CN100388925C - Medication composition of replenishing qi to invigorate the spleen, preparation method and usage - Google Patents
Medication composition of replenishing qi to invigorate the spleen, preparation method and usage Download PDFInfo
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- CN100388925C CN100388925C CNB2005100210208A CN200510021020A CN100388925C CN 100388925 C CN100388925 C CN 100388925C CN B2005100210208 A CNB2005100210208 A CN B2005100210208A CN 200510021020 A CN200510021020 A CN 200510021020A CN 100388925 C CN100388925 C CN 100388925C
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Abstract
The present invention provides a medical composition for replenishing qi to invigorate the spleen, which is a medicine prepared from total saponin of ginseng, white atractylodes rhizome oil, red ginseng and the complex polysaccharide of white atractylodes rhizome as raw materials. The present invention also provides a preparation method for the medical composition. The medicine of the present invention has the function of treating gastric precancerous lesion, and the medicine combining the four effective components has specific medical effects, perfroms cooperative effects, has strong controllability and stable quality and provides a new selection in clinic.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition of replenishing QI to invigorate the spleen, particularly, is to be that the effective site that the raw material extraction prepares is the pharmaceutical composition that feedstock production forms with the Chinese crude drug, belongs to drug world.
Background technology
Malignant tumor is well-known to human physical and mental health's serious threat.The patient of malignant tumor is died from the whole world every year, and 35% is the tumor of digestive system, wherein, and the mortality rate tool first place of gastric cancer.China's cancer feelings in 1997 announce that the case fatality rate of cancer rises to 18.14% of the nineties by 11.7% of the seventies, and gastric cancer occupies first of 10 kinds of malignant tumor.Recent study shows that gastric cancer is developed into by normal structure before the malignant tumor, be through quite long evolving stage, i.e. a preventing precancerous lesion of stomach cancer (Precancerous Lesions of gastric cancer).Because the cause of disease of gastric cancer still imperfectly understands, implement relatively difficulty of etiotropic primary prevention, therefore, Chinese scholars preventing precancerous lesion of stomach cancer as the important content of carrying out the gastric cancer secondary prevention.World Health Organization (WHO) has unified the understanding of disease (or state) and precancerous lesion (or sick damage) before the cancer in 1978.Disease is meant and the stomach benign disease that substantial connection is arranged of gastric cancer before the cancer, comprises atrophic gastritis, residual stomach, polyp of stomach, pernicious anemia and gastric ulcer etc.And preventing precancerous lesion of stomach cancer be meant gastric mucosa occur in, the severe atypical hyperplasia or/and imperfection colon type intestinalization give birth to.Chinese scholars has been carried out research widely to preventing precancerous lesion of stomach cancer.As discoveries such as Levin B., calcium, selenium, folic acid etc. are suitable for the chemoprophylaxis of gastric cancer precancerosis, and chemoprophylactic mechanism may comprise corrects carcinogenic activity and cell absorption, suppresses the conduction of abnormal signal, and the inducing cell accent is died and suppressed carcinogenesis; Foreign scholar Fang JY.NenseyYM. domestic scholars Yang Jia fine jade, results of study such as Fang Jingyuan, Li Chunqi show, folic acid, retinoic acid to the gastric cancer cancer before disease and preventing precancerous lesion of stomach cancer certain drug action is arranged, its mechanism is may a cancer cell specific induction of apoptosis relevant.But above achievement in research, it is to be solved to also have many problems to have, not high such as the potency ratio of medicine, suppresses the late result of precancerous lesion and does not affirm that the dosage of medicine and the effect relation of reverse are unclear, and the mechanism that suppresses precancerous lesion is imprecise.In a word, up to the present, still lack the sure chemotherapy medicine of curative effect.
In recent years, Chinese scholar has been carried out some researchs to preventing precancerous lesion of stomach cancer under Chinese medical theory instructs.Professor thinks as Dong Jianhua, and preventing precancerous lesion of stomach cancer belongs to the category of the traditional Chinese medical science " stuffy sensation of deftciency type ", is divided into deficiency of both QI and YIN, the bright stomach of asthenic fire, empty cloudy weak three types of taste.Professor Zhang Jingren thinks, atrophic gastritis merge imperfection colon type intestinal epithelial metaplasia or in, the severe atypical hyperplasia, be exactly preventing precancerous lesion of stomach cancer, morbidity is with weakness of the spleen and stomach, just QI-insufficiency is relevant.It is generally acknowledged that at present preventing precancerous lesion of stomach cancer belongs to the category of traditional Chinese medical science feeling of fullness, gastric abscess, see more that insufficiency of the spleen is the basic pathogenesis of preventing precancerous lesion of stomach cancer, be the grade positive correlation between the weight of insufficiency of the spleen symptom and preventing precancerous lesion of stomach cancer and the gastric cancer generation stages of development pathological changes with deficiency syndrome.
By Radix Ginseng, the Rhizoma Atractylodis Macrocephalae is pharmaceutical preparation beginning of feedstock production to be stated from bright " Standards of Diagnosis and Treatment ", forms by Radix Ginseng, the Rhizoma Atractylodis Macrocephalae, and be the basic representative prescription for the treatment of syndrome of deficiency of spleen qi.It springs from Han dynasty Zhang Zhongjing treatise on Febrile Diseases lizhong decoction, the Tang Dynasty Wang Tao " medical secrets of official " Radix Ginseng drink, Song dynasty " prescription of peaceful benevolent dispensary ", and Cheng Fang is in the Ming Dynasty " Standards of Diagnosis and Treatment ".Ming Dynasty's Wang Kentang " Standards of Diagnosis and Treatment. class side " volume one year " medicine cream of the present invention ... Radix Ginseng, each five equilibrium of the Rhizoma Atractylodis Macrocephalae, the thick soup clothes of decocting it ".Because technology is simple, volatile oil wherein is volatile, and effective ingredient easily loses, and drug effect is indeterminate.
Summary of the invention
In order to address the above problem, technical scheme of the present invention has provided a kind of pharmaceutical composition of replenishing QI to invigorate the spleen, and another technical scheme of the present invention has provided this preparation of drug combination method and purposes.
The invention provides a kind of pharmaceutical composition of replenishing QI to invigorate the spleen, it is to contain the medicament that is prepared from by the following weight proportion raw material:
Radix Ginseng total saponins 5-50 part, Rhizoma Atractylodis Macrocephalae oil 1-10 part, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide 5-50 part.
Further, it is the medicament that is prepared from by the following weight proportion raw material:
Radix Ginseng total saponins 5-50 part, Rhizoma Atractylodis Macrocephalae oil 1-10 part, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide 5-50 part.
Wherein, ginsenoside Rg1 and ginsenoside Re's gross weight percentage composition accounts for 0.5-5% in the described Radix Ginseng total saponins, and the percentage by weight of volatile oil 〉=60% in the described Rhizoma Atractylodis Macrocephalae oil contains linoleic acid 〉=30mg in every milliliter of Rhizoma Atractylodis Macrocephalae oil.
Described medicament is: pill, capsule, granule, tablet, oral liquid, injection.
The present invention also provides the method for this pharmaceutical composition, and it comprises the steps:
A, get the Radix Ginseng Rubra medical material, add 50%～80% soak with ethanol, reflux, extract, filters, merging filtrate, filtrate is collected eluent by macroporous adsorbent resin, reclaims ethanol, concentrate, Radix Ginseng total saponins;
C, get the Rhizoma Atractylodis Macrocephalae, edible vegetable oil is solvent, extracts, and filters, and filtrate is reclaimed the solvent vegetable oil, and products therefrom is a Rhizoma Atractylodis Macrocephalae oil;
D, get Rhizoma Atractylodis Macrocephalae medicinal residues, add ethanol extraction and filter, medicinal residues mix with the Radix Ginseng Rubra medicinal residues again, decoct with water, and filter, and are refining, standby, must Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide;
E, with the Radix Ginseng total saponins of b, c, d preparation, from art oil, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide by following weight proportion, add the preparation that acceptable accessories or complementary composition are prepared from.
Radix Ginseng total saponins 5-50 part, Rhizoma Atractylodis Macrocephalae oil 1-10 part, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide 5-50 part.
Wherein, the described Rhizoma Atractylodis Macrocephalae oil of step c can join in the beta-schardinger dextrin-aqueous solution, ultrasonic enclose, and cold preservation, centrifugal, precipitation promptly gets and get Rhizoma Atractylodis Macrocephalae oil behind enclose.
Wherein, weight percentage 〉=60% of Radix Ginseng total saponins in the described Radix Ginseng Rubra dry extract, ginsenoside Rg1 and ginsenoside Re's gross weight percentage composition 〉=6%, the weight percentage of volatile oil 〉=60% in the Rhizoma Atractylodis Macrocephalae oil; The weight percentage of the complex polysaccharide in d step Radix Ginseng Rubra and the Rhizoma Atractylodis Macrocephalae dry extract is with glucose meter 〉=60%.
The present invention also provides the purposes of this pharmaceutical composition in the medicine of preparation replenishing QI to invigorate the spleen.
The present invention also provides the purposes of this pharmaceutical composition in the medicine of preparation treatment gastric cancer.Further, described medicine is the medicine that suppresses preventing precancerous lesion of stomach cancer.
Motherland's medical science thinks that preventing precancerous lesion of stomach cancer belongs to categories such as " feeling of fullness ", " gastric abscess ", and deficiency of spleen-QI is its basic pathogenesis, clinical mainly with loss of appetite, glutted or the epigastrium dull pain in food back, and big loose stool is thin, deficiency of the spleen causing weakness of QI symptoms such as physical fatigue and lassitude of spirit show as feature.So we are at above-mentioned insufficiency of the spleen symptom and pathogenesis, take the Therapeutic Principle of " treatment of deficiency-syndrome by reinforcement ", " damage person's benefit ", reuse Radix Ginseng Rubra and be monarch, Gan Wen goes into spleen, " QI invigorating in the accent " (Materia Medica of Decoction), " with middle spleen invigorating " (" book on Chinese herbal medicine converges and says "), " digesting and appetizing " (" Japan hanako materia medica "), " promoting blood circulation; broken hard long-pending " (" Mingyi Bielu "), all deficiency of spleen-QI decline, and see diseases such as the tired lazy speech of lack of appetite, abdominal distention, loose stool, body, and that uses all has certain curative effect.Minister is with the Rhizoma Atractylodis Macrocephalae, dispelling the damp pathogen with bitter and warm drugs spleen invigorating, " it is full to remove spasm in the epigastrium " (" Mingyi Bielu ").Two medicines share, and play the merit of QI invigorating and spleen invigorating altogether.Make empty must the benefit of temper and wet do not stagnate, strong the stagnating again of fortuneization disappears.So can be to spleen-deficient preventing precancerous lesion of stomach cancer and general deficiency of spleen-QI and stomach-QI and fortune function decay person, the state of an illness of hitting, evident in efficacy.Prove through pharmacological evaluation, the pharmaceutical composition of being made up of drug effective region red ginseng saponin of the present invention, Rhizoma Atractylodis Macrocephalae oil, Radix Ginseng Rubra compound polysaccharide, Rhizoma Atractylodis Macrocephalae coumound polysaccharide has the obvious insufficiency of the spleen symptom of improving the vinegar Rats with Spleen-deficiency, increase the content of urine D-xylose, the quantity of rising thymus corpuscle, enlarge splenic T, B cellular regions, improve T-lymphocyte conversion ratio, correct the sick damage of gastrointestinal tissue, improve the ability of mice with spleen deficiency gastrointestinal motility; Prolong the swimming time of mice with spleen deficiency, increase anoxia enduring, cold-resistant, resistant to elevated temperatures ability; Correct the insufficiency of the spleen symptom of preventing precancerous lesion of stomach cancer rat, mice, improve gastric mucosa atrophy, congestion and edema, the pathological change of the expansion of gastric gland somatocyst, dysplasia, intestinal gland metaplasia, correct the pathological lesion of intestinal tissue, correct the insufficiency of the spleen symptom of stomach cancer metastasis Mus, suppress the gastric cancer pathological changes, regulation and control gastric cancer gene.
Medicine of the present invention has the effect of the insufficiency of the spleen preventing precancerous lesion of stomach cancer of treatment, and the combination of four effective sites, and drug effect is clear and definite, has brought into play the effect of Synergistic, and controllability is strong, and steady quality provides a kind of new selection for clinical.
Obviously, according to foregoing of the present invention,,, can also make modification, replacement or the change of other various ways not breaking away under the above-mentioned basic fundamental thought of the present invention prerequisite according to the ordinary skill knowledge and the customary means of this area.
The specific embodiment of form is described in further detail foregoing of the present invention again by the following examples.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
The specific embodiment
The extraction process of embodiment 1 raw material of the present invention
Get the Radix Ginseng Rubra medical material, add 50% soak with ethanol, reflux, extract, filters merging filtrate; Medicinal liquid reclaims ethanol, concentrates, and last macroporous adsorbent resin is used earlier and 20% ethanol elution remove impurity, and 70% ethanol elution of the usefulness that continues is collected 70% ethanol elution, reclaims ethanol, concentrates, and is standby.The Rhizoma Atractylodis Macrocephalae soaks with solvent naphtha (or vegetable oil), and reflux, extract, filters, and filtrate is reclaimed solvent, join in the beta-schardinger dextrin-aqueous solution, and ultrasonic enclose, cold preservation, centrifugal, precipitate standby; Get Rhizoma Atractylodis Macrocephalae medicinal residues, add ethanol extraction and filter, medicinal residues mix with the Radix Ginseng Rubra medicinal residues again, decoct with water, and filter, and filtrate concentrates, and is centrifugal, and the supernatant precipitate with ethanol is centrifugal, precipitates standby.The red ginseng saponin concentrated solution adds Rhizoma Atractylodis Macrocephalae oil Benexate Hydrochloride and complex polysaccharide precipitation, and mix homogeneously is a parent nucleus with starch, granulates, and granulate incapsulates, promptly.
The extraction process of embodiment 2 raw materials of the present invention
Get the Radix Ginseng Rubra medical material, add 80% soak with ethanol, reflux, extract, filters merging filtrate; Medicinal liquid reclaims ethanol, concentrates, and last macroporous adsorbent resin is used earlier and 20% ethanol elution remove impurity, and 70% ethanol elution of the usefulness that continues is collected 70% ethanol elution, reclaims ethanol, concentrates, and is standby.Rhizoma Atractylodis Macrocephalae solvent naphtha (or vegetable oil) soaks, and reflux, extract, filters, and filtrate is reclaimed solvent, concentrates, and is standby; Get Rhizoma Atractylodis Macrocephalae medicinal residues, add ethanol extraction and filter, medicinal residues mix with the Radix Ginseng Rubra medicinal residues again, decoct with water, and filter, and filtrate concentrates, and is centrifugal, and the supernatant precipitate with ethanol is centrifugal, precipitates standby.The red ginseng saponin concentrated solution adds Rhizoma Atractylodis Macrocephalae oil and complex polysaccharide precipitation, and mix homogeneously is a parent nucleus with starch, granulates, and granulate incapsulates, promptly.
The preparation of embodiment 3 medicines of the present invention
Method by embodiment 1 is prepared into Radix Ginseng total saponins 50g, Rhizoma Atractylodis Macrocephalae volatile oil 10g, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide 50g, adds starch granulation, granulate, adds magnesium stearate, and tabletting gets tablet.
The preparation of embodiment 4 medicines of the present invention
Be prepared into Radix Ginseng total saponins 5g, Rhizoma Atractylodis Macrocephalae volatile oil 1g, Radix Ginseng Rubra and from art complex polysaccharide 5g by the method for embodiment 1, add that starch is granulated, granulate, add magnesium stearate, tabletting, tablet.
The preparation of embodiment 5 medicines of the present invention
Method by embodiment 1 is prepared into Radix Ginseng total saponins 25g, Rhizoma Atractylodis Macrocephalae volatile oil 10g, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide 25g, and adding starch is granulated, granulate is encapsulated, gets capsule.
The preparation of embodiment 6 medicines of the present invention
Method by embodiment 1 is prepared into Radix Ginseng total saponins 40g, Rhizoma Atractylodis Macrocephalae volatile oil 10g, Radix Ginseng Rubra and the granulation of Rhizoma Atractylodis Macrocephalae coumound polysaccharide 50g adding starch, granulate is encapsulated, gets capsule.
The preparation of embodiment 7 medicines of the present invention
Method by embodiment 1 is prepared into Radix Ginseng total saponins 10g, Rhizoma Atractylodis Macrocephalae volatile oil 10g, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide 10g, with Rhizoma Atractylodis Macrocephalae oil emulsifying, adding Radix Ginseng total saponins, complex polysaccharide, adds the injection additives, and adjust pH and osmotic pressure are prepared into injection.
The preparation of embodiment 8 medicines of the present invention
Method by embodiment 1 is prepared into Radix Ginseng total saponins 50g, Rhizoma Atractylodis Macrocephalae volatile oil 1g, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide 5g, and adding starch is granulated, granulate is encapsulated, gets capsule.
Embodiment 9 drug quality control methods of the present invention
(1) infrared spectrum is differentiated
It is an amount of to get medicament capsule content of the present invention, and the polysaccharide that is dissolved in water filters, and filtrate adds 95% ethanol after contain the alcohol amount and reach about 80%, leaves standstill 12 hours, and is centrifugal, collecting precipitation, vacuum drying, complex polysaccharide.The IR collection of illustrative plates unanimity that polysaccharide is closed in two replies.And the complex polysaccharide got in the same batch sample relatively removed IR collection of illustrative plates before and after the albumen, and the result, two collection of illustrative plates are closely similar.Radix Ginseng Rubra polysaccharide that again will identical method for making with complex polysaccharide makes and Rhizoma Atractylodis polysaccharide are made the IR collection of illustrative plates relatively under the proteic situation that do not eliminate, the IR collection of illustrative plates of Radix Ginseng Rubra polysaccharide and Rhizoma Atractylodis polysaccharide is very close from the position of absworption peak, but the size at peak has some differences.At 1740.5cm
-1The place, Radix Ginseng Rubra polysaccharide and complex polysaccharide have a small peak and Rhizoma Atractylodis polysaccharide does not have.
(2) high performance capillary electrophoresis
1. instrument: BeckmanHPCE capillary electrophoresis apparatus; Diode array detector; Capillary column (56cm * 75 μ m).
The investigation of chromatographic condition: influence is separated and the condition that detects such as column temperature, mobile phase, detection wavelength are investigated.Final definite chromatographic condition is: L boric acid-sodium borate; Column temperature: 30 ℃; Voltage: 15KV.
2. the investigation of sample solution preparation method
To each factor of influencing the sample hydrolysis such as hydrolysis time, hydrolysis method, H
2SO
4The selection of addition, neutralization reagent etc. is investigated.Final definite hydrolysising condition adds H for taking by weighing the 30mg polysaccharide
2SO
45ml, 100 ℃ of hydrolysis are with Ba (OH)
2A neutralization part earlier, when closing on terminal point, reuse BaCO
3Be neutralized to neutrality.With 12000 rev/mins rotating speed centrifugal 10 minutes respectively, supernatant filtered with 0.45 μ m filter membrane, promptly.
The preparation of Rhizoma Atractylodis polysaccharide sample: precision takes by weighing the about 30mg of Rhizoma Atractylodis polysaccharide, preparation as stated above, promptly.
The preparation of Radix Ginseng Rubra polysaccharide sample: precision takes by weighing the about 30mg of Radix Ginseng Rubra polysaccharide, preparation as stated above, promptly.
The preparation of Radix Ginseng Rubra, Rhizoma Atractylodis Macrocephalae coumound polysaccharide sample: precision takes by weighing Radix Ginseng Rubra, the about 30mg of Rhizoma Atractylodis Macrocephalae coumound polysaccharide, preparation as stated above, promptly.
The preparation of beta-schardinger dextrin-sample: precision takes by weighing the about 30mg of beta-schardinger dextrin-, preparation as stated above, promptly.
3. the preparation of starch: precision takes by weighing the about 30mg of starch, preparation as stated above, promptly.
4. assay method: draw each 1 μ l of above-mentioned sample solution respectively, inject efficient capillary electrophoresis apparatus and measure, the result is as follows:
Under fixed wave length 204 and 214nm mensuration, the appearance time basically identical of Rhizoma Atractylodis polysaccharide and Radix Ginseng Rubra polysaccharide, spectrogram basically identical.But under full wavelength scanner, obtain three-dimensional spectrum, find that Rhizoma Atractylodis polysaccharide has a peak at the 284nm place, the Radix Ginseng Rubra polysaccharide does not have this peak; And the Radix Ginseng Rubra polysaccharide has an absworption peak at the 190nm place, and Rhizoma Atractylodis polysaccharide does not have this peak; Complex polysaccharide all has absworption peak at 284nm and 190nm place.
In addition, beta-schardinger dextrin-and starch are measured, found that their spectrogram does not have absorption at 284nm and 190nm place.Illustrate that they do not influence the capillary electrophoresis analysis of polysaccharide.
Conclusion: adopt high performance capillary electrophoresis can distinguish ginseng polysaccharide and Rhizoma Atractylodis polysaccharide, and beta-schardinger dextrin-and starch are noiseless in the side.
The quality control of embodiment 10 medicine material Rhizoma Atractylodis Macrocephalae oils of the present invention
(1) mensuration of total volatile oil: precision is measured Rhizoma Atractylodis Macrocephalae oil 1ml, by " volatile oil content is measured in the operation of first method in Chinese pharmacopoeia version pharmacopeia in 2000 the appendix XD determination of volatile oil method.
This product contains the volatilization oil mass must not be less than 60%.
(2) mensuration of linoleic acid content in the Rhizoma Atractylodis Macrocephalae oil: the photograph gas chromatography (" appendix VIE of Chinese pharmacopoeia version in 2000) measure.
The test of chromatographic condition and system suitability: 10%DEGS is a carrier, and Chromsorb w Aw-DMCS is immobile phase (60～80 orders, the glass column of 2.1m * 3mm).Column temperature: 165 ℃.Injector temperature: 220 ℃.Detector: FID, 230 ℃.H
2: 30ml/ minute.Air:100ml/ minute.N
2: 60ml/ minute.Theoretical cam curve (N) is not less than 1000 in methyl linoleate.
The preparation of reference substance solution: get the about 1g of methyl linoleate reference substance, the accurate title, decide, and puts in the 50ml measuring bottle, adds n-hexane dissolution and be settled to scale.Precision is measured 3.0ml and is put the 10ml measuring bottle, adds normal hexane to scale, promptly.
The preparation of need testing solution and mensuration: measure the about 0.25ml of this product, the accurate title, get surely, puts in the tool plug scale test tube, potassium hydroxide-methanol solution the 2.0ml that adds 0.5mol/L, shake up, put in 70 ℃ of water-baths saponification 15 minutes, take out, be chilled to room temperature, add 12.5% boron trifluoride-methanol solution 2ml again, put in 70 ℃ of water-baths esterification 5 minutes, take out, be chilled to room temperature, the accurate normal hexane 2.0ml that adds shakes up, and adds saturated nacl aqueous solution 2.0ml again, shake up, leave standstill, shake up, leave standstill, write down the cumulative volume of supernatant, accurate reference substance solution and each 2 μ l of need testing solution of drawing, inject gas chromatograph is measured the methyl linoleate peak area, calculate content with external standard method, promptly.
The every ml of this product contains linoleic acid (C
18H
32O
2) must not be less than 30.0mg.
(3) stability
By " Chinese pharmacopoeia version in 2000 reaches relevant stability techniques requirement in " medicine registration management way ", test is according to the method for " medicament capsule quality standard draft of the present invention ", through 6 months study on the stability of room temperature, the result shows that this product is basicly stable to Rhizoma Atractylodis Macrocephalae oil clathrate and finished product.
Below prove beneficial effect of the present invention by concrete pharmacodynamics test.Prove that medicine of the present invention has the inhibition preventing precancerous lesion of stomach cancer.
Test example 1 medicine of the present invention is to the therapeutical effect of insufficiency of the spleen preventing precancerous lesion of stomach cancer rat
One, experiment material
1, medicine
1. be subjected to the reagent thing: medicine capsule of the present invention, the method preparation of embodiment 1, every g dry extract contains crude drug in whole 10g.Be mixed with 1%, 2%, 4% medicinal liquid (being that every ml contains crude drug in whole 0.1g, 0.2g, 0.4g) with distilled water standby.
2. positive drug: ZHENGDA YANGWEI CHONGJI, to produce by Zhengda Qingchunbao Pharmaceutical Co., Ltd, authentication code: the accurate word of medicine (1993) 0376-1 number defended in Zhejiang.Bismuth potassium citrate granule is produced by beautiful pearl pharmaceutical Co. Ltd, and authentication code: (90) are defended the accurate word X-91-3 of medicine number.
2, animal
The SD rat, male and female all have, and body weight 180～220g is provided by Chengdu University of Traditional Chinese Medicine Animal Experimental Study center, the quality certification number: No. the 8th, the real moving pipe matter (97) in river.The quarantine back is standby.
Two, experimental technique
Get 70 of SD rats, be divided into dosage group (N=10), medicament capsule low dose group of the present invention (N=10) in blank group (N=10), insufficiency of the spleen preventing precancerous lesion of stomach cancer model group (N=10), ZHENGDA YANGWEI CHONGJI group (N=10), bismuth potassium citrate group (N=10), medicament capsule high dose group of the present invention (N=10), the medicament capsule of the present invention at random.Except that the blank group, active immunity, hungry full inequality add drinks the ammonia method and duplicates insufficiency of the spleen preventing precancerous lesion of stomach cancer animal model 90d.91d irritates stomach isometric(al) distilled water, stomach nourishing granule medicinal liquid 15g/kg, bismuth potassium citrate medicinal liquid 1.00g/kg, 4%, 2%, 1% medicament capsule medicinal liquid 10ml/kg of the present invention, successive administration 30d respectively by group.Measure the weight of animals, body temperature, food ration and spontaneous activity before the experiment, after the modeling, later every 10d surveys body weight 1 time, so that adjust the administration volume; The weight of animals, body temperature, food ration and spontaneous activity are measured with method in the treatment back.121d, femoral vein is got blood, separation of serum, by the requirement of gastrin radioimmunoassay determination test kit, the FJ-2003r enumerator is measured serum gastrin content; Put to death animal, get each one in stomach, jejunum tissue after cuing open inspection, drop into 10% formalin internal fixation immediately, paraffin embedding, HE dyes, observation by light microscope.
Three, experimental result
1. to the influence of the insufficiency of the spleen symptom of insufficiency of the spleen preventing precancerous lesion of stomach cancer rat
Insufficiency of the spleen preventing precancerous lesion of stomach cancer model group rat food ration obviously reduces, and spontaneous activity reduces, perpendicular hair, and abdominal distention, stool abnormity, the dirt of part animal crissum moves, and compares obvious difference with the blank group.After Drug therapy of the present invention, food ration, spontaneous activity all obviously raise, and the high dose group the weight of animals obviously increases, compare with insufficiency of the spleen preventing precancerous lesion of stomach cancer model group, notable difference is arranged, compare with the ZHENGDA YANGWEI CHONGJI group, no significant difference, but obviously be better than the bismuth potassium citrate group.The result sees table 1 for details.
The influence of the table 1 pair insufficiency of the spleen symptom of Rats with Spleen-deficiency (X ± SD)
Annotate: compare with the blank group
*P＜0.01;
Compare Δ Δ P＜0.01 with model group;
2, to the morphologic influence of insufficiency of the spleen preventing precancerous lesion of stomach cancer rat gastroenteric pathology
1. cuing open inspection observes: insufficiency of the spleen preventing precancerous lesion of stomach cancer model group animal intestines and stomach flatulence, gastric mucosa attenuation, jejunum congestion and edema.After Drug therapy, medicine of the present invention senior middle school low dose group, bismuth potassium citrate group, ZHENGDA YANGWEI CHONGJI group recover normal substantially.
2. pathological study: the downright bad atrophy of insufficiency of the spleen preventing precancerous lesion of stomach cancer model group animal gastric epithelial cell, the expansion of gastric gland somatocyst, dysplasia, intestinal gland metaplasia, congestion and edema under the gastric mucosa; Jejunum congestion and edema, epithelial cell necrosis come off.Medicine of the present invention senior middle school low dose group, bismuth potassium citrate group rat stomach enteropathy are decreased and are recovered normal substantially, with blank group there was no significant difference.The result sees table 2 for details.
Table 2 medicament capsule of the present invention is to the influence of insufficiency of the spleen preventing precancerous lesion of stomach cancer rat stomach, jejunum tissue morphology (X ± SD)
Annotate: compare with the blank group
*P＜0.05;
Compare Δ P＜0.05 with the model of spleen deficiency group;
Four, conclusion
Medicine of the present invention can improve insufficiency of the spleen atrophic symptom of rats, the content of rising serum gastrin, the pathological change that congestion and edema and jejunum congestion and edema, epithelial cell necrosis come off under the downright bad atrophy of correction gastric mucosa, the expansion of gastric gland somatocyst, dysplasia, intestinal gland metaplasia, the gastric mucosa, its effect is better than bismuth potassium citrate and ZHENGDA YANGWEI CHONGJI.
Test example 2 medicines of the present invention are to the therapeutical effect of insufficiency of the spleen preventing precancerous lesion of stomach cancer mice
One, experiment material
1, medicine
1. be subjected to the reagent thing: medicament capsule of the present invention, by the preparation of embodiment 2 methods, every g dry extract contains crude drug in whole 10g.Be mixed with 1%, 2%, 4% medicinal liquid (being that every ml contains crude drug in whole 0.1g, 0.2g, 0.4g) with distilled water standby.
2. positive drug: WEIFUCHUN PIAN, specification: the 0.37g/ sheet, produce by Huqingyutang Pharmaceutical Co., Ltd., Hangzhou City, lot number 000102, preparation: 29 (10.73g) grinds and is dissolved as 72.5ml for smalls adds sterilized water.
3. 2-ethyl nitrosamine, medical university institute of oncology, West China provides; The Shanxi white vinegar is by Shanxi Province U.S. and occupy the production of old vinegar company limited, acidity 3.5g/100ml, lot number 200012.
2, animal
The NIH mice, male and female all have, and body weight 18～22g is provided by Chengdu University of Traditional Chinese Medicine Animal Experimental Study center, the quality certification number: No. the 6th, the real moving pipe matter (99) in river.The quarantine back is standby.
Two, experimental technique
Get 60 of NIH mices, be divided into dosage group (N=10), medicament capsule low dose group of the present invention (N=10) in blank group (N=10), insufficiency of the spleen preventing precancerous lesion of stomach cancer model group (N=10), WEIFUCHUN group (N=10), medicament capsule high dose group of the present invention (N=10), the medicament capsule of the present invention at random.Except that the blank group, all the other each groups are pressed document
(11.12.)Irritate stomach Shanxi white vinegar 15ml/kg after the fasting, irritate stomach Shanxi white vinegar 10ml/kg 2-10 day, irritate stomach 2-ethyl nitrosamine 2.8mg/kg 11-130 day, duplicate insufficiency of the spleen preventing precancerous lesion of stomach cancer animal model.131-160 day, irritate stomach isometric(al) distilled water, WEIFUCHUN medicinal liquid 1.48g/kg, medicament capsule medicinal liquid 0.4g/kg of the present invention, 0.2g/kg, 0.1g/kg, successive administration 30d respectively by group.Measure the weight of animals, body temperature, food ration and spontaneous activity before the experiment, after the modeling, later every 10d surveys body weight 1 time, so that adjust the administration volume; The weight of animals, body temperature, food ration and spontaneous activity are measured with method in the treatment back.161d puts to death animal, gets one of gastric tissue after cuing open inspection, drops into 10% formalin internal fixation immediately, paraffin embedding, HE dyeing, observation by light microscope.
Three, experimental result
1. to the influence of insufficiency of the spleen preventing precancerous lesion of stomach cancer Mice with Spleen symptom
Insufficiency of the spleen preventing precancerous lesion of stomach cancer model group mice erect hair, hogback, spontaneous activity reduce, ingest obviously reduce, lose weight, abdominal distention, stool abnormity, the dirt of part animal crissum are moved, become thin, by the fluffy tarnishing of hair.After medicament capsule treatment of the present invention, insufficiency of the spleen transference cures such as food ration, spontaneous activity, body weight, abdomen shape, quilt hair.
2, to the morphologic influence of insufficiency of the spleen preventing precancerous lesion of stomach cancer mice gastroenteric pathology
1. cuing open inspection observes: insufficiency of the spleen preventing precancerous lesion of stomach cancer model group animal intestines and stomach flatulence, gastric mucosa attenuation, jejunum congestion and edema.After Drug therapy, medicament capsule of the present invention senior middle school low dose group, WEIFUCHUN group recover normal substantially.
2. pathological study: severe dysplasia is obvious in the insufficiency of the spleen preventing precancerous lesion of stomach cancer model group animal stomach, with the blank group significant difference is arranged relatively.Medicament capsule of the present invention senior middle school low dose group, WEIFUCHUN group mice gastric tissue recover normally with model control group significant difference is arranged relatively substantially.The result sees table 3 for details.
Table 3 medicament capsule of the present invention is to the influence of insufficiency of the spleen preventing precancerous lesion of stomach cancer mice gastric tissue form (X ± SD)
Annotate: compare with the blank group
*P＜0.05;
Compare Δ P＜0.05 with the model of spleen deficiency group;
Four, conclusion
Medicine of the present invention can improve the symptom of mice with spleen deficiency, improves the preventing precancerous lesion of stomach cancer of gastric tissue, can reverse dysplasia.
Test example 3 medicines of the present invention are to the therapeutical effect of insufficiency of the spleen stomach cancer metastasis Mus
One, experiment material
1, medicine
1. be subjected to the reagent thing: medicament capsule of the present invention, by embodiment 1 preparation, every g dry extract contains crude drug in whole 10g.Be mixed with 1%, 2%, 4% medicinal liquid (being that every ml contains crude drug in whole 0.1g, 0.2g, 0.4g) with distilled water standby.
2. positive drug: WEIFUCHUN PIAN, specification: the 0.37g/ sheet, produce by Huqingyutang Pharmaceutical Co., Ltd., Hangzhou City, lot number 020717, preparation: 29 (10.73g) grinds and is dissolved as 72.5ml for smalls adds sterilized water.5-fluorouracil, specification: 0.25g/10ml is produced by Tianjin gold credit aminoacid company limited, lot number 0207012, preparation: 12ml5-Fu adds sterilized water to 100ml (100ml:300mg).
2, animal
108 of BALB/c-nu/nu nude mices, the SPF animal, male and female all have, 8 ages in week.Provide by Chengdu University of Traditional Chinese Medicine Animal Experimental Study center.The animal quality certification: 99-6, the quarantine back is standby.
3, tumor source
Human stomach cancer cell line SGC-7901 suspension (3 * 10
6) provide by the institute of oncology, West China.
4, reagent
TIMP1 (H-150), rabbit polyclonal IgG is available from Santa Cruz company.
TIMP2 (H-140), rabbit polyclonal IgG is available from Santa Cruz company.
MMP2 (C-19), goat polyclonal IgG is available from Santa Cruz company.
UPA (M-20), goat polyclonal IgG is available from Santa Cruz company.
PAI-(H-135), rabbit polyclonal IgG is available from Santa Cruz company.
MMP9, rabbit anti MMP-9 (Ab1), BA0573 is available from Wuhan doctor's moral Bioisystech Co., Ltd.
The corresponding two anti-test kits that reach are available from Beijing Zhong Shan Bioisystech Co., Ltd.
5, equipment
The CB3201 numeral is traced the formula toy activity observation instrument China river sub-industrial group of NORTEL
Electronic balance PA2003 Shanghai balance equipment factory
Olympus BX50 optical microscope Japan
MIAS-2000 type image analysis system Sichuan University image graphics institute
6, facility
SPF (specific pathogen free condition) laboratory animal barrier system is provided by Chengdu University of Traditional Chinese Medicine Animal Experimental Study center, and the Laboratory Animal Facility environment meets the SPF grade standard, the zoopery quality certification number: 99-5.
Two, experimental technique
1, the foundation of insufficiency of the spleen stomach cancer metastasis rat animal model and packet transaction
Get 108 of BALB/c-nu/nu nude mices.Test 1d with the SGC-7901 suspension inoculation in 2 nude mice oxters (0.2ml/ only), cultivate the stomach organization piece, treat after the stomach organization block length becomes 1cm * 1cm big this stomach organization piece to be gone down to posterity and cultivate, treat when this experiment reached for the 4th generation that it grows up to 1cm * 1cm standby greatly (8 week) to solid tumor.Its cofree aseptic full-valence pellet feed of ingesting began all the other 100 nude mices are made the syndrome of spleen-deficiency animal model in the 9th week, before the experiment behind the fasting 36h, irritate first stomach with the Shanxi white vinegar by 15ml/kg/d, press 10mI/kg/d filling stomach 6d later on; During the 10th week, under strict aseptic technique, undergo surgery and make the stomach cancer metastasis animal model, promptly the operation the 1st day with pentobarbital sodium 40mg/kg intraperitoneal anesthesia under, the surgical field of view skin degerming, and under the nude mice xiphoid-process, open the long longitudinal cut of a 0.7cm-1cm, find stomach, after cut serous coat and sews up the aforementioned stomach organization piece 0.1 * 0.1cm that reached for the 4th generation with the purse string suture method at the glandular stomach place, close the abdominal cavity.Experimentation carries out in SPF Animal Lab, postoperative is divided into 6 groups at random with these 100 nude mices, be 20 of insufficiency of the spleen model of gastric carcinoma groups, 20 of western medicine group, 15 of Chinese medicine matched groups, 15 of medicament capsule low dose group of the present invention, the dosage group is 15 in the medicament capsule of the present invention, 15 of medicament capsule high dose group of the present invention, perform the operation and began in the 2nd day freely to drink the sterilized water except that insufficiency of the spleen model of gastric carcinoma group, the Western medicine group is irritated stomach and is given 5-fluorouracil 30mg/kg/d, the Chinese medicine matched group is irritated stomach and is given WEIFUCHUN 1.48g/kg/d, medicament capsule low dose group of the present invention is irritated stomach and is given medicament capsule medicinal liquid 100mg/kg/d of the present invention, the dosage group is irritated stomach and is given medicament capsule medicinal liquid 200mg/kg/d of the present invention in the medicament capsule of the present invention, medicament capsule high dose group of the present invention is irritated stomach and is given medicament capsule medicinal liquid 400mg/kg/d of the present invention, except that model group, all treat 21 days (3 week) by this ruling by law, fasting 24 hours, taking off cervical vertebra puts to death, take out stomach, vertically cut open along the greater gastric curvature side, behind the clean food debris of normal saline, that gastric tissue is liquid-solid fixed to be measured with 10% neutral formalin.
In the experiment weekly electronic balance claim body weight, toy activity observation instrument to measure the food ration that spontaneous activity, electronic balance in 10 minutes claims the 24h animal, observe other performances of stool character, abdomen shape and body surface.
2, the morphologic observation of histopathology staining procedure and stomach cancer cell
With liquid-solid fixed 48 hours of aforementioned gastric tissue 10% neutral formalin, dehydration of alcohol step by step, dimethylbenzene is transparent, it is cured to soak, paraffin embedding, conventional section 4um, HE dyeing. each specimen under 10 * 40 high power lenses, 20 cancerous cell nuclear morphology of random observation, and (pixel that the area in cross section equals to drop in the cross section in is counted the test area (a=d that multiply by each test pixel point representative to measure the area of nucleus and corresponding kernel under MIAS-2000 type image analysis system
2k
2Pc)), girth (referring to length) around tested object periphery, maximum gauge, (the calliper diameter of planar graph is meant the distance between two parallel lines of this figure of tangent to minimum diameter, its available maximum gauge, minimum diameter and average calliper diameter are described) and form factor (refer generally to the two-dimensional shapes factor, the variation of concentrated expression two dimension morphosis, dimensionless group (promptly not having unit), when testee is the garden, form factor equals 1, irregular or when being ellipse when object, form factor is less than 1, the degree of irregularity in reflection testee cross section) 5 kinds of morphological parameters.
3, gastric cancer is invaded the detection of profit, metastasis related gene expression
Immunohistochemical staining step (SP method): adopt peroxidase that biotin labeled second antibody is connected with the streptomycete avidin and stromatin mixed liquor to measure determined antigen in cell and the tissue.Painted main process is as follows:
1. paraffin section de-waxing is to water, with 3 times * 5min. of PBS liquid (0.01mol/L PH7.4) flushing
23%H
2O
2Incubated at room 10 minutes to eliminate the activity of endogenous peroxydase, is hatched 10min. under the room temperature
3. distilled water flushing, PBS soaked 5 minutes.
4.10% normal goats serum (PBS dilution) sealing, incubated at room 10 minutes.The serum deprivation that inclines is not washed, and drips the anti-of 1: 100 dilution proportion, hatches 1 hour for 37 ℃.
5.PBS flushing, 5 minutes * 3 times.
6. drip second filial generation biotin labeling two anti-working solutions, 37 ℃ or incubated at room 30 minutes.
7.PBS flushing, 5 minutes * 3 times.
8. drip second filial generation Radix Cochleariae officinalis enzyme labelling strepto-avidin working solution, hatched 30 minutes for 37 ℃.
9.PBS flushing, 5 minutes * 3 times.
10.DAB colour developing.
11. tap water fully washes, haematoxylin redyeing, mounting.
Positive findings is judged: MMP2, MMP9, TIMP1, TIMP2, uPA and PAI-1 positive material sepia are to faint yellow, be filament shape or fine particulate, MMP2, MMP9, the main expressive site of TIMP1, TIMP2 are at the tumor cell endochylema, and part is expressed in mesenchyma stroma of tumors cell (comprising vascular endothelial cell) endochylema and substrate; UPA and PAI-1 expressive site are mainly at the vascular endothelial cell endochylema, and part is in tumor cell endochylema and substrate.
MMP2, MMP9, TIMP1, TIMP2, the detection of uPA and PAI-1 gene expression: at first SP method stained is carried out complete observation under 10 * 10 low power fields, seek MMP2 in the tumor, MMP9, TIMP1, TIMP2, (promptly " focus " distinguished for uPA and the abundantest zone of PAI-1 positive material, the darkest companion of dyed color/or the dyeing area is maximum), in 10 * 20, select 20 hot zones then under times visual field, adopt MIAS-2000 type image analysis system to measure the area of each index positive material, average optical, integral optical density, average blackness is represented their content.
Statistical procedures: all measurement datas represent that with mean ± standard deviation (x ± SD), adopt the SPSS11.0 statistical package to handle relatively uses LSD (variance is neat) and S-N-K or Tamhane (heterogeneity of variance) between group.
Three, experimental result
1, insufficiency of the spleen performance
After irritating stomach 6d with the Shanxi white vinegar, variation has all taken place in the ordinary circumstance of experiment nude mice, food ration, spontaneous activity, weight loss, abdominal part swells, stool becomes half congealed or is little dried graininess, with make model of spleen deficiency before utmost point significant difference is arranged.Dead 60 animals altogether in the experimentation, wherein irritating under stomach and the operation double blow at vinegar has 47 animal deads, back in irritating the stomach treatment dead again 13, particularly the Western medicine group is because the side effect of 5-fu is big, and animal becomes thin dead nearly 7, and postmortem does not have special.When experiment finished, animal was respectively 10 of insufficiency of the spleen model of gastric carcinoma groups, 6 of western medicine group, and 6 of Chinese medicine matched groups, 6 of medicament capsule low dose group of the present invention, the dosage group is 6 in the medicament capsule of the present invention, 6 of medicament capsule high dose group of the present invention.
2, histopathology is observed
Observe insufficiency of the spleen model of gastric carcinoma group cancerous tissue structure disturbance under the HE stained, be infiltrative growth, unclear with the surrounding tissue boundary, cancerous cell is medium bigger than normal, rounded, oval, polygon and irregular shape, and endochylema is less, nuclear staining is darker, ovalize, triangle and irregular shape, and majority is the cavity shape, karyokinesis is as more, kernel is thick, is positioned at nuclear central authorities or off normal, and majority has a kernel, minority have two or more than, dyeing is dark.5 treatment group cancerous cell sizes are similar to model group, but shape is in the majority with circle, ellipse, and polygon and irregular shape minimizing, especially He area reduce, shape obviously becomes, and rule, kernel number and area all reduce than model group.On the nuclear area, the Western medicine group has significant difference, and all the other 4 group differences do not have significance; Nuclear girth and nuclear maximum gauge reduce than model group, and Western medicine group and medicine high dose group of the present invention have significant difference, and all the other 3 groups of differences do not have significance; The nuclear minimum diameter reduces than model group, but there are no significant difference; Represent the nuclear form factor of nuclear shape to increase than model group, its Chinese medicine and western medicine group and Chinese medicine matched group have significant difference, and the basic, normal, high dosage group of medicine of the present invention has utmost point significant difference; The comparison model group of kernel area, kernel and nuclear area reduces, and 5 treatment groups all have utmost point significant difference; The kernel form factor increases than model group, and except that medicine high dose group of the present invention has the significant difference, all the other respectively organize equal difference does not have significance.The result sees table 4, table 5 for details.
Cancerous cell kernel metamorphosis in the table 4 BALB/c-nu/nu nude mouse stomach organization (x ± SD)
Annotate: compare with insufficiency of the spleen model of gastric carcinoma group:
*P＜0.05,
*P＜0.01.
Cancerous cell nuclear morphology variation in the table 5 BALB/c-nu/nu nude mouse stomach organization (x ± SD)
Annotate: compare with insufficiency of the spleen model of gastric carcinoma group:
*P＜0.05,
*P＜0.01.
3, gastric cancer is invaded the expression of profit, metastasis related gene
3.1TIMP1 expression of gene
The more insufficiency of the spleen model of gastric carcinoma group of expression, western medicine group and the Chinese medicine matched group of TIMP1 gene Senior Three dosage group in medicament capsule of the present invention is low strengthen to some extent, but difference does not have significance.The result sees table 6 for details.
TIMP1 changes of contents in the table 6 BALB/c-nu/nu nude mouse stomach organization cell (x ± SD)
3.2.TIMP2 expression of gene
The TIMP2 gene strengthens in all more insufficiency of the spleen model of gastric carcinoma group of the expression of five treatment groups, and wherein WEIFUCHUN group, the high, medium and low dosage group of medicament capsule of the present invention and insufficiency of the spleen gastric cancer group relatively have significance (p＜0.05) or utmost point significant difference (p＜0.01).The result sees table 7 for details.
The comparison of TIMP2 content in the table 7BALB/c-nu/nu nude mouse stomach organization cell (x ± SD)
Annotate: compare with insufficiency of the spleen model of gastric carcinoma group:
*P＜0.05,
*P＜0.01.
3.3MMP2 expression of gene
The all more insufficiency of the spleen model of gastric carcinoma group of the expression of MMP2 gene in five treatment groups weakens, wherein dosage group and the more insufficiency of the spleen model of gastric carcinoma group of high dose group have utmost point significant difference (p＜0.01) in Chinese medicine matched group, the medicament capsule of the present invention, the more insufficiency of the spleen model of gastric carcinoma group of medicament capsule low dose group of the present invention has significant difference (p＜0.05), and the more insufficiency of the spleen model of gastric carcinoma group of Western medicine group difference does not have significance.The result sees table 8 for details.
The comparison of MMP2 content in the table 8 BALB/c-nu/nu nude mouse stomach organization cell (x ± SD)
Annotate: compare with insufficiency of the spleen model of gastric carcinoma group:
*P＜0.05,
*P＜0.01.
3.4MMP9 expression of gene
The expression of MMP9 gene in western medicine group is more insufficiency of the spleen, and the model of gastric carcinoma group has certain enhancing, but difference does not have significance; The more insufficiency of the spleen model of gastric carcinoma group of expression of Senior Three dosage group weakens in Chinese medicine matched group, medicament capsule of the present invention hang down, and the more insufficiency of the spleen model of gastric carcinoma group of medicament capsule high dose group of the present invention has significant difference (p＜0.05).The result sees table 9 for details.
The comparison of MMP9 content in the table 9 BALB/c-nu/nu nude mouse stomach organization cell (x ± SD)
Annotate: compare with insufficiency of the spleen model of gastric carcinoma group:
*P＜0.05,
*P＜0.01.
3.5uPA expression of gene
The uPA gene weakens in all more insufficiency of the spleen model of gastric carcinoma group of the expression of five treatment groups, and the more insufficiency of the spleen model of gastric carcinoma group of dosage group has significant difference (p＜0.05) in the medicine of the present invention, and the more insufficiency of the spleen model of gastric carcinoma group of medicine high dose group of the present invention has utmost point significant difference (p＜0.01).The result sees table 10 for details.
The comparison of uPA content in the table 10 BALB/c-nu/nu nude mouse stomach organization cell (x ± SD)
Annotate: compare with insufficiency of the spleen model of gastric carcinoma group:
*P＜0.05,
*P＜0.01.
3.6PAI-1 expression of gene
The PAI-1 gene strengthens in all more insufficiency of the spleen model of gastric carcinoma group of the expression of five treatment groups, wherein Chinese medicine matched group, the low middle more insufficiency of the spleen model of gastric carcinoma group of Senior Three dosage group of medicine of the present invention have significant difference (p＜0.05), and western medicine group is more insufficiency of the spleen, and model of gastric carcinoma group difference does not have significance.The result sees table 11 for details.
The comparison of PAI-1 content in the table 11 BALB/c-nu/nu nude mouse stomach organization cell (x ± SD)
Annotate: compare with insufficiency of the spleen model of gastric carcinoma group:
*P＜0.05,
*P＜0.01.
Four, conclusion
After Drug therapy of the present invention, insufficiency of the spleen stomach cancer metastasis Mus cancerous cell breaks up again, shows as atypia and reduces, and differentiation degree increases; Insufficiency of the spleen stomach cancer metastasis Mus cancer cell infiltration, transfer reduce, and show that the ECM degraded reduces in the cancerous tissue, and one of its mechanism is that digestive enzyme and activator thereof are lower, and while ECM digestive enzyme inhibitor and activator mortifier are higher.Results suggest: medicine of the present invention not only has the effect of inducing insufficiency of the spleen stomach cancer metastasis Mus stomach cancer cell to break up again, but also it is molten to disappear by the secretion performance antagonism ECM that regulates ECM degradation enzyme system inhibitor corresponding with it, plays the effect that anti-stomach cancer cell soaks into, shifts.
4, brief summary
Medicine of the present invention is made up of Radix Ginseng (Radix Ginseng Rubra), the Rhizoma Atractylodis Macrocephalae, has the effect of spleen invigorating, QI invigorating, inhibition preventing precancerous lesion of stomach cancer, cures mainly the effect of preventing precancerous lesion of stomach cancer syndrome of deficiency of spleen qi.Through Pharmacodynamic test of active extract, the result is as follows:
1. medicine of the present invention can improve the insufficiency of the spleen symptom of vinegar Rats with Spleen-deficiency, increases the content of urine D-xylose, the quantity of rising thymus corpuscle, enlarge splenic T, B cellular regions, improve T-lymphocyte conversion ratio, correct the sick damage of gastrointestinal tissue, improve the ability of mice with spleen deficiency gastrointestinal motility.
2. medicine of the present invention can prolong the swimming time of mice with spleen deficiency, increases anoxia enduring, cold-resistant, resistant to elevated temperatures ability.
3. medicine of the present invention can obviously be corrected the insufficiency of the spleen symptom of preventing precancerous lesion of stomach cancer rat, mice, improves gastric mucosa atrophy, congestion and edema, the pathological change of the expansion of gastric gland somatocyst, dysplasia, intestinal gland metaplasia, the pathological lesion of correction intestinal tissue.
4. medicine of the present invention has the insufficiency of the spleen symptom of obvious correction stomach cancer metastasis Mus, suppresses the gastric cancer pathological changes, the effect of regulation and control gastric cancer gene.
Above-mentioned result of the test shows that medicine of the present invention has spleen invigorating, QI invigorating, inhibition preventing precancerous lesion of stomach cancer, treats the effect of insufficiency of the spleen preventing precancerous lesion of stomach cancer.
Claims (8)
1. the pharmaceutical composition of a replenishing QI to invigorate the spleen, it is characterized in that: it is the medicament that is prepared from by following raw materials by weight proportions:
Radix Ginseng total saponins 5-50 part, Rhizoma Atractylodis Macrocephalae oil 1-10 part, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide 5-50 part.
2. the pharmaceutical composition of replenishing QI to invigorate the spleen according to claim 1, it is characterized in that: ginsenoside Rg1 and ginsenoside Re's gross weight percentage composition accounts for 0.5-5% in the described Radix Ginseng total saponins, the percentage by weight of volatile oil 〉=60% in the described Rhizoma Atractylodis Macrocephalae oil contains linoleic acid 〉=30mg in every milliliter of Rhizoma Atractylodis Macrocephalae oil.
3. the pharmaceutical composition of replenishing QI to invigorate the spleen according to claim 1 and 2, it is characterized in that: described medicament is: pill, capsule, granule, tablet, oral liquid or injection.
4. method for preparing the pharmaceutical composition of the described replenishing QI to invigorate the spleen of claim 1, it comprises the steps:
A, get the Radix Ginseng Rubra medical material, add 50%～80%7 alcohol and soak, reflux, extract, filters, merging filtrate, filtrate is collected eluent by macroporous adsorbent resin, reclaims ethanol, concentrate, Radix Ginseng total saponins;
B, get the Rhizoma Atractylodis Macrocephalae, edible vegetable oil is solvent, extracts, and filters, and filtrate is reclaimed the solvent vegetable oil, and products therefrom is a Rhizoma Atractylodis Macrocephalae oil;
C, get Rhizoma Atractylodis Macrocephalae medicinal residues, add ethanol extraction and filter, medicinal residues mix with the Radix Ginseng Rubra medicinal residues again, decoct with water, and filter, and are refining, standby, must Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide;
D, with Radix Ginseng total saponins, Rhizoma Atractylodis Macrocephalae oil, Radix Ginseng Rubra and the Rhizoma Atractylodis Macrocephalae coumound polysaccharide of a, b, c preparation by following weight proportion, add the preparation that acceptable accessories or complementary composition are prepared from:
Radix Ginseng total saponins 5-50 part, Rhizoma Atractylodis Macrocephalae oil 1-10 part, Radix Ginseng Rubra and Rhizoma Atractylodis Macrocephalae coumound polysaccharide 5-50 part.
5. the preparation of drug combination method of replenishing QI to invigorate the spleen according to claim 4, it is characterized in that: the described Rhizoma Atractylodis Macrocephalae oil of step b joins in the beta-schardinger dextrin-aqueous solution, ultrasonic enclose, cold preservation, centrifugal, precipitation promptly gets the Rhizoma Atractylodis Macrocephalae oil behind enclose.
The described pharmaceutical composition of claim 1 the preparation replenishing QI to invigorate the spleen medicine in purposes.
7. the purposes of the described pharmaceutical composition of claim 1 in the medicine of preparation treatment gastric cancer.
8. purposes according to claim 7 is characterized in that: described medicine is to suppress/or the medicine of treatment preventing precancerous lesion of stomach cancer.
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| CN1559541A (en) * | 2004-03-10 | 2005-01-05 | 李莛岐 | Spleen fortifying qi boosting capsule and its manufacturing method |
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