Summary of the invention
Technical problem to be solved by the present invention lies in provide a kind of anti-lung-cancer medicament suitable for obstruction of the circulation of vital energy matter and its preparation
Method, the drug dissipate the stasis of blood as the rules for the treatment of according to theory of traditional Chinese medical science, using strengthening vital QI to eliminate pathogenic factors, relieve stagnation, can obviously inhibit obstruction of the circulation of vital energy matter lung cancer tumor
Growth and transfer.
In order to solve the above-mentioned technical problems, the present invention provides a kind of anti-lung-cancer medicament suitable for obstruction of the circulation of vital energy matter, the medicines
Object includes following raw medicinal material: Radix Codonopsis, oldenlandia diffusa, Sculellaria barbata, Prunella vulgaris, edible tulip, semen litchi, raw pearl barley, bulb of thunberg fritillary, perfume (or spice)
Attached, cortex albiziae and Radix Glycyrrhizae;Wherein, all raw medicinal materials are the medicine materical crude slice after processing;
The drug is made of the raw medicinal material of following parts by weight: Radix Codonopsis 10-20 parts by weight, oldenlandia diffusa 25-35
Parts by weight, Sculellaria barbata 25-35 parts by weight, Prunella vulgaris 20-28 parts by weight, edible tulip 20-28 parts by weight, semen litchi 20-28 weight
Part, raw pearl barley 25-35 parts by weight, bulb of thunberg fritillary 13-23 parts by weight, rhizoma cyperi 7-17 parts by weight, cortex albiziae 13-23 parts by weight and Radix Glycyrrhizae 4-
14 parts by weight.
The drug is preferably made of the raw medicinal material of following parts by weight: Radix Codonopsis 12-18 parts by weight, oldenlandia diffusa
27-32 parts by weight, Sculellaria barbata 28-32 parts by weight, Prunella vulgaris 23-25 parts by weight, edible tulip 22-27 parts by weight, semen litchi 22-26
Parts by weight, raw pearl barley 27-33 parts by weight, bulb of thunberg fritillary 15-20 parts by weight, rhizoma cyperi 10-15 parts by weight, cortex albiziae 15-20 parts by weight and
Radix Glycyrrhizae 6-12 parts by weight.
The drug can also preferably be made of the raw medicinal material of following parts by weight: 15 parts by weight of Radix Codonopsis, agkistrodon
30 parts by weight of tongue grass, 30 parts by weight of Sculellaria barbata, 24 parts by weight of Prunella vulgaris, 24 parts by weight of edible tulip, 24 parts by weight of semen litchi, the raw heart of a lotus seed
Rice 30 parts by weight, 18 parts by weight of bulb of thunberg fritillary, 9 parts by weight of 12 parts by weight of rhizoma cyperi, 18 parts by weight of cortex albiziae and Radix Glycyrrhizae.
The drug is made of the raw medicinal material of following parts by weight again: 10 parts by weight of Radix Codonopsis, 35 weight of oldenlandia diffusa
Part, 35 parts by weight of Sculellaria barbata, 20 parts by weight of Prunella vulgaris, 28 parts by weight of edible tulip, 28 parts by weight of semen litchi, 35 weight of raw pearl barley
Part, 13 parts by weight of bulb of thunberg fritillary, 4 parts by weight of 7 parts by weight of rhizoma cyperi, 13 parts by weight of cortex albiziae and Radix Glycyrrhizae.
The drug can also preferably be made of the raw medicinal material of following parts by weight: 20 parts by weight of Radix Codonopsis, agkistrodon
25 parts by weight of tongue grass, 25 parts by weight of Sculellaria barbata, 28 parts by weight of Prunella vulgaris, 20 parts by weight of edible tulip, 20 parts by weight of semen litchi, the raw heart of a lotus seed
Rice 25 parts by weight, 23 parts by weight of bulb of thunberg fritillary, 14 parts by weight of 17 parts by weight of rhizoma cyperi, 23 parts by weight of cortex albiziae and Radix Glycyrrhizae.
In order to preferably realize foregoing invention purpose, the present invention also provides a kind of anti-lung-cancer medicaments suitable for obstruction of the circulation of vital energy matter
Preparation method, preparation method includes:
(1) all raw medicinal materials are weighed by the parts by weight, cold water is added, impregnated 2-3 hours, it is small then to boil 2-3
When, filtering obtains medical fluid one, the dregs of a decoction are spare;
(2) dregs of a decoction warm water in step (1) decocts 1-2 hours, and filtering obtains medical fluid two and the dregs of a decoction;
(3) it is filtered after merging medical fluid one and medical fluid two, continues heating and decoct, in decoction process and donkey-hide gelatin, Huang is added
Wine and honey, and be stirred continuously, it decocts to the complete molten of donkey-hide gelatin, continues heating after filtering until stirring into cream taste;Wherein, Ah
The additive amount of glue, yellow rice wine and honey is respectively 10-14 parts by weight, 9-11 parts by weight and 12-17 parts by weight;Preferably, donkey-hide gelatin, Huang
The additive amount of wine and honey is respectively 12 parts by weight, 10 parts by weight and 15 parts by weight;
(4) cream taste is poured into while hot and carries out packing work in automatic packer, dispensed, obtained into according to every bag of 20g
Product are placed on dry environment and are on the waiting list.
The beneficial effects of the present invention are: drug of the invention is according to traditional Chinese medical theory, compatibility is reasonable, and prescription is rigorous, to help
It is the rules for the treatment of that positive eliminating evil, relieve stagnation, which dissipate the stasis of blood, has extraordinary anti-tumor effect of suppression cancer to obstruction of the circulation of vital energy matter lung cancer.
Specific embodiment
Tcm constitution is the important component of Basic Theories of Chinese Medicine, is a new branch of science.Phenomenon of constitution is that the mankind are raw
A kind of movable important behaviour form is ordered, is referred in human vital process, in native endowment and institute's shape of acquired on the basis of
At morphosis, comprehensive, metastable intrinsic speciality in terms of physiological function and psychological condition, be the mankind in growth, hair
It is formed by during educating and human body personal characteristics natural, that social environment is adaptable.Constitution and card had both had the difference of essence,
There are close ties again, constitution decides body to the tendency in the easy property suffered from and pathological process of certain diseases in many cases
Property.The difference of constitution leads to the variability of disease: the cause of disease is identical, and constitution is different, and card is also different;Disease is identical, and constitution is different, card
Also different;Disease is different, and constitution is identical, and card is also identical, i.e., constitution is with the different card of disease, different disease with the basis demonstrate,proved.
For the present invention exactly from constitution, providing for obstruction of the circulation of vital energy constitution lung cancer a kind of has effects that the medicine of tumor-inhibiting anticancer
Object, including following raw medicinal material: Radix Codonopsis, oldenlandia diffusa, Sculellaria barbata, Prunella vulgaris, edible tulip, semen litchi, raw pearl barley, bulb of thunberg fritillary,
Rhizoma cyperi, cortex albiziae and Radix Glycyrrhizae.Wherein, Radix Codonopsis is monarch drug in a prescription.Radix Codonopsis is sweet, flat, and returns spleen, lung two pass through, and energy tonifying spleen lung qi enriches blood, promotes the production of body fluid.
" materia medica ": " tonifying spleen nourishing the stomach, moistening lung and producing body fluid are good for gas in fortune, this is not very mutually remote with ginseng ";" new compilation of materia medica ": " bowl spares benefit
Gas and taste, relieving thirst and restlessness.The middle micro- void of gas is mended, very safety to adjust ".White flower oldenlandia diffusa, Sculellaria barbata, semen coicis, perfume (or spice)
Attached, cortex albiziae is ministerial drug.It is white flower oldenlandia diffusa slight bitter, sweet, it trembles with fear, returns stomach, large intestine, small intestinl channel.Kind removing toxic substances heat-clearing, dampness removing are logical
Leaching.Sculellaria barbata acrid flavour, slight bitter are put down, and the heart, small intestine, lung channel are entered, and have the benefits of clearing heat and detoxicating, inducing diuresis for removing edema, activating microcirculation and removing stasis medicinal.Semen coicis
Sweet, light, cool, returns spleen, stomach, lung channel have the benefits of invigorating spleen for diuresis, detumescence, eliminating impediment, clearing away heat and eliminating pus.Compendium of Material Medica: " semen coicis, sun
Bright medicine, can invigorating spleen and reinforcing stomach.Reinforcing the mother-organ to treat deficiency, thus consumptive lung disease, pulmonary abscess with it ".Rhizoma cyperi is pungent, slight bitter, micro- sweet and neutral.Return liver, spleen, three
Jiao Jing.Can soothing liver-qi stagnation, menstruction regulating and pain relieving, in qi-regulating tune.Cortex albiziae is sweet, flat, return heart, liver, lung channel.Resolving stagnation for tranquilization, activating blood circulation and reducing swelling.
Bulb of thunberg fritillary, Prunella vulgaris, edible tulip, semen litchi are adjutant.Bulb of thunberg fritillary is bitter, cold, return lung, the heart channel of Hang-Shaoyin, has clearing heat and detoxicating, dissipating bind to disappear the function of carbuncle
Effect.Supplementary Amplifications of the Compendium of Materia Medica: " detoxify sharp phlegm, open the inhibited lung-energy, all lung man folder wind fire have phlegm person preferably this." Prunella vulgaris is pungent, bitter, it trembles with fear,
Liver, gallbladder two is returned to pass through, clearing heat-fire, mass dissipating and swelling eliminating." Chongqing hall notes ": " Prunella vulgaris, acrid flavour and it is sweet, therefore among dissipating bind, have conjunction concurrently
The function of positive yin-nourishing." edible tulip is sweet, micro-pungent, cool, and liver, spleen two is returned to pass through, clearing heat and detoxicating, dissipating bind disappears carbuncle.Semen litchi is pungent, slight bitter, temperature.
Return liver, stomach meridian.Promoting the circulation of qi dissipating bind, eliminating cold to stop pain.Radix Glycyrrhizae is sweet, flat, effect tonifying middle-Jiao and Qi, expelling phlegm and arresting coughing, relieving spasm to stop pain, heat-clearing solution
Poison.Raw the benefits of taking its clearing heat and detoxicating with Radix Glycyrrhizae master, and coordinating the drug actions of a prescription, to make medicine.Full side is made a general survey of, compatibility is reasonable, and prescription is rigorous, medicine
Energy is special, meets traditional Chinese medical theory, all medicines share, plays strengthening vital QI to eliminate pathogenic factors altogether, relieve stagnation dissipates the stasis of blood, has the benefits of pressing down cancer anti-tumor.
The present invention will be described in detail below with reference to the drawings of preferred embodiments, whereby to the present invention how applied technology method
Technical problem is solved, and the realization process for reaching technical effect can fully understand and implement.
Embodiment 3
The embodiment of the invention provides a kind of for treating the drug of obstruction of the circulation of vital energy matter lung cancer, by the raw medicinal material system of following g number
At: Radix Codonopsis 200g, oldenlandia diffusa 250g, Sculellaria barbata 250g, Prunella vulgaris 280g, edible tulip 200g, semen litchi 200g, raw pearl barley
250g, bulb of thunberg fritillary 230g, rhizoma cyperi 170g, cortex albiziae 230g and Radix Glycyrrhizae 140g;All raw medicinal materials are the medicine materical crude slice after processing;
Preparation method are as follows:
(1) all raw medicinal materials are weighed by above-mentioned weight, cold water are added, impregnated 2 hours, then boil 3 hours, filtered,
Medical fluid one is obtained, the dregs of a decoction are spare;
(2) dregs of a decoction warm water in step (1) decocts 2 hours, and filtering obtains medical fluid two and the dregs of a decoction;
(3) it is filtered after merging medical fluid one and medical fluid two, continues heating and decoct, in decoction process and donkey-hide gelatin, Huang is added
Wine and honey, and be stirred continuously, it decocts to the complete molten of donkey-hide gelatin, continues heating after filtering until stirring into cream taste;Wherein, Ah
The additive amount of glue, yellow rice wine and honey is respectively 140g, 110g and 120g;
(4) cream taste is poured into while hot and carries out packing work in automatic packer, dispensed, obtained into according to every bag of 20g
Product are placed on dry environment and are on the waiting list.
Syndrome is the core and soul of Chinese medicine, and zoopery is to connect basic medical and clinical bridge and tie,
Therefore carry out with disease combination principle be instruct zoopery be TCM basis research the only way which must be passed.But due to animal and
Otherness (such as language expression symptom, tongue picture, pulse condition, the colour of skin and feelings will symptom) between human body, determines that tcm syndrome exists
It is simulated with animal larger with the difficulty of judgement.Therefore, have using TCM syndrome diagnostic criteria to be distinctive with guiding principle, with animal
The information characteristics of diagnostic significance carry out integration comparison, equivalent association, and accurate, quantization modern physiological pathological biochemistry is combined to refer to
Mark is to evaluate Chinese medicine to the effective way of Animal diseases and therapeutic effect of syndrome.Therefore, the present invention is carried out following right by animal model
Than test, for proving that drug of the invention has a clear superiority in treatment obstruction of the circulation of vital energy matter lung cancer.
Comparative test
Main agents and drug are as follows:
Interferon-γ (IFN-γ) enzyme-linked immunoassay kit: Bioengineering Research Institute is built up in Nanjing;Leucocyte is situated between
Element -2(IL-2) enzyme-linked immunoassay kit: Nanjing is built up in Bioengineering Research Institute;Southern Sea Brand cigarette: BJ Tobacco Factory is raw
It produces, tar content 8mg, nicotine content in smoke: 0.8mg, carbon monoxide in flue gas content: 10mg;Hydrocortisone injection is purchased from day
Saliva medicine company Jiaozhuo Co., Ltd;Adrenalin hydrochloride injection, specification 1g:1mg are purchased from Shanghai Hefeng Pharmaceutical Co., Ltd.;It is small
Mouse thyroid-stimulating hormone (TSH) enzyme-linked immunoassay kit;Corticotropin (ACTH) enzyme-linked assay kit, by south
Bioengineering Research Institute's offer is built up in capital;Testosterone (T) radioimmunoassay kitss, by Tianjin Jiuding Medical Biological Engineering Co., Ltd
It provides;Cyclic adenosine monophosphate (cAMP) enzyme-linked immunoassay kit: Bioengineering Research Institute is built up in Nanjing;Follicle-stimulating hormone (FSH)
Enzyme-linked immunoassay kit: Bioengineering Research Institute is built up in Nanjing;High lipid food is purchased from Shandong Province's disease control and prevention
The heart;Interleukins radio-immunity test kit (IL-6), builds up Bioengineering Research Institute purchased from Nanjing;Vascular endothelial growth
Factor C(VEgF-C) enzyme-linked immunoassay kit: Bioengineering Research Institute is built up in Nanjing;DAB color developing agent: it is purchased from China fir in Beijing
Biotechnology Co., Ltd, Golden Bridge;OVA (grade), AI (OH) 3 are purchased from Sigma, USA;0.9% sodium chloride injection, is purchased from
The tomb Guo great pharmaceutical Co. Ltd.
Experiment equipment is as follows:
Superclean bench: Beijing Great Wall air purification engineering company
Animal contaminated case: Shanghai Yu Yan scientific instrument Co., Ltd
High-pressure sterilizing pot: Japanese Sanyo company
Microplate reader: Thermo company, the U.S.
Inverted microscope: Japanese Olympus company
Disecting microscope: Japanese Nikon company
CO2 cell incubator: German Teraeus company
Vernier caliper: Shanghai experimental measurement company
Superspeed refrigerated centrifuge: German Herge company
Analytical precision balances: Mettler company, Switzerland
- 80 DEG C of refrigerators: Japanese Sanyo company
General refrigerator: Qingdao HaiEr Co., Ltd
Cell counting board: Shanghai precision instrumentation Co., Ltd
Micropipettor: German Eppendor company
Automatic blood rheology test instrument is provided by Center for Disease Control, Shandong Province.
One, gentle constitution and obstruction of the circulation of vital energy constitution model are established
1 experimental animal
Animal strains C57BL/6 inbred strais black rat, grade SPF rank, male, 6-8 week old, weight 18 ± 2g, 80,
Quality testing unit: Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences sells unit: Beijing China Fukang biotechnology share
Co., Ltd, credit number: SCXK (capital) 2009-0004.
2 test methods
2.1 feeding method
It feeds in SPF grades of animal houses (Center for Disease Control, Shandong Province animal house), room temperature (22 ± 2) DEG C, humidity (55 ±
5) %, each 12h of artificial lighting light and shade, well-ventilated;Mouse maintains feed, pulls together feed corporation,Ltd purchased from Beijing Austria, section, license
Card number: the capital SCXK() 2009-0012;Free water;Padding is replaced in time according to padding situation.
2.2 groupings and modeling method
2.2.1 group technology:
80 Lewis mouse are randomly divided into two experimental groups, each experimental group 40, respectively gentle matter group and the obstruction of the circulation of vital energy
Matter group.
2.2.2 modeling method:
Two groups of mouse are raised respectively two weeks with different rearing conditions, the rearing conditions of two experimental groups are specific as follows:
(1) it gentle matter group: feeds with normal diet, free water, room temperature (22 ± 2) DEG C, humidity (55 ± 5) %, artificial light
Illuminate dark each 12h.It is continuous to feed 14 days under the conditions of well-ventilated.
(2) obstruction of the circulation of vital energy matter group: feeding 7 days under the conditions of above-mentioned gentle matter group, then from the 8th day to the 14th day, is tied up with band
Firmly rat four limbs, and continue to feed using the feed mode of gentle matter group.
3 experimental evaluation indexs and result
The observation of 3.1 mouse weights
Each test group of animals respectively surveys a weight after before experiment starts with experiment.The measured body weight time is 9:
00am-10:00am calculates the average weight of two experimental mice experiment front and backs, and the results are shown in Table 1.
1 liang of experimental mice experiment forward backward averaging weight of table () table of comparisons
By above-mentioned data it is found that obstruction of the circulation of vital energy matter group is compared with gentle matter group mouse weight, the T inspection through two independent samples
It tests, P < 0.05 is statistically analyzed, and each group of data weight has differences, and has statistical significance.Wherein, obstruction of the circulation of vital energy matter group is small
Mouse average weight is significantly less than gentle matter group average mice body weight.
The observation of 3.2 mouse macroscopic tokens
Life habit, the state of mind, food-intake, the hair situation, size of two experimental mices are observed and recorded daily
Just situations such as situation, skin and train of thought color, extravasated blood change.It is as follows to observe result:
(1) gentle matter group: mouse shows as active active, is quick on the draw, back hair color is glossy, and feed water is normal;
(2) obstruction of the circulation of vital energy matter group: mouse gradually appears the performance of obstruction of the circulation of vital energy constitution, and such as burnout lazy move, apathetic, four limbs are crispaturaed,
Being irritable and getting angry easily, emotional, food-intake is reduced.
3.3 Evaluation of blood test and method
In experiment the 15th day, 5 mouse are respectively randomly selected from two groups of mouse, blood are taken using eye socket blood extracting method, after taking blood
It is added in the test tube without anti-coagulants, room temperature blood natural coagulation 10-20 minutes, is centrifuged 25 minutes (2500 revs/min).Carefully collect
Upper serum should be centrifuged again as precipitated during preservation.Serum is subjected to following Indexs measure respectively:
(1) ACTH content in serum: ACTH is detected using enzyme-linked immunosorbent assay instrument (microplate reader).The results are shown in Table 2.
The ACTH content table of comparisons in 2 two groups of mice serums of table (± S)
Group |
ACTH(pg/g) |
Gentle matter group |
31.51±10.12* |
Obstruction of the circulation of vital energy matter group |
58.67±6.81 |
It is independent through two by above-mentioned data it is found that obstruction of the circulation of vital energy matter group is compared with ACTH content in gentle matter group mice serum
The T of sample is examined, and P < 0.05 is statistically analyzed, having differences property of each group of data, and obstruction of the circulation of vital energy matter group mouse ACTH content is obvious
Higher than gentle matter group mouse ACTH content.
(2) it whole blood viscosity: is carried out respectively with tri- shear rates of 10S-1,50S-1,200S-1 with lectin from hemolymph automatic tester
Whole blood viscosity detection.The results are shown in Table 3.
3 two groups of mice serum whole blood viscosity tables of comparisons of table (± SmPaS)
Group |
10S-1(mPaS) |
50S-1(mPaS) |
200S-1(mPaS) |
Gentle matter group |
11.31±1.03 |
5.49±1.19 |
3.88±0.62 |
Obstruction of the circulation of vital energy matter group |
13.82±1.21* |
8.12±1.11** |
6.41±0.83*** |
Note: * P < 0.05;* P < 0.05;* * P < 0.05.
By above-mentioned data it is found that obstruction of the circulation of vital energy matter group is compared with gentle matter group Mouse whole blood viscosity, the T through two independent samples
It examines, P < 0.05, each group of data has differences, and has statistical significance, the whole blood under obstruction of the circulation of vital energy matter group three groups of shear rates of mouse
Viscosity is obviously higher than gentle matter group Mouse whole blood viscosity.
To sum up, by the observation of table 1, the data of table 2 and table 3 and macroscopic token it is found that two groups of mouse are in weight and characterization
There is notable difference, there is also significant differences for ACTH content and whole blood viscosity in serum, and obstruction of the circulation of vital energy matter group mouse from characterization and is surveyed
Fixed number illustrates that this constitution modeling experiment is stablized effectively, models successfully according to matching with obstruction of the circulation of vital energy constitutive character.
Two, the lung cancer model of gentle constitution and obstruction of the circulation of vital energy constitution is established
1 experimental animal and cell strain
Experimental animal of two experimental mices (every group 40) as this modeling after the modeling of above-mentioned constitution, and respectively
As gentle matter group and obstruction of the circulation of vital energy matter group.
Lewis lung cancer cell line, is provided by Shandong Academy of Medical Sciences and is inoculated with, and is inoculated with random from mouse kind with mouse
Extract 5 progress lotus knurls.It is sent to laboratory after being inoculated with successfully and carries out conventinal breeding.
2 experimental methods and step
2.1 tumor cell inoculations for the first time
It takes the tumour of the C57BL/6 mouse of lotus Lewis lung carcinoma cell as knurl source, mouse cervical dislocation is put to death, ultra-clean work
Make to separate tumor tissues of the well-grown pale pink without necrosis under platform aseptic condition.It is cut into small pieces with tissue shear, by tumor (g):
The ratio of physiological saline (g) 1:3 is unidirectionally ground into homogenate with glass homogenizer, crosses 200 mesh nylon wires, at single cell suspension, receives
It combines in centrifuge tube.With superspeed refrigerated centrifuge, 1000r/min is centrifuged 3min, abandons supernatant, and physiological saline is added, and is centrifuged with method
It washed once.Tumor cell suspension is taken, 0.4% trypan blue dye liquor is added, trypan blue exclusion experiment shows that living cells is greater than 95%, adjustment
Cell concentration is 1.0*107/g。
Aseptically, with syringe inoculated tumour cell suspension 0.2g/ at the armpit of two experimental mices
(a containing about oncocyte 2.0*106) mixes suspension before being inoculated with every time, cell infusion is slightly suitable after the completion to press pin hole, prevents liquid
It overflows.
The 5th day after inoculation, two test group mouse-borne tumor success or not are observed, lotus knurl success mouse is that lung cancer is modeled as
Function.
Three, drug study
The successful mouse of lotus knurl 30 in above-mentioned two groups is chosen respectively, as the animal model of drug study, and by every group
30 mouse are randomly divided into Liang Ge group, and respectively gentle matter control group, gentle matter are to drug, obstruction of the circulation of vital energy matter control group and obstruction of the circulation of vital energy matter
Administration group, 15 mouse of each group.Specific medication is as shown in table 4:
Table 4 respectively tests the medication of group mouse
Note: above-mentioned dosage is equivalent to 70kg adult by the equivalent dose of body surface area conversion dosage, is equivalent to simultaneously
10 times of per weight people's dosage.
Composition analyzed index and result:
(1) weight: after above-mentioned administration mode continuous use four weeks, claim each group mouse weight with electronic balance and remember
The results are shown in Table 5 for record;
Table 5 respectively tests group's average mice body weight table of comparisons after being administered
Group |
Quantity (only) |
Average weight (g) |
Gentle matter control group |
15 |
27.06±2.10 |
Gentle matter administration group |
15 |
32.37±1.62 |
Obstruction of the circulation of vital energy matter control group |
15 |
25.03±2.81 |
Obstruction of the circulation of vital energy matter administration group |
15 |
27.35±2.73 |
From the above data, it can be seen that one-way analysis of variance, big group difference shows: P < 0.05, has significant difference
Property;Further progress compares two-by-two, and the administration group of every kind of constitution grouping is compared with non-medication group, and: P < 0.05 has significant difference
Property;One-way analysis of variance: P > 0.05 is carried out between different constitution administration groups, does not have otherness.
(2) gross tumor volume: the 7th day after inoculation, 10 days, 13 days, 16 days, 19 days, 22 days, 25 days, 28 days, precision is used
For 0.1mm vernier caliper measurement and record mouse subcutaneous tumors tubercle size, calculate gross tumor volume, calculation formula: V (cm3)=
0.52×ab2(a is knurl longest diameter, and b is shortest diameter).The average value of every group's mouse tumor volume is calculated, as a result such as table 6
It is shown.
Each group's mouse tumor average external volume change situation (cm3) during being administered of table 6
It is swollen according to the mouse of group to can be seen that gentle matter control group, gentle matter administration group and obstruction of the circulation of vital energy confrontation by above-mentioned data
Knurl product growth rate is essentially identical, and under drug effect, administration group volume growth rate gradually slows down since the mid-term, rather than
Administration group, tumour growth rate have the tendency that speeding earlier above.Every kind of constitution administration group and non-medication group carry out statistical procedures, P
< 0.05.
(3) knurl weight and tumour inhibiting rate: each group mouse is put to death in dislocation of craning one after 28 days after inoculation, removes mouse skin rapidly
Lower tumour claims knurl weight, calculates each group tumor control rate, the results are shown in Table 7.
Wherein, the calculation formula of tumor control rate are as follows:
Tumor control rate=(the gentle average knurl weight of matter control group average knurl weight-to be measured group)/gentle matter control group is averaged tumor
Weight × 100%.
Each experiment group mouse knurl final weight table of comparisons of table 7 (± S, g)
Group |
Quantity (only) |
Knurl weight (g) |
Tumour inhibiting rate (%) |
Gentle matter control group |
15 |
7.86±1.27 |
0 |
Gentle matter administration group |
15 |
5.91±1.43 |
24.81 |
Obstruction of the circulation of vital energy matter control group |
15 |
9.72±0.47 |
-23.66 |
Obstruction of the circulation of vital energy matter administration group |
15 |
5.59±0.74 |
28.88 |
It can be seen that every kind of constitution Liang Ge group by above-mentioned data to compare through row: gentle matter administration group and gentle confrontation
Comparing: P < 0.05 according to group has extremely significant otherness;Obstruction of the circulation of vital energy matter administration group is compared with obstruction of the circulation of vital energy matter control group, and: P < 0.05 has
Significance difference is anisotropic.Various difference constitution administration groups compare P > 0.05, no significant difference.Illustrate drug of the invention to mouse tumor
Growth have apparent inhibiting effect, and two kinds of constitution model mice tumor inhibitory effects are showed well, two kinds of constitutions are small
Mouse tumour inhibiting rate no significant difference.
(4) after mouse is put to death, two sides lungs Lung metastases inhibiting rate: are separated.It is fixed for 24 hours with 4% paraformaldehyde solution.In
Pulmonary metastases stove is counted under disecting microscope, transfer stove is the protuberance of white or milky nodal-like.This test is using counting
It is completed under double-blind conditions, the results are shown in Table 8.
Wherein, the calculation formula of Lung metastases inhibiting rate are as follows:
Lung metastases inhibiting rate (%)=(gentle matter control group be averaged lung surface metastatic nodules-to be measured group respectively be averaged lung table
Face metastatic nodules)/gentle matter control group is averaged Lung metastases tubercle number × 100%.
Table 8 respectively experiment group mouse Pulmonary metastasis focuses number and Lung metastases inhibiting rate (± S)
Group |
Quantity (only) |
Pulmonary metastasis focuses number (a) |
Lung metastases inhibiting rate (%) |
Gentle matter control group |
15 |
11.30±2.95 |
0 |
Gentle matter administration group |
15 |
8.35±1.78 |
26.10 |
Obstruction of the circulation of vital energy matter control group |
15 |
11.10±1.20 |
1.77 |
Obstruction of the circulation of vital energy matter administration group |
15 |
7.70±2.36 |
31.85 |
It can be seen that every kind of constitution Liang Ge group by above-mentioned data to be compared: gentle matter administration group and gentle confrontation
Comparing: P < 0.05 according to group has extremely significant otherness;Obstruction of the circulation of vital energy matter administration group is compared with obstruction of the circulation of vital energy matter control group, and: P < 0.05 has
Significant difference;Various difference constitution administration groups compare P > 0.05, no significant difference.Illustrate that drug of the invention can reduce mouse
Neoplasm lung metastasis rate, and it is directed to above two constitution no significant difference.
(5) the VEGF immunohistochemistry detection of tumor tissues
Sample is made in the tumor tissues that each experiment group randomly selects 10 mouse, and is respectively put into 10% neutral good fortune
Fixed in your Malin's solution 4 μM of serial section 2 of row are opened for 24 hours, row VEGF dyeing, microwave antigen retrieval, conventional dehydration, transparent, leaching
Wax, embedding.According to the operation for illustrating to carry out immunohistochemistry of SABC method staining kit.
It is expressed using immunohistochemical method SP method count detection VEGF-C, all samples are normal after fixation, embedding, slice
Advise row HE dyeing.Primary antibody is rabbit-anti mouse monoclonal antibody, and 1:100 dilution, biotinylation secondary antibody is goat anti-mouse Igg.Each reality
It is as shown in table 9 to test group's VEGF ImmunohistochemistryResults Results.
Table 9 respectively the positive cell VEGE-C average optical density table of comparisons in experiment group's tumor tissues (± S)
Group |
Quantity (only) |
Average optical density |
Gentle matter control group |
10 |
0.158±0.033 |
Gentle matter administration group |
10 |
0.131±0.040 |
Obstruction of the circulation of vital energy matter control group |
10 |
0.197±0.034 |
Obstruction of the circulation of vital energy matter administration group |
10 |
0.154±0.023 |
It can be seen that every kind of constitution Liang Ge group by above-mentioned data to be compared: gentle matter administration group and gentle confrontation
Comparing: P < 0.05 according to group has extremely significant otherness;Obstruction of the circulation of vital energy matter administration group is compared with obstruction of the circulation of vital energy matter control group, and: P < 0.05 has
Significance difference is anisotropic.Comparison among groups: P > 0.05, no significant difference are administered in two kinds of different constitutions.Illustrate that drug of the present invention can reduce
Vegf expression reduces Tumor Angiongesis, and then inhibits tumour growth.
(6) serum Testing index (ACTH and whole blood viscosity)
Before putting to death after the end of the experiment, under a clean environment respectively to obstruction of the circulation of vital energy matter control group and obstruction of the circulation of vital energy matter administration group
Mouse eyeground takes blood, and blood sample is placed in anticoagulation test tube.With the content of ACTH in enzyme-linked immunization measurement peripheral blood serum
And whole blood viscosity, as a result as shown in Table 10 and Table 11.
ACTH content in 10 liang of experiment group mice serums of table (± S, pg/g)
Group |
ACTH(pg/g) |
Obstruction of the circulation of vital energy matter control group |
59.93±6.29 |
Obstruction of the circulation of vital energy matter administration group |
34.58±8.32* |
11 liang of experiment group Mouse whole blood viscosity measurements of table (±S ,mPa·S)
Group |
Quantity (only) |
10S-1 |
50S-1 |
200S-1 |
Obstruction of the circulation of vital energy matter control group |
15 |
16.82±1.21 |
11.18±1.11 |
8.41±0.83 |
Obstruction of the circulation of vital energy matter administration group |
15 |
12.74±0.56 |
7.82±1.21 |
4.21±1.21 |
By above-mentioned data it is found that ACTH content is significantly lower than obstruction of the circulation of vital energy matter control group in the mice serum of obstruction of the circulation of vital energy matter administration group,
And closer to the ACTH content value of normal mouse.Illustrate that drug of the invention has the effect for adjusting corticotropin
Fruit, and then by adjusting biased constitution, play the role of anticancer.
To sum up, pass through above-mentioned animal model test, it can be seen that drug of the invention can be improved the body of obstruction of the circulation of vital energy matter mouse
Weight, while having the effect of reducing tumour growth rate and reducing mouse tumor Lung metastases rate, and can reduce vegf expression and subtract
Few Tumor Angiongesis, adjusts the effect of corticotropin, to adjust biased constitution, and then enhances tumor-inhibiting anticancer
Effect, the i.e. present invention can have the effect of extraordinary tumor-inhibiting anticancer to obstruction of the circulation of vital energy matter lung cancer population.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and
Within principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.