CN106474207B - Nourishing lung and activating blood capsule for treating or the purposes for preventing pulmonary lesion medicine caused by PM2.5 - Google Patents
Nourishing lung and activating blood capsule for treating or the purposes for preventing pulmonary lesion medicine caused by PM2.5 Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/481—Astragalus (milkvetch)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/487—Psoralea
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/4841—Filling excipients; Inactive ingredients
- A61K9/485—Inorganic compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention belongs to technical field of traditional Chinese medicines, and in particular to a kind of nourishing lung and activating blood capsule is used to treat or prevent the purposes of pulmonary lesion medicine, and the Chinese medicine composition is suitable from medicinal material compatibility, meet traditional Chinese medicine and modern medicine study theory, good drug efficacy and toxic side effect is low.Described Chinese medicine composition is mainly made up of the Radix Astragali, the radix paeoniae rubrathe and psoralea corylifolia.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicines, it is related to the use that nourishing lung and activating blood capsule is used to treat or prevent pulmonary lesion medicine
On the way, and in particular to nourishing lung and activating blood capsule is used for the purposes for the medicine for treating and/or preventing pulmonary lesion caused by PM2.5.
Background technology
In recent years, atmosphere pollution constitutes serious threat to health, in various atmosphere pollution generally acknowledged at present
In thing, the Epidemiological Correlation of particulate matter and each terminal of health effect is the closest, and particulate matter has become evaluation greatly
The significant pollutant of the quantitative health hazard of gas pollution.Increasing epidemiology, toxicological information research have proven to big
Aerated particle thing especially diameter<2.5um particulate matter, the increase with death and the incidence of disease is relevant, can cause asthma, lung work(
Can decline, inflammation in respiratory system, even involve cardiovascular system, promote cancer.
" PM2.5 is to the injury of lungs of rat and its research of mechanism ", Qu Hongmei, Lanzhou University's Master dissertation,
Research in 2006 shows to cause the rise of ACP, AKP, ALB, TP, LDH content in lung-douching fluid after PM2.5 contaminations；IgG, IgA liter
Height, and dose-dependent relationship is presented；Alveolar macrophages phagocytic activity is reduced, and points out PM2.5 particulate matters to draw after entering body
Injury of lungs is played, disease causes the infringement of pulmonary parenchyma and biomembrane, excite body local immunity and mucosa-immune and from different degrees of
Reduce non-specific immune function.At present, there is not the medicine of pulmonary lesion disease caused by effective treatment PM2.5 temporarily clinically
Thing.
Nourishing lung and activating blood capsule is made up of the Radix Astragali, the radix paeoniae rubrathe, psoralea corylifolia, and qi and activate blood circulation, tonifying lung is reinforced the kidney, and (is alleviated for pulmonary heart disease
Phase) category syndrome of blood stasis due to qi deficiency, symptoms include：Cough shortness of breath, or cough and asthma are uncomfortable in chest, and palpitation, limb is cold weak, soreness and weakness of waist and knees, die Blausucht,
Pale tongue with white fur or tongue are purple dark etc.." effect of nourishing lung and activating blood capsule and clinical efficacy ", Wu Di etc., the practical medicine of China, volume 7 the
8 phases, the 148-149 pages, point out that nourishing lung and activating blood capsule has effects that qi and activate blood circulation, tonifying lung reinforce the kidney in March, 2012, it is possible to decrease complete
Carbon dioxide partial pressure, raising partial pressure of oxygen, oxygen saturation, increase cardiac output, coronary blood flow, drop in blood viscosity, reduction vim and vigour
Low peripheral vascular resistance, coronary resistance, myocardial oxygen consumption, treatment bronchial astehma, pneumoconiosis, pulmonary heart disease effect are preferable.
《Chinese Pharmacopoeia》2015 editions first describes nourishing lung and activating blood capsule prescription and preparation method thereof, Radix Astragali 720g, red
Chinese herbaceous peony 720g, psoralea corylifolia 360g, take radix paeoniae rubrathe 180g to be ground into fine powder, standby；Remaining flavour of a drug add water to cook it is secondary, for the first time plus 8 times
Water is measured, 6 times of amount water are added for the second time, 1 hour every time, collecting decoction was filtered, and it is 1.05~1.15 that filtrate, which is concentrated into relative density,
(80 DEG C), plus ethanol make alcohol content up to 60%, are sufficiently stirred for, and stand 24 hours, leaching supernatant, reclaim ethanol extremely without alcohol taste,
Continue to be concentrated into relative density for 1.35~1.40 (80 DEG C), add above-mentioned radix paeoniae rubrathe fine powder, mix, dry, pulverize, use 90% second
Alcohol is pelletized, and is dried, plus right amount of auxiliary materials, is mixed, and is loaded capsule, is made 1000, produces.And the content is only described to the Radix Astragali
The content of the principle active component Paeoniflorin of principle active component Astragaloside IV and the radix paeoniae rubrathe is defined, not to psoralea corylifolia
The content of active ingredient psoralen and isopsorapen be defined, it has been found that the Psoralen obtained by the preparation method
The content of fat element and Isopsoralen is relatively low.
Furthermore it has been found that existing nourishing lung and activating blood capsule easily draws moist during storage at present, cause medicine not
Shelf-stable, influences the stability of medicine.
In view of this, it is clinical in the urgent need to curative effect is clear and definite, toxic side effect is small, convenient to take and easily stored medicine can be used for controlling
The medicine treated and/or prevent PM2.5 to cause injury of lungs, to meet the need in market and patient.
The content of the invention
The present invention is intended to provide nourishing lung and activating blood capsule is used to prepare pulmonary lesion caused by treatment and/or prevention PM2.5
The purposes of medicine, the Chinese medicine composition is suitable from medicinal material compatibility, meets traditional Chinese medicine and modern medicine study theory, good drug efficacy and
Toxic side effect is low.
In order to achieve the above object, the present invention uses following technical scheme：
Nourishing lung and activating blood capsule is used for the purposes for preparing the medicine of pulmonary lesion caused by treatment and/or prevention PM2.5, described
Nourishing lung and activating blood capsule be mainly made up of the Radix Astragali, the radix paeoniae rubrathe and psoralea corylifolia.It is preferred that, the Radix Astragali is that 50~100 parts of (weight), radix paeoniae rubrathe are
50~100 parts (weight), psoralea corylifolia are 25~50 parts (weight).It is furthermore preferred that it is 72 parts that the Radix Astragali, which is 72 parts (weight), the radix paeoniae rubrathe,
(weight), psoralea corylifolia are 36 parts (weight).
Nourishing lung and activating blood capsule is used for the purposes for preparing the medicine of pulmonary lesion caused by treatment and/or prevention PM2.5, described
Nourishing lung and activating blood capsule contain the following active material of percentage by weight：>=0.18% Astragaloside IV, >=2.0% Paeoniflorin
With >=0.75% psoralen and isopsorapen.It is preferred that, contain >=0.20% Astragaloside IV, >=2.3% Chinese herbaceous peony
Glycosides and >=0.85% psoralen and isopsorapen.
Nourishing lung and activating blood capsule is used for the purposes for preparing the medicine of pulmonary lesion caused by treatment and/or prevention PM2.5, described
Nourishing lung and activating blood capsule reduction PM2.5 caused by mouse lung damage lung airway resistance, improve lung compliance, improve lung lead to
Airway dysfunction.
Nourishing lung and activating blood capsule is used for the purposes for preparing the medicine of pulmonary lesion caused by treatment and/or prevention PM2.5, described
Nourishing lung and activating blood capsule reduction PM2.5 caused by mouse lung damage lung tissue in inflammatory cytokine concentration.
Nourishing lung and activating blood capsule is used for the purposes for preparing the medicine of pulmonary lesion caused by treatment and/or prevention PM2.5, described
Nourishing lung and activating blood capsule repair PM2.5 caused by mouse pulmonary lesion.
Nourishing lung and activating blood capsule is used for the purposes for preparing the medicine of pulmonary lesion caused by treatment and/or prevention PM2.5, described
The preparation method of nourishing lung and activating blood capsule be：(1) Radix Astragali is cut into slices, the radix paeoniae rubrathe and psoralea corylifolia are ground into coarse powder；(2) 60%-70%
Alcohol reflux is extracted 2 times, and 5 times of amount 60%-70% ethanol are added for the first time, extracts 2h, and second of 4 times of amount 60%-70% ethanol is carried
Take 1h；(3) merge extract solution twice, filtration, filtrate decompression is recycled to no alcohol taste, continue to be concentrated under reduced pressure into relative density be
1.30-1.40 clear cream；(4) Jia 1.0~starch of 2.0 times of parts by weight, mix, 90% alcohol granulation, dry, addition is no more than
5% talcum powder and 0.1%-2% magnesium stearate, are mixed, and are loaded capsule, are produced.
Nourishing lung and activating blood capsule is used for the purposes for preparing the medicine of pulmonary lesion caused by treatment and/or prevention PM2.5, described
The preparation method of nourishing lung and activating blood capsule be：(1) Radix Astragali, the radix paeoniae rubrathe and psoralea corylifolia are crushed, are placed in extraction kettle, with 70%~
90% ethanol is entrainer；(2) supercritical CO is adjusted2Fluid temperature (F.T.), pressure；(3) extraction temperature is 30 DEG C~50 DEG C, extraction pressure
Power is 20MPa~40MPa, and extraction time is 1h~3h；(4) extract is collected, centrifugal concentrating is evaporated extract；(5) Jia 1.0~
The starch of 2.0 times of parts by weight, is mixed, 90% alcohol granulation, is dried, and adds the talcum powder no more than 5% and 0.1%~2%
Magnesium stearate, is mixed, and is loaded capsule, is produced.
It is preferred that, the preparation method of nourishing lung and activating blood capsule is：(1) Radix Astragali, the radix paeoniae rubrathe and psoralea corylifolia are crushed, is placed in extraction kettle
In, using 80% ethanol as entrainer；(2) supercritical CO is adjusted2Fluid temperature (F.T.), pressure；(3) 30 DEG C of extraction temperature, extracting pressure
20MPa, extraction time 2h；(4) extract is collected, centrifugal concentrating is evaporated extract；(5) Jia 1.0~shallow lakes of 2.0 times of parts by weight
Powder, is mixed, 90% alcohol granulation, is dried, and adds the talcum powder and 0.1%~1% magnesium stearate no more than 5%, is mixed, dress
Enter capsule, produce.
The present invention has advantages below：
(1) nourishing lung and activating blood capsule of the present invention is made from high-quality Chinese medicine, with low cost and compatibility is suitable, meets traditional Chinese medicine
And modern medicine study theory, and related formulation is made using modern medicine pharmaceutical technology, safe toxic side effect is low.
(2) in PM2.5 mouse models, nourishing lung and activating blood capsule of the present invention can be significantly improved compared with model control group
The PFT of PM2.5 mouse, pathological section and injury of lungs repair index result and show that Chinese medicine composition of the present invention can make model small
Mouse injury of lungs is obviously improved, and can significantly reduce inflammatory cytokine amount.
(3) nourishing lung and activating blood capsule of the present invention is extracted using alcohol extracting method, and preparation technology is simple to operation, active material Radix Astragali first
The content height of glycosides, Paeoniflorin and psoralen, Isopsoralen, the content of especially psoralen and isopsorapen is substantially high
In reference composition.
(4) nourishing lung and activating blood capsule of the present invention is extracted using super critical extraction, active material Astragaloside IV, Paeoniflorin and benefit
The content height of bone fat element, Isopsoralen.
(5) nourishing lung and activating blood capsule of the present invention is moist almost without drawing, and stability is good, shelf-stable.
(6) nourishing lung and activating blood capsule of the present invention prepares simple, it is easy to clinical practice.
(7) nourishing lung and activating blood capsule of the present invention is to active material Astragaloside IV, Paeoniflorin and psoralen, different benefit
Brief description of the drawings
Fig. 1 cuts into slices for PM2.5 model mices lung pathology.
Fig. 2 is PM2.5 model mice injury of lungs pathological sections.
Embodiment
The embodiment of form, further specifically to the above work of the present invention by the following examples
It is bright.But the scope that this should not be interpreted as to above-mentioned theme of the invention is only limitted to following examples.
The composition of raw materials of the nourishing lung and activating blood capsule of the present invention of embodiment 1
Composition 1：Radix Astragali 500g, radix paeoniae rubrathe 1000g, psoralea corylifolia 360g.
Composition 2：Radix Astragali 1000g, radix paeoniae rubrathe 1000g, psoralea corylifolia 500g.
Composition 3：Radix Astragali 500g, radix paeoniae rubrathe 500g, psoralea corylifolia 250g.
Composition 4：Radix Astragali 720g, radix paeoniae rubrathe 720g, psoralea corylifolia 360g.
The preparation of the nourishing lung and activating blood capsule extract of the present invention of embodiment 2 and the content of active material
The preparation of nourishing lung and activating blood capsule of the present invention：
Radix Astragali section, the radix paeoniae rubrathe and the psoralea corylifolia of the composition of raw materials of the composition 4 of Example 1 are ground into coarse powder, 60%-70%
Ethanol alcohol reflux is extracted 2 times, and Jia 5 for the first time measures 60%-70% ethanol, extracts 2h, and second of 4 times of amount 60%-70% ethanol is carried
1h is taken, merges extract solution twice, filtration, filtrate decompression is recycled to no alcohol taste, continues to be concentrated under reduced pressure into relative density for 1.30-
1.40 clear cream, is produced.
Contain following active material in the nourishing lung and activating blood capsule extract：Astragaloside IV, Paeoniflorin, psoralen and different
Psoralen, the results are shown in Table 1, assay method according to《Chinese Pharmacopoeia》The method that (2015 editions) first.
The nourishing lung and activating blood capsule Active substance content of the present invention of table 1
Comparative composition preparation method：
Radix Astragali 720g, radix paeoniae rubrathe 720g, psoralea corylifolia 360g, take radix paeoniae rubrathe 180g to be ground into fine powder, standby；Remaining flavour of a drug adds water pan-fried
Boil secondary, 8 times of amount water are added for the first time, 6 times of amount water are added for the second time, 1 hour every time, collecting decoction was filtered, and filtrate is concentrated into relatively
Density is 1.05~1.15 (80 DEG C), plus ethanol makes alcohol content up to 60%, is sufficiently stirred for, and stands 24 hours, and leaching supernatant is returned
Ethanol is received to without alcohol taste, continuing to be concentrated into relative density for 1.35~1.40 (80 DEG C), above-mentioned radix paeoniae rubrathe fine powder is added, mixes, do
It is dry, crush.
The nourishing lung and activating blood capsule of the present invention of table 2 is compared with reference composition activity substance content
The preparation of the nourishing lung and activating blood capsule extract of the present invention of embodiment 3 and the content of active material
This experiment is to supercritical CO2The technological parameter of extraction equipment carries out comprehensive study, finds various extraction conditions (extraction
Take temperature, pressure, entrainer and time) influence to extraction quantity and active principle content, through investigating entrainer concentration of alcohol shadow
Smaller (concentration of alcohol is 70%~90%, and 80% is more excellent) is rung, extracting pressure, extraction temperature, extraction time and entrainer are main
The influence factor wanted, therefore four factors are investigated to extracting by index orthogonal experiment of the yield of Astragaloside IV and extract
The influence of effect.Investigation factor and level are shown in Table 3.
The nourishing lung and activating blood capsule extraction process of the present invention of table 3 investigates factor level table
Test method weighs Radix Astragali 720g, radix paeoniae rubrathe 720g, psoralea corylifolia 360g, is placed in after crushing in extraction kettle, adjusts CO2Stream
Temperature, regulation extraction temperature and pressure, are extracted, parallel three parts by the requirement of orthogonal test table 4, merge extract solution.
The nourishing lung and activating blood capsule process for super-critical extracting of the present invention of table 4 investigates orthogonal arrage
The analysis of variance table of table 5
F0.05(2,2)=19.00F0.01(2,2)=99.00
From the variance analysis of table 5：Optimal selection technique should be A3B2C2.That is 30 DEG C of extraction temperature, extracting pressure 40MPa,
Extraction time 1h.
According to above-mentioned selection process, the activity substance content of obtained nourishing lung and activating blood capsule is as shown in table 6 below.
The nourishing lung and activating blood capsule Active substance content of the present invention of table 6
The preparation of the nourishing lung and activating blood capsule of the present invention of embodiment 4
The clear cream according to made from the preparation method of embodiment 2 or 3, plus 1.0~starch of 2.0 times of parts by weight, mixes, 90%
Alcohol granulation, is dried, and is added the magnesium stearate of the talcum powder and 0.1%-2% no more than 5%, is mixed, loads capsule, be made
1000, produce.
Nourishing lung and activating blood capsule of the present invention is hygroscopic to be determined
Experimental method：Dry tool plug glass measuring cup is taken, experiment the previous day is placed in ± 1 DEG C of freeze-day with constant temperature of suitable 25 DEG C
Device (design temperature is 25 DEG C ± 1 DEG C, relative humidity is 80% ± 2%) in, it is accurate into determining weight.Take test sample appropriate, tiling
In above-mentioned measuring cup, test sample thickness is about 1mm, accurate into determining weight.Measuring cup is open, and be placed in together with bottle cap
State under the conditions of constant temperature and humidity 24 hours, cover measuring cup lid, it is accurate into determining weight.
Experimental result：Nourishing lung and activating blood capsule percentage weight increase of the present invention is no more than 2%, shows nourishing lung and activating blood capsule of the present invention
It is moist almost without drawing, it is shown in Table 7.
Table 7 nourishing lung and activating blood capsule of the present invention draws moist
Influence of the nourishing lung and activating blood capsule of the present invention of embodiment 5 to PM2.5 mouse models
Test objective：
By PM2.5 mouse model experiments, observation nourishing lung and activating blood capsule is to animal pattern lung functions, lung injury
Effect.
Animal：
Test animal is SPF grades of health ICR female mices 60, and body weight 22-26g is (by the magnificent Fukang biotechnology in Beijing
Limited company provides).Raised in 22 DEG C -28 DEG C of temperature, relative humidity 50%-60% cleaning ambient, using 12/12
Illumination.The using and nurse of animal meets the human care of animal welfare association of Beijing China-Japan Friendship Hospital.
Medicine：
By the composition 4 of the embodiment of the present invention 1, according to its consumption of being grown up, 0.41g crude drugs/ml, gastric infusion is made into.
Establishment of mouse model and pharmaceutical intervention：
Healthy ICR female mices 60,3 groups are randomly divided into by body weight：In Normal group, PM2.5 model groups, the present invention
Drug composition group, every group 20.Blank control group：The 1st, 8,15 and 22 day via intranasal application, which is set up, respectively at model instills 0.9% life
Salt solution (20 μ L/ are only) is managed, remaining time normally raises.PM2.5 model groups：Using nasal cavity instillation PM2.5 suspension methods, respectively at
Model sets up the 1st, 8,15 and 22 day via intranasal application instillation PM2.5 suspension (40mg/kgbw, 20 μ L/ are only), the 1st day to the 22nd day,
0.2ml//the d of physiological saline of administered by oral gavage 0.9%.Chinese medicine composition group of the present invention：The same mould of PM2.5 Establishment of mouse model methods
Type control group, the 1st day to the 22nd day, administered by oral gavage nourishing lung and activating blood Chinese Medicines (0.82g/Kg, 0.2ml//d).Each experiment
Group is put to death after last time is contaminated in 48 hours.
Result of the test and analysis：
1st, mouse model analysis of Pulmonary Function (high dose contamination control group)
2% amobarbital intraperitoneal injection of anesthesia mouse (0.4mL/40g), takes dorsal position to be fixed on operating desk, cuts off neck
Fur, exposure tracheae, and blunt separation tracheae, a kerf is cut at the nearly head of tracheae, gas will be inserted at the gas-tpe fitting of intubation
Pipe, and fixed with cotton thread, transfer mouse to body retouches instrument platform, connection lung ventilator and gas-tpe fitting, records the PFT phase of mouse
Close the change of index.And with the acute analysis (lung function instrument of AniRes2005 animal lung function analysis systems：Beijing Bei Lan Boke skill
Co., Ltd).It the results are shown in Table 8.
Influence of the nourishing lung and activating blood capsule of table 8 to PM2.5 model mice PFTs
* represents the P compared with model group<0.01, * represents the P compared with model group<0.05
As seen from the results in Table 8, nourishing lung and activating blood Capsules group significantly drops to the airway resistance (RL, RE) of mouse compared with model group
Low (p<0.01), lung compliance (Cdyn) significantly improves (p<0.01), pulmonary ventilation function (PEF) is significantly improved.Show, the present invention
Chinese medicine composition can significantly improve the PFT of PM2.5 mouse.
2nd, pathological analysis (three kinds of poisoning dosages)
De- neck puts to death mouse, opens chest exposure thoracic cavity：Upper lobe of left lung of the separation at hilus pulumonis is fixed on 10% formalin
24h, conventional dehydration FFPE, slice row HE dyeing, light Microscopic observation；The right side lobe of the lung is taken in case homogenate.As a result accompanying drawing 1 is seen.
Interpretation of result：
Blank control group mouse lung tissue has no obvious inflammatory cell infiltration；Bronchium mucous membrane and bronchial wall structure are complete
It is whole；Alveolar is evenly distributed, structural integrity, steeps a small amount of inflammatory cell infiltration of intracavitary；Interstitial lung inflammatory conditions are lighter；Pulmonary capillaries
Structural integrity, has no bleeding.
The main pathological manifestations of PM2.5 model mice lungs change for lung tissue inflammatory, the inflammation such as neutrophil leucocyte, lymphocyte
Property cellular infiltration showed increased.Bronchium mucous membrane partial injury, is mainly shown as bronchium wall thickening, with going out on a small quantity
Blood, bleeding；Alveolar space increases, and the fracture of part alveolar, fusion, intracavitary have secretion, mainly has monocyte, neutral grain thin
Born of the same parents infiltrate, it is seen that consolidation phenomenon occurs in lymphocytosis, part alveolar, and alveolar septum is broadening；Interstitial lung has a large amount of inflammatories thin
Born of the same parents infiltrate；There is bleeding in pulmonary capillaries part, is oozed out with inflammatory, tube wall is thickened in various degree.
Nourishing lung and activating blood Capsules group can be such that the inflammatory change of model mice lung tissue substantially subtracts compared with PM2.5 model mices
Gently；Bronchium mucosa injury lesser extent；Alveolar structure is relatively complete, and luminal sectetion is reduced, and inflammatory cell infiltration is obvious
Mitigate；Interstitial lung inflammatory cell infiltration makes moderate progress；Pulmonary capillaries structure is more complete, it is seen that a small amount of bleeding.
3rd, ELISA detects (three kinds of poisoning dosages)
Take 0.1g lung tissues to be put into 1.5mlEP pipes, be prepared into 10% lung homogenate liquid, centrifuge 4 DEG C, 12000r/
Min, 20min, take supernatant.The operating procedure explanation pointed out by ELISA kit carries out related scorching in lung homogenate supernatant
Property medium content.It the results are shown in Table 9-14.
Influence of the nourishing lung and activating blood Capsules group of table 9 to IL-1 in various concentrations PM2.5 model mice lung tissues
| Low dosage contamination group | Middle dosage contamination group | High dose contamination group | |
| Blank group | 3616.25±1176.73** | 3616.25±1176.73** | 3616.25±1176.73** |
| Contamination group | 5253.98±1350.61 | 5327.92±1274.07 | 6005.13±1815.30 |
| Chinese medicine composition group | 4457.90±1425.27** | 4187.91±998.28** | 4782.04±1187.23** |
Quantitative analysis is usedRepresent, * * represent the P compared with model group<0.01, * represents the P compared with model group<
0.05。
Influence of the nourishing lung and activating blood Capsules group of table 10 to IL-4 in various concentrations PM2.5 model mice lung tissues
| Low dosage contamination group | Middle dosage contamination group | High dose contamination group | |
| Blank group | 252.51±37.50** | 252.51±37.50** | 252.51±37.50** |
| Contamination group | 358.45±32.57 | 371.97±37.063 | 395.20±68.29 |
| Chinese medicine composition group | 256.42±65.80** | 233.86±50.61** | 299.63±53.85** |
Quantitative analysis is usedRepresent, * * represent the P compared with model group<0.01, * represents the P compared with model group<
0.05。
Influence of the nourishing lung and activating blood Capsules group of table 11 to IL-6 in various concentrations PM2.5 model mice lung tissues
| Low dosage contamination group | Middle dosage contamination group | High dose contamination group | |
| Blank group | 3890.26±1015.36** | 3890.26±1015.36** | 3890.26±1015.36** |
| Contamination group | 5008.03±1518.92 | 5560.60±1738.32 | 5985.88±2086.78 |
| Chinese medicine composition group | 4466.32±1440.85** | 4179.10±1136.73** | 5226.43±1973.88** |
Quantitative analysis is usedRepresent, * * represent the P compared with model group<0.01, * represents the P compared with model group<
0.05。
Influence of the nourishing lung and activating blood Capsules group of table 12 to IL-10 in various concentrations PM2.5 model mice lung tissues
| Low dosage contamination group | Middle dosage contamination group | High dose contamination group | |
| Blank group | 2930.63±632.28** | 2930.63±632.28** | 2930.63±632.28** |
| Contamination group | 4194.15±487.22 | 4741.14±1082.31 | 6219±724.16 |
| Chinese medicine composition group | 3120.58±691.67** | 3248.64±786.94** | 3647.24±1336.03** |
Quantitative analysis is usedRepresent, * * represent the P compared with model group<0.01, * represents the P compared with model group<
0.05。
Influence of the nourishing lung and activating blood Capsules group of table 13 to IL-17 in various concentrations PM2.5 model mice lung tissues
| Low dosage contamination group | Middle dosage contamination group | High dose contamination group | |
| Blank group | 591.90±139.92** | 591.90±139.92** | 591.90±139.92** |
| Contamination group | 761.42±126.19 | 883.38±116.30 | 1008.89±187.59 |
| Chinese medicine composition group | 583.11±136.36** | 643.62±101.51** | 616.28±169.70** |
Quantitative analysis is usedRepresent, * * represent the P compared with model group<0.01, * represents the P compared with model group<
0.05。
Influence of the nourishing lung and activating blood Capsules group of table 14 to TNF-α in various concentrations PM2.5 model mice lung tissues
| Low dosage contamination group | Middle dosage contamination group | High dose contamination group | |
| Blank group | 10365.69±1530.90** | 10365.69±1530.90** | 10365.69±1530.90** |
| Contamination group | 15017.8±1284.77 | 18838.35±716.01 | 20278.18±1896.80 |
| Chinese medicine composition group | 12732.06±1184.12** | 14130.99±3103.43** | 13216.5±1332.48** |
Quantitative analysis is usedRepresent, * * represent the P compared with model group<0.01, * represents the P compared with model group<
0.05。
From table 9-14 results, compared with model group, nourishing lung and activating blood Capsules group is to various concentrations PM2.5 model mice lungs
Significantly, each inflammatory cytokine content significantly drops inflammatory cytokine (IL-1,4,6,10,17, TNF-α) inhibitory action in tissue
Low (P<0.01).
4th, Masson three-color process step：(high dose contamination group)
(1) paraffin section de-waxing is to water；(2) running water and distillation are washed successively；(3) with Regaud haematine dye liquors 5-
10min；(4) fully washing, such as crossing dye can hydrochloride alcohol differentiation；(5) distillation washing；(6) Masson Ponceaux acid fuchsins are used
Liquid 5-10min；(7) 2% glacial acetic acid aqueous solutions embathe a moment；(8) the 1% phosphomolybdic acid aqueous solution break up 3-5min；(9) without water
Wash, directly with aniline blue liquid dye 5min；(10) a moment is embathed with 0.2% glacial acetic acid aqueous solution；(11) 95% alcohol, without watery wine
Essence, dimethylbenzene are transparent, neutral gum sealing.Every section choose 6 unduplicated visuals field (× 200 times), MASSON dyeing with
Blue collagen deposition is positive signal, is analyzed with Image Proplus multi-media color Pathologic image analysis softwares.Calculate
The ratio of mouse lung tissue collagen deposition area and the lung tissue gross area in the visual field, and average.As a result accompanying drawing 2 is seen.By attached
Fig. 2 understands that visible HMBG1 is mainly seen in visible KGF in alveolar space, B and mainly appears on tracheal wall and alveolar tissue gap in A
In.Influence of the visible nourishing lung and activating blood capsule to collagen deposition and tissue fibrosis in PM2.5 model mice lung tissues in C, wherein
Collagenous fibres, mucus, cartilage take on a red color in blueness, endochylema, muscle, cellulose, neuroglia, karyon black and blue color.
5. SABC (high dose contamination group)
Lung tissue section routinely after dewaxing, is detected according to immunohistochemical kit concrete operation step.Using the U.S.
The Image-Pro Plus image analysis softwares of Media Cybernetics companies production are analyzed, with accumulation optical density
(Integrated Optical Density, IOD) reflects protein expression total amount.
The nourishing lung and activating blood Capsules group of table 15 repairs Index Influence to PM2.5 model mices injury of lungs
| MASSON | HMGB1 | KGF | |
| Blank group | 7.43±1.42** | 9804.32±5175.90** | 18232.15±8342.99** |
| Contamination group | 22.44±5.43 | 21655.11±5053.67 | 25278.79±10469.46 |
| Chinese medicine composition group | 10.0±1.98** | 10323.82±5580.02** | 17697.89±6162.62** |
Related quantitative analysis is usedRepresent, * * represent the P compared with model group<0.01, * represents the P compared with model group<
0.05。
As shown in Table 15, compared with model group, nourishing lung and activating blood capsule forms a team to repair index to PM2.5 model mices injury of lungs
Significantly improve (P<0.01).
6.PCR is detected：(high dose contamination group)
Take mouse lung tissue 100mg, Trizol one-step method extracted total RNA, through reverse transcription synthesize cDNA (wherein 2 μ g RNA,
1 μ l oligo (dT), DEPC water to 12 μ l).Mouse lung tissue β-actin primer sequences：Forward primer 5 '-
GTGACGTTGACATCCGTAAAGA-3 ' ,-GTAACAGTCCGCCTAGAAGCAC-3 ' of reverse primer 5 '；IL-8 primer sequences：
- the CATCTTCGTCCGTCCCTGTG-3 ' of forward primer 5 ' ,-GCCAACAGTAGCCTTCACCCA-3 ' of reverse primer 5 '；SIgA draws
Thing sequence：- the GCTACAGTGTGTCCAGCGTCCT-3 ' of forward primer 5 ', reverse primer 5 '-
TGCCAGACTCAGGATGGGTAAC-3’.Method is researched and analysed using relative quantification, with 2-△△CtMake index to be analyzed.
Expression of the IL-8 of the table 16 and sIgA mRNA in different group mouse lung tissue homogenates
| IL-8 | sIgA | |
| Blank group | 0.58±0.43** | 0.039±0.029** |
| Contamination group | 1.80±0.70 | 0.347±0.101 |
| Chinese medicine composition group | 1.14±0.36** | 0.157±0.104** |
The expression quantity of IL-8 and sIgA in mouse is usedRepresent, * * represent the P compared with model group<0.01, n >=8
Expression of the IL-8 and sIgA mRNA in different group mouse lung tissue homogenates.Blank control group and model group
Expression compared to IL-8 and sIgA is substantially reduced, and nourishing lung and activating blood capsule thing group intervention group inflammatory mediator content is significantly lower than model
Group.
Claims (2)
1. nourishing lung and activating blood capsule is used for the purposes for preparing the medicine of pulmonary lesion caused by treatment and/or prevention PM2.5, its feature
It is, described nourishing lung and activating blood capsule is by 72 parts of the Radix Astragali（Weight）, 72 parts of the radix paeoniae rubrathe（Weight）With 36 parts of psoralea corylifolia（Weight）Composition,
The preparation method of described nourishing lung and activating blood capsule is：
（1）The Radix Astragali, the radix paeoniae rubrathe and psoralea corylifolia are crushed, are placed in extraction kettle, using 70% ~ 90% ethanol as entrainer；
（2）Adjust supercritical CO2Fluid temperature (F.T.), pressure；
（3）Extraction temperature is 30 DEG C ~ 50 DEG C, and extracting pressure is 20MPa ~ 40MPa, and extraction time is 1h ~ 3h；
（4）Extract is collected, centrifugal concentrating is evaporated extract；
（5）Plus 1.0 ~ starch of 2.0 times of parts by weight, mix, 90% alcohol granulation, dry, add talcum powder no more than 5% and
0.1% ~ 2% magnesium stearate, is mixed, and is loaded capsule, is produced；
Described nourishing lung and activating blood capsule contains the following active material of percentage by weight：>=0.20% Astragaloside IV, >=2.3%
Paeoniflorin and >=0.85% psoralen and isopsorapen.
2. nourishing lung and activating blood capsule as claimed in claim 1 is used to prepare pulmonary lesion caused by treatment and/or prevention PM2.5
The purposes of medicine, it is characterised in that the air flue of the lung of mouse lung damage caused by described nourishing lung and activating blood capsule reduction PM2.5
Resistance, improves lung compliance, improves the inflammation in pulmonary ventilation function, the lung tissue of mouse lung damage caused by reduction PM2.5
Cytokine concentrations, repair the pulmonary lesion of mouse caused by PM2.5.
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Denomination of invention: The use of Bufei Huoxue Capsules for the treatment or prevention of lung injury caused by PM2.5 Effective date of registration: 20230329 Granted publication date: 20170929 Pledgee: Industrial and Commercial Bank of China Limited Yunfu Yun'an Branch Pledgor: GUANGDONG LEIYUNSHANG PHARMACEUTICAL Co.,Ltd. Registration number: Y2023980036720 |