CN105148031A - Product for preventing or repairing PM2.5 caused lung injury and preparation method of product - Google Patents
Product for preventing or repairing PM2.5 caused lung injury and preparation method of product Download PDFInfo
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Abstract
The invention discloses a product for preventing or repairing PM2.5 caused lung injury and a preparation method of the product, and belongs to the field of traditional Chinese medicine; the product is characterized by consisting of raw materials of dendrobe and/or astragalus membranaceus, wherein preferably, the product further consists of scutellaria baicalensis; and preferably, the product is prepared into instant granules. Upon model mice experiments, the product can be used for significantly improving PM2.5 caused mice lung tissue and airway injury; the product is the first internal product at home and abroad for preventing or repairing PM2.5 caused lung/respiratory system injury at present; and the product, which is medicinal and edible, is suitable for long-term drinking and is applicable to both prevention and treatment.
Description
Technical field
The present invention relates to tcm field, particularly a kind of prevention or reparation PM2.5 cause the goods of pulmonary lesion.
Background technology
Atmospheric particulates (PM) are the common names of aerial atmospheric pollution particulate matter of suspending, and composition is complicated, form of diverse, are the significant pollutant of the quantitative health hazard evaluating atmospheric pollution.Particulate matter is fine particle especially, has been acknowledged as very harmful, representative extremely strong atmosphere pollution, and according to the large young pathbreaker of particulate matter, it is divided into coarseparticulate, fine particle, superfine particulate matter.PM2.5 (ParticulateMatter2.5) is the particulate matter of aerodynamic diameter≤2.5 μm.Chemical composition mainly comprises two large classes, and one is primary particulate, mainly contains organic carbon, carbon black, dust; Two is secondary aerosol species, mainly contains ammonium sulfate (ammonium sulfite), ammonium nitrate etc.The basic feature of PM2.5 is that volume is little, lightweight, and the holdup time is long in an atmosphere, and can propagate, and causes large-scale air pollution.The size of microgranule determines its position in respiratory tract deposition, the particulate matter of PM10 and PM2.5 to be aerodynamic diameter be respectively 10 μm and 2.5 μm, they can enter respiratory tract by breathing, and a part is stopped by nasal cavity fine hair, and the particulate matter that diameter is less than 2.5 μm enters human lung.That is PM10 major sedimentary is at upper respiratory tract, PM2.5 is then deposited on alveolar by breathing, Churg etc. analyze the lungs dissec of 10 routine Vancouver residents, and in pulmonary parenchyma, the particle 96% of deposition is PM2.5, confirms that PM2.5 is deposited on the viewpoint of alveolar.PM2.5 enters blood by interior ventilation and acts on other organs of human body, and its surface adsorption has a lot of poisonous and harmful substance, comprises heavy metal as Pb, Cd, Cu, Ni, multiring aromatic hydrocarbon, formaldehyde etc.
The epidemiology of nearly 20 years and toxicological study confirm, Atmospheric particulates can cause prevalence and the sickness rate, particularly PM2.5 of multisystem disease can cause serious respiratory system damage, cause M & M obviously to raise.Zanobetti studies discovery, and the share a common fate rising of systemic disease of the rising of PM2.5 daily mean of concentration becomes obvious positive correlation, every 10ug/m
3by the risk factor of rising 2.07%.Dominici research shows, the mean concentration change of PM2.5 obviously will affect rising respiratory system disease admission rate every day, such as often raise 10ug/m
3the rising of admission rate 8.0% will be caused.Epidemiology statistics shows: with increasing of Atmospheric Particulate Matter concentration, the prevalence of each systemic disease such as respiratory system, blood circulation will be caused, M & M all significantly rises, there is certain positive correlation in the two, this association especially significant reaction is having in the Susceptible population such as heart and lung diseases medical history person, old man, child.Toxic component in PM2.5 and the harmful substance carried, by stimulating bronchial epithelial cell, thus produce ROS, pulmonary alveolar macrophage starts defense mechanism Phagocytic granules thing, discharge a large amount of reactive oxygen free radical or active oxygen simultaneously, cause oxidative system and antioxidant system unbalance, cause tissue injury.The physicochemical property prompting of PM2.5 has potential toxicity to pulmonary's immune system, not only affect the specific immune response of body, and affect nonspecific immune reaction, cause immune system disequilibrium, immunologic cellular activity reduces, add the susceptibility of body to antibacterial, virus etc., the prevalence of pulmonary disease is increased.Atmospheric particulates attack lung tissue as foreign body, and body immune system excessively discharges inflammatory mediator and causes inflammatory factor excessively to assemble, thus upset the stable of lung tissue local microenvironment, reduce repair ability and defense function, and then cause tissue injury.
Current doctor trained in Western medicine not yet provides prophylactico-therapeutic measures prevention PM2.5 to the infringement of human respiratory, and primary protection method is by reducing PM2.5 discharge, wear antidusting and anti-poisoning mask, carry out Personal hygiene, regulate emotion, increase and greenly to plant, strengthen the aspects such as diet care and prevent.
Upper respiratory tract infection is clinical common, multiple diseases, clinical treatment is based on antibiotic, the drawback that antibiotic therapy upper respiratory tract infection causes is dysbacteriosis and superinfection, also causes the drug resistance of human pathogen to improve gradually, and the raw plain insensitive fastbacteria that creates antagonism.
Theory of Chinese medical science is emphasized " preventive treatment of disease ", the thought just having strong physique to prevent trouble before it happens in the idea that China is ancient.Ancients are particular about living with peacefully of man and nature, adhere to all the time " harmony between man and nature ", follow the dynamic equilibrium between man and nature, social three, keep the coordination of self negative and positive.Because mostly each disease is wind, cold, summer-heat, wet etc. multifactorly causes a disease, need determination for the treatment of based on pathogenesis obtained through differentiation of symptoms and signs to use multiple therapy methods to carry out Comprehensive Treatment, reach the functional status of equilibrium between yin and yang.The thought of " preventive treatment of disease " is taked in inventor's guess, takes some pre-anti-air pollutions, improves the medicine of immunity, may improve pulmonary function, and defence Atmospheric particulates PM2.5, to the damage of lung tissue and air flue, benefits healthy.
Summary of the invention
Inventor is not based on when having effective scheme to solve air environmental pollution at present, in line with the principle of the traditional Chinese medical science " organic conception ", through research and development screening, find a kind of can the square medicine of integration of edible and medicinal herbs of long-term taking, it has antibacterial, to control the inflammatory reaction of injury of lung effect, strengthening vital QI to eliminate pathogenic factors, treating both the principal and secondary aspects of a disease, PM2.5 is to the function of the infringement of respiratory system in enhancing body defence.
The scheme of request protection of the present invention is as follows:
Prevention or reparation PM2.5 cause goods for pulmonary lesion, and its raw material comprises Herba Dendrobii and/or the Radix Astragali.
Described raw material also comprises Radix Scutellariae.
The dosage form of described goods is granule.
Acceptable adjuvant is also comprised in described goods.
Prevention or reparation PM2.5 cause goods for pulmonary lesion, and its raw material is made up of Herba Dendrobii, the Radix Astragali and Radix Scutellariae.
Described Herba Dendrobii is selected from Herba Dendrobii, Dendrobium nobile, purple dendrobium, Herba Dendrobii.
Prevention or reparation PM2.5 cause a food for pulmonary lesion, it is characterized in that, include above-mentioned arbitrary goods.
Preparation prevention or reparation PM2.5 cause the method for the goods of pulmonary lesion, and it is characterized in that, the raw material of employing comprises Herba Dendrobii and/or the Radix Astragali.
Described raw material also comprises Radix Scutellariae.
The present invention is in line with the principle of the traditional Chinese medical science " organic conception ", " harmony between man and nature ", meticulous selection, assembly and checking, determine to adopt Herba Dendrobii and/or the Radix Astragali and Radix Scutellariae to form prescription, three kinds of raw materials are can the safe medical material of integration of edible and medicinal herbs, can long-term taking, three medicine combination strengthening vital QI to eliminate pathogenic factors, treating both the principal and secondary aspects of a disease, PM2.5 is to the function of the infringement of respiratory system in enhancing body defence.Experimental data shows, this prescription can repair the injury of lung (see Fig. 5) of PM2.5 model mice, and obviously improve the ventilatory function of model mice, the pulmonary function of mice improves.
The research of the present invention to selected raw material is as follows:
The Mucosal Immunity of Herba Dendrobii regulates active
Herba Dendrobii (Classification system: DendrobiumnobileLindl), has another name called lofty or bottomless palpus, bracketplant, Lin Lan, Dendrobium nobile Lindl China etc.Stem is upright, and meat shape is plump, and slightly flat is cylindrical, long 10 ~ 60 centimetres, slightly reaches 1.3 centimetres.Medicinal part: stem; Property slightly sweet flavor micro-salty, cold, return stomach, kidney, lung meridian.Reinforcing stomach reg fluid, nourishing YIN and clearing away heat.Medicinal dendrobium refers generally to: Dendrobium nobile, purple dendrobium, Herba Dendrobii, Herba Dendrobii.Preferably these are several in the present invention, but are not limited thereto.
The polysaccharide composition of Herba Dendrobii has the effect of enhancing human body immunity power, mouse experiment proves, Dendrobium officinale polysaccharide can offset the violent decline adding caused peripheral leukocytes of Immunosuppressant Cyclophosphamide under experiment condition forcefully, eliminates its destructive side effect.Also can produce migration inhibitory factor by Promote immunity systemic lymphoid cell simultaneously, effectively offset the side effect adding caused lifting mobile inhibition index of cyclophosphamide.Herba Dendrobii also can promote the phagocytic function of tumor animal macrophage.Strengthen the activity of the lymphocytic propagation of T and differentiation and NK cell.And the serum hemolysin value of tumor animal can be significantly improved, no matter prompting Herba Dendrobii is to non-specific immunity, or specific cellular immunity and humoral immune function, and all improve effect.
Astragalus polysaccharides affects intestinal immunity
The Radix Astragali (Classification system: Astragalusmembranaceus (Fisch.) Bunge.), the history of medicinal existing more than 2000 year so far, it has enhancing human body immunity function, protects the liver, diuresis, defying age, anti-stress, blood pressure lowering and antibacterial action more widely.Nature and flavor: sweet, tepor.Return lung, spleen, liver, kidney channel.Astragalus polysaccharides (astragaluspolysaccharides, APS) is the primary bioactive components of the Radix Astragali, has raising immunity of organisms, antitumor, antioxidation, alleviates the multiple pharmacological effect such as diabetic complication, antiviral.
APS is by improving the secretory volume of Intestinal Mucosa Microvascular Endothelial Cells carbon monoxide (NO); maintain the constant and protection body non-specific immunity of microcirculation environment in body, in the adjustment etc. of nerve immunity function, there is important biological function.Result of study shows, APS also can improve vascular permeability by changing capillary endothelium NO secretion, and maintenance microcirculation environment is constant and protect body non-specific immunity, plays immunological enhancement.
The application of Radix Scutellariae in respiratory system disease
Radix Scutellariae (Classification system: ScutellariabaicalensisGeorgi), another name Radix Scutellariae, tsuchikane tea root, be Labiatae Scutellaria herbaceos perennial, the root of Radix Scutellariae is used as medicine, bitter in the mouth, cold in nature, have heat clearing and damp drying, eliminating fire and detoxication, etc. effect.Cure mainly epidemic febrile disease, upper respiratory tract infection, cough due to lung-heat, damp and hot jaundice, pneumonia, dysentery, hemoptysis, etc. disease.Enter the heart, lung, gallbladder, large intestine channel.
Radix Scutellariae meridian distribution of property and flavor: bitter, cold.Return lung, gallbladder, stomach, large intestine channel.Removing the relative excess fire, except damp and hot, heat-phlegm on diaphragm, falls in purte the white, is usually used in controlling chronic tracheitis, upper respiratory tract infection, and the lung moistening that especially Folium Scutellariae is special is reduced internal heat function, has very effective prophylactic treatment effect to cough due to lung-heat, chronic pharyngitis etc.Modern pharmacological research display Radix Scutellariae has antipyretic, bacteriostasis, also has the effect such as relaxing smooth muscle, anti-hypoxia.The multiple alkaloids such as the berberine in Radix Scutellariae, berberine, 13-methyl-.psi.-coptisine., have inhibitory action to dysentery bacterium, staphylococcus aureus, streptococcus, Diplococcus pneumoniae.Bacteriostatic experiment proves inhibited to diphtheria village bacterium, bacillus pyocyaneus, Hemolytic streptococcus, escherichia coli etc. of Radix Scutellariae.
The present invention adopts the method for via intranasal application instillation PM2.5 suspension successfully to establish the model of mouse lung tissue and Airway damage.
The present invention shows the result of study of model mice, and the pulmonary ventilation function that PM2.5 causes model mice is limited, and after Chinese medicine of the present invention is intervened, obviously improve the ventilatory function of model mice, the pulmonary function of mice improves (see table 3).
The result of study of the morphologic change of mouse lung tissue is shown:
The main pathology that PM2.5 causes model of lung injury mice shows as inflammatory change, the inflammatory cell infiltration such as neutrophilic granulocyte, lymphocyte showed increased; Bronchia mucosa partial injury, with on a small quantity hemorrhage, ooze out phenomenon; Alveolar septum is broadening, and alveolar space increases, the fracture of part alveolar, fusion, and intracavity has secretions, mainly contains mononuclear cell, neutrophil infiltration, and consolidation phenomenon appears in part alveolar; Interstitial lung has massive inflammatory cells infiltrated.
Blank group mouse lung tissue has a small amount of inflammatory cell infiltration; Alveolar is evenly distributed, structural integrity, a small amount of inflammatory cell infiltration of bubble intracavity; Interstitial lung inflammatory conditions obviously improves.
The HE coloration result display of Chinese medicinal granule group mice, the inflammatory that obviously can be alleviated lung tissue by Chinese medicine intervention is changed; Alveolar structure is more complete, and luminal sectetion thing reduces to some extent, and inflammatory cell infiltration obviously alleviates; Interstitial lung inflammatory cell infiltration makes moderate progress; Pulmonary capillary structure is comparatively complete, visible a small amount of bleeding.
To sum up, experimental data shows, Chinese medicine composition Tetramune of the present invention obviously can improve the damage that PM2.5 causes mouse lung tissue and air flue.It is the product for oral administration that can be used for preventing or repairing pulmonary caused by PM2.5/respiratory system damage pioneering both at home and abroad at present.
The raw material of Chinese medicine that goods of the present invention adopt, comprises Herba Dendrobii, the Radix Astragali, Radix Scutellariae, and based on the research of the present invention to the drug effect principle of selected raw material, the raw material of goods of the present invention can be only Herba Dendrobii or the Radix Astragali, also can the two have.In a preferred embodiment, preferably adopt Radix Scutellariae as raw material further, lung meridian returned by Radix Scutellariae, is conducive to the drug action of goods of the present invention in pulmonary, and also make goods of the present invention have moistening and cleaning throat and go the effects such as lung-fire, specimen is taken in concurrently.
Due to selected three kinds of raw materials be all this area know altogether can the crude drug of integration of edible and medicinal herbs, the crowd be under haze environment can eat for a long time, prevention and/or repair PM2.5 to the damage of respiratory tract, pulmonary.
For convenience of long-term taking, described goods are preferably made into granule by the present invention, and bath is just drinkable.But be not limited to granule.
In some embodiments of the invention, described goods do not add any auxiliary material.And preferably directly make the product of the forms such as decoction pieces, beverage, tea bag further.
Goods of the present invention, when making some special forms, can add edible adjuvant or excipient.The confection that such as child likes, can add and prepare confection edible auxiliary material used.
The present invention also asks protection package to contain the food for resisting PM2.5 of goods of the present invention.
The present invention also asks the preparation method protecting goods of the present invention; any scale manufacture behavior based on commercial object; all invaded the patent right of preparation method of the present invention, described behavior mainly comprises the process adopting the raw material of goods of the present invention to produce the product for resisting PM2.5.
The Herba Dendrobii adopted in the solution of the present invention is the various Herba Dendrobiis known in prior art, such as Herba Dendrobii, Herba Dendrobii, purple dendrobium etc.
List of abbreviations
The English full name (Chinese) of abbreviation
PM2.5Particleswithanaerodynamicoflessthan2.5umorinhaledp articles (particulate matter that diameter is less than 2.5 microns)
PMl0Particleswithanaerodynamicoflessthan10umorinhaledPar ticles (diameter is less than the particulate matter of 10 microns also known as pellet)
FVCforcedvitalcapacity (forced vital capacity)
ICRInstituteofCancerResearch (american cancer institute)
RLMaxMaximuminspiratoryairwayresistance (maximum inhale airway resistance)
ReMaxMaximalexpiratoryairwayresistance (maximum expiration airway resistance)
RLAreaInspiratoryairwayresistancearea (air-breathing airway resistance area)
ReAreaExpiratoryairwayresistancearea (expiration airway resistance area)
CdynMaxthelungmaximumdynamiccompliance (the maximum Cdgn dyanamic compliance of lung)
CdynAreathelungdynamiccompliancearea (pulmonary dynamic compliance area)
PEFPeekexpiratoryflow (firmly maximum expiratory flow rate)
HEhematoxylinandeosin (h and E)
COPDchronicobstructivepulmonarydisease (chronic obstructive pulmonary disease)
Accompanying drawing explanation
Fig. 1. modeling mice is compared with the airway resistance of healthy mice,
Wherein vertical coordinate represents airway resistance, unit " cmH
2o × s ÷ ml ";
Abscissa is often organized and is from left to right followed successively by blank group, model group.
Fig. 2. modeling mice is compared with healthy mice compliance and maximum expiratory flow rate,
Wherein vertical coordinate represents air flue compliance, unit " ml ÷ cmH
2o ";
Abscissa is often organized and is from left to right followed successively by blank group, model group.
Fig. 3. each experimental mice airway resistance RLMax, ReMax, RLArea, ReArea compare,
Wherein vertical coordinate represents airway resistance, unit " cmH
2o × s ÷ ml ";
Abscissa is often organized from left to right be followed successively by blank group, model group, Chinese drug-treated group.
Three groups are compared, and have significant difference P<0.05, RL and RE of model control group is all obviously greater than blank group
(P<0.05), and higher than Chinese medicine intervention group (P<0.05).
Fig. 4. each experimental mice lung compliance CdynMax, CdynArea and maximum expiratory flow rate PEF compare,
Wherein vertical coordinate represents air flue compliance, unit " ml ÷ cmH
2o ";
Abscissa is often organized from left to right be followed successively by blank group, model group, Chinese drug-treated group.
More blank group of Cdyn and PEF obviously declines (P<0.05), and Chinese medicine intervention group comparatively model control group raises (P<0.05) to some extent;
Fig. 5. each experimental mice superior lobe of left lung lung tissue HE coloration result
Wherein A1 (× 200), A2 (× 400) are blank group, and mouse lung tissue has a small amount of inflammatory cell infiltration; Bronchia mucosa and bronchial wall structural integrity; Alveolar is evenly distributed, structural integrity, a small amount of inflammatory cell infiltration of bubble intracavity; Interstitial lung light inflammation wet face state;
B1 (× 200), B2 (× 400) are model control group, and lung tissue inflammatory changes obviously, the inflammatory cell infiltration such as neutrophilic granulocyte, lymphocyte showed increased; Bronchia mucosa partial injury, with on a small quantity hemorrhage, ooze out phenomenon; Alveolar septum is broadening, and alveolar space increases, the fracture of part alveolar, fusion, and intracavity has secretions, and consolidation phenomenon appears in part alveolar; Interstitial lung has massive inflammatory cells infiltrated;
C1 (× 200), C2 (× 400) are Chinese medicine intervention group, and mouse lung essence and interstitial have a small amount of inflammatory cell infiltration, and alveolar structure is more complete, and luminal sectetion thing reduces to some extent, and inflammatory cell infiltration obviously alleviates.
Detailed description of the invention
The foundation of embodiment 1. experimental mouse model
Experimental animal: healthy ICR female mice, body weight 28-30g, is provided by Beijing HFK Bio-Technology Co., Ltd..
PM2.5 suspension :-2015 years 01 month winters of in December, 2014, gather the Atmospheric particulates PM2.5 between stage casing, North 4th Ring Road, Beijing Bao Fu temple bridge and institute's bridge, adopt Anderson type particle sampler HBL-GUV and PM2.5 sickle with the use of, clean quartz fibre filter membrane is adopted to sample, and carry out retting conditions process with deionized water, ultrasonic vibration machine, cross leaching filtrate and carry out freezing, vacuum drying treatment, get the solution that powder normal saline becomes desired concn 45mg/ml.
Animal Model:
Blank group: modeling same day is 0 day, instill 0.9% normal saline (40 μ L/ only) respectively at the 0th, 7,14 and 21 day via intranasal application, all the other times normally raise, totally 3 weeks.
Model control group: adopt nasal cavity instillation PM2.5 suspension method, the 0th is set up respectively at model, 7,14 and 21 days via intranasal application instil the PM2.5 suspension prepared, PM2.5 suspension is given according to 60mg/kg.bw, calculate by 30g body weight, about 1.8mg/40 μ L/ only, put to death in latter 48 hours by last contamination.
Chinese medicine intervention group: the same model control group of method for establishing model.
Mice after modeling has been carried out pathological staining observation, and the main pathology that discovery PM2.5 causes model of lung injury mice shows as inflammatory change, the inflammatory cell infiltration such as neutrophilic granulocyte, lymphocyte showed increased; Bronchia mucosa partial injury, with on a small quantity hemorrhage, ooze out phenomenon; Alveolar septum is broadening, and alveolar space increases, the fracture of part alveolar, fusion, and intracavity has secretions, mainly contains mononuclear cell, neutrophil infiltration, and consolidation phenomenon appears in part alveolar; Interstitial lung has massive inflammatory cells infiltrated, sees B1, B2 in Fig. 5.Blank group mouse lung tissue has a small amount of inflammatory cell infiltration; Alveolar is evenly distributed, structural integrity, a small amount of inflammatory cell infiltration of bubble intracavity; Interstitial lung inflammatory conditions obviously improves, and sees A1, A2 in Fig. 5.
In addition, we also have detected the lung function index of mice, and the inspiratory resistance of discovery model group and expiratory resistance are all obviously greater than blank group and see Fig. 1 and Fig. 2.
Result shows, the present invention adopts the method for via intranasal application instillation PM2.5 suspension successfully to establish the mouse model of mouse lung tissue and Airway damage.
Embodiment 2. goods of the present invention are on the impact of model mice
One, materials and methods:
Normal healthy controls mice: healthy ICR female mice, body weight 28-30g, is provided by Beijing HFK Bio-Technology Co., Ltd..
Model mice: with embodiment 1
Raw material of Chinese medicine:
Herba Dendrobii: general Chinese medicine pharmacy can buy.
The Radix Astragali: general Chinese medicine pharmacy can buy.
Radix Scutellariae: general Chinese medicine pharmacy can buy
Granule is made according to the formula of table 1.This method of granulating is this area conventional method, and in large-scale production preparation, those skilled in the art can adopt other technique, or adjust according to general knowledge.
Raw material of Chinese medicine carries out water extraction, extracting solution concentrate gained extractum and adjuvant dextrin by weight 2:1 mix and make hands and pinch agglomerating, namely the loose soft material of pressure, 14-22 mesh sieve web plate is passed through again with fashion of extrusion, make uniform granule, then water content is dried within 2% in 60-80 degrees celsius, after sieving to obtain uniform granule.
Table 1. is for the crude drug formula of the goods of the present embodiment Chinese medicine intervention group
| Chinese medicine product is numbered | Raw material | Proportioning |
| 1 | Herba Dendrobii+the Radix Astragali+Radix Scutellariae | 3:3:2 |
| 1 | Herba Dendrobii+the Radix Astragali+Radix Scutellariae | 1:1:1 |
| 2 | Herba Dendrobii | 100％ |
| 3 | The Radix Astragali | 100％ |
| 4 | Herba Dendrobii+Radix Astragali | 1:1 |
Mouse experiment:
Blank group, model control group: from modeling final day, administered by oral gavage 0.9% normal saline, every day every 0.2ml, continuous three weeks.
Chinese medicine intervention group: convert according to the per kilogram of body weight of animals and human beings body metering conversion factor table, from modeling final day, administered by oral gavage Chinese medicine granules, every day every 0.2ml, the amount of gavaging is equivalent to every day adult and measures 40g crude drug, continuous three weeks.
Two, result detects
The first step. lung function
1) 1% pentobarbital intraperitoneal injection of anesthesia mice (0.3ml/30g), makes its deep anesthesia as far as possible, in order to avoid autonomous respiration is resisted mutually with respirator, affects result;
2) get dorsal position and be fixed on operating board, cut off skin of neck with tissue shear, tissue clamps chorista exposes trachea, and blunt separation trachea;
3) cut a kerf, about 1.0mm in the nearly head place of trachea, the gas-tpe fitting place of intubate is inserted trachea, and fixes with cotton thread;
4) shift mice to body and retouch instrument platform;
5) assisted respiartion machine and gas-tpe fitting is connected;
6) change of the indexs such as pulmonary function RLMax, ReMax, RLArea, ReArea, CdynMax, CdynArea, PEF of mice is recorded.(air-breathing/breath pressure of laboratory animal provides by respirator, makes maintenance steady state).
Second step. animal lung tissue's Morphology sample gathers
1) mice moves to operating board after finishing lung function, and from xiphoid-process, left border of sternum cuts off rib, exposes breast chamber completely;
2) taken off by left lung with tissue shear, isolate near hilus pulumonis place superior lobe of left lung with tissue clamps, size is about 2mm × 2mm × 2mm;
3) the clean gauze of superior lobe of left lung is covered, and fix gauze both sides (not damaging lung tissue) with paper clip, make lung tissue be immersed in the neutral formalin solution fixative of 10% completely, fixing 24h-72h.
3rd step. Morphology sample process
1) tissue dewatering, infiltration and paraffin embedding
1. ethanol successively 50% → 70% → 85% → 95%, each 90min; Rear ethanol 100% → 100%, each 60min, dewaters;
2. ethanol, each equal portions → dimethylbenzene → dimethylbenzene of dimethylbenzene, often step carries out transparent processing, equal 60min;
3.1/2 dimethylbenzene+1/2 paraffin (90min) → paraffin I (120min) → paraffin II (120min), carries out infiltration process by 62 DEG C;
4. tissue is put into paraffin mould, set tissue location in order to avoid affect chipping qualities, in the cold bench of paraffin wax embedding, carry out embedding treatment again after cooling;
5. with microtome, embedded piece of tissue is cut into the tissue of thick about 5 μm, is put in drift sheet machine and launches, then section is dragged on the poly-D-lysine membrane microscope slide of the number of finishing, carry out paster and toast.
2) HE stained preparation process
1. paraffin section is placed in dimethylbenzene I, II, each 15min;
2. dehydrated alcohol I, II → 95% ethanol I, II → each 2min of 90% → 80% → 70% → 60% → 50% ethanol;
3. tap water 5min;
4.Harris haematoxylin dyeing 8-15min;
5. the unnecessary dye liquor of tap water;
6.1% hydrochloride alcohol differentiation 30sec;
7. fully wash oil blackeite 10min in tap water;
8.1% Yihong aqueous solution dyeing 10min;
9. wash 1min from the beginning;
10. dehydration of alcohol: 50% → 60% → 70% → 80% → 90% → 95% ethanol I, II → each 2min of dehydrated alcohol I, II step by step;
The each transparent processing 10min of 11. dimethylbenzene I, II;
12. neutral gum mountings.
4th step. statistical analysis
All data all adopt SPSS20.0 statistical analysis software to carry out statistical procedures.
Three, experimental result
1. ordinary circumstance
Blank group mice is without death, and hair luster degree is good, and body weight slowly increases gradually, and breathing and thorax activity are normally.
Model control group mice during modeling has the phenomena of mortality, successively dead 2, mortality rate is 5.3% (1/19, because gavage misoperation causes dead mouse, this kind of situation belongs to experimental subject and normally comes off), existence mouse hair low in glossiness, body weight increasess slowly, there is weight loss in part, rapid breathing, thorax are obviously movable, spiritual restlessness.
Chinese medicine intervention group (in table 1 No. 1 goods) during modeling mice without the phenomena of mortality (because gavage misoperation causes dead mouse 2, this kind of situation belongs to experimental subject and normally comes off), existence mouse hair glossiness slightly gloss, body weight slowly increases, respiratory movement is comparatively mild, and mice is content to retain sovereignty over a part of the country quiet.Body weight change is in table 2.In table 1, other goods also demonstrate similar result.
The each experimental mice body weight change (± s, n=18) of table 2
Note: compare with blank group, * P<0.05; Compare with model control group, #P<0.05.First time body weight and the 4th body weight difference d meet normality and distribute, adopt paired t-test, t=-12.773, P=0.00<0.05, difference has statistical significance, and the body weight according to Average value compare the 4th each experimental mice of body weight is increased significantly comparatively for the first time.
2. obviously improve the ventilatory function of model mice
The inspiratory resistance of result display model matched group and expiratory resistance are all obviously greater than blank group (P<0.05), and higher than Chinese medicine intervention group (P<0.05), see Fig. 3; Air flue compliance and firmly more blank group of maximum expiratory flow rate obviously decline (P<0.05), and No. 1 goods Chinese medicine intervention group comparatively model control group significantly raises (P<0.05), sees Fig. 4.
Illustrate and cause model mice by PM2.5 to deposit ventilatory function limited, after Chinese medicine is intervened, the pulmonary function of mice improves (see table 3), almost returns to the level identical with blank mice and healthy mice.
In table 1, other goods also demonstrate similar result.
The each experimental mice pulmonary function change of table 3. is compared (n=8)
Note: use Kruskal-Wallis inspection,
RLMax：x
2＝12.660，P＝0.002<0.05；
ReMax：x
2＝12.260，P＝0.002<0.05；
CdynMax：x
2＝20.165，P＝0.000<0.05；
RLArea：x
2＝8.685，P＝0.013<0.05；
ReArea：x
2＝5.315，P＝0.070>0.05；
CdynArea：x
2＝6.585，P＝0.037<0.05；
PEF：x
2＝15.140，P＝0.001<0.05.
3. to the morphologic change of mouse lung tissue
This experimentation shows, and the main pathology that PM2.5 causes model of lung injury mice shows as inflammatory change, the inflammatory cell infiltration such as neutrophilic granulocyte, lymphocyte showed increased; Bronchia mucosa partial injury, with on a small quantity hemorrhage, ooze out phenomenon; Alveolar septum is broadening, and alveolar space increases, the fracture of part alveolar, fusion, and intracavity has secretions, mainly contains mononuclear cell, neutrophil infiltration, and consolidation phenomenon appears in part alveolar; Interstitial lung has massive inflammatory cells infiltrated, sees B1B2 in Fig. 5.Blank group mouse lung tissue has a small amount of inflammatory cell infiltration; Alveolar is evenly distributed, structural integrity, a small amount of inflammatory cell infiltration of bubble intracavity; Interstitial lung inflammatory conditions obviously improves, and sees A1, A2 in Fig. 5.The HE coloration result display of Chinese medicinal granule group (No. 1 goods) mice, the inflammatory that obviously can be alleviated lung tissue by Chinese medicine intervention is changed; Alveolar structure is more complete, and luminal sectetion thing reduces to some extent, and inflammatory cell infiltration obviously alleviates; Interstitial lung inflammatory cell infiltration makes moderate progress; Pulmonary capillary structure is comparatively complete, and visible a small amount of bleeding, is shown in that in Fig. 5, in C1 with C2. table 1, other goods also demonstrate similar result.
To sum up show, via intranasal application instillation PM2.5 suspension successfully can set up the model of mouse lung tissue and Airway damage, and experimental result shows Chinese medicine product of the present invention obviously can improve the damage that PM2.5 causes mouse lung tissue and air flue.
Claims (9)
1. prevention or reparation PM2.5 cause goods for pulmonary lesion, and its raw material comprises Herba Dendrobii and/or the Radix Astragali.
2. goods according to claim 1, described raw material also comprises Radix Scutellariae.
3. goods according to claim 1 and 2, the dosage form of described goods is granule.
4., according to the arbitrary described goods of claim 1-2, in described goods, also comprise acceptable adjuvant.
5. prevention or reparation PM2.5 cause goods for pulmonary lesion, and its raw material is made up of Herba Dendrobii, the Radix Astragali and Radix Scutellariae.
6., according to the arbitrary described goods of claim 1-5, described Herba Dendrobii is selected from Herba Dendrobii, Dendrobium nobile, purple dendrobium, Herba Dendrobii.
7. prevention or reparation PM2.5 cause a food for pulmonary lesion, it is characterized in that, include the arbitrary described goods of claim 1-6.
8. preparation prevention or reparation PM2.5 cause the method for the goods of pulmonary lesion, and it is characterized in that, the raw material of employing comprises Herba Dendrobii and/or the Radix Astragali.
9. method according to claim 8, described raw material also comprises Radix Scutellariae.
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| CN108283692A (en) * | 2018-04-23 | 2018-07-17 | 张国敏 | A kind of Chinese medicine composition and preparation method thereof for treating virus flu |
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