CN1316999C - Medicine for treating wind warmth and lung heat pathocondition and its preparation method - Google Patents
Medicine for treating wind warmth and lung heat pathocondition and its preparation method Download PDFInfo
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Abstract
The present invention discloses a new medicine for treating wind warmth and lung heat pathocondition, which is mainly used for treating patients with wind warmth and lung heat, such as pneumonitis, bronchitis, pulmonary abscess, wind-heat type common cold, etc. The present invention uses a composition containing negundo chastetree fruit, baikal skullcap root, rhizoma anemarrhenae, radix trichosanthis, bitter apricot seed, tatarian aster root, common coltsfoot flower and pepperweed seed as raw materials; the raw materials are proportionally prepared and are then manufactured into various preparations after the raw materials are respectively processed according to the different characteristics of various medicines, such as, pelletized granules, mixtures (comprising oral medicinal liquid), etc. The present invention has the advantages of novel formulas, high creativity, refined formulas, the treatment of manifestation and the root causes of diseases, conspicuous curative effects and high safety. The present invention is suitable for both the young and the old.
Description
Technical field
The present invention relates to a kind of medicine that belongs to traditional Chinese medical science wind-warm and pulmonary heat syndrome for the treatment of, specifically a kind of is a kind of Chinese patent medicine that belongs to traditional Chinese medical science wind-warm and pulmonary heat syndrome for the treatment of that raw material is made with the Chinese herbal medicine, the invention still further relates to the preparation method of this medicine.
Background technology
Wind-warm and pulmonary heat syndrome is a kind of common, multiple respiratory tract disease, is common in diseases such as various pneumonia, bronchitis, pulmonary abscess and anemopyretic cold.Symptoms such as clinical normal appearance heating, cough, asthma, pulmonary lesion.Bring very big misery for patient's body and mind, seriously endangering people's physical and mental health.Medical circles adopt the Western medicine antibiotic therapy for this type of disease more at present, as: ceftriaxone sodium, imipenum, cefuroxime, erythromycin and gentamycin etc.These medicines carry out heavy dose of disease controlling when severe infections appears in patient have certain advantage, but because bacterial drug resistance makes these antibiotic effective doses increasing, thereby toxic and side effects also increases thereupon, expense is also very expensive, the antibiotic utilization rate of China hospital is higher than 30% international average level far away up to 80%.If the long-term heavy dose of abuse of antibiotic will make its sensitive bacterial produce drug resistance quickly, thereby make antibiotic curative effect decline, service life shorten in advance, thereby make the big increase of the National People's Congress that dies from these sensitive bacterials.Therefore antibiotic only is adapted at the state of an illness and uses with disease controlling when heavier, and medicine of the present invention is treated in early days and cooperated antibiotic particularly to have good advantages aspect the treatment after stable disease.
Summary of the invention
The object of the present invention is to provide a kind of medicine that belongs to traditional Chinese medical science wind-warm and pulmonary heat syndrome for the treatment of, be mainly used in the diseases such as cough, heating, asthma and pulmonary lesion that treatment pneumonia, bronchitis outbreak, pulmonary abscess and anemopyretic cold etc. cause.Treat with animal economy from many aspects.Pharmacological action: have certain inhibition pathogenic microorganism effect, the refrigeration function that Quan Fangyou is stronger, immunoregulation effect, relieving cough and resolving phlegm antiasthmatic effect, expansion bronchus effect.Instant effect, the curative effect height, safe and reliable, expense is low.The present invention's prescription derives from hereditary recipe among the people.Be used for always among the people that pneumonia, bronchitis, bacterial virus sexuality are emitted, the existing 100 years history of the treatment of pulmonary abscess.The health that in the rural area of shortage of medical services and medicines is the numerous common people has been made significant contribution.Demonstrate better curative effect.This side has stronger novelty and distinctive feature, and the prescription flavour of a drug are few, has only 8 flavors, does not have the sense of accumulation, no wonder, inclined to one side, difficult flavour of a drug.There is not roughly the same part fully with the existing prescription that is used for upper respiratory tract infection.This Herba indigoferae Pseudotinctoriae of Fructus Viticis Negundo is the medical material of using among the people in the side, derives from the fruit of Verenaceae plant five-leaved chaste tree Vitexnegundo L., and the medicine source is very abundant.
Another object of the present invention provides a kind of preparation method of this drug agents.
The epidemic febrile disease that solution of the present invention is based on Chinese medicine to affection due to external wind and heat (or folder damp) attack lung,, the mechanism understanding of the wind-warm and pulmonary heat syndrome due to the hot stasis of blood of the expectorant poison resistance lung, from traditional Chinese herbal medicine, filtered out one group have heat clearing and damp drying, eliminating fire and detoxication, the natural drug composition of the cough-relieving of promoting the production of body fluid forms.Chen Pingbai " an epidemic febrile disease caused by exogenous pathogens piece of writing ": " pathogenic wind-warm is a disease, and the moon in spring and winter are in the majority, or aversion to wind or aversion to wind not, must fever of the body, and cough, excessive thirst." often hold under the arm wetly for suffering from for wind heat, the key of human body morbidity depends on the ability of body disease-resistant heresy; interior through day: " in healthy energy deposits, heresy can not be done " " institute of heresy gathers, and its gas must be empty " when the human body incoordination between cold and warm, daily life is not normal; eating and drinking without temperance; healthy energy, body fluid are impaired, defend outer ability and descend, and wind heat pathogenic factor; invade body in an opponent's defence causes the primary disease generation.Treatment is many based on heat clearing and damp drying, eliminating fire and detoxication, and the cough-relieving of promoting the production of body fluid is auxilliary.
Medicine of the present invention is (consumption is a weight ratio) of being made by the compositions that contains following composition of medicine:
Fructus Viticis Negundo 4%-30% Radix Scutellariae 4%-30% Rhizoma Anemarrhenae 4%-25% Radix Trichosanthis 4%-25%
Flos Farfarae 4%-25% Radix Asteris 2%-25% Semen Armeniacae Amarum 2%-25% Semen Lepidii (Semen Descurainiae) 2%-25%
The formula optimization of preparation medicine of the present invention is that weight (part) ratio range is:
Fructus Viticis Negundo 10%-20% Radix Scutellariae 10%-20% Rhizoma Anemarrhenae 8%-18% Radix Trichosanthis 8%-18%
Flos Farfarae 8%-18% Radix Asteris 6%-16% Semen Armeniacae Amarum 6%-16% Semen Lepidii (Semen Descurainiae) 6%-16%
Monarch drug Fructus Viticis Negundo heat-clearing and toxic substances removing among the side of the present invention, expelling phlegm for arresting cough is relievingd asthma, and is the upper respiratory tract infection that is usually used among the people, and resisting pathogenic microbes effect is arranged, and repairs the alveolar cell function, to expectorant, cough, breathe heavily all effective, so be one of monarch drug; Another monarch drug Radix Scutellariae is heat clearing and damp drying, eliminating fire and detoxication, and clearing heat in QI system moves back excess-heat, the heat of the clear part of the body cavity above the diaphragm housing the heart and lungs, first key medicine of being longer than clearing away lung-heat especially; The Radix Trichosanthis clearing away heat and promoting production of body fluid cures mainly the disease of consumption of body fluid caused by febrile disease, lung-heat type cough, with the Radix Scutellariae mutual reinforcement between for using; Rhizoma Anemarrhenae clearing away heat-fire, nourshing Yin and drynsessmoistening prescription is used for epidemic febrile disease cough due to lung-heat, deficiency of YIN cough caused by dryness, helps Radix Scutellariae, Fructus Viticis Negundo two monarch drug heat clearing away, helps Radix Trichosanthis to promote the production of body fluid, lung being a delicate viscus, make clearing away heat-fire and do not stay dry, so Radix Trichosanthis, the Rhizoma Anemarrhenae are ministerial drug altogether; Assistant is relievingd asthma with almond cough-relieving; Assistant is with the Radix Asteris preventing phlegm from forming and stopping coughing; Assistant is with Flos Farfarae nourishing the lung to keep the adverse QI downward, and relieving cough and resolving phlegm and Radix Asteris mutual reinforcement between are for using; The Semen Lepidii (Semen Descurainiae) eliminating pathogen from the lung for relieving asthma.Four adjuvant drugs help the Fructus Viticis Negundo relieving cough and resolving phlegm to relieving asthma altogether.All medicines are made a concerted effort, and play heat clearing and damp drying, eliminating fire and detoxication altogether, the effect of the relieving cough and asthma that promotes the production of body fluid; Diseases such as pneumonia, bronchitis, bacterial virus sexuality are emitted, upper respiratory tract infection such as pulmonary abscess causes heating, cough there is good therapeutical effect.Its pharmacological action has: have certain inhibition pathogenic microorganism effect, the refrigeration function that Quan Fangyou is stronger, immunoregulation effect, antiinflammation, relieving cough and resolving phlegm antiasthmatic effect, expansion bronchus effect.
Medicine of the present invention can be made various dosage forms according to conventional method.Wherein preferred particulates agent, mixture (containing oral liquid).
Process for producing granula:
The Fructus Viticis Negundo full dose is made fine powder (crossing 80 mesh sieves), and surplus medicine extracts and is condensed into extractum according to the decocting method of preparation granule, mix with Fructus Viticis Negundo medicated powder, the adding Icing Sugar (extractum: mix homogeneously Icing Sugar=1: 0.15), make the suspension ability granule.
Medicine treating both the principal and secondary aspects of a disease of the present invention can reduce body temperature quickly, eliminates cough and expectoration, Zhichuan, antiinflammatory.Taking convenience, patient easily receives, and side effect is little, and expense is low, instant effect.
Study effectiveness of the present invention and pharmacological action from the animal pharmacodynamics test below.
The extractum animal experiment research that the present invention makes
The effect of 1 antibacterium
1.1 in-vitro antibacterial test
1.1.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001.Take by weighing 20 grams extractum of the present invention during test, add to 50 ℃ of M-H agar culture medium mixings that 55.8ml melts after, 100mg/ml concentration, take out this medicinal liquid of 40ml again and be used for test.Promptly sucking-off 20ml pastille culture medium is toppled over plate from 40ml, add 20ml in the remaining 20ml pastille culture medium again and do not have the dilution of medicine culture medium, behind the mixing again sucking-off 20ml topple over plate, and the like, promptly containing extractum final concentration of the present invention (with the crude drug amount) in the preparation of this doubling dilution is that a series of pastille plates of 1000mg/ml, 500,250,125,62.5,31.2,15.6,8,42,1,0.5,0.25,0.125,0.06,0.03mg/ml are stand-by.
Erythromycin: specification: the 0.2g/ sheet, lot number: 65835061, Beijing Pharmaceutical Factory No.3 produces.With sterilized water dissolving preparation, final concentration is that the serial pastille plate of 128,64,32,16,8,4,2,1,0.5,0.25,0.125,0.06,0.03,0.015,0.008 μ g/ml is stand-by during test.
Antibacterial: clinical isolates strain: the test bacterial strain uses therefor is 2000.1 ~ 2002.4 and clinically causes antibacterial from what the attached First Academy of Chengdu Huaxi Medical Univ, Annex II institute collected, and identifies again through the conventional method of inspection.
Staphylococcus aureus 26 strains, block its coccus 10 strain at pneumonia streptococcus scorching 26 strains, Jia Xingrongxuexinglianqiujun 10 strains, beta hemolytic streptococcus 10 strains, 10 strains of Ke Shi pneumobacillus, escherichia coli 10 strains.
Standard Quality Control bacterial strain: staphylococcus aureus ATCC25923, escherichia coli ATCC25922, the scorching NCTC10662 of pneumonia streptococcus are that Sichuan Industrial Institute of Antibiotics preserves bacterial strain.
Culture medium: M-H culture medium: caseinhydrolysate 17.5g, beef powder 5g, soluble starch 1.5g, adding distil water 1000ml, pH7.2.Add the 15g agar powder in the solid medium.Be used for Gram-positive, negative aerobe.
1.1.2 experimental technique
Adopt the agar doubling dilution to measure the minimum inhibitory concentration of extractum of the present invention.With multiple spot inoculation instrument with microbionation on the agar plate surface that contains different pharmaceutical concentration, every some bacteria containing amount is about 10
5CFU/ml is hatched 18 ~ 20 hours observed results for 37 ℃, crosses the minimum inhibitory concentration (MIC value) of medicine to this bacterium with the least concentration of contained drug in the no bacterial growth plate culture medium.
1.1.3 experimental result
Experimental result sees Table 1.
Table 1 extractum antibacterial activity in vitro test of the present invention
| Antibacterial (strain) | Extractum of the present invention (mg/ml) | Erythromycin (μ g/ml) | |
| Staphylococcus aureus (26) | MICRange MIC 50 MIC 90 | 0.05-60.5 15.6 62.5 | 0.06-128 32 64 |
| Pneumonia streptococcus inflammation (26) | MICRange MIC 50 MIC 90 | 0.06-60.5 15.6 125 | 0.5-64 4 32 |
| Jia Xingrongxuexinglianqiujun (10) | MICRange MIC 50 MIC 90 | 2-165 16.6 135 | 0.5-65 5 30 |
| Beta hemolytic streptococcus (10) | MICRange MIC 50 MIC 90 | 2-145 16.6 135 | 0.5-64 6 31 |
| Ke Shi pneumobacillus (10) | MICRange MIC 50 MIC 90 | 0.06-62.5 62.5 62.5 | 0.06-128 0.5 128 |
| Escherichia coli (10) | MICRange MIC 50 MIC 90 | 15.6-62.5 31.2 62.5 | 0.5-4 1 4 |
| Block its coccus (10) | MICRange MIC 50 MIC 90 | 0.06-65.5 11.2 62.5 | 0.5-45 2 12 |
| The Quality Control bacterial strain | MIC | |
| Extractum of the present invention (mg/ml) | Erythromycin (μ g/ml) | |
| The scorching NCTC10662 staphylococcus aureus of escherichia coli ATCC25922 pneumonia streptococcus ATCC25923 | 15.6 0.06 15.6 | 0.25 0.5 32 |
1.1.4 conclusion: by in the table 1 as seen, extractum of the present invention all has certain antibacterial vigor to examination Gram-positive, negative bacteria.Extractum of the present invention is stronger to the antibacterial vigor of staphylococcus aureus, and the MIC scope is 0.05-60.5mg/ml; To Jia Xingrongxuexinglianqiujun MIC value scope at 2-165mg/ml; To beta hemolytic streptococcus MIC value scope at 2-145mg/ml; Extractum of the present invention is stronger to pneumonia streptococcus inflammation, the antibacterial vigor of Ke Shi pneumobacillus, and MIC value scope is at 0.06 ~ 62.5mg/ml; To colibacillary MIC value scope at 15.6-62.5mg/ml; Stronger to the antibacterial vigor that blocks its coccus, MIC value 0.06-65.5mg/ml.
1.2 endogenous protective test
1.2.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001.
Erythromycin tablets: specification: the 0.2g/ sheet, lot number: 65835061, Beijing Pharmaceutical Factory No.3 produces.
Experimental strain: the antibacterial that is used for infection animal is that staphylococcus aureus 011915, pneumonia connect coccus, is the clinical separation pathogenic bacterium of collecting from the area, Chengdu calendar year 2001.
Laboratory animal: select healthy Kunming kind white mice for use, body weight 18-22 gram, male and female half and half are provided by Sichuan Industrial Institute of Antibiotics animal reproduction chamber.Raise No. 85 in animal quality certification river.
1.2.2 experimental technique
The preparation of test organisms liquid: test strain is inoculated in the MH meat soup, cultivated 18 hours for 37 ℃, with the suitable diluted for use of 5% sterilization dry yeast liquid.
The mensuration of the minimum bacterium amount (MLD) that causes death: get healthy Kunming mouse, body weight 18-22 gram, random packet, every group of 10 mices, male and female half and half, draw the dilution bacterium liquid of above-mentioned difference respectively lumbar injection go in the mice body, every Mus 0.5ml, infect the continuous observation in back 7 days, and record dead mouse number, measure (MLD) with the minimum bacterium amount that causes mice 100% death as minimum deadly bacterium.With the infection dosage of this bacterium amount as the endogenous protective test.
The preparation of medicinal liquid: all with 0.5%CMC and normal saline preparation, extractum of the present invention is mixed with the medicinal liquid that concentration is 235.8mg/ml, 157.2mg/ml, 78.6mg/ml, 39.3mg/ml, 19.65mg/ml (in the crude drug amount) in the test medication.The concentration of erythromycin is 2.80mg/ml, 2.24mg/ml, 1.80mg/ml, 1.44mg/ml, 1.15mg/ml, 0.92mg/ml, 0.74mg/ml.
Infect and the treatment experiment: laboratory animal is evenly divided into groups at random by sex, body weight, male and female half and half, every group of 10 mices, each organizes respectively, and dosage is: 5895mg/kg, 3930mg/kg, 1965mg/ml, 983mg/kg, 491mg/kg (in the crude drug amount, are respectively 30,20,10,5,2.5 times of clinical Coming-of-Age Day consumption.Clinical consumption per day: 196.5mg (crude drug)/kg, the dosage of positive control drug erythromycin is: 70mg/kg, 56mg.kg, 45mg/kg, 36mg/kg, 28.8mg/kg, 23.0mg/kg, 18.5mg/kg.
Mouse stomach extract powder medicinal liquid of the present invention, once a day, successive administration 3 days, every mice 0.5ml/20g respectively organizes the mouse peritoneal infection and is tried bacterium liquid after administration in the 4th day, and every Mus infection dosage is 0.5ml (1MLD).Erythromycin each gastric infusion after reaching 4 hours at once behind the infectious bacteria liquid is once established the infection matched group simultaneously, continues to be administered to the 10th day, and record infects dead mouse number in back seven days, calculates its median effective dose (ED according to infecting back seven days animals survived numbers
50) and 95% fiducial limit.
Data Processing in Experiment: extractum of the present invention and the median effective dose ED of contrast medicine erythromycin to escherichia coli ATCC25922, the effect of the scorching NCTC10662 infecting mouse of pneumonia streptococcus endogenous protective
50And 95% fiducial limit all by the program software of Bliss method utilization Chinese Academy of Medical Sciences establishment, use a computer and statistical procedures.
1.2.3 experimental result
The results are shown in Table 2 and table 3
The protective effect (streptococcus pneumoniae) of table 2 extractum of the present invention and erythromycin to infecting in the mice body
| Infection strain (bacterial strain number) intoxicating bacterium amount (CFU/ml) | Medicine | Dosage (mg/kg) | Death toll (only) | Mortality rate % | ED 50And 95% fiducial limit (mg/kg) |
| Number of animals (only) | |||||
| Streptococcus pneumoniae No.011915 (5.2 * 10 7) | Extractum of the present invention (pressing the crude drug amount calculates) | 5895 3930 1965 983 491 | 1/10 4/10 5/10 7/10 9/10 | 10 40 50 70 90 | 2027 (1208-3466) |
| Erythromycin | 45.00 36.00 28.80 23.0 18.5 | 1/10 3/10 5/10 7/10 9/10 | 10 30 50 70 90 | 28.67 (24.22-33.97) | |
| Negative control | - | 10/10 | 100 |
The protective effect (escherichia coli) of table 3 extractum of the present invention and erythromycin to infecting in the mice body
| Infection strain (bacterial strain number) intoxicating bacterium amount (CFU/ml) | Medicine | Dosage (mg/kg) | Death toll (only) | Mortality rate % | ED 50And 95% fiducial limit (mg/kg) |
| Number of animals (only) | |||||
| Escherichia coli No.00125 (5.2 * 10 4) | Extractum of the present invention (pressing the crude drug amount calculates) | 5895 3930 1965 983 491 | 3/10 6/10 8/10 9/10 10/10 | 30 60 80 90 100 | 4162 (2782-9010) |
| Erythromycin | 70.00 56.00 45.00 36.0 28.8 | 1/10 3/10 6/10 7/10 9/10 | 10 30 60 70 90 | 45.87 (38.94-54.61) | |
| Negative control | - | 10/10 | 100 |
The result shows: extractum of the present invention, erythromycin are irritated stomach respectively to the ED of administration to the scorching NCTC10662 infecting mouse of pneumonia streptococcus
50Value is respectively 2027mg/kg and 28.67mg/kg, to the ED of escherichia coli 9912 infecting mouses
50Be respectively 4162mg/kg and 45.87mg/kg.
1.2.4 conclusion: can find out that from above experimental result extractum of the present invention also presents certain vivo bacteria corrosion action, to the ED of pneumonia streptococcus inflammation and coli-infection mice
50Be respectively 2027mg (crude drug)/kg and 4162mg (crude drug)/kg.
2 extractum extracorporeal antivirus effect experimentation laboratory reports of the present invention
2.1 experiment material
2.1.1 be subjected to reagent: extractum of the present invention.Brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001.
2.1.2 positive control drug: virazole (ribavirin) injection: specification 100mg/ml/ props up, and lot number is
2003101; The Yichang three gorges pharmaceutical factory produces, and purchases the western medical group company in Chengdu.
2.1.3 Strain: pneumonitis virus (F.m100) and respiratory syncytial virus (RSV) are provided by Shoudu Inst. of Pediatrics and this chamber (clinical separation strain) respectively.
1.4 cell strain
2.1.4:Hep-2 cell: bio-engineering corporation provides by the eternal brightness in Shantou City, and growth nutrient solution is 10% calf serum Eagles liquid, before the experiment my chamber recover, go down to posterity, the packing test tube.
2.2 experimental technique
2.2.1 drug toxicity is measured
With Hanks liquid with extractum of the present invention and positive control drug virazole respectively from 1: 10-1: 80 make doubling dilution, get above 2 kinds of medicinal liquids and 1: 10-1: 80 medicinal liquids are that 0.1ml inoculates each 2 of Hep-2 cell pipes respectively, after waiting to adsorb 45 minutes, add 2% calf serum Eagles liquid 0.9ml, establish cell contrast, blank synchronously, put 37 ℃ of incubators.Every day the observation of cell toxic reaction, and the record result.
2.2.2 virus virulence is measured
With Hanks liquid with pneumonitis virus (F.m100) and respiratory syncytial virus (RSV) from 10
-1-10
-6Dilution.Get the viral liquid of the various concentration of above 10 times of dilutions and inoculate 0.1ml respectively in 4 Hep-2 cell pipes, wait to adsorb 45 minutes after, add 2% calf serum Eagles liquid 0.9ml, establish cell contrast synchronously, put 37 ℃ of incubators.Observation of cell pathological changes every day (CPE), with high concentration virus pipe appearance 〉=3+ cytopathy (CPE), the cell contrast is normal, but termination test.With least concentration virus pipe appearance 〉=2+ cytopathy (CPE) is the virus virulence terminal point, calculates this virus TCID
50
2.2.3 antivirus test
Get 100TCID
50/ 0.1ml pneumonitis virus (F.m100) and respiratory syncytial virus (RSV) infect each 2 of Hep-2 cell pipes, after waiting to adsorb 60 minutes, flush away virus liquid, the stock solution and 1 that adds 0.1ml extractum of the present invention and positive control drug virazole eye drop respectively: 10-1: 80 dilute liquid medicines, after waiting to adsorb 45 minutes, add 2% calf serum Eagles liquid 0.8ml respectively at each viral Hep-2 cell pipe, establish cell contrast, virus control and blank (replacing medicinal liquid) synchronously, put 37 ℃ of incubators with Hanks liquid.Every day the observation of cell pathological changes, and the record result.
2.3 experimental result
2.3.1 cell toxicity test result
The stock solution of extractum of the present invention and positive control drug virazole and 1: 10-1: 80 medicinal liquids all do not have any toxic reaction to the Hep-2 cell.The results are shown in Table 4
Table 4 cell toxicity test result
| Medicinal liquid | Cell | Stock solution | 1∶10 | 1∶20 | 1∶40 | 1∶80 | The cell contrast | Blank |
| Extractum of the present invention | Hep-2 | 0/2 | 0/2 | 0/2 | 0/2 | 0/2 | 0/2 | 0/2 |
| Virazole | Hep-2 | 0/2 | 0/2 | 0/2 | 0/2 | 0/2 | 0/2 | 0/2 |
Annotate: " 0 " expression cell does not have pathological changes; " 2 " expression cell pipe number.
2.3.2 virus virulence measurement result
Virus is selected responsive permissive cell for use, adds the viral liquid of the variable concentrations of 10 times of dilutions respectively, with Reed, Muench method, calculates viral half infection cell amount (TCID according to the cytopathy result
50).The results are shown in
Table 5
Each viroid TCID of table 5
50The result
| Virus | Pneumonitis virus (F.m100) | Respiratory syncytial virus (RSV) |
| TCID 50 | 10 -4.66 | 10 -8.22 |
2.3.3 antivirus test result
The stock solution of extractum of the present invention (1: 1) and other concentration liquids all have pathological changes effect in the cell of inhibition to pneumonitis virus (F.m100) and respiratory syncytial virus (RSV).
Positive control drug, virazole stock solution (1: 1) all has pathological changes effect in the cell of inhibition to pneumonitis virus (F.m100) and respiratory syncytial virus (RSV).
The results are shown in Table 6
Table 6 extractum of the present invention and ribavirin injection antiviral result
| Medicinal liquid | Virus | Liquor strength | The cell contrast | Virus control | ||||
| Stock solution | 1∶10 | 1∶20 | 1∶40 | 1∶80 | ||||
| Extractum of the present invention | F.m100 RSV | 0/2 0/2 | 0/2 0/2 | 0/2 0/2 | 2/2 2/2 | 2/2 2/2 | 0/2 0/2 | 2/2 2/2 |
| Virazole | F.m100 RSV | 0/2 0/2 | 2/2 2/2 | 2/2 2/2 | 2/2 2/2 | 2/2 2/2 | 0/2 0/2 | 2/2 2/2 |
Annotate: 0/2 expression, 2 solencytes do not have pathological changes.2/2 expression, 2 solencytes all have pathological changes.
Extractum and contrast medicine virazole are pressed Reed, Muench method in intracellular antiviral result according to the present invention, calculate and suppress cell virus infective dose (TCID
50), minimum effective drug concentration (MTC), medium effective concentration (IC
50) and therapeutic index (TI), the results are shown in Table 7.
Table 7 extractum of the present invention and virazole relevant data result
| Medicinal liquid | Virus | TCID 50 | MTC | IC 50 | TI |
| Extractum of the present invention | F.m100 RSV | 10 -4.66 10 -4.66 | 1∶20 1∶20 | 1∶28.16 1∶28.16 | 20 20 |
| Virazole | F.m100 RSV | 10 -4.66 10 -8.22 | 1∶1 1∶1 | 1∶1.4 1∶1.4 | 1 1 |
2.4 conclusion
Extractum of the present invention and pneumonitis virus (F.m100) and respiratory syncytial virus (RSV) all had in various degree inhibitory action.The stock solution of positive control drug virazole (1: 1) has inhibitory action to pneumonitis virus (F.m100) and respiratory syncytial virus (RSV) type.Extractum of the present invention all is better than virazole to the inhibitory action of pneumonitis virus (F.m100) and respiratory syncytial virus (RSV).
3 immunoregulation effects
3.1 the present invention is to the influence of cyclophosphamide mice reticuloendothelial system (RES) phagocytic function
3.1.1 test material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Blank product: normal saline.
Positive reference substance: levamisole hydrochloride, the 25mg/ sheet is produced by guilin pharmacy factory.Lot number: 980901, be mixed with 0.025% concentration, dosage 5mg/kg.
Cyclophosphamide: Hualian Pharmaceutical Co., Ltd., Shanghai produces, lot number 981120.Dosage 80mg/kg.
Reagent: Na
2CO
3(AR), reach the production of evolution material Fine Chemical Works, lot number: 010604 by the sky, Tianjin.
-get the pavilion ink, produce by Beijing Ink Factory.
Instrument: UVIKON-N type ultraviolet-uisible spectrophotometer, make by BIO-TEK company.
Animal: Kunming mouse, body weight 18-20g, male and female half and half.Provide the quality certification by Sichuan Academy of Medical Sciences's Experimental Animal Center: No. the 24101102nd, the moving word of doctor.
3.1.2 test method
60 of Kunming mouses, group is divided into 6 groups (seeing Table 8) at random, 10 every group, is divided into the basic, normal, high dosage group of blank group, model group, levamisole hydrochloride group and extract powder of the present invention.Wherein blank group and model group are irritated stomach and are given normal saline; The basic, normal, high dosage of extract powder of the present invention irritates respectively that stomach gives 98.2,196.3,393.0mg crude drug/kg (be equivalent to respectively the clinical consumption per day of people 5 times, 10 times, 20 times), the administration volume is the 0.2ml/10g the weight of animals, once a day, successive administration is 10 days; The levamisole hydrochloride group is irritated stomach 5mg/kg, successive administration 7 days.Simultaneously in the 6th, 7 two day all the other equal intraperitoneal injection of cyclophosphamide (dosage 80mg/kg) except that the blank group.In last administration 30min, weigh, tail vein injection one the pavilion ink (0.1ml/10g.bw of 16mg carbon granule/ml), respectively after injecting ink 5,10min gets blood 0.025ml with the special blood suction pipe of getting from the mouse orbit rear vein beard, and is blown into 0.1%NaCO immediately
3Among the liquid 2ml, suction pipe is drawn repeatedly, in order to avoid blood invests tube wall, on ultraviolet-uisible spectrophotometer in 675nm place colorimetric.Cut open the belly, get the spleen liver and weigh, be calculated as follows phagocytic index K and engulf coefficient (correction phagocytic index) α.
OD
1, OD
2Be the optical density of different time institute blood sampling, T
2-T
1Be the time difference of blood sampling,
3.1.3 result of the test
Result of the test sees Table 8:
Table 8 the present invention is to the influence of cyclophosphamide mice reticuloendothelial system (RES) phagocytic function (x ± SD)
| Group | Dosage (the g crude drug/kg) | Number of animals (only) | Phagocytic index (K) * 10 -2 | Engulf coefficient (α) |
| Blank | Deng the capacity distilled water | 10 | 5.11±1.02 * | 6.58±0.84 * |
| Model group | Deng the capacity distilled water | 10 | 3.10±1.12 | 4.48±0.74 |
| Levamisole | 5mg/kg | 10 | 6.18±1.67 * | 7.02±1.10 * |
| Low dose group of the present invention | 98.2 | 10 | 6.31±1.17 * | 7.16±0.83 * |
| Dosage group among the present invention | 196.3 | 10 | 6.51±0.98 * | 7.98±0.85 * |
| High dose group of the present invention | 393.0 | 10 | 6.23±2.01 * | 7.09±1.44 * |
Annotate: compare with model group:
*P＜0.05.
Table 8 result shows that the OD value calculates K and α after measured, and the K value and the α value of the basic, normal, high dosage group of levamisole and the present invention all have increase, have compared significant difference p＜0.05 with model group.
3.1.4 conclusion (of pressure testing): after the white mice per os gavages the present invention, can increase mice phagocytic index K and engulf factor alpha.Illustrate that the present invention has the effect of the reticuloendothelial system phagocytic activity of enhancing.
3.2 granule is done the influence that immunogenic hemolysin produces to chicken red blood cell
3.2.1 test material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Blank product: normal saline.
Positive reference substance: levamisole hydrochloride, the 25mg/ sheet is produced by guilin pharmacy factory.Lot number: 980901, be mixed with 0.025% concentration, dosage 5mg/kg.
Cyclophosphamide: Hualian Pharmaceutical Co., Ltd., Shanghai produces, lot number 981120.Dosage 80mg/kg.
Reagent: Na
2CO
3(AR), reach the production of evolution material Fine Chemical Works, lot number: 010604 by the sky, Tianjin.
-get the pavilion ink, produce by Beijing Ink Factory.
Alsever ' s solution: citric acid trisodium, 2H
2O 8.0g, citric acid 0.5g, anhydrous glucose 18.7g, sodium chloride 4.2g, distilled water add to 1000ml.
Chicken red blood cell is preserved in AlseverShi liquid at 4 ℃ of refrigerators, faces the time spent and gives a baby a bath on the third day after its birth time with normal saline, is made into 5% suspension and gets final product.
Complement is made into 10% with guinea pig serum with normal saline.Be incubated 30min in the calorstat, stopped reaction in 0 ℃ of refrigerator
Instrument: UV-75 type spectrophotometer.
Animal: Kunming mouse, body weight 18-20g, male and female half and half.Provide the quality certification by Sichuan Academy of Medical Sciences's Experimental Animal Center: No. the 24101102nd, the moving word of doctor.
3.2.2 test method
60 mices are divided into 6 groups (seeing Table 9) at random, 10 every group, are divided into the basic, normal, high dosage group of blank group, model group, levamisole hydrochloride group and extract powder of the present invention.Every group every animal ip5% chicken red blood cell stock solution 0.2ml/ only carried out immunity in first day.Administration after 1 hour, wherein blank group and model group are irritated stomach and are given normal saline; The basic, normal, high dosage of extract powder of the present invention irritates respectively that stomach gives 98.2,196.3,393.0mg crude drug/kg (be equivalent to respectively the clinical consumption per day of people 5 times, 10 times, 20 times), the administration volume is the 0.2ml/10g the weight of animals, once a day, successive administration is 10 days; The levamisole hydrochloride group is irritated stomach 5mg/kg, successive administration 7 days.In began in the 7th day except that the blank group all the other after administration half an hour simultaneously every day subcutaneous injection 0.1% cyclophosphamide (dosage 80mg/kg).
1h after the last administration and behind the cyclophosphamide of subcutaneous injection 0.1%, every animal is plucked eyeball and gets blood 1ml, centrifugal, get serum and dilute 100 times with normal saline, get dilute serum 1m1, with 5% chicken erythrocyte suspension 0.5ml, 10% complement 0.5ml mixes, and 37 ℃ of water bath heat preservation half an hour, puts cessation reaction in 0 degree centigrade of refrigerator again, centrifugal, get supernatant in spectrophotometer 540nm place colorimetric, photometry density (OD) is calculated hemolysin value (OD value * 100), and win spleen, calculate index and spleen index (spleen weight/body weight * 10).Other establishes the not blank group of increase serum.Relatively each the group difference.
3.2.3 result of the test
The results are shown in Table 9
Table 9 granule is made the influence that immunogenic hemolysin produces (n=10, x ± SD) to chicken red blood cell
| Group | Dosage (the g crude drug/kg) | Hemolysin content | Index and spleen index (mg/10g) |
| Blank group | The equivalent normal saline | 4.78±0.49 ** | 65.14±7.34 ** |
| Model group | The equivalent normal saline | 3.18±0.65 | 40.20±8.92 |
| Levamisole hydrochloride | 5mg/kg | 4.12±0.54 ** | 53.08±10.19 * |
| Extractum low dose group of the present invention | 98.2 | 3.94±0.69 | 53.36±8.02 * |
| Dosage group in the extractum of the present invention | 196.3 | 4.23±0.70 ** | 53.10±8.13 ** |
| Extractum high dose group of the present invention | 393.0 | 4.75±0.35 ** | 64.39±5.98 ** |
Annotate: compare with model group:
*P＜0.05,
*P＜0.01
The result shows the existing model group of the hemolysin content of blank group, levamisole hydrochloride and the middle and high dosage group of the present invention relatively, through the t check, significant difference (P＜0.01) is arranged; Index and spleen index index and spleen index and model group be levamisole hydrochloride and low dose group of the present invention relatively, through the t check, variant (P＜0.05), blank group and the middle and high dosage group of the present invention are checked variant P＜0.01 through t.
3.2.4 conclusion (of pressure testing): the present invention has obvious rising effect to mice hemolysin content and the index and spleen index that cyclophosphamide causes immunocompromised.Illustrate that the present invention can strengthen humoral immune function.
4 antitussive actions: the present invention causes the influence of cough latent period and cough number of times to mice ammonia
4.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Codeine phosphate inj: specification is that 30mg/ props up, and lot number 20010601 is by Shenyang No. 1 Pharmaceutical Factory production.
Strong aqua ammonia: analytical pure, lot number 20001211, chemical reagent factory in Chengdu produces.
Animal: Kunming mouse, body weight 18-20g, male and female half and half are provided by Sichuan Academy of Medical Sciences's Experimental Animal Center.The quality certification: No. the 24101102nd, the moving word of doctor.
4.2 experimental technique
Choose before the experiment 50 of the mices of ammonia irritative cough sensitivity, be divided into 5 groups (seeing Table 10) at random, every group 10, the basic, normal, high dosage of extract powder of the present invention irritates respectively that stomach gives 98.2,196.3,393.0mg crude drug/kg (be equivalent to respectively the clinical consumption per day of people 5 times, 10 times, 20 times), the administration volume is the 0.2ml/10g the weight of animals, negative group replaces with the distilled water of isometric(al) amount, once a day, and successive administration 10 days.Codeine phosphate group dosage is 0.03g/kg, and route of administration is a lumbar injection, and administration time is back 3 days.In the dry wide mouthed bottle of 500ml is put mice in last administration 30 minutes, the built-in cotton balls of bottle, only inject strong aqua ammonia 0.5ml/ to cotton balls, seal bottleneck immediately, after 5 seconds mice is taken out rapidly, observe mouse cough incubation period (injecting strong aqua ammonia certainly and begin, is incubation period to cough institute takes place through the time) and inject strong aqua ammonia certainly and begin to cough in the kind in 5 minutes that (mouse cough shows as the abdominal muscle contraction to number of times, magnify mouth simultaneously, cough sound sometimes).Comparison between organizing with each group mouse cough incubation period and cough number of times and negative control group.
4.3 experimental result
The results are shown in Table 10
Table 10 the present invention causes the influence of cough latent period and cough number of times to mice ammonia
| Group | Number of animals | Drug dose (the mg crude drug/kg) | Cough latent period (second, x ± s) | Cough number of times in 5 fens kinds (x ± s) |
| Dosage group extractum high dose group of the present invention in the distilled water group codeine phosphate group extractum low dose group of the present invention extractum of the present invention | 10 10 10 10 10 | Deng capacity distilled water 0.03g/kg 98.2 196.3 393.0 | 31.6±17.2 215.8±71.2 ** 208.2±57.4 ** 155.2±52.6 ** 133.2±58.4 ** | 17.4±6.8 2.6±1.8 ** 3.4±2.5 ** 6.5±3.9 ** 9.5±5.4 ** |
Annotate: compare with the distilled water group:
*P＜0.01.
The result shows that cough all has obvious antitussive action to the strong aqua ammonia induced mice for codeine phosphate group, the basic, normal, high dosage group of extract powder of the present invention in the table.
4.4 conclusion: cough has obvious antitussive action to extractum of the present invention to the strong aqua ammonia induced mice.
5 phlegm-dispelling functions
5.1 the present invention is to the influence of the phenol red excretion of mice trachea section
5.1.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Ammonium chloride: specification 0.3g/ sheet, lot number 980409 is produced by Chongqing pharmacy six factories.
Phenol red: analytical pure, lot number 980327, Tianjin chemical reagent one factory.
Spectrophotometer: UVIKON-N type ultraviolet-uisible spectrophotometer, make by BIO-TEK company.
Animal: Kunming mouse, body weight 18-20g, male and female half and half are provided by Sichuan Academy of Medical Sciences's Experimental Animal Center.The quality certification: No. the 24101102nd, the moving word of doctor.
5.1.2 experimental technique
50 of Kunming mouses, group is divided into 5 groups (seeing Table 11) at random, every group 10, the basic, normal, high dosage of extract powder of the present invention irritates respectively that stomach gives 98.2,196.3,393.0mg crude drug/kg (be equivalent to respectively the clinical consumption per day of people 5 times, 10 times, 20 times), the administration volume is the 0.2ml/10g the weight of animals, negative group replaces with the distilled water of isometric(al) amount, once a day, and successive administration 10 days; The ammonium chloride group is irritated stomach 1.0g/kg, successive administration 3 days.In last administration 60 minutes, lumbar injection 5% phenol red solution 0.2ml/ only injected and took off cervical vertebra execution mice in back 30 minutes.It is lain on the back be fixed on the operation plate, cut off neck center skin, separate trachea, No. 7 syringe needles that will polish down in larynx insert about 0.3cm in the tracheas, with the silk thread ligation fixing after, with 1ml syringe absorption 5%NaHCO
3Solution 0.5ml by syringe needle aeration pipe 3 times back and forth, collects and mixes irrigating solution 3 times, puts in the test tube centrifugally then, gets supernatant and measures the OD value in spectrophotometer 546nm wavelength place.Use phenol red work one standard curve simultaneously, calculate phenol red content (m/ml) according to standard curve.With each organize phenol red content and negative control group organize between relatively.
5.1.3 experimental result
The results are shown in Table 11
Table 11 the present invention to the influence of the phenol red excretion of mice trachea section (N=10, second, x ± s)
| Group | Drug dose (the mg crude drug/kg) | The phenol red excretion amount of trachea (m/ml) |
| The distilled water group | Deng capacity distilled water 0.2ml/10g | 2.87±0.67 |
| The ammonium chloride group | 1.0g/kg | 5.72±1.06 ** |
| Extract powder low dose group of the present invention | 98.2 | 4.66±0.96 * |
| Dosage group in the extract powder of the present invention | 196.3 | 4.25±0.94 ** |
| Extract powder high dose group of the present invention | 393.0 | 2.76±0.76 ** |
Annotate: compare with the distilled water group:
*P＜0.01.
The result shows that ammonium chloride group, the middle and high dosage group of extract powder of the present invention have obvious facilitation to the amount of the phenol red excretion of mice trachea section in the table.
5.1.4 conclusion: the middle and high dosage of extractum of the present invention has obvious facilitation to the amount of the phenol red excretion of mice trachea section, thereby has phlegm-dispelling functions.
5.2 the present invention is to the influence of rabbit isolated tracheal cilium mucus stream motion
5.2.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Ammonium chloride: specification 0.3g/ sheet, lot number 980409 is produced by Chongqing pharmacy six factories.
Animal: 50 of rabbit, body weight 2.1-2.2kg, male and female half and half are provided by Sichuan Academy of Medical Sciences's Experimental Animal Center.The quality certification: No. the 24101102nd, the moving word of doctor.
5.2.2 experimental technique
Get 50 of rabbit, be divided into 5 groups (seeing Table 12) at random, every group 10, the basic, normal, high dosage of extract powder of the present invention irritates respectively that stomach gives 98.2,196.3,393.0mg crude drug/kg (be equivalent to respectively the clinical consumption per day of people 5 times, 10 times, 20 times), the administration volume is the 0.2ml/10g the weight of animals, negative group replaces with the distilled water of isometric(al) amount, once a day, and successive administration 10 days; The ammonium chloride group is irritated stomach 1.0g/kg, successive administration 3 days.In the last administration after 1 hour, with the hammer rabbit of hitting unconsciously, the femoral artery sacrificed by exsanguination, peel off and take out the whole section trachea of larynx rapidly, put into the tyrode's solution of oxygenation immediately, one section trachea of clip to the trachea crotch, vertically be divided into two sections, get one section and put on the moistening Cotton Gossypii of 37 ℃ of tyrode's solutions, intima of the trachea upwards, trachea is placed in the culture fluid with Cotton Gossypii together, keep 30 ° of inclinations, the larynx end makes progress, and periphery is shone with cold light source on it with 37 ℃ of constant temperature water baths, stablized 5 minutes, dip in a little india ink with syringe needle and be put in the lower end, aim at trachea, observe the prepared Chinese ink granule operation required time of 2mm with the anatomical lens that amplifies 20 times.Each sample view 3 times is got the meansigma methodss of 3 times.The each observation finishes, and the trachea section is put rinsing in the tyrode's solution, removing prepared Chinese ink,
Survey again after 5 minutes 1 time, to carrying out statistical disposition running time.
5.2.3 experimental result
The results are shown in Table 12
The influence of table 12 pair rabbit isolated tracheal cilium mucus stream motion (N=10, second, x ± s)
| Group | Dosage (g/kg) | Running time (second) |
| The distilled water group | Deng capacity distilled water 0.2ml/10g | 260.00±50.62 |
| The ammonium chloride group | 1.0g/kg | 37.50±8.20 ** |
| Low dose group of the present invention | 98.2 | 39.00±15.55 ** |
| Dosage group among the present invention | 196.3 | 26.12±10.03 ** |
| High dose group of the present invention | 393.0 | 23.00±5.15 ** |
Annotate: compare with distilled water group group,
*P＜0.01
The result shows ammonium chloride group, the effect that motion has obvious promotion to accelerate to rabbit isolated tracheal cilium mucus stream of the basic, normal, high dosage group of extract powder of the present invention in the table.
5.2.1 conclusion: the basic, normal, high dosage of extractum of the present invention has phlegm-dispelling functions.
6 antiasthmatic effects
6.1 the present invention causes the influence of Cavia porcellus asthma to the salty and histamine phosphate of acetyl chloride gallbladder
6.1.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Aminophylline: specification 0.3g/ sheet, lot number 001017, Sichuan Province Deyang Huakang Pharmaceutical Co., ltd. produces.
Cause and breathe heavily liquid: the salty and 0.1% histamine phosphate mixed liquor autogamy in 1: 1 of 2% acetyl chloride gallbladder.
Animal: Cavia porcellus, body weight 150-200g, male and female half and half are provided by Sichuan Academy of Medical Sciences's Experimental Animal Center.The quality certification: No. the 25103102nd, the moving word of doctor.
6.1.2 experimental technique
Before the experiment Cavia porcellus is put in the airtight bell jar one by one, in bell jar, spray to cause by jet atomization device with the 53kPa constant voltage and breathed heavily liquid (salty and 1: 1 mixed liquor of 0.1% histamine phosphate of 2% acetyl chloride gallbladder) 15 seconds, after spraying stops, whether the observation Cavia porcellus tic of panting property occurred in 2.5 minutes, if insensitive animal do not occur being considered as, abandon it.Select 50 satisfactory responsive Cavia porcelluss altogether, be divided into 5 groups (seeing Table 13) at random, every group 10, the basic, normal, high dosage of extract powder of the present invention irritates respectively that stomach gives 98.2,196.3,393.0mg crude drug/kg (be equivalent to respectively the clinical consumption per day of people 5 times, 10 times, 20 times), the administration volume is the 0.2ml/10g the weight of animals, negative group replaces with the distilled water of isometric(al) amount, once a day, and successive administration 10 days.Aminophylline group dosage is 86mg/kg, and route of administration is for irritating stomach, and administration time is back 3 days.In last administration 30 minutes Cavia porcellus is put in the volumetrical airtight bell jar of 4L, with WM-6 type gas compressor, in bell jar, spray to cause by jet atomization device with the 53kPa constant voltage and breathed heavily liquid (salty and 1: 1 mixed liquor of 0.1% histamine phosphate of 2% acetyl chloride gallbladder) 15 seconds, after spraying stops, observed Cavia porcellus 5 minutes, writing down each Cavia porcellus is asthma incubation period from stopping after the spraying to the tic of panting property occurring, if not the panting property tic of kind above 5 minutes, then its cough latent period is still by 300 seconds.Comparison between organizing with each group Cavia porcellus asthma time incubation period and negative control group.
6.1.3 experimental result
The results are shown in Table 13
Table 13 the present invention to the salty and histamine phosphate of acetyl chloride gallbladder cause the preclinical influence of Cavia porcellus asthma (N=10, second, x ± s)
| Group | Drug dose (the mg crude drug/kg) | Panting property tic asthma incubation period |
| The distilled water group | Deng capacity distilled water 0.2ml/10g | 142.6±28.8 |
| The aminophylline group | 0.086g/kg | 267.6±32.6 ** |
| Extract powder low dose group of the present invention | 98.2 | 221.3±46.9 ** |
| Dosage group in the extract powder of the present invention | 196.3 | 239.0±48.8 ** |
| Extract powder high dose group of the present invention | 393.0 | 280.9±20.5 ** |
Annotate: compare with the distilled water group:
*P＜0.01.
The result shows that salty and histamine phosphate causes that Cavia porcellus asthma has the prolongation effect incubation period to the acetyl chloride gallbladder for aminophylline group, the basic, normal, high dosage group of extract powder of the present invention in the table.
6.1.4 conclusion: extractum of the present invention causes that with histamine phosphate Cavia porcellus asthma has tangible antiasthmatic effect to the acetyl chloride gallbladder is salty.
6.2 extractum of the present invention causes the influence of bronchoconstriction to acetylcholine
6.2.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Positive drug: CHUANBEI PIPA GAO, natural drug pharmaceutical factory in Jiu Zhaigou produces, lot number 010408,100ml/ bottle.
Reagent: acetylcholine: Shanghai chemical reagent head factory is produced.
K-H (Krebs-Henseleit) liquid, (95%O
2+ 5%CO
2) mist,
The desk-top balance recorder of instrument: XWTD-264, the big magnificent instrument plant in Shanghai produces
Ultrathermostat: experimental facilities factory in Chongqing produces
Animal: Cavia porcellus, body weight 150-200g, male and female half and half are provided by Sichuan Academy of Medical Sciences's Experimental Animal Center.The quality certification: No. the 28103002nd, the moving word of doctor.
6.2.2 experimental technique
Get 50 of 400-500g Cavia porcelluss, be divided into 5 groups (seeing Table 14), shoot dead, isolate trachea, preparation trachea spiral bar places to fill 37 ℃ of Magnus' baths of 15ml K-H (Krebs-Henseleit) nutritional solution constant temperature in K-H (Krebs-Henseleit) nutritional solution, and the lower end is fixed, the upper end connects desk-top self-balancing recorder by tonotransducer, continues to feed (95%O
2+ 5%CO
2) mist, tracheal strip load 3g, specimen balance 90min begins test after stablizing, and adds acetylcholine earlier, and final concentration is 10
-5Mol, after waiting to act on peaking, the basic, normal, high dosage of extract powder of the present invention that adds variable concentrations respectively irritates respectively that stomach gives 98.2,196.3,393.0mg crude drug/kg (be equivalent to respectively the clinical consumption per day of people 5 times, 10 times, 20 times), CHUANBEI PIPA GAO group dosage is QQQmg/kg, negative group with the distilled water of isometric(al) amount replace (make that the bath final concentration is respectively 27,54,108mg crude drug/ml), observe the spasmolysis of medicine to tracheal smooth muscle
6.2.3 result of the test
The results are shown in Table 14
Table 14 extractum of the present invention causes influence (N=10, the x ± s) of bronchoconstriction to acetylcholine
| Group | Dosage (g/kg) | Spasmolytic percentage rate (%) |
| Dosage group extract powder high dose group of the present invention in the distilled water group CHUANBEI PIPA GAO group extract powder low dose group of the present invention extract powder of the present invention | Equivalent distilled water 12.3mg cream weight/ml 98.2 196.3 393.0 | 2.52±0.82 82.55±7.04 ** 27.89±5.17 ** 50.39±10.64 ** 72.57±10.33 ** |
Annotate: compare with distilled water group group,
*P＜0.01
But the result shows all contraction bronchus effect P＜0.01 of the antagonism acetylcholine of concentration dependent of CHUANBEI PIPA GAO group and the basic, normal, high dosage group of extract powder of the present invention.And the distilled water group does not have this effect.
6.2.4 conclusion: medicine of the present invention has antiasthmatic effect.
7 refrigeration functions
7.1 the present invention is to the influence of fever in rabbits due to the triple vaccine
7.1.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Aspirin Enteric-coated Tablets: white tablet, the 0.3g/ sheet, the northwest second synthesis pharmaceutical factory produces, lot number 981201, it is standby to be mixed with 0.06g/ml with distilled water.
Typhoid fever, paratyphoid fever, second triple vaccine: dosage 1.0ml/kg body weight is provided by Chengdu Inst. of Biological Products.
Instrument: WMZ-03 temperature indicator, Shanghai Medical Instrument and Meter Factory.
Animal: 50 of rabbit, body weight 2.1-2.2kg, male and female half and half are provided by Sichuan Academy of Medical Sciences's Experimental Animal Center.The quality certification: No. the 24101102nd, the moving word of doctor
7.1.2 experimental technique
Select 38.5-39.6 ℃ of anus temperature with the temperature indicator before the experiment, and the temperature difference is no more than some of 0.3 ℃ of rabbit, under 20 ℃ of conditions of room temperature, each rabbit ear edge intravenous injection typhoid fever, paratyphoid fever, second triple vaccine 1.0ml/kg body weight, survey the anus temperature after 1 hour, choose intensification 50 of the fever in rabbit more than 0.6 ℃, be divided into 5 groups at random by the intensification situation, every group 10, disposable administration extract powder of the present invention is low, in, high dose is irritated stomach respectively and is given 98.2,196.3,393.0mg crude drug/kg (is equivalent to 5 times of the clinical consumption per day of people respectively, 10 times, 20 times), the administration volume is the 0.2ml/10g the weight of animals, and negative group replaces with the distilled water of isometric(al) amount.Aspirin group dosage is 86mg/kg, and route of administration was surveyed anus temperature in 0.5,1,2,3,4,5,6 hour for irritating stomach after the administration, write down each time period animal anus temperature, and the feminine gender group organize between relatively
7.1.3 experimental result
The results are shown in Table 15
Table 15 the present invention to the influence of fever in rabbits due to the triple vaccine (N=10, ℃: x ± SD)
| Group | Dosage | Normal body temperature | Pyrogenicity body temperature | Body temperature after the administration | ||||||
| 0.5h | 1h | 2h | 3h | 4h | 5h | 6h | ||||
| Blank | Water | 39.2± 0.3 | 40.0 ±0.3 | 39.9± 0.3 | 39.8± 0.7 | 40.0± 0.3 | 40.0± 0.4 | 39.3± 0.5 | 39.4± 0.6 | 39.3± 0.5 |
| The aspirin group | 0.6 | 39.2± 0.3 | 40.2 ±0.4 | 39.4± 0.5 * | 39.4± 0.3 * | 39.6± 0.5 * | 39.6± 0.5 * | 39.2± 0.4 * | 38.7± 0.6 * | 38.7± 0.6 * |
| Low dose group of the present invention | 98.2 | 39.0± 0.3 | 40.0 ±0.2 | 39.3± 0.6 * | 39.4± 0.6 * | 39.5± 0.5 * | 39.4± 0.4 * | 39.0± 0.5 * | 38.9± 0.8 * | 39.0± 0.8 * |
| Dosage group among the present invention | 196. 3 | 39.0± 0.3 | 40.1 ±0.2 | 39.3± 0.5 * | 39.3± 0.4 * | 39.4± 0.3 * | 39.4± 0.4 * | 39.1± 0.5 * | 38.6± 0.4 * | 39.1± 0.7 * |
| High dose group of the present invention | 393. 0 | 39.1± 0.3 | 40.2 ±0.3 | 39.1± 0.5 * | 39.1± 0.4 * | 39.2± 0.2 * | 39.2± 0.2 * | 39.0± 0.4 * | 38.2± 0.2 * | 39.0± 0.3 * |
Annotate: compare with distilled water group group,
*P＜0.05
The result shows: aspirin group, the basic, normal, high dosage of extract powder of the present invention and blank group relatively have significant difference, P＜0.05
7.1.4 conclusion
The basic, normal, high dosage of extract powder of the present invention has the effect of fever in rabbits due to the remarkable inhibition triple vaccine, and this refrigeration function is sustainable more than 3 hours.
7.2 the present invention causes the influence of rat fever effect to the young beer yeast
7.2.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Aspirin Enteric-coated Tablets: white tablet, the 0.3g/ sheet, the northwest second synthesis pharmaceutical factory produces, lot number 981201, it is standby to be mixed with 0.06g/ml with distilled water.
The young beer yeast: Chengdu medicated beer company limited provides.Centrifugal with the normal saline cyclic washing, calculate with the yeast weight in wet base of supernatant, be mixed with 15% outstanding solution with normal saline and in 4 ℃ of refrigerators, preserve standby.
Animal: the Wistar rat, male, 140-250g is provided by Chengdu University of Traditional Chinese Medicine's Experimental Animal Center, the quality certification number: No. 8, the real moving Guan Zhidi in river.
Instrument: Omron MC-3B type computer numeral formula clinical thermometer, Dalian, Omron company limited is produced.
7.2.2 experimental technique
Twice of temperature of experiment before measurement rat anus (temperature indicator probe inserts anus 2cm place), select the young beer yeast 1ml/100g pyrogenicity of normal body temperature rat back subcutaneous injection 15%, 50 of rats that choosing after the pyrogenicity heated up in 6 hours surpasses 0.8 ℃ are divided into 5 groups (seeing Table 16) at random according to body weight, every group 10, immediately by following disposable administration: extract powder of the present invention is low, in, high dose is irritated stomach respectively and is given 98.2,196.3,393.0mg crude drug/kg (is equivalent to 5 times of the clinical consumption per day of people respectively, 10 times, 20 times), the administration volume is the 0.2ml/10g the weight of animals, and negative group replaces with the distilled water of isometric(al) amount; Positive drug aspirin 0.6g/kg (be equivalent to the clinical consumption per day of people 20 times), the administration volume is 1ml/100g, measures 0.5,1,2,3,4 hour anus temperature after the administration respectively, calculates the difference with normal anus temperature, relatively the difference of each group and negative group.
7.2.3 experimental result
The results are shown in Table 16
Table 16 the present invention to the young beer yeast cause the rat fever effect influence (n=10, ℃, x ± SD)
| Group | Dosage g/kg | Normal body temperature | The temperature difference behind the pyrogenicity 6h | Anus temperature approach after the administration | ||||
| 0.5h | 1h | 2h | 3h | 4h | ||||
| The distilled water group | Deng the capacity distilled water | 36.9 ±0.4 | 1.5 ±06 | 1.6 ±0.4 | 1.8 ±0.4 | 1.5 ±0.4 | 1.4 ±0.6 | 1.1 ±0.5 |
| The aspirin group | 0.6 | 37.0 ±0.4 | 1.3 ±0.2 | 0.8 ±0.3 ** | 0.3 ±0.4 *** | 0.2 ±0.5 *** | 0.1 ±0.5 *** | 0.2 ±0.5 ** |
| The test sample low dose group | 98.2 | 36.9 ±0.5 | 1.3 ±0.6 | 0.9 ±0.5 * | 0.5 ±0.7 ** | 0.2 ±0.9 *** | 0.2 ±0.7 *** | 0.1 ±0.7 *** |
| Dosage group in the test sample | 196.3 | 36.9 ±0.4 | 1.4 ±0.4 | 0.9 ±0.6 * | 0.7 ±0.6 ** | 0.3 ±0.4 *** | 0.3 ±0.6 *** | 0.2 ±0.5 *** |
| The test sample high dose group | 393.0 | 36.9 ±0.5 | 1.5 ±0.4 | 1.0 ±0.4 ** | 0.9 ±0.6 ** | 0.5 ±0.7 *** | 0.4 ±0.4 *** | 0.2 ±0.6 *** |
Annotate: compare with the distilled water group
*P＜0.05,
*P＜0.01,
* *P＜0.001
The result shows the significantly reduction of rat anus temperature after aspirin can make young beer yeast pyrogenicity, show tangible refrigeration function, the high, medium and low dosage of extractum of the present invention also can make the rat anus temperature after the young beer yeast pyrogenicity significantly reduce, and shows tangible refrigeration function.
7.2.4 conclusion: the high, medium and low dosage of extractum of the present invention can make the rat anus temperature after the young beer yeast pyrogenicity significantly reduce, and has significant refrigeration function.
8 antiinflammatory actions: the influence that the present invention increases lumbar injection acetic acid induced mice abdominal cavity capillary permeability
8.1 experiment material
Medicine: extract powder of the present invention, brown ceramic powder, every gram is equivalent to the 2.789g crude drug, by preparation teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine on behalf of preparation.Lot number 021001, with distilled water preparation 35.2,70.4,140.9mg/ml (be equivalent to 98.2,196.3 respectively, 393.0mg crude drug/ml) for basic, normal, high dosage group use.
Aspirin Enteric-coated Tablets: white tablet, the 0.3g/ sheet, the northwest second synthesis pharmaceutical factory produces, lot number 981201, it is standby to be mixed with 0.06g/ml with distilled water.
Azovan blue: reagent three factories in Shanghai are mixed with 0.5% solution for standby with normal saline.
0.6% acetate solution: add the normal saline preparation with Glacial acetic acid.
Instrument: UVIKON-N type ultraviolet-uisible spectrophotometer, make by BIO-TEK company
Animal: Kunming mouse, body weight 18-20g, male and female half and half are provided by Sichuan Academy of Medical Sciences's Experimental Animal Center.The quality certification: No. the 24101102nd, the moving word of doctor.
8.2 experimental technique
Select 50 of healthy mices to be divided into 5 groups (seeing Table 17) at random according to the sex body weight, every group 10, the basic, normal, high dosage of extract powder of the present invention irritates respectively that stomach gives 98.2,196.3,393.0mg crude drug/kg (be equivalent to respectively the clinical consumption per day of people 5 times, 10 times, 20 times), the administration volume is the 0.2ml/10g the weight of animals, negative group replaces with the distilled water of isometric(al) amount, once a day, successive administration is 10 days; Positive drug aspirin 0.6g/kg (be equivalent to the clinical consumption per day of people 20 times), the administration volume is 1ml/100g.Positive drug aspirin 0.6g/kg (be equivalent to the clinical consumption per day of people 20 times), the administration volume is 1ml/100g, the administration cycle is 3 days, half an hour after the last administration, the azovan blue 0.1ml/10g of every tail vein injection 0.5%, the acetate solution of lumbar injection 0.6% (dosage 0.2ml/ only) immediately, taking off cervical vertebra after 20 minutes puts to death, (divide three times with the 6ml normal saline, 2ml/ time) the washing abdominal cavity, (number) in test tube with suction pipe sucking-off washing liquid, it is fixed molten to 10ml to add normal saline, 3000rpm got supernatant in centrifugal 15 minutes in 590nm punishment light photometer colorimetric, respectively organized optical density value (OD) and the negative difference of organizing.
8.3 experimental result
The results are shown in Table 17
The influence that table 17 the present invention increases lumbar injection acetic acid induced mice abdominal cavity capillary permeability (N=10, x ± SD)
| Group | Drug dose (the mg crude drug/kg) | Optical density value (OD) |
| The distilled water group | Deng the capacity distilled water | 0.370±0.137 |
| The aspirin group | 0.06g/kg | 0.211±0.111 ** |
| Extract powder low dose group of the present invention | 98.2 | 0.214±0.102 ** |
| Dosage group in the extract powder of the present invention | 196.3 | 0.190±0.106 ** |
| Extract powder high dose group of the present invention | 393.0. | 0.196±0.112 ** |
Annotate: compare with the distilled water group:
*P＜0.05;
*P＜0.01.
The result shows that aspirin and the high, medium and low dosage of extractum of the present invention all increase lumbar injection acetic acid induced mice abdominal cavity capillary permeability and has the obvious suppression effect.
8.4 conclusion: the high, medium and low dosage of extractum of the present invention has tangible antiinflammatory action.
The embodiment of preparation aspect further specifies the present invention for example below, and this embodiment only is used to the present invention is described and the present invention is not had any restriction.
The specific embodiment
Embodiment:
By following proportioning take by weighing raw material make one Coming-of-Age Day consumption:
Fructus Viticis Negundo 20g Radix Scutellariae 20g Rhizoma Anemarrhenae 15g Radix Trichosanthis 15g
Flos Farfarae 15g Radix Asteris 12g Semen Armeniacae Amarum 12g Semen Lepidii (Semen Descurainiae) 12g
Production method is as follows:
The Fructus Viticis Negundo full dose is made fine powder (crossing 80 mesh sieves), and surplus medicine extracts and is condensed into extractum according to the decocting method of preparation granule, mix with Fructus Viticis Negundo medicated powder, the adding Icing Sugar (extractum: mix homogeneously Icing Sugar=1: 0.15), make the suspension ability granule.
Claims (5)
1. treat the medicine that belongs to traditional Chinese medical science wind-warm and pulmonary heat syndrome for one kind, it is characterized in that wherein making the raw materials of effective components medicine and consist of:
Fructus Viticis Negundo 10%-20% Radix Scutellariae 10%-20% Rhizoma Anemarrhenae 8%-18% Radix Trichosanthis 8%-18%
Flos Farfarae 8%-18% Radix Asteris 6%-16% Semen Armeniacae Amarum 6%-16% Semen Lepidii (Semen Descurainiae) 6%-16%.
2. treatment according to claim 1 belongs to the medicine of traditional Chinese medical science wind-warm and pulmonary heat syndrome, wherein makes the raw materials of effective components medicine and consists of:
Fructus Viticis Negundo 20g Radix Scutellariae 20g Rhizoma Anemarrhenae 15g Radix Trichosanthis 15g
Flos Farfarae 15g Radix Asteris 12g Semen Armeniacae Amarum 12g Semen Lepidii (Semen Descurainiae) 12g.
3. treatment according to claim 1 and 2 belongs to the medicine of traditional Chinese medical science wind-warm and pulmonary heat syndrome, it is characterized in that said traditional Chinese medical science wind-warm and pulmonary heat syndrome is the sick genus of pneumonia, bronchitis, pulmonary abscess and anemopyretic cold traditional Chinese medical science wind-warm and pulmonary heat syndrome person.
4. treatment according to claim 1 and 2 belongs to the medicine of traditional Chinese medical science wind-warm and pulmonary heat syndrome, it is characterized in that wherein making that the raw materials of effective components medicine makes is granule or mixture or oral liquid.
5. treatment according to claim 1 and 2 belongs to the medicine of traditional Chinese medical science wind-warm and pulmonary heat syndrome, it is characterized in that: the fine powder that Fructus Viticis Negundo was ground into 80 mesh sieves, in addition Radix Scutellariae, the Rhizoma Anemarrhenae, Radix Trichosanthis, Semen Armeniacae Amarum, Radix Asteris, Flos Farfarae, Semen Lepidii (Semen Descurainiae) are added decocting in water and carry three times, be condensed into thick paste after three decocting liquids are merged; The Fructus Viticis Negundo fine powder is added mixing in the thick paste, granulate granulate, drying, packing.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1063819A (en) * | 1991-02-05 | 1992-08-26 | 王柱亭 | Formula of Kezeng ointment |
| CN1365710A (en) * | 2001-01-15 | 2002-08-28 | 杨孟君 | Nano inflammation-relieving and antitussive medicine and its preparing process |
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2003
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1063819A (en) * | 1991-02-05 | 1992-08-26 | 王柱亭 | Formula of Kezeng ointment |
| CN1365710A (en) * | 2001-01-15 | 2002-08-28 | 杨孟君 | Nano inflammation-relieving and antitussive medicine and its preparing process |
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