychological pain to patient, and its late result is also not to the utmost
People's will.The non-specific effect for killing tumour cell of radiation and chemotherapy causes normal cell and organ injury not can avoid, with
Cancer itself is not died of as some patients, but indirectly dies of toxicity brought by radiation and chemotherapy drug.By
It is not difficult to find out that, all there are some drawbacks in the big conventional therapy of the three of malignant tumour above, and people expect there is some new methods and hand
Duan Laiyu cancer is struggled.1994, Canadian schipper professor proposed in the new model about malignant tumour concept,
Effective treatment does not need the complete recession of tumour, and the reactivity of body is mostly important to cancer therapeutic.This viewpoint mentions
It matches out with the characteristics of righting reinforcing of traditional Chinese medicine treatment malignant tumour.Chinese medicine with its righting (enhancing immunity of organisms), dispel
Evil (killing tumor cell) and few side effects are increasingly taken seriously.
Summary of the invention
For existing issue, technical problem to be solved by the invention is to provide a kind of for treating in malignant tumour
Drug composition.Full side is clearing heat and detoxicating, strengthens the body resistance to consolidate the constitution and gets rid of evils, the function of the powdered repellent-antalgesic medicine stasis of blood, according to clinical verification satisfactory effect.The present invention
Further investigated is carried out to Chinese medicine composition antitumor action and mechanism with advanced experimental technique means.The present invention will be to develop
The clear safely and effectively anti-tumor Chinese medicine new drug of mechanism of action lays the foundation, before important theory significance and wide application
Scape.
For this purpose, the present invention provides a kind of Chinese medicine composition for treating malignant tumour and application.The Chinese medicine composition be by
Following bulk pharmaceutical chemicals are prepared: rhizoma menispermi, Radix Notoginseng, Radix Astragali.
Specifically, the Chinese medicine composition is prepared by each bulk pharmaceutical chemicals of following parts by weight: 5-15 parts of rhizoma menispermi, three
Seven 5-10 parts, 15-25 parts of Radix Astragali.
Preferably, the Chinese medicine composition is prepared by each bulk pharmaceutical chemicals of following parts by weight: 10 parts of rhizoma menispermi, Radix Notoginseng
7.5 parts, 20 parts of Radix Astragali.
Auxiliary material needed for preparations shaping is added after crushing in the bulk pharmaceutical chemicals, is made into clinic according to pharmaceutical preparation conventional method
Upper suitable various preparations, the preparation includes capsule, pill, granule, tablet or oral solution.The system of invention formulation
Preparation Method belongs to conventional preparation method, and there is no need to go into details herein.
Chinese medicine composition of the invention can be used for preparing the drug for the treatment of malignant tumour.
Chinese medicine composition of the invention its full side is clearing heat and detoxicating, strengthens the body resistance to consolidate the constitution and gets rid of evils, the function of the powdered repellent-antalgesic medicine stasis of blood, through clinic
Verify satisfactory effect.Meanwhile the present invention is by studying the Chinese medicine composition to the antitumor action of internal and external tumor model
And mechanism of action, the new Chinese medicine for further research and development treatment malignant tumour provide objective basis.
Detailed description of the invention
Fig. 1 is cancer of pancreas BxPC-3 cell growth curve；
Fig. 2 is the traditional chinese medicine composition of the invention to the morphologic influence diagram of P2 group cancer of pancreas mouse heterotopic transplantation tumor；
Fig. 3 is the traditional chinese medicine composition of the invention to the morphologic influence diagram of P1 group cancer of pancreas mouse heterotopic transplantation tumor；
Fig. 4 is the traditional chinese medicine composition of the invention to the morphologic influence diagram of 5-FU group cancer of pancreas mouse heterotopic transplantation tumor；
Fig. 5 is the traditional chinese medicine composition of the invention to the morphologic influence diagram of M group cancer of pancreas mouse heterotopic transplantation tumor.
Specific embodiment
Below with reference to embodiment, the invention will be further described, it should be understood that these embodiments are only used for illustration
Purpose, be never limited in protection scope of the present invention.
Chinese medicine composition used is prepared in accordance with the following methods:
By the weighed each bulk pharmaceutical chemicals of following weight: 10 parts of rhizoma menispermi, 7.5 parts of Radix Notoginseng, 20 parts of Radix Astragali.The medicinal distilled water of raw material
1h is impregnated, decocting 2 times, each 30min, decocting liquid mixes twice, and Rotary Evaporators are concentrated into 50mL respectively, and water is made in 4 DEG C of preservations
Decoction is used for following research:
1. data and method
1.1 patients of inclusion criteria 75 are in September, 2011 in August, -2013 in Heilongjiang University of Chinese Medicine attached first
The malignant tumor of digestive tract patient of hospital treatment.The overwhelming majority is that doctor trained in Western medicine diagnosis is definite when all cases patient assessment, hand
Recidivist after art, chemotherapy.
1.2 general information set control group and treatment group.It is shown in Table 1
1 two groups of clinical datas of table
1.3 clinical stages, carried out according to International Union Against Cancer (UICC) 1987 about the TNM standard of malignant tumour clinical
By stages.In 75 malignant tumor patients, majority is mid-term (accounting for 85%), and has the transfer of different situations, such as enlargement of lymph nodes,
Stomach, brain, abdominal metastas, with chest, ascites and fever, bleeding, pain etc..
1.4 clinical criterias are all to make a definite diagnosis tumour for positive according to modern medicine inspection: 1. rabat, x line, cr,
The instruments such as MRI, dry plate check positive (+)；2. the fiberoptic bronchoscopy positive (+)；3. living tissue is drawn materials through pathological tissue
Check positive (+)；4. secretion exfoliative cytology checks positive (+)；5. Celom liquid loading, which draws water, checks positive (+)；6. all distal ends
There is transferrer at organ or position；7. clinical symptoms show it is related to the above diagnose combine clarify a diagnosis.
1.5 treatment method treatment groups apply 1 dose/day of Chinese medicine composition, 20 days as one therapeutic course, totally 3 courses for the treatment of；Control group is not
Use Chinese medicine composition.Two groups are assisted in the treatment of simultaneously, such as hydrothorax of dispelling, ascites, analgesic, are stopped blooding, improved a poor appetite and defaecation.
1.6 criterions of therapeutical effect carry out blood according to the state of an illness, urine, feces routine test and specific index are chemically examined, the film making of x line, B ultrasound,
CT etc. is checked, observes clinical symptoms, such as weight, cough, feed, pain, bleeding, body temperature, breathing, heart rate, hydrothorax, ascites, is swollen
Tumor size etc..
2. observing result
2 two groups of patient clinical total effects of table compare
It is analyzed through Ridit, there is significant difference (P < 0.05) between two groups.
2 pharmacological experiment of test example
Chinese medicine composition used is prepared in accordance with the following methods:
By the weighed each bulk pharmaceutical chemicals of following weight: 10 parts of rhizoma menispermi, 7.5 parts of Radix Notoginseng, 20 parts of Radix Astragali.The medicinal distilled water of raw material
1h is impregnated, decocting 2 times, each 30min, decocting liquid mixes twice, and Rotary Evaporators are concentrated into 50mL respectively, and water is made in 4 DEG C of preservations
Decoction is used for following research:
1. experiment in vitro: in vitro culture is carried out to human pancreas cancer BxPC-3 cell, with Chinese medicine composition high and low dose group,
The culture solution intervention of 5-FU (5 FU 5 fluorouracil), the normal culture solution culture of blank control group carry out following experiment: utilizing placenta
Orchid dyeing measurement BxPC-3 cell growth cycle simultaneously draws growth curve；After detecting pharmaceutical intervention using cell scratch experiment
The locomitivity of BxPC-3 cell；With BxPC-3 cel l proliferation after MTT method detection pharmaceutical intervention；Using fluidic cell
Art detects BxPC-3 cell cycle and Apoptosis after pharmaceutical intervention.
2. experiment in vivo: preparing Nude Mouse Model, weight and tumour growth feelings of the observation Chinese medicine composition to mouse
The influence of condition, and measure spleen index, tumour inhibiting rate；Tumour cell ultra microstructure is observed under transmission electron microscope, is inquired into the present invention with this
The antitumor action and its mechanism of action of drug composition are carried out especially by following experiment:
Experiment in vitro:
1. experimental material
Chinese medicine composition is prepared according to the above method, according to a conventional method be concentrated decocting liquid at 2g/ml crude drug concentration, it is cold after sterilizing
Spare preparation decocting liquid is hidden, 4 DEG C save backup.People original position pancreas cancer cell strain BxPC-3 is purchased from Chinese Academy of Sciences Shanghai life
Research institute's cell resource center.Urea pyrimidine Tegafur Tablet, Qilu Pharmaceutical Co., Ltd., lot number 0502002；Culture solution:
RPMI1640, IMDM are purchased from Gibco company；Fetal calf serum: Hangzhou Chinese holly bio-engineering corporation product.
2. key instrument equipment
CO2Incubator (2323 type of TC), superclean bench, digital display Constant Temp. Oven (202-2A type), inverted phase contrast
Microscope (LH50A type), enzyme-linked immunosorbent assay instrument (DG3022A type), high speed tabletop centrifuge (TGL-16C type), centrifuge
(LD42 type), research microscope (BH-2 type), flow cytometer etc..
3. experimental method
3.1 cancer of pancreas BxPC-3 cell culture
3.1.1 the recovery of cell: being removed from liquid nitrogen freeze-stored cell, and cryopreservation tube investment has been warmed up to 37 DEG C of water rapidly
It thaws in bath, and constantly shakes, make to freeze liquid in pipe and melt rapidly in 1~2min.With 75% alcohol disinfecting cryopreservation tube
Outer wall moves back in superclean bench.By in liquid in pipe sucking the EP pipe of sterilized mistake, 1000rpm/min is centrifuged 3min,
Supernatant is abandoned, 10ml culture medium is added, being blown and beaten becomes single cell suspension, cell count, and adjustment concentration is that 5 × 105/ml connects
Culture bottle is placed in 37 DEG C, 5%CO in culture bottle by kind2Incubator in cultivate, next day changes liquid.
3.1.2 the passage of cell: can carry out cell passage when the adherent 80-90% of cell, careful to draw former culture medium, use
PBS is rinsed twice, discards PBS, appropriate trypsin solution is added, jiggles culture bottle, bottom of bottle is made to come into full contact with digestion pancreatin,
Bottle cap is covered tightly, is placed in 37 DEG C of incubators after 3~5min, when discovery cell has a small amount of fall off, addition culture medium termination is taken out and disappears
Change, collect liquid in EP pipe, 1500rpm/min is centrifuged 5min, abandons supernatant, is cleaned twice with PBS, discards PBS, and training is added
Base is supported to blow and beat cell for single cell suspension and adjust cell concentration 1 × 105/ ml is inoculated in culture bottle, and culture bottle is placed in
37 DEG C, 5%CO2Incubator in continue to cultivate.
3.1.3 cell freezes: will change liquid on the day before cell cryopreservation, according to the operation that cell passes on, cell concentration adjusted
It is 3~9 × 106/ ml, 1500 rpm/min are centrifuged 5min, abandon supernatant, and appropriate pre-configured frozen stock solution (10% is added
DMSO is sufficiently mixed with 90%FBS, matching while using), keep mixing with cells uniform, is sub-packed in marked cryopreservation tube, every pipe
1ml.Then cell is placed in 4 DEG C of 10min, -20 DEG C of 30min, -80 DEG C overnight, are finally putting into liquid nitrogen and save for a long time.
3.1.4 the counting of cell: single cell suspension is made in the postdigestive cell of pancreatin, and 100 μ l cell suspensions is taken to be added
100 μ l tires expect blue dye liquor (0.4%), mix well.A set of clean blood counting chamber is taken, coverslip is covered on blood count slot
On, it draws the above-mentioned cell count liquid of 10 μ l and is squeezed into along coverslip edge, record under inverted microscope and be not colored in four quadrants
Cell number.The cell concentration of cell suspension is the sum of four quadrant cell numbers/4 × 2 × 104。
3.1.5 pharmaceutical intervention: logarithmic growth phase human pancreas cancer BxPC-3 cell, with trypsin digestion, PBS cleaning 3
It is secondary, single cell suspension is made, cell concentration is adjusted to 1 × 105/ ml, is inoculated in 25cm2In culture bottle, it is put into 37 DEG C of 5%CO2
After incubator culture for 24 hours, former culture medium is abandoned, the culture medium of final concentration of 80, the 40 μ g/ml containing Chinese medicine composition is separately added into, sun
Property control group the culture medium of the final concentration of 50 μ g/ml containing 5-FU is added, blank control group adds normal incubation medium, is placed in 37 DEG C 5%
CO2Incubator culture 48h.
3.2 placenta indigo plant decoration methods detect cell growth curve
Logarithmic growth phase human pancreas cancer BxPC-3 cell, with trypsin digestion, PBS is cleaned 3 times, is made unicellular outstanding
Cell concentration is adjusted to 1 × 10 by liquid4A/ml is inoculated in 24 well culture plates with the 300 every holes μ l, is put into 37 DEG C of 5%CO2Culture
Case continues to cultivate, 1 day (for 24 hours), 2 days (48h), 3 days (72h), 4 days (96h), 5 days (120h), 6 days (144h) daily it is same
Time takes four holes to be counted respectively.Experimental data in the past few days is collected, the growth curve of human pancreas cancer BxPC-3 cell is drawn.
3.3MTT method detects cell Proliferation
Logarithmic growth phase human pancreas cancer BxPC-3 cell, with trypsin digestion, PBS is cleaned 3 times, is made unicellular outstanding
Cell concentration is adjusted to 2 × 10 by liquid5A/ml is inoculated in 96 well culture plates with the 100 every holes μ l.It cultivates in the incubator for 24 hours
Afterwards, culture medium is replaced, the culture medium of final concentration of 80, the 40 μ g/ml containing Chinese medicine composition is separately added into, positive controls addition contains
The culture medium of the final concentration of 50 μ g/ml of 5-FU, every 100 μ l of hole, blank control group add normal incubation medium, and every 100 μ l of hole, every group sets
6 multiple holes continue to cultivate 48h, and culture terminates preceding 4h, and MTT (5mg/ml) 10 μ l is added in each culture hole, continue after cultivating 4h,
Careful to draw culture medium, every hole is added DMSO100 μ l, in wavelength is at 490nm with microplate reader after 37 DEG C of shaking tables concussion 10min
Detect the absorbance value (OD) in each hole.The above experiment is repeated 3 times.Receipt is collected, calculates drug to BxPC-3 cell inhibitory rate.
3.4 scratch experiments detect cell motility
Compare ruler in 6 orifice plates behind with marker first, equably draws horizontal line, it is standardized per every about 0.5~1cm
Road crosses via hole.Every hole is at least across 3 lines.About 5 × 10 are added in every hole5A BxPC-3 cell is placed in training in incubator
After supporting for 24 hours, compares ruler with 20 μ l pipette tips, draw scratch perpendicular to the horizontal line of behind as far as possible.It is washed cell per well 3 times with PBS, to go
Except lower cell is drawn, it is separately added into the culture medium of final concentration of 80, the 40 μ g/ml containing Chinese medicine composition, control group is added containing 5-FU
The culture medium of final concentration of 50 μ g/ml, blank control group add normal incubation medium, and every group sets 4 multiple holes, are put into 37 DEG C of 5%CO2Training
Case is supported, continues to cultivate 48h.At the shooting scratch of sampling in 0,24,48 hour.The distance of scratch is measured using Photoshop software,
Every hole randomly selects 3 positions.Cell motility can be judged according to the relative distance that scoring position cell heals.
Scratch healing relative distance (%)=(l-48h distance/0h distance) × 100%
3.5 detection apoptosis rates: the cancer of pancreas BxPC-3 cell of every group of logarithmic growth phase is with 2 × l05/ ml, 4ml connect
Kind is in 25cm2In culture bottle, after culture 24 hours, final concentration of 80 μ g/ml of Chinese medicine composition, 40 μ g/ml is added in administration group
The culture medium of the final concentration of 50 μ g/ml containing 5-FU is added in culture medium, positive controls, and blank control group supplies isometric culture
Base is placed after cultivating 48 hours in incubator, and PBS is cleaned 2 times, and pancreatin digests 3~5min, and culture medium is added and terminates digestion, receives
Collect cell in 15ml centrifuge tube, 1500rpm is centrifuged 5min, and PBS is washed twice, and single cell suspension is made.5 μ l Annexin are added
V-FITC is mixed gently.Room temperature (20~25 DEG C), which is protected from light, is incubated for 10min (aluminium foil can be used to be protected from light).1000g centrifugation
5min abandons supernatant, 190 μ l Annexin V-FITC combination liquid is added, cell is carefully resuspended.10 μ l propidium iodide stain liquid are added
(Propidium iodide, PI), mixes gently, ice bath avoid light place.Flow cytometer detection should be carried out interior for 24 hours after the completion of dyeing,
Preferably completed on the same day.Flow cytometer is the red burst light of detection at 488 n m wavelength in excitation wavelength, while detection light
The case where scattering.Software is analyzed using instrument itself to analyze cell DNA content and light scattering situation.Above data passes through
CellQuest software is collected and is handled.
3.6 detection cell cycles: the cancer of pancreas BxPC-3 cell of every group of logarithmic growth phase is with 2 × l05/ ml, 4ml inoculation
In 25cm2In culture bottle, after culture 24 hours, the training of Chinese medicine composition final concentration of 80 μ g/ml, 40 μ g/ml is added in administration group
Base is supported, the culture medium of the final concentration of 50 μ g/ml containing 5-FU is added in positive controls, and blank control group supplies isometric culture
Base is placed and is cultivated 48 hours in incubator.PBS is cleaned 2 times, and pancreatin digests 3~5min, and diploid product culture solution termination is added and disappears
Change, collect cell in 15ml centrifuge tube, 1500rpm is centrifuged 5min, and PBS is washed twice, and single cell suspension is made.1500rpm from
The heart is removed except cell fragment, is fixed with -20 DEG C of 70% ethyl alcohol of pre-cooling, and 4 DEG C overnight, and 1500rpm is centrifuged 5min, abandons supernatant, often
Propidium iodide stain liquid is added to final concentration of 50mg/L in pipe, is protected from light, 37 DEG C of incubation 30min use argon with flow cytomery
Ion laser excites fluorescence, and excitation light wave wavelength is 488nm, and transmitting optical wavelength is 650nm, each sample counting 20000
Cell carries out cell cycle analysis using Multicycle software.
The analysis of 3.7 data
T is examined between the statistical analysis of group difference uses SPSS software to carry out group, and each group of data is usedIndicate own
P value indicates to compare two-by-two, and P < 0.05 is significant difference, and P < 0.01 is that difference is extremely significant, all has statistical significance.
4 experimental results
The growth curve of 4.1 cancer of pancreas BxPC-3 cells
Cancer of pancreas BxPC-3 cell growth curve is drawn as shown in Figure 1, with cell concentration 1 according to different time viable count
×104After/ml inoculation, logarithmic growth phase was entered from the 4th day, the growth inhibition phase is entered after the 5th day.
4.2 cancer of pancreas BxPC-3 cell motility situations
The case where 1 cancer of pancreas BxPC-3 cell motility of table
Note: compared with blank group, P < 0.05 is expressed as *, and P < 0.01 is expressed as * *, and compared with 5-FU group, P < 0.05 is indicated
For #, P < 0.01 is expressed as ##.
4.3 cancer of pancreas BxPC-3 cell proliferative conditions
The case where 2 cancer of pancreas BxPC-3 cell Proliferation of table
Note: compared with blank group, P < 0.05 is expressed as *, and P < 0.01 is expressed as * *, and compared with 5-FU group, P < 0.05 is indicated
For #, P < 0.01 is expressed as ##.
4.4 cancer of pancreas BxPC-3 apoptosis rate situations
The case where 3 cancer of pancreas BxPC-3 apoptosis rate of table
Note: P < 0.05 is expressed as * compared with blank group, and P < 0.01 is expressed as * *.
4.5 cancer of pancreas BxPC-3 cell cycle stages
The case where 4 cancer of pancreas BxPC-3 cell cycle of table
Note: P < 0.05 is expressed as * compared with blank group, and P < 0.01 is expressed as * *.
Experiment in vivo part:
1 experimental material
SPF grades of BALB/c mouses of 1.1 experimental animals and tumor strain (4 week old) are purchased from the limited public affairs of Shanghai Si Laike experimental animal
Department, people original position pancreas cancer cell strain BxPC-3 are purchased from Shanghai Inst. of Life Science, CAS cell resource center.
1.2 experimental drugs and reagent Chinese medicine composition are prepared by above-mentioned side, and decocting liquid is concentrated according to a conventional method into the life of 2g/ml
Concentration refrigerates spare preparation decocting liquid after sterilizing, 4 DEG C save backup.5-FU (sigma company, the U.S.), modified form RPMI-
1640 culture mediums, fetal calf serum, dual anti-(100 μ g/ml of 100 μ g/ml of penicillin and streptomysin), digestion pancreatin (U.S. HyClone
Company), Trizol kit, DEPC (the raw work bioengineering in Shanghai), Raf-1 antibody (SANTA CRUZ), MEK1 antibody, ERK1
Antibody (Cell Signaling), goat anti-rabbit igg secondary antibody, goat anti-rat IgG secondary antibody, goat anti-mouse IgG secondary antibody,
GAPDH mouse monoclonal antibody (abcam company, the U.S.), Protein Extraction Reagent box, BCA determination of protein concentration kit (green skies biology skill
Art research institute), pre-dyed albumen Marker (Fermentas), other reagents are domestic or the pure grade of Import Analysis.
1.3 instrument and equipments: medical type clean bench (Beijing Dong Lianhaer instrument manufacturing Co., Ltd)；CO2 incubator
(Li Kang Development Co., Ltd)；Inverted microscope (philippines company, the U.S.)；Refrigerator (the limited public affairs of Hefei Rongshida refrigerator
Department)；Liquid nitrogen biological container (Cengdu Jinfeng Liquid Nitrogen Container Co. Ltd.)；Super constant temperature trough (the limited public affairs of the permanent scientific instrument in Shanghai one
Department)；Supercentrifuge (Hunan Hunan instrument H1650-W)；Real-time quantitative PCR detector (Bio Rad Laboratories)；Multipurpose decoloration table concentrator
(Suzhou victory beauty SYC-2101)；Labworks image acquisition and analysis software (Alpha Innotech company, the U.S.)；Vertical electrophoresis apparatus and transferring film
System (Bio Rod company, the U.S.).
2. experimental method
2.1 animal models, grouping and administration logarithmic growth phase BxPC-3 cell, it is 5 × 10 that concentration, which is made,6A/ml's
Cell suspension is spare.It is subcutaneous to extract the nude mice right fore armpit that cell suspension 0.2ml/ is only inoculated in after conventional transdermal disinfection.It is dynamic
Object, which is weighed in, is randomly divided into five groups, model group (M), physiological saline group (C), positive controls (5-FU), and Chinese medicine composition is high
Dosage group (P2) and Chinese medicine composition low dose group (P1), every group 6, totally 30.C group is not inoculated with BxPC-3 cell, other four
Group is inoculated with.Animal inoculation pvaccination carries out pharmaceutical intervention afterwards for 24 hours, and successive administration 21 days, 0.9% physiology was injected intraperitoneally in C group, M group daily
Only, only, i.e., daily administration dosage is 20mg/kg to intraperitoneal injection 2mg/ml 5-FU 0.2ml/ to 5-FU group to salt water 0.2ml/ daily；
Only, i.e., daily administration dosage is 20mg/kg to intraperitoneal injection 2mg/ml Chinese medicine composition 0.2ml/ to P20 group daily；The daily abdomen of P10 group
Chamber injects 1mg/ml Chinese medicine composition 0.2ml/ only, i.e., daily administration dosage is 10mg/kg.
After above 5 groups of nude mice tumor formations (gross tumor volume about 100mm3), weight is measured, and swollen from in-vitro measurements with vernier caliper
The longest diameter and shortest diameter of tumor, and gross tumor volume, relative tumour volume and relative body weight are calculated according to formula, 2~3 times weekly,
With dynamic observation tumour growth situation；After being observed continuously 8 weeks, nude mice is put to death after weighing, tumor tissues are taken under aseptic condition, is claimed
Knurl weight calculates tumour inhibiting rate；Changed using tumor cell morphology in transmission electron microscope observing transplantable tumor.
2.2 statistical analysis
The data obtained is analyzed using SPSS17.0 analysis software, all data with mean ± standard deviation (x ± s),
Group difference compares to be examined with t, and otherness compares using variance analysis between multiple groups.Evaluation criteria P < 0.05 is to have statistics
Meaning, P < 0.01 are to have conspicuousness statistical significance.
3. experimental result
3.1 BxPC-3 cancer of pancreas mouse model body weights
5 BxPC-3 cancer of pancreas mouse model body weights of table
Note: P2 indicates Chinese medicine composition 20mg/kg group, and P1 indicates Chinese medicine composition 10mg/kg group, and 5-FU indicates positive right
According to group, M group indicates model control group, and C indicates blank control group.
Experimental result is as shown in Table 5, and experiment each group mouse weight has increase, and wherein the increase of 5-FU group mouse weight is the most
Slowly, the increase of C group mouse weight is most fast, and P2 group, and P1 group and the increase of M group mouse weight are slowly but slightly fast compared with 5-fU group compared with C group.
Each group is administered compared with C group without statistical difference, each group is administered compared with M group without statistical difference.
3.2 BxPC-3 cancer of pancreas mouse model spleen weights and spleen index situation
6 BxPC-3 cancer of pancreas mouse model spleen weight of table and spleen index situation
Note: P2 indicates Chinese medicine composition 20mg/kg group, and P1 indicates Chinese medicine composition 10mg/kg group, and 5-FU indicates positive right
According to group, M group indicates model control group, and C indicates blank control group.5-FU * compared with C group indicates P < 0.05.
As shown in table 6, P2 group, P1 group equal no difference of science of statistics compared with C group spleen weight, P2 group, P1 group is compared with M group spleen weight
No difference of science of statistics, 5-FU group is compared with C group and M group spleen weight with statistical difference.Each administration group is equal compared with C group spleen index
No difference of science of statistics, P2 group P1 group no difference of science of statistics compared with M group spleen index, 5-FU group have compared with C group and M group spleen index
There is statistical difference.
3.3 BxPC-3 cancer of pancreas mouse model knurl weights and tumour inhibiting rate situation
7 BxPC-3 cancer of pancreas mouse model knurl weight of table and tumour inhibiting rate situation
Note: P2 indicates Chinese medicine composition 20mg/kg group, and P1 indicates Chinese medicine composition 10mg/kg group, and 5-FU indicates positive right
According to group, M group indicates model control group, and C indicates blank control group.Each group * compared with M group indicates that P < 0.05, * * indicate P <
0.01。
Shown in table 7, P2, P1 and 5-FU group knurl weight have a degree of mitigation, P2, P1 and 5-FU group knurl weight and M group ratio
More statistically significant, P2 group is statistically significant compared with M group knurl weight；Each group tumor control rate is respectively
55.88%, 36.14%, 30.88%.
3.4 Chinese medicine compositions are on the morphologic influence of cancer of pancreas BxPC-3 mouse heterotopic transplantation tumor tissue
Fig. 2-5 is respectively that Chinese medicine composition is morphologic to P2, P1,5-FU and M group cancer of pancreas mouse heterotopic transplantation tumor
Influence diagram.
The foregoing is merely presently preferred embodiments of the present invention, is merely illustrative for the purpose of the present invention, and not restrictive
's.Those skilled in the art understand that in the spirit and scope defined by the claims in the present invention many changes can be carried out to it,
It modifies or even equivalent, but falls in protection scope of the present invention.
Claims (5)
1. a kind of Chinese medicine composition for treating malignant tumour is to be prepared by each bulk pharmaceutical chemicals of following parts by weight: rhizoma menispermi 5-
15 parts, 5-10 parts of Radix Notoginseng, 15-25 parts of Radix Astragali；
Wherein, the malignant tumour is malignant tumor of digestive tract.
2. Chinese medicine composition according to claim 1 is to be prepared by each bulk pharmaceutical chemicals of following parts by weight: rhizoma menispermi 10
Part, 7.5 parts of Radix Notoginseng, 20 parts of Radix Astragali.
3. the preparation method of Chinese medicine composition of any of claims 1 or 2 is that preparations shaping institute is added after crushing each bulk pharmaceutical chemicals
The auxiliary material needed is made into clinically suitable various preparations according to pharmaceutical preparation conventional method.
4. preparation method according to claim 3, the preparation includes capsule, pill, granule, tablet or oral
Liquid.
5. Chinese medicine composition of any of claims 1 or 2, the application in preparation treatment malignant tumor of digestive tract drug.
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