CN110433181A - Application of the hayashishita Shanxi American ginseng in preparation treatment Chronic Obstructive Pulmonary Disease drug - Google Patents
Application of the hayashishita Shanxi American ginseng in preparation treatment Chronic Obstructive Pulmonary Disease drug Download PDFInfo
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Abstract
The present invention relates to a kind of application of hayashishita Shanxi American ginseng in preparation treatment Chronic Obstructive Pulmonary Disease drug, belong to the new medicinal usage field of hayashishita Shanxi American ginseng.The application of hayashishita Shanxi American ginseng and its ethanol extract in preparation treatment Chronic Obstructive Pulmonary Disease drug.Hayashishita Shanxi American ginseng and its ethanol extract can improve experimental model lung function, inhibit inflammatory cell infiltration, inhibit the release of inflammatory factor TNF-α, IL-1 β and IL-6, improve total SOD vigor, repair oxidativestress damage.Mitigate slight illness for patient, improves the quality of living.
Description
Technical field
The invention belongs to fields of biomedicine, are related to the new medicinal usage of hayashishita Shanxi American ginseng, and in particular to hayashishita Shanxi American ginseng
Application of the ethanol extract in preparation prevention and treatment Chronic Obstructive Pulmonary Disease drug.
Background technique
Chronic Obstructive Pulmonary Disease (Chronic Obstructive Pulmonary Disease, COPD) is a kind of chronic inflammation
Disease property lung disease, feature are duration, the respiratory symptom and flow limitation for carrying out sexual development, and clinical manifestation is chronic cough
It coughs, expectoration, wheeze, shortness of breath uncomfortable in chest, expiratory dyspnea, serious person and with symptoms such as tired thin anxieties.40 years old Chinese or more people
For the disease incidence of group COPD up to 8.2%, foreign countries are 9-10%；The COPD death rate is high, before the World Health Organization statistics whole world in 2016
In ten causes of death, COPD is number three position.
Generally believe that COPD is a kind of disease that can be prevented and treated at present, but without effective treatment means.It is main
Illness the reason is that smoking and the factors such as environmental pollution, the primary treatments of COPD be long-term administration and in daily life oneself
I constrains, and drug treatment relies primarily on anticholinergic drug, β2Receptor stimulating agent, glucocorticoid or a variety of drugs connection
With, however at present, these types of drug in the market has certain side effect and tolerance, causes conditions of patients can not
Effectively alleviated, and from the development process that can not reverse COPD without constraint, it can not also prevent the chronic inflammatory reaction of patient
Development.Clinically also there are many cases of Chinese patent drug and chemical drug drug combination treatment COPD at present.
Research finds inflammatory reaction and oxidativestress damage is two main pathogenesis of COPD, in the air flue and lung of COPD patient
There is chronic inflammation in tissue, the noxious material such as bacteria lipopolysaccharide (LPS) in air enter during human body latter aspect can activate
Property granulocyte, macrophage and Dendritic Cells etc., while can be thin by activation air flue inner cell such as human airway epithelial cells, alveolar
Born of the same parents etc. activate the damage associated molecular pattern of inflammation, so as to cause body discharge various cell factors, acute phase reactive protein
With the substances such as antibacterial peptide, lead to the lung tissue chronic inflammation of duration.In the numerous inflammatory factors for participating in inflammatory reaction process
Three kinds of tumor necrosis factor-alpha (TNF-α), interleukin-1 ' beta ' (IL-1 β), interleukin-6 (IL-6) factors have played weight
It acts on, their contents level reflects the degree of inflammatory reaction.
Oxidativestress damage is the generation and antioxidase due to activity in vivo oxygen (Reactive Oxygen Species, ROS)
Between the ability of ROS of degrading it is unbalance caused by, in healthy human body, it is equilibrium state that ROS, which is generated between antioxidant system, one
Denier then generates oxidativestress damage since endogenous or external source causes balance to be broken.Research finds that smoking patients lung is easily introduced
External source ROS causes lung to occur oxidativestress damage, and studies have found that oxidativestress damage and inflammatory reaction in mutually according to
The relationship rely, mutually promoted.Lipid oxidation reaction can occur during oxidativestress damage, generate malonaldehyde (MDA), therefore
The content of MDA can be used to judge the process of oxidative stress, while the vigor of superoxide dismutase (SOD) reflects people in vivo
The oxidation resistant ability of body.
American Ginseng (Panax quinquefolius) is that Araliaceae (Araliaceae) Panax (Panax Linn.) is perennial
Herbaceous plant, alias American ginseng, American ginseng, Western ginseng, originates in the Wisconsin State of Canadian big Quebec and the U.S..It passes
System Chinese medicine is used as medicine with American Ginseng root, cures mainly deficiency of vital energy yin deficiency, interior heat, cough and asthma phlegm blood, and abnormal heat is tired of tired, is quenched one's thirst, the dry larynx of mouth is dry.The West
The main active of ginseng is ginsenoside, research shows that ginsenoside has anti-inflammatory, antitumor, anti-oxidant, raising immunity
Isoreactivity.Hayashishita Shanxi American ginseng is that seeds of American ginseng is manually sowed to the obtained American Ginseng in wild mountain forest environment, with this
Achieve the purpose that the wild American Ginseng growing environment of simulation.Have no American Ginseng for preventing and treating the relevant report of COPD at present.
Summary of the invention
The present invention provides a kind of application of hayashishita Shanxi American ginseng in preparation treatment Chronic Obstructive Pulmonary Disease drug.
The technical solution adopted by the present invention is that: a kind of hayashishita Shanxi American ginseng is in preparation treatment Chronic Obstructive Pulmonary Disease drug
Using.
Application of the hayashishita Shanxi American ginseng ethanol extract in preparation treatment Chronic Obstructive Pulmonary Disease drug.
Hayashishita Shanxi American ginseng and its ethanol extract can improve experimental model lung function, inhibit inflammatory cell infiltration, inhibit scorching
The release of inflammation factor TNF-α, IL-1 β and IL-6 improves total SOD vigor, repairs oxidativestress damage.
Drug of the present invention is made of a effective amount of hayashishita Shanxi American ginseng ethanol extract and pharmaceutically acceptable auxiliary material, and auxiliary material is
Disintegrating agent, filler, lubricant, wetting agent, emulsifier, adhesive.The pharmaceutical preparation is tablet, capsule, pill, molten
Liquor, injection, powder, sucking preparation, spray, aerosol, syrup, oral solution formulation.
Application of the hayashishita Shanxi American ginseng ethanol extract in preparation treatment Chronic Obstructive Pulmonary Disease drug.
The present invention carries out zoopery using hayashishita Shanxi American ginseng and its ethanol extract, and experimental model is smoke from cigarette exposure institute
The COPD experimental model of cause.Smoking is to lead to the principal element of COPD, therefore cigarette induction simulation mouse model is used to study hayashishita
The relaxation effect that Shanxi American ginseng ethanol extract induces cigarette.Using smoke from cigarette exposure induced mice model, and use woods
American Ginseng ethanol extract of going down the hill intervenes animal, and experimental result is shown: 1, the hayashishita Shanxi American ginseng ethanol extract that stomach-filling is given
There is protective effect to the lung function of COPD mouse caused by smoke from cigarette exposure；2, to COPD model mice caused by smoke from cigarette exposure
The infiltration of inflammatory cell, the expression of inflammatory factor have inhibiting effect；3, have to the intracorporal oxidativestress damage of model mice
Alleviate, repair.
Detailed description of the invention
Fig. 1 is influence diagram of the hayashishita Shanxi American ginseng to COPD model mice weight.
Specific embodiment
Embodiment 1: the preparation of hayashishita Shanxi American ginseng tablet
5g hayashishita Shanxi American ginseng is taken, starch 30g is added and is uniformly mixed, spraying into 70% ethyl alcohol is that softwood is made in wetting agent；Cross 20 mesh
Sieve, obtains wet granular, 60 DEG C of dryings, and every 30min is stirred one time, and moisture is controlled 1% -3%.Magnesium stearate powder is added, mixes
It is even, it is pressed into 1000.
Embodiment 2: hayashishita Shanxi American ginseng ethanol extract the preparation method is as follows:
Fresh hayashishita Shanxi American ginseng sample is taken, dries and crosses 40 meshes after grinding, takes gained powder 8g 80% ethyl alcohol (v/ of 200ml
V) refluxing extraction, is extracted 3 times by 80 DEG C of Extracting temperature, each 2h, and vacuum-concentrcted obtains medicinal extract, sufficiently dry powder
2.16g。
Embodiment 3: the preparation of hayashishita Shanxi American ginseng ethanol extract tablet
5g hayashishita Shanxi American ginseng ethanol extract is taken, starch 30g is added and is uniformly mixed, 70% ethyl alcohol of penetrating is made soft for wetting agent
Material；20 meshes are crossed, wet granular, 60 DEG C of dryings are obtained, every 30min is stirred one time, and moisture is controlled 1% -3%.Magnesium stearate is added
Powder mixes, is pressed into 1000.
Embodiment 4: the preparation of hayashishita Shanxi American ginseng ethanol extract capsule
Supplementary material is crossed to No. five sieves, in favor of mixing completely, uniformly, weighs hayashishita Shanxi American ginseng ethanol extract 5g,
Starch 28g is uniformly mixed, encapsulated 1000.
Effect of the invention is described further by pharmacodynamic experiment below,
Experimental example 1: influence of the hayashishita Shanxi American ginseng to COPD model mice weight.
After being exposed to smoke from cigarette for a long time, first check for directly affecting mouse weight, mouse is weighed weekly once.Such as
Shown in Fig. 1, compared with blank control group, the significant mitigation of mouse weight in model group shows that COPD model is successfully established, and
And mouse shows COPD related symptoms relevant to loss of appetite.However, compared with model group, dexamethasone and hayashishita Shanxi
The intervention of American ginseng can dramatically increase mouse weight.Wherein, hayashishita Shanxi American ginseng high dose group is shown more better than positive drug group
Effect.
Conclusion: hayashishita Shanxi American ginseng can effectively restore the weight of COPD model mice.
Experimental example 2: hayashishita Shanxi American ginseng makees the improvement of Chronic Obstructive Pulmonary Disease (COPD) model mice pathologic state colony
With.
The main reason for passive sucking smoke from cigarette is COPD formation, has become the basis for establishing mouse COPD inflammatory model.
Therefore, the passive smoking experiment plastic device of 18 centimetres of 40 cm x of 60 cm x has been made in this laboratory by oneself, on chest top
Portion has 12 ventilation openings (diameter=5 millimeter) for divulging information, and side has 1 ventilation hole for connecting mist source.Mouse is divided at random
It is 6 groups: Normal group, model group, positive drug group and the high, medium and low dosage group of hayashishita Shanxi American ginseng (40,20,10mg/kg).
It other 5 groups of mouse will be placed in self-control sealing device in addition to Normal group.The smoke from cigarette lighted is by inflating pump through side
Venthole is pumped into case, while the venthole at top is by rubber belt sealing.The smog of 4 cigarettes is continuously pumped into sealing experiment every time
In device, total suspended particles concentration is made to reach 300mg/m3, continue 10min.Later, lid is removed, makes mouse in air environment
Middle rest 10 minutes.Above-mentioned steps carry out 6 times (cigarette exposure total time: 1h) daily, and continuous 6 days weekly, total duration was
13 weeks.After modeling success, the 9th week starts, and pure water (10mL/kg), positive drug are given in stomach-filling respectively for Normal group, model group
Group stomach-filling is given dexamethasone (2.0mg/kg), and the hayashishita mountain of corresponding dosage is given in stomach-filling to each dosage group of hayashishita Shanxi American ginseng respectively
American Ginseng, successive administration 36 days.At the 92nd day, mouse acquired blood and puts to death, fill immediately after after yellow Jackets are anaesthetized
Lung tissue is infused, left lung tissue is taken out from thoracic cavity, is fixed on 4% paraformaldehyde, and be embedded in paraffin, Hematoxylin-eosin (HE)
Dyeing observation pathologic state colony changes.
As a result: epithelial cell is complete in the lung tissue of blank control group mouse, changes without apparent inflammatory cell infiltration and pathology
Become；Model group mouse bronchial lung tissue has the pathological change of Chronic Obstructive Pulmonary Disease, intrapulmonary bronchiole mucociliary column
The visible denaturation of columnar epithelium, necrosis, visible a large amount of cell infiltrations around bronchiole wall, alveolar number substantially reduce；It is positive right
Substantially reduced according to bronchiole wall damage in group mouse lung, peribronchiolar alveolar structure is complete；Hayashishita Shanxi American ginseng is high, in
Dosage group bronchus cilium is slight adhesion, denaturation, bronchial smooth muscle thickness slightly increase；Tracheal mucosa lower layer, muscle layer are visible
A small amount of acidophic cell；Lymphocyte and a small amount of plasmocyte infiltrating.
Experimental example 3: influence of the hayashishita Shanxi American ginseng to Chronic Obstructive Pulmonary Disease (COPD) model mice lung function.
BALA/c mouse, 20 ± 2g of weight are randomly divided into 6 groups: Normal group, model group, positive drug group and hayashishita Shan Xiyang
Join high, medium and low dosage group (40,20,10mg/kg).Modeling method is the same as example 1.After modeling success, the 9th week starts, normal control
Stomach-filling is given pure water (10mL/kg) respectively for group, model group, and dexamethasone (2.0mg/kg), hayashishita are given in positive drug group stomach-filling
The hayashishita Shanxi American ginseng of corresponding dosage is given in stomach-filling to each dosage group of Shanxi American ginseng respectively, and successive administration 36 days.
Lung function tests: mouse is anaesthetized using 1% yellow Jackets, and after deep anaesthesia, mouse is fixed on trachea cannula plate
On, tracheae is peeled away and cuts off a small notch between tracheae goes in ring cartilage, intubation is inserted by notch and passes through operation lines
Fixed, a low level of lying on the back is placed in toy pulmonary function test apparatus enclosure volume and traces in case.In correctly connection animal, Pneumatic sensing
After device and pressure sensor, tranquil breathing is recorded to obtain baseline value.Then, lung volume, static and dynamic lung function are obtained
The parameter of energy, forced expiratory volume/forced vital capacity (FEV100/FVC) including 100ms, function residual capacity (FRC), air flue
Resistance (RI), static compliance (Cchord) and lung volume (Lung volume).
As a result: compared with Normal group, model group mouse lung function parameter FRC, RI and Cchord are dramatically increased, parameter
FEV100/FVC and Lung volume is remarkably decreased, five parameter differences statistically significant (P < 0.05 or P <
0.01), illustrate modeling success.Compared with model group, high, medium and low dosage group lung function parameter FRC, the RI of hayashishita Shanxi American ginseng and
Cchord is remarkably decreased, and parameter FEV100/FVC and Lung volume, which are dramatically increased, levels off to normal group, and difference has system
Meter learns meaning (P < 0.05 or P < 0.01).Compared to model group, positive drug group is in addition to significantly improving RI, and other parameters are without bright
It is aobvious to change, it the results are shown in Table 1.
Influence (n=10, X ± SD) of the 1 hayashishita Shanxi American ginseng of table to COPD model mice lung function
##Compared with Normal group, P < 0.01
*Compared with model group, P < 0.05；**Compared with model group, P < 0.01
Conclusion: the hayashishita Shanxi middle and high dosage of American ginseng can significantly improve the lung function of Chronic Obstructive Pulmonary Disease (COPD) model mice
Energy.
Experimental example 4: hayashishita Shanxi American ginseng to proinflammatory factor IL-6 in Chronic Obstructive Pulmonary Disease (COPD) model mice serum,
IL-1 β, the influence of TNF-α expression.
Modeling method and medication are with experimental example 1, after administration, by mouse anesthesia, take blood, execution.Centrifugal blood 10min
(3000r/min, 4 DEG C), Aspirate supernatant obtain serum be placed in -20 DEG C it is to be measured.Enzyme-linked immunosorbent assay (ELISA) detects serum
Interleukins is horizontal；Interleukins (IL-6, IL-1 β) and tumor necrosis factor-alpha (TNF-α) detection kit are by the U.S.
R&D company provides.As a result: model group IL-6, IL-1 β, TNF-α expression more normally organize significantly raised (P < 0.01)；Hayashishita
The high, medium and low dosage group IL-6 of Shanxi American ginseng, IL-1 β, TNF-α expression is compared with model group significant decrease (P < 0.05).
1 hayashishita Shanxi American ginseng of table to IL-6 in COPD model mice serum, the influence of IL-1 β and TNF-α expression (n=10, X ±
SD)
##Compared with Normal group, P < 0.01
*Compared with model group, P < 0.05；**Compared with model group, P < 0.01
Conclusion: the table of proinflammatory factor IL-6, IL-1 β and TNF-α in the COPD mice serum that hayashishita Shanxi American ginseng induces cigarette
Up to there is inhibiting effect.
Embodiment 5: hayashishita Shanxi American ginseng is to superoxides in Chronic Obstructive Pulmonary Disease (COPD) model mice lung homogenate
The influence of mutase (SOD) and malonaldehyde (MDA) expression.
Modeling method and medication are the same as experimental example 1.Mouse is put to death, immediately after perfusion lung group after yellow Jackets are anaesthetized
Knit, take out right lung tissue from thoracic cavity, lung weighing rinses in cold saline, and wiped with filter paper it is dry, then cut about 1.0cm ×
The weighing of 0.5cm lung tissue, is added 9 times of lung tissue weight of physiological saline, is placed in homogenizer and is homogenized in low temperature environment, then 3
000r min, 4 DEG C of centrifugation 10min obtain supernatant.Enzyme-linked immunosorbent assay (ELISA) detects SOD and MDA in lung homogenate
It is horizontal；Detection kit is provided by R&D company, the U.S..As a result: more normal group of model group SOD expression be substantially reduced (P <
0.01), MDA expression more normally organizes significantly raised (P < 0.01)；American ginseng high, medium and low dosage group SOD in hayashishita Shanxi expresses water
Flat significantly to increase (P < 0.05) compared with model group, more normal group of MDA expression is substantially reduced (P < 0.05).
The influence (n=10, X ± SD) that 1 hayashishita Shanxi American ginseng of table expresses SOD in COPD model mice lung homogenate and MDA
##Compared with Normal group, P < 0.01
*Compared with model group, P < 0.05；**Compared with model group, P < 0.01
Conclusion: the table of superoxide dismutase (SOD) in the COPD mouse lung tissue homogenate that hayashishita Shanxi American ginseng induces cigarette
Up to there is facilitation, and there is inhibiting effect to the expression of malonaldehyde (MDA).
Experimental example 6: hayashishita Shanxi American ginseng is to Chronic Obstructive Pulmonary Disease (COPD) model mice metabolic profiling analysis.
Modeling method and medication are the same as experimental example 1.After last time is administered, by mouse anesthesia, blood, execution are taken.Centrifugal blood
10min (3000r/min, 4 DEG C), Aspirate supernatant obtain serum be placed in -20 DEG C it is to be measured.By using UPLC-Q/TOF-MS with just,
The metabolism spectrogram of negative mode acquisition blood serum sample.It is analyzed by data, identifies difference metabolin and metabolic pathway.
Conclusion: hayashishita Shanxi American ginseng shows apparent Difference of Metabolism to the COPD mouse that cigarette induces, and passes through metabolic pathway
It is more intuitive to illustrate the anti-COPD mechanism of action of hayashishita Shanxi American ginseng.
Therefore the COPD for using cigarette inducing mouse simulation COPD model research hayashishita Shanxi American ginseng ethanol extract to induce cigarette
Relaxation effect.
In conclusion the correlation of TNF-α, the variation of IL-1 β and IL-6 contents level and COPD facilitates under drug effect
It is further understood that the specific mechanism of action of inflammatory factor, while judging drug to oxidation by detection MDA content and SOD vigor
The intervention effect of stress damage provides clinical guidance and foundation for the prevention and treatment of COPD, mitigates ailing, raising life for patient
Bioplasm amount.
Claims (5)
1. a kind of application of hayashishita Shanxi American ginseng in preparation treatment Chronic Obstructive Pulmonary Disease drug.
2. application of the hayashishita Shanxi according to claim 1 American ginseng in preparation treatment Chronic Obstructive Pulmonary Disease drug,
It is characterized by: proinflammatory factor IL-6, IL-1 β and TNF-α in the COPD mice serum that hayashishita Shanxi American ginseng induces cigarette
Expression has inhibiting effect.
3. application of the hayashishita Shanxi according to claim 1 American ginseng in preparation treatment Chronic Obstructive Pulmonary Disease drug,
It is characterized by: in the COPD mouse lung tissue homogenate that hayashishita Shanxi American ginseng induces cigarette superoxide dismutase SOD table
Up to there is facilitation, and there is inhibiting effect to the expression of malonaldehyde MDA.
4. a kind of application of hayashishita Shanxi American ginseng ethanol extract in preparation treatment Chronic Obstructive Pulmonary Disease drug.
5. a kind of hayashishita Shanxi American ginseng ethanol extract according to claim 4 treats Chronic Obstructive Pulmonary Disease in preparation
Application in drug is held sign and is: the hayashishita Shanxi American ginseng ethanol extract the preparation method is as follows: taking fresh hayashishita mountain
American Ginseng sample dries and crosses 40 meshes after grinding, and gained powder is taken to be extracted with 25 times of 80% alcohol reflux, and 80 DEG C of Extracting temperature,
Each 2h is extracted 3 times, and vacuum-concentrcted obtains medicinal extract, sufficiently dry powder.
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| CN112162100A (en) * | 2020-09-30 | 2021-01-01 | 锦州医科大学附属第一医院 | Application of TNF-alpha detection substance in preparation of chronic obstructive pulmonary disease diagnosis or treatment reagent |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112162100A (en) * | 2020-09-30 | 2021-01-01 | 锦州医科大学附属第一医院 | Application of TNF-alpha detection substance in preparation of chronic obstructive pulmonary disease diagnosis or treatment reagent |
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| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20191112 |