CN108524766A - A kind of Chinese medicine composition and its application - Google Patents
A kind of Chinese medicine composition and its application Download PDFInfo
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/894—Dioscoreaceae (Yam family)
- A61K36/8945—Dioscorea, e.g. yam, Chinese yam or water yam
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/60—Moraceae (Mulberry family), e.g. breadfruit or fig
- A61K36/605—Morus (mulberry)
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8969—Polygonatum (Solomon's seal)
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Abstract
The present invention provides a kind of Chinese medicine composition, bulk pharmaceutical chemicals include two kinds or more of combination in Chinese yam, arillus longan, mulberry leaf, Exocarpium Citri Rubrum, radix polygonati officinalis, bulbus fritillariae cirrhosae, Rhizoma Atractylodis Macrocephalae, Semen Lablab Album, lotus leaf.The present invention also provides a kind of pharmaceutical preparations including above-mentioned Chinese medicine composition and above-mentioned Chinese medicine composition to prepare the application in preventing or alleviating the pharmaceutical preparation of injury of lungs caused by pellet.Chinese medicine composition provided by the invention is accumulated by reducing active oxygen, malonaldehyde, interleukin-6, interleukin 8 and tumor necrosis factor, increase superoxide dismutase, glutathione peroxidase content, to improve, body is anti-oxidant and anti-inflammatory power, enhance immunity of organisms, and then achievees the purpose that resist pellet damage body.
Description
Technical field
The present invention relates to pharmaceutical fields, specifically, a kind of Chinese medicine composition and its application.
Background technology
Effect of air pollution on human health has become the social concern of everybody general concern.According to China Environmental Science
Research institute's expert statistics share 2000 multinomial researchs and point out, in inhalable particles, maximum to human health damage is diameter
Particulate matter (fine particulate matter, PM2.5) less than or equal to 2.5 microns.PM2.5 has grain size small, in body
The features such as interior residence time is long, is not easy to be metabolized, and levels of toxic substances is high.As a kind of pellet, PM2.5 can pass through
Respiratory system enters human lung, is deposited in bronchus and alveolar, wherein more tiny ingredient even can penetrate alveolar, interferes
The gas exchanges of intrapulmonary cause the bioprocess such as lung cells oxidativestress damage, inflammatory reaction, Apoptosis, strong to human body
Health makes a big impact.Therefore, preventing or alleviate PM causes the situation of disease very urgent.
Declare that prevention or alleviation pellet cause the product of injury of lungs all not play and make us full currently on the market
The effect of meaning.Research shows that some commercially available haze-proof masks cannot play the role of obstructing PM2.5 at all；Indoor air purification
Device can adsorb some particles, but its ozone generated seriously endangers human health.Therefore, by traditional Chinese medicine theory and modern times
Pharmacology is combined, in injury of lungs caused by rationally utilizing Chinese medicine composition resource, exploitation to prevent or alleviate pellet
Drug composition has important practical significance.
Invention content
For the problems in the relevant technologies, the present invention has studied a kind of Chinese medicine composition, which is characterized in that the Chinese medicine group
The bulk pharmaceutical chemicals for closing object include two kinds in Chinese yam, arillus longan, mulberry leaf, Exocarpium Citri Rubrum, radix polygonati officinalis, bulbus fritillariae cirrhosae, Rhizoma Atractylodis Macrocephalae, Semen Lablab Album, lotus leaf or
Above combination.
The bulk pharmaceutical chemicals of above-mentioned Chinese medicine composition are by Chinese yam, arillus longan, mulberry leaf, Exocarpium Citri Rubrum, radix polygonati officinalis, bulbus fritillariae cirrhosae, Rhizoma Atractylodis Macrocephalae, white flat
Two kinds or more of group in beans, lotus leaf is combined into.
The bulk pharmaceutical chemicals of above-mentioned Chinese medicine composition by two kinds in Chinese yam, arillus longan, mulberry leaf, Exocarpium Citri Rubrum, radix polygonati officinalis, bulbus fritillariae cirrhosae or with
On group be combined into.
The bulk pharmaceutical chemicals of above-mentioned Chinese medicine composition are combined by two or three of group in Rhizoma Atractylodis Macrocephalae, Semen Lablab Album, mulberry leaf.
The bulk pharmaceutical chemicals of above-mentioned Chinese medicine composition are combined by two or three of group in radix polygonati officinalis, Semen Lablab Album, lotus leaf.
The bulk pharmaceutical chemicals of above-mentioned Chinese medicine composition are made of radix polygonati officinalis and mulberry leaf.
The bulk pharmaceutical chemicals of above-mentioned Chinese medicine composition are by two kinds or more of group in Exocarpium Citri Rubrum, Rhizoma Atractylodis Macrocephalae, mulberry leaf, Semen Lablab Album, radix polygonati officinalis
It is combined into.
In above-mentioned Chinese medicine composition, Exocarpium Citri Rubrum extract, Rhizoma Atractylodis Macrocephalae extract, mulberry-leaf extract, Semen Lablab Album extract, radix polygonati officinalis
The mass ratio of extract is 1-2:1-4:2-6:6-10:30-34.
The present invention also provides a kind of pharmaceutical preparations of injury of lungs caused by prevention or alleviation pellet, wherein
The pharmaceutical preparation includes above-mentioned Chinese medicine composition.
In said medicine preparation, the dosage form of the pharmaceutical preparation is selected from tablet, capsule, pill, aerosol, oral solution
Body preparation, granule, powder, syrup or combination thereof.
The present invention also provides above-mentioned Chinese medicine compositions in injury of lungs caused by preparation prevention or alleviation pellet
Pharmaceutical preparation in application.
The present invention provides a kind of Chinese medicine composition and including the pharmaceutical preparation of the Chinese medicine composition, the Chinese medicine composition
Active oxygen, malonaldehyde, interleukin-6, interleukin 8 and tumor necrosis factor accumulation can be reduced, superoxides is increased
Mutase, glutathione peroxidase content, to improve, body is anti-oxidant and anti-inflammatory power, enhancing immunity of organisms reach
To the effect for resisting pellet damage body, which can be used for preventing or alleviates pellet to make
At injury of lungs.
Description of the drawings
It in order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, below will be to institute in embodiment
Attached drawing to be used is needed to be briefly described, it should be apparent that, the accompanying drawings in the following description is only some implementations of the present invention
Example, for those of ordinary skill in the art, without creative efforts, can also obtain according to these attached drawings
Obtain other attached drawings.
Fig. 1 .1 show the particle size distribution figure of particulate matter (PM).
Fig. 2 .1a show the influence diagram that the PM of various concentration acts on the different time to A549 cellular oxidation stress situations,
In, " * " is indicated compared with blank group, P<0.05；" * * " is indicated compared with blank group, P<0.01；" * * * " is indicated and blank group phase
Than P<0.001.
Fig. 2 .1b show the influence diagram that the PM of various concentration acts on the different time to A549 cellular inflammation states, wherein
" * " is indicated compared with blank group, P<0.05；" * * " is indicated compared with blank group, P<0.01；" * * * " is indicated compared with blank group,
P<0.001。
Fig. 2 .2a show the influence diagram that the PM of various concentration acts on the different time to A549 cell survival rates, wherein " * "
It indicates compared with blank group, P<0.05；" * * " is indicated compared with blank group, P<0.01；" * * * " is indicated compared with blank group, P<
0.001。
Fig. 2 .2b show influence diagrams of the PM to A549 apoptosis status.
Fig. 2 .2c show influence diagrams of the PM to A549 apoptosis status, wherein " * * " is indicated compared with the control group, P<
0.01。
Fig. 2 .2d show influence diagrams of the PM to A549 apoptosis status, wherein " * * " is indicated compared with the control group, P<
0.01。
Fig. 2 .2e show the influence diagram that PM damages A549 cell mitochondrials, wherein in two row figures below JC-1, the left side
Light-colored part in one row figure (i.e. first row figure) is green, and the light-colored part in the row figure of the right one (i.e. secondary series figure) is red
Color, wherein " * * " is indicated compared with the control group, P<0.01.
Fig. 2 .2f show influence diagrams of the PM to A549 cell TNF-a expression quantity, wherein " * " is indicated compared with the control group, P<
0.05；" * * " is indicated compared with the control group, P<0.01.
Fig. 3 .1 show the standardization extraction flow of 105 Chinese medical extracts.
Fig. 4 .1a show the influence diagram of 105 Chinese medical extracts A549 cellular oxidations stress situation caused by PM.
Fig. 4 .1b show the influence diagram of 21 Chinese medical extracts A549 cellular oxidations stress situation caused by PM, wherein
" ## " is indicated compared with blank control group, P<0.01；" * " is indicated compared with model group, P<0.05；" * * " is indicated and model group phase
Than P<0.01；" * * * " is indicated compared with model group, P<0.001.
Fig. 4 .1c show the influence diagram of 6 Chinese medical extracts A549 cellular oxidations stress situation caused by PM, wherein
" ### " is indicated compared with blank control group, P<0.001；" * * * " is indicated compared with model group, P<0.001.
Fig. 4 .1d show the influence diagram of 6 Chinese medical extracts A549 cellular oxidations stress situation caused by PM, wherein
" ## " is indicated compared with blank control group, P<0.01；" * " is indicated compared with model group, P<0.05；" * * " is indicated and model group phase
Than P<0.01.
Fig. 4 .1e show the influence diagram of 6 Chinese medical extracts A549 cellular oxidations stress situation caused by PM, wherein
" ### " is indicated compared with blank control group, P<0.001；" * * * " is indicated compared with model group, P<0.001.
Fig. 4 .1f show 6 Chinese medical extracts caused by PM A549 cellular oxidations stress correlative protein expression exception shadow
Ring figure, wherein " # " expression is compared with blank control group, P<0.05；" * " is indicated compared with model group, P<0.05；" * * " is indicated
Compared with model group, P<0.01；" * * * " is indicated compared with model group, P<0.001.
Fig. 4 .2a show the influence diagram of 105 Chinese medical extracts A549 cellular inflammations state caused by PM.
Fig. 4 .2b show the influence diagram of 10 Chinese medical extracts A549 cellular inflammations state caused by PM, wherein " ### "
It indicates compared with blank control group, P<0.001；" * * * " is indicated compared with model group, P<0.001.
Fig. 4 .2c show the influence diagram of 3 Chinese medical extracts A549 cellular inflammations state caused by PM, wherein " ### "
It indicates compared with blank control group, P<0.001；" * * " is indicated compared with model group, P<0.01.
Fig. 4 .2d show the influence diagram of 3 Chinese medical extracts A549 cellular inflammations state caused by PM, wherein " ### "
It indicates compared with blank control group, P<0.001；" * * " is indicated compared with model group, P<0.01.
Fig. 4 .2e show the influence diagram of 3 Chinese medical extracts A549 cellular inflammations state caused by PM, wherein " ### "
It indicates compared with blank control group, P<0.001；" * * " is indicated compared with model group, P<0.01.
Fig. 4 .2f show the influence of 3 Chinese medical extracts A549 cellular inflammations correlative protein expression exception caused by PM
Figure, " ### " expression is compared with blank control group, P<0.001；" * " is indicated compared with model group, P<0.05；" * * " is indicated and mould
Type group is compared, P<0.01；" * * * " is indicated compared with model group, P<0.001.
Fig. 4 .3a show the influence diagram of 3 Chinese medical extract A549 apoptosis status caused by PM.
Fig. 4 .3b show the influence diagram of 3 Chinese medical extract A549 apoptosis status caused by PM, wherein " ## " table
Show compared with blank control group, P<0.01；" * " is indicated compared with model group, P<0.05, " * * " is indicated compared with model group, P<
0.01。
Fig. 4 .3c show the influence diagram that 3 Chinese medical extracts A549 cell mitochondrials caused by PM damage, wherein JC-1
In two following row figures, the light-colored part in one row figure of the left side is green, and the light-colored part in one row figure of the right is red,
In, " ## " is indicated compared with blank control group, P<0.01；" * " is indicated compared with model group, P<0.05；" * * " is indicated and model
Group is compared, P<0.01.
Fig. 4 .3d show the influence diagram of 3 Chinese medical extract A549 apoptosis status caused by PM, wherein " ## " table
Show compared with blank control group, P<0.01；" * " is indicated compared with model group, P<0.05；" * * " is indicated compared with model group, P<
0.01。
Fig. 4 .3e show the influence of 3 Chinese medical extracts and the inhibitors of apoptosis low survival rate of A549 cells caused by PM
Figure, wherein wherein, " ## " is indicated compared with blank control group, P<0.01；" * " is indicated compared with model group, P<0.05；“**”
It indicates compared with model group, P<0.01.
Fig. 4 .3f show the influence of 3 Chinese medical extracts A549 cell death related proteins abnormal expression caused by PM
Figure, wherein wherein, " ## " is indicated compared with blank control group, P<0.01；" * " is indicated compared with model group, P<0.05；“**”
It indicates compared with model group, P<0.01.
Fig. 5 .1 show that traditional Chinese medicine compatibility theory instructs prescription design drawing.
Fig. 6 .1 show influence diagram of the Chinese medicine composition to PM drainage rates in zebra fish body.
Fig. 7 .1 show the influence diagram of Chinese medicine composition mouse injury of lungs caused by PM.
Fig. 7 .2 show the influence diagram of Chinese medicine composition mouse oxidative stress status caused by PM, wherein " ## " indicate with
Blank control group is compared, P<0.01；" * " is indicated compared with model group, P<0.05；" * * " is indicated compared with model group, P<0.01.
Fig. 7 .3 show the influence diagram of Chinese medicine composition mouse inflammatory conditions caused by PM, wherein " ### " is indicated and sky
White control group is compared, P<0.001；" * * " is indicated compared with model group, P<0.01；" * * * " is indicated compared with model group, P<
0.001。
Specific implementation mode
Below in conjunction in the embodiment of the present invention, technical solution in the embodiment of the present invention carries out clear, complete
Ground describes, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on this
Embodiment in invention, the every other embodiment that those of ordinary skill in the art are obtained belong to the model that the present invention protects
It encloses.
According to some embodiments of the present invention, the present invention provides a kind of Chinese medicine composition of integration of drinking and medicinal herbs, bulk pharmaceutical chemicals
Including arbitrary two kinds or more of group in Chinese yam, arillus longan, mulberry leaf, Exocarpium Citri Rubrum, radix polygonati officinalis, bulbus fritillariae cirrhosae, Rhizoma Atractylodis Macrocephalae, Semen Lablab Album, lotus leaf
It closes.Injury of lungs caused by the Chinese medicine composition can be used for preparing prevention or alleviate pellet (such as caused by PM2.5
Injury of lungs) pharmaceutical preparation.In some embodiments, which can also include not influencing it to pellet
Caused by injury of lungs therapeutic effect other Chinese medicine composition components.
According to some embodiments of the present invention, the bulk pharmaceutical chemicals of Chinese medicine composition are by Chinese yam, arillus longan, mulberry leaf, Exocarpium Citri Rubrum, jade
Two kinds or more of group in bamboo, bulbus fritillariae cirrhosae, Rhizoma Atractylodis Macrocephalae, Semen Lablab Album, lotus leaf is combined into.
According to some embodiments of the present invention, the bulk pharmaceutical chemicals of Chinese medicine composition are by Chinese yam, arillus longan, mulberry leaf, Exocarpium Citri Rubrum, jade
Two kinds or more of group in bamboo, bulbus fritillariae cirrhosae is combined into, and is mainly prevented or is alleviated by improving antioxidant activity and is inhalable
Injury of lungs caused by particulate matter, in the Chinese medicine composition, yam extract, Arillus Longan extract, mulberry-leaf extract, Exocarpium Citri Rubrum carry
Take object, Rhizoma Polygonati Odorati extract, Bulbus Fritillariae Cirrhosae extract mass ratio be 2:16:4:1:32:32.
According to some embodiments of the present invention, the bulk pharmaceutical chemicals of Chinese medicine composition can be by two in Rhizoma Atractylodis Macrocephalae, Semen Lablab Album, mulberry leaf
Kind or three kinds of group are combined into, and lung caused by mainly preventing or alleviating pellet by improving anti-inflammatory activity damages
Wound, in the Chinese medicine composition, Rhizoma Atractylodis Macrocephalae extract, Semen Lablab Album extract, mulberry-leaf extract mass ratio be 1:4:2.
According to some embodiments of the present invention, the bulk pharmaceutical chemicals of Chinese medicine composition can be by two in radix polygonati officinalis, Semen Lablab Album, lotus leaf
Kind or three kinds of group are combined into, caused by mainly preventing by improving anti-apoptotic activity or alleviate pellet
Injury of lungs, in the Chinese medicine composition, Rhizoma Polygonati Odorati extract, Semen Lablab Album extract, Lotus Leafextract mass ratio be 4:1:2.
According to some embodiments of the present invention, the bulk pharmaceutical chemicals of Chinese medicine composition can be made of radix polygonati officinalis and mulberry leaf, main
Injury of lungs caused by preventing by improving macrophage phagocytosis effect or treat pellet, in the Chinese medicine composition
In, the mass ratio of Rhizoma Polygonati Odorati extract and mulberry-leaf extract is 8:1.
According to some embodiments of the present invention, the bulk pharmaceutical chemicals of Chinese medicine composition may include Exocarpium Citri Rubrum, it is Rhizoma Atractylodis Macrocephalae, mulberry leaf, white flat
One or more combinations in beans, radix polygonati officinalis, it is preferable that the bulk pharmaceutical chemicals of Chinese medicine composition can be by Exocarpium Citri Rubrum, Rhizoma Atractylodis Macrocephalae, mulberry leaf, white
One or more groups in hyacinth bean, radix polygonati officinalis are combined into, wherein Exocarpium Citri Rubrum extract, Rhizoma Atractylodis Macrocephalae extract, mulberry-leaf extract, white flat
Beans extract, Rhizoma Polygonati Odorati extract mass ratio be 1-2:1-4:2-6:6-10:30-34, it is preferable that Exocarpium Citri Rubrum extract, Rhizoma Atractylodis Macrocephalae carry
Take object, mulberry-leaf extract, Semen Lablab Album extract, Rhizoma Polygonati Odorati extract mass ratio be 1:2:4:8:32.
According to some embodiments of the present invention, the present invention also provides caused by a kind of prevention or alleviation pellet
The pharmaceutical preparation of injury of lungs, the pharmaceutical preparation include above-mentioned Chinese medicine composition.
According to some embodiments of the present invention, prevent in preparation the present invention also provides above-mentioned Chinese medicine composition or alleviation can
The dosage form of application in the pharmaceutical preparation of injury of lungs caused by sucking particulate matter, the pharmaceutical preparation is selected from tablet, capsule, ball
Agent, aerosol, oral liquid, granule, powder, syrup or combination thereof.
According to some embodiments of the present invention, the present invention also provides a kind of preparation method of Chinese medicine composition, specific steps
It is as follows：By Exocarpium Citri Rubrum, Rhizoma Atractylodis Macrocephalae, mulberry leaf, Semen Lablab Album, radix polygonati officinalis respectively with 10 times of amount distilled water heating and refluxing extractions 2 times, each 2h, mistake
Filter, merging filtrate are concentrated under reduced pressure at 50-60 DEG C, then freeze-dried, obtain the extract of this 5 taste Chinese medicine；Exocarpium Citri Rubrum is extracted
Object, Rhizoma Atractylodis Macrocephalae extract, mulberry-leaf extract, Semen Lablab Album extract and Rhizoma Polygonati Odorati extract in mass ratio 1:2:4:8:32 weigh and mix
It is even, obtain powdered Chinese medicine composition.
The test material and its source that the present invention uses are as follows：
1. Chinese medicine：105 kinds of Chinese medicines are purchased from Beijing three and pharmaceutcal corporation, Ltd.
2. reagent：Superoxide dismutase (SOD) kit, Bioengineering Research Institute is built up purchased from Nanjing；Polyethylene glycol
Octyl phenyl ether Triton-X 100 is purchased from Shanghai Sheng Gong bioengineering Co., Ltd；Mouse malonaldehyde (MDA) ELISA reagents
Box, people's malonaldehyde (MDA) ELISA kit, mouse active oxygen (ROS) ELISA kit, people's active oxygen (ROS) ELISA reagents
Box, mouse glutathione peroxidase (GSH-Px) ELISA detection kit, glutathione peroxidase (GSH-Px)
ELISA detection kit, RIPA tissue/cells lysate, mouse interleukin 8 (IL-8) ELISA kit, human leukocytes
Interleukin 8 (IL-8) ELISA kit, mouse interleukin 6 (IL-6) ELISA kit, human interleukin 6 (IL-6)
ELISA kit, 1 β of mouse interleukin (IL-1 β) ELISA kit, Human interleukin 1 beta (IL-1 β) ELISA reagents
Box, murine tumor necrosis factor ELISA detection kit and human tumour necrosis factor ELISA detection kit, are purchased from Shanghai
Enzyme-linked bio tech ltd；Tetramethyl azo azoles salt (MTT) is purchased from Shanghai Sheng Gong bioengineering Co., Ltd；One-step method
TUNEL Apoptosis in-situ detection reagents box, apoptotic mitochondrial film potential detection kit (JC-1), Annexin V-
The bis- dye cell apoptosis detection kits of kFluor488/PI, Caspase inhibitor Z-VAD-FMK, purchased from triumphant base biology；
Caspase3 Antibody, Caspase8 Antibody, Caspase9 Antibody, it is safe purchased from Tianjin profit.
3. cell line：Non-small cell lung carcinoma cell line (A549) is purchased from Nanjing KaiJi Biology Science Development Co., Ltd
4. experimental animal：SPF grades of Kunming 18~22g of male mice, while buying feed (national standard rodent is dry
Dry feed).
5. instrument：AR2202CN electronic balances are purchased from Shanghai Ao Haosi Instrument Ltd.；Tabletop refrigerated centrifuge,
Thermo LEGEND MICRO ZIR；BCD-192KJ4 degree refrigerators are purchased from Qingdao HaiEr Co., Ltd；DW-86L286 is vertical
Body ultra-low temp is purchased from Qingdao Haire Special Electrical Appliances Co., Ltd；Enzyme-linked immunosorbent assay instrument, Thermo fisher
scientific；Japanese Shimadzu UV-1800 ultraviolet specrophotometers are purchased from Shanghai Milky Way Environmental Technology AS；YSL-1A
Multifunctional mouse autonomic activities recorder is purchased from Jinan Yi Yan development in science and technology Co., Ltd；Common thermometer, purchased from Shaanxi medical treatment
Instrument factory；0.22 μm of filter, Millipore；Bio II A Biohazard Safety Equipments, Thailand of Spain thing reach (Telstar)；STIK
IL-161HI CO2Incubator is purchased from Shi Doukai instrument and equipments (Shanghai) Co., Ltd.；CKX41 inverted phase contrast microscopes, Olympic
Bath；96 orifice plates of Thermo Nunc are purchased from Thermo Fisher Scientific Inc.；BX53 just sets fluorescence microscope, purchased from Austria
Woods Bath.
6. particulate matter PM：The PM in air is acquired using PM high power capacity automatic sampling machines.
The preparation method of drug and reagent used in 7.：(1) preparation of single medicinal material extract test sample：It is distilled with 10 times of amounts
Water heating and refluxing extraction 2 times, each 2h, filtering, merging filtrate are concentrated under reduced pressure at 50-60 DEG C, then freeze-dried, in obtaining
Drug composition extract；Chinese medicine composition extract 100mg is dissolved in (10mg/ml) ultrasonic dissolution in 1640 culture medium 10ml to obtain
To mother liquid medicine, with being used after 0.22 μm of filter filtration sterilization.Matching while using, the preparation and use of solution should all be in sterilized bios
It is operated in safety cabinet；(2) preparation of Chinese medicine composition test sample：Using distilled water as solvent in mouse test, by powdered Chinese medicine
Composition is configured to 0.5g/ml, 1g/ml and 2g/ml3 concentration；Using fertilized eggs culture solution as solvent (fertilization in zebra fish test
Ovum culture solution configuration method：Sodium chloride 3.5g, potassium chloride 0.05g, sodium bicarbonate 0.025g, calcium chloride 0.1g, are dissolved in distilled water
In, it is used in combination distilled water to mend to 1L), powdered Chinese medicine composition is configured to 5mg/ml；(3) test cell line PM suspension is matched
It sets：20mg PM particulate matters are weighed loaded in 4ml EP pipes, sterilizes in high steam pot mesohigh, is added in superclean bench
1640 culture mediums of 2ml dissolve, and the PM Particulate suspension mother liquors of a concentration of 10mg/ml are made, are sealed with sealed membrane, at ultrasound
1h is managed, matching while using is configured to required concentration with 1640 culture mediums and is ultrasonically treated 10min when handling cell；(4) animal experiment
With the configuration of PM suspension：Weigh 0.5g PM be dissolved in be made into 0.9% physiological saline of 20mL a concentration of 25mg/ml PM it is outstanding
Supernatant liquid is ultrasonically treated 1h, makes particle is mixed to dissipate uniformly, it is spare to be positioned over 4 DEG C of refrigerators.
The characterization of 1 pellet of embodiment
Analyze particles used particle size using Particle Size Analyzer and obtain its particle diameter distribution, by inductive coupling etc. from
Element in daughter mass spectrum (ICP-MS) Atomic Emission Spectral Analysis sample and its content are analyzed using X-ray diffraction in sample
Compound and its content, the result is shown in Figure 1 .1, table 1 and 2, wherein Fig. 1 .1 show used in particulate matter size distribution, to the greatest extent
Use the particulate matter with the size distribution in Fig. 1 .1 in the pipe present invention, but the present invention is directed used
Injury of lungs caused by pellet (i.e. grain size is in 10 microns of particulate matters (i.e. PM10) below) in grain object.
The element of 1 particulate matter of table forms and content
The compound of 2 particulate matter of table forms and content
Embodiment 2 establishes PM and causes alveolar epithelium A549 cellular damages --- oxidative stress, inflammation and Apoptosis Model
1. establish PM cause A549 cellular oxidations stress and inflammatory model
Using 1640 culture mediums by PM mother liquors be diluted to successively 400 μ g/ml, 200 μ g/ml, 100 μ g/ml, 50 μ g/ml and
The PM suspensions of 25 μ g/ml act on A549 cells 6h, 12h, for 24 hours and 48h respectively, measure superoxide dismutase in cell pyrolysis liquid
(SOD) interleukin-6 (IL-6) content in content and cell culture fluid is used in combination mtt assay to measure cell survival rate, is contained with SOD
The ratio of amount and cell survival rate is index, and evaluation various concentration PM acts on different time to A549 cellular oxidation stress situations
It influences；Using the ratio of IL-6 contents and cell survival rate as index, evaluation various concentration PM acts on different time to A549 cells
The influence of inflammatory conditions.
As a result see Fig. 2 .1a and 2.1b.Fig. 2 .1a show the PM effect 48h of 200 μ g/ml and 400 μ g/ml, A549 cell oxygen
Change stress be damaged the most seriously, because the PM of 400 μ g/ml keeps cell survival rate relatively low, to ensure the lasting progress of follow-up test, choosing
The PM effects 48h of fixed 200 μ g/ml is to cause A549 cellular oxidation Stress model conditions；
Fig. 2 .1b show the PM effect 6h of 200 μ g/ml and 400 μ g/ml, the content of A549 cell secretes interleukins 6
At most, the PM of 200 μ g/ml is unlikely to too big to the survival rate of cell again, and the PM effects 6h of selected 200 μ g/ml is to cause
A549 cellular inflammation Model Conditions.
2. establishing PM causes A549 model of cell apoptosis
Using 1640 culture mediums by PM mother liquors be diluted to successively 400 μ g/ml, 200 μ g/ml, 100 μ g/ml, 50 μ g/ml and
The PM suspensions of 25 μ g/ml act on respectively A549 cells for 24 hours, 48h, 72h and 96h, utilize mtt assay measure cell survival rate, screening
Go out has the PM concentration significantly affected and action time to A549 cell survival rates；Using the bis- dyeing of Annexin V-FITC/PI
The PM that method detects 200 μ g/ml acts on the apoptosis situation with the A549 cells of 48h for 24 hours；Using TUNEL immunostainings experiment (according to
It is two above experimental result, selected that cell is caused the PM concentration of apparent apoptotic state and action time occur) verification PM causes
The condition of A549 Apoptosis；Since numerous studies show two classical pathway mitochondrial approach and the ways TNF of Apoptosis
Diameter, thus it is thin with the A549 of 48h for 24 hours by 200 μ g/ml PM effects using mitochondrial membrane potential detection kit (JC-1) detection
Born of the same parents' injury of mitochondria situation is different using solid phase sandwich method enzyme-linked immunosorbent assay (ELISA) detection various concentration PM effects
The expression of the tumor necrosis factor (TNF-α) of time A549 cell, thus illustrates that PM causes the approach of A549 Apoptosis,
The optimum condition that PM causes A549 Apoptosis is further verified simultaneously.
As a result see Fig. 2 .2a, 2.2b, 2.2c, 2.2d, 2.2e, 2.2f.Fig. 2 .2a show PM with time and dose-dependent
Mode reduces A549 cell survival rates, and compared with the control group, when the PM of 200 μ g/ml acts on 48h, A549 cell survival rates are notable
It reduces.Fig. 2 .2b show that control group A nnexinV-FITC/PI is negative, and illustrate that cell membrane is complete, outside no phosphatidylserine
It turns over, cell is normal；There are the Annexin V-FITC positives in part cell for 24 hours for PM effects, and PI is negative, illustrates the cell of some cells
Membrane phospholipid acyl serine turns up but cell is touched and still preserved completely, this is early apoptosis of cells feature；PM acts on 48h cells and presents
Annexin V-FITC are positive, and PI is positive, illustrates that cell membrane is imperfect, cell is in late apoptic.Fig. 2 .2c are shown by 200 μ
It is Q1+Q2=5.17% that the A549 cells of the PM effects of g/ml for 24 hours, which generate the ratio of apoptosis, and the A549 cells for acting on 48h generate
The ratio of apoptosis is Q1+Q2=17.85%, apoptosis ratio significantly raised (the AnnexinV positives, Propidium iodide negative (lower right)
Reflect the ratio of early apoptosis；AnnexinV is positive, and propidium iodide positive (upper right side) reflects late apoptic and the ratio of necrosis
Rate).Fig. 2 .2d show that the A549 cell cores concentration that 48h is acted on by the PM of 200 μ g/ml, dyeing are deepened, and nucleus is in crescent
Shape starts apoptosis.Fig. 2 .2e show the increase of the PM action times with 200 μ g/ml, and mitochondrial membrane potential, which depolarizes, to be made
Red fluorescence gradually weakens, and illustrates that mitochondrial transmembrane potentials destroy, and cell starts apoptosis, i.e. endogenous apoptosis approach is induced in PM
A549 Apoptosis in exist.Fig. 2 .2f show with the concentration of PM and action time increase, the expression quantity of TNF-α in concentration and
Dose dependent rises, and shows that exogenous apoptosis pathway exists in the A549 Apoptosis that PM is induced, and compares blank group, 200 μ
The expression quantity of TNF-α has significant difference when the PM effect 48h of g/ml.In conclusion the PM of selected 200 μ g/ml acts on 48h
To cause the condition of A549 model of cell apoptosis.
Embodiment 3 establishes Chinese medicine composition active ingredient entity library
It requires to be standardized extraction, concentration, freeze-drying and Quick Response Code mark, profit to 105 taste Chinese medicines according to food-grade
Finger-print is established with 105 extracts of efficient liquid phase (HPLC) and thin-layer chromatography (TLC) method pair.Chinese medical extract preparation flow
See Fig. 3 .1.
The screening of embodiment 4 causes Chinese medicine composition of the A549 cellular damages with mitigation to PM
1. screening the best administration concentration of 105 Chinese medical extracts
A549 cells to be tested are divided into 632 groups, respectively blank control group (Control), model group (Model) and
The 6 of 0.3125mg/ml, 0.625mg/ml, 1.25mg/ml, 2.5mg/ml, 5mg/ml, 10mg/ml of 105 Chinese medical extracts
A concentration group, after the Chinese medical extracts of 632 Chinese medical extract group respective concentrations intervenes 1h, model group and Chinese medical extract group
48h is acted on the PM2.5 of 200 μ g/ml, cell survival rate is measured with mtt assay, goes out 105 by index screening of cell survival rate
The best administration concentration of Chinese medical extract, the results are shown in Table 3.
The best administration concentration of 3 105 Chinese medical extracts of table
Cause A549 cellular oxidations stress be with the Chinese medical extract of mitigation PM 2. screening
(1) screening and verification
A549 cells to be tested are divided into 107 groups, respectively blank control group (Control), model group (Model), 105
A Chinese medical extract group (administration concentration reference table 3).After 105 Chinese medical extract groups intervene 1h with corresponding Chinese medical extract, mould
Type group and Chinese medical extract group act on 48h with the PM2.5 of 200 μ g/ml, measure superoxide dismutase (SOD) content, are used in combination
Mtt assay measures cell survival rate, and using the ratio of SOD contents and cell survival rate as index, SOD contents can be significantly improved by filtering out
Chinese medical extract；Anti-oxidation stress activity to screening obtained Chinese medical extract does repeated verification；In conjunction with Chinese medicine pharmacology
By, further filter out with tonifying lung benefit spleen, the Chinese medical extract of tonifying Qi nourishing yin effect, and pass through measure active oxygen (ROS), paddy
The sweet peptide peroxidase (GSH-Px) of Guang, malonaldehyde (MDA) cause oxidative stress status in the PM of Chinese medical extract intervention
Content (administration concentration is with drug treating time with the screening experiment for measuring ROS) in A549 cell culture mediums deeply evaluates and screens
The anti-oxidation stress activity of obtained Chinese medical extract；In addition, testing (principle using ABTS：ABTS2,2- azines-(3- ethyls
Benzothiazoline -6- sulfonic acid) di-ammonium salts] it is not oxidized before be it is white, can be by K2S2O8Etc. reagent oxidations, generate it is glaucous
Radical cation ABTS+.In the presence of antioxidant (such as aldehydes matter) for having hydrogen supply capacity, ABTS+ is reacted,
Become do not have coloured ABTS.The ability of antioxidant for clearing ABTS+ can use the measure of the change of absorbance at 414 or 734nm)
In the antioxidant activity for the Chinese medical extract that the verification screening of chemical level obtains.
Table 4 ABTS experiments measure the antioxidant activity of Chinese medical extract
Note：The sample concentration that EC50 is free radical scavenging activity when being 50%, value is smaller to show that antioxidant activity is stronger；
TEAC values show that more greatly antioxidant activity is stronger.
As a result see Fig. 4 .1a, 4.1b, 4.1c, 4.1d, 4.1e and table 4.The most of Chinese medicine composition of Fig. 4 .1a displays all has
There is antioxidant activity, but antioxidant activity size has otherness, therefrom filters out 21 more significant Chinese medical extracts of activity；
Fig. 4 .1b show that the antioxidant activity of 21 taste Chinese medical extracts has repeatability, and in conjunction with Fig. 4 .3a, it is cytotoxic to exclude tool
Chinese medical extract obtains 6 Chinese medical extracts；Fig. 4 .1c, 4.1d, 4.1e and table 4 are by measuring biology index of correlation and change
Method proves that last screening obtains Chinese yam, arillus longan, mulberry leaf, Exocarpium Citri Rubrum, radix polygonati officinalis and bulbus fritillariae cirrhosae this 6 Chinese medical extracts and all has
Significant antioxidant activity.According to some embodiments, yam extract, Arillus Longan extract, mulberry-leaf extract, Exocarpium Citri Rubrum extraction
Object, Rhizoma Polygonati Odorati extract, Bulbus Fritillariae Cirrhosae extract mass ratio be 2:16:4:1:32:32.
(2) Study on Molecular Mechanism
A549 cells to be tested are divided into 9 groups, respectively blank control group (Control), model group (Model), the positive
Medicine group (N-acetyl-L-cysteine (NAC)) and 6 Chinese medical extract groups (administration concentration reference table 3), positive drug group and 6
After Chinese medical extract group intervenes 1h with relative medicine, the PM2.5 of 200 μ g/ml of model group, positive drug group and Chinese medical extract group
48h is acted on, using protein immunoblot technology, 6 Chinese medical extracts that detection screening obtains are logical to endogenous anti-oxidation stress
Total Nrf2 in road, Nrf2 in endochylema, in core Nrf2 and heme oxygenase 1 (HO-1) protein expression level influence, into one
Step, which is inquired into, causes A549 cellular oxidations stress be with the mechanism of action of the Chinese medical extract of mitigation PM.
As a result see Fig. 4 .1f, the Chinese medical extract that display screening obtains can improve total Nrf2 to some extent, Nrf2 in core
With the expression of heme oxygenase 1 (HO-1), the expression of Nrf2 in endochylema is reduced to some extent, thus infers sieve
It selects 6 obtained Chinese medical extracts may be by promoting Nrf2 core transpositions, and then promotes the expression of heme oxygenase 1 (HO-1)
And play alleviate PM cause A549 cellular oxidations stress effect.
3. screening the Chinese medical extract for causing A549 cellular inflammations that there is mitigation PM
(1) screening and verification
A549 cells to be tested are divided into 107 groups, respectively blank control group (Control), model group (Model), 105
A Chinese medical extract group (administration concentration reference table 3), after 105 Chinese medical extract groups intervene 1h with corresponding Chinese medicine composition, mould
Type group and Chinese medical extract group act on 6h with the PM2.5 of 200 μ g/ml, measure interleukin-6 (IL-6) content, mtt assay is used in combination
Cell survival rate is measured, using the ratio of IL-6 contents and cell survival rate as index, IL-6 contents can be significantly improved by filtering out
Chinese medical extract；Anti-inflammatory activity to screening obtained Chinese medical extract does repeated verification；In conjunction with traditional Chinese medical theory, further
It filters out with tonifying lung benefit spleen, the Chinese medical extract of tonifying Qi nourishing yin effect, and by measuring interleukin 8 (IL-8), it is white thin
Born of the same parents' interleukin 1 beta (IL-1 β), tumor necrosis factor (TNF-α) cause the A549 cells of inflammatory conditions in the PM of Chinese medical extract intervention
Deeply evaluating and screening obtains content (administration concentration is with drug treating time with the screening experiment for measuring IL-6) in culture medium
The anti-inflammatory activity of Chinese medical extract.
As a result see Fig. 4 .2a, 4.2b, 4.2c, 4.2d, 4.2e.Fig. 4 .2a display portion Chinese medical extracts have anti-inflammatory work
Property, and active size has otherness, therefrom filters out 10 more significant Chinese medical extracts of activity；Fig. 4 .2b are shown in 10
The anti-inflammatory activity of medicament extract has repeatability, in conjunction with Fig. 4 .2a, excludes have cytotoxic Chinese medical extract, obtains 3 tastes
Chinese medical extract；Fig. 4 .2c, 4.2d, 4.2e prove that last screening obtains Rhizoma Atractylodis Macrocephalae, Semen Lablab Album by measuring biology index of correlation
This 3 taste Chinese medical extract all has significant anti-inflammatory activity with mulberry leaf.According to some embodiments, Rhizoma Atractylodis Macrocephalae extract, Semen Lablab Album carry
Take object, the mass ratio of mulberry-leaf extract is 1:4:2.
(2) Study on Molecular Mechanism
A549 cells to be tested are divided into 5 groups, respectively blank control group (Control), model group (Model), in 3
Medicament extract group (administration concentration reference table 3), 3 Chinese medical extract groups with corresponding Chinese medicine composition intervene 1h after, model group and
Chinese medical extract group acts on 6h with the PM2.5 of 200 μ g/ml, extracts and detects NF- in (polyacrylamide gel electrophoresis method) endochylema
In κ B and I κ B α, karyon in NF- κ B and total protein STAT3 content, further inquire into and cause A549 cellular inflammations to have PM
The mechanism of action of the Chinese medical extract of mitigation.
As a result see Fig. 4 .2f, show that the Chinese medical extract screened can be substantially reduced expression of the NF- κ B in core, rise
Expression of high NF- κ B and I κ the B α in endochylema, reduces the expression of STAT3, thus infers that the 3 taste Chinese medical extracts that screening obtains can
It can be the expression by increasing I κ B α in endochylema, and then inhibit NF- κ B nuclear imports and play anti-inflammatory effect, while in 3 tastes
Medicament extract may inhibit increased inflammation by reducing the expression of STAT3.
4. screening the Chinese medical extract for causing A549 Apoptosis that there is mitigation PM
(1) screening and verification
A549 cells to be tested are divided into 632 groups, respectively blank control group (Control), model group (Model) and
The 0.3125mg/ml of 105 Chinese medical extracts, 0.625mg/ml, 1.25mg/ml, 2.5mg/ml, the 6 of 5mg/ml, 10mg/ml
A concentration group, after the Chinese medicine compositions of 630 Chinese medical extract group respective concentrations intervenes 1h, model group and Chinese medical extract group
48h is acted on the PM2.5 of 200 μ g/ml, cell survival rate is measured with mtt assay, goes out to improve using cell survival rate as index screening thin
The Chinese medical extract of born of the same parents' survival rate best results；The Chinese medicine obtained using the verification screening of Annexin V-FITC/PI double-stainings
The anti-apoptotic activities of extract；The Chinese medical extract pair obtained using mitochondrial membrane potential detection kit (JC-1) detection screening
PM causes the influence that A549 cell mitochondrials damage, and further verifies its anti-apoptotic activities；It is verified using TUNEL immunostainings
Screen the anti-apoptotic activities (administration concentration reference table 3) of obtained Chinese medical extract.
As a result see Fig. 4 .3a, 4.3b, 4.3c, 4.3d.Cell survival rate experiment display portion Chinese medicine is measured using mtt assay to carry
It takes object that there is the activity for improving cell survival rate, therefrom filters out 3 more significant Chinese medical extracts of activity；Fig. 4 .3a show 3
A Chinese medical extract makes the Annexin V-FITC positives occur, and the cell of the PI positives is substantially reduced, and illustrates 3 Chinese medical extracts
A549 apoptosis status caused by can alleviating PM；Fig. 4 .3b show that 3 taste Chinese medical extracts can be substantially reduced PM and cause
The incidence of A549 Apoptosis；4.3c shows that the ratio of red fluorescence/green fluorescence of 3 Chinese medical extract groups is apparently higher than
Model group illustrates that 3 Chinese medical extracts can be relieved the incidence that PM causes A549 cell mitochondrials to damage；4.3d display
The TUNE positive quantity of 3 Chinese medical extract groups significantly reduces, and illustrates that 3 Chinese medical extracts have anti-apoptotic activities.To sum up institute
It states, this 3 Chinese medical extracts of radix polygonati officinalis, Semen Lablab Album and lotus leaf all have significant anti-apoptotic activity.According to some embodiments,
Rhizoma Polygonati Odorati extract, Semen Lablab Album extract, Lotus Leafextract mass ratio be 4:1:2.
(2) Study on Molecular Mechanism
A549 cells to be tested are divided into 6 groups, respectively blank control group (Control), model group (Model), apoptosis
Inhibitor group (Z-VAD), 3 Chinese medical extract groups (administration concentration reference table 3), inhibitors of apoptosis group and 3 Chinese medical extracts
After group uses z-VAD-fmk (a concentration of 0.25uM, solvent are cell culture fluid) and respective concentration Chinese medical extract to intervene 1h respectively,
Model group, inhibitors of apoptosis group and 3 Chinese medical extract groups act on 48h with the PM2.5 of 200 μ g/ml, are detected using mtt assay thin
Born of the same parents' survival rate；A549 cells to be tested are divided into 5 groups, respectively blank control group (Control), model group (Model), 3
Chinese medical extract group (administration concentration reference table 3), after 3 Chinese medical extract groups intervene 1h with corresponding Chinese medical extract, model group
48h is acted on the Chinese medical extract group PM2.5 of 200 μ g/ml, the weight of apoptosis relational approach is detected using Western Blot methods
The expression of albumen is wanted, the caspase-8 albumen of the caspase-9 albumen of mitochondria pathway and TNF approach is included, is further inquired into
The mechanism of action of Chinese medical extract of the A549 Apoptosis with mitigation is caused to PM.
As a result see Fig. 4 .3e and Fig. 4 .3f.Fig. 4 .3e show that inhibitors of apoptosis group and 3 Chinese medical extract groups can be shown
The phenomenon that A549 cell survival rates reduce caused by reversing PM is write, since z-VAD-fmk is extensive caspase inhibitor, outside
The downstream albumen of source property and endogenous apoptosis approach belongs to caspase families, thus it is speculated that 3 Chinese medical extracts may pass through inhibition
The relevant caspase protein expressions of apoptosis and play a role；The Chinese medical extract that Fig. 4 .3f display screenings obtain can inhibit
Caspase-9 and caspase-8 protein activations, and then inhibit caspase-3 protein activations, wherein radix polygonati officinalis inhibits with lotus leaf
The effect of caspase-9 protein activations inhibits caspase-9 better than the inhibiting effect to caspase-8 protein activations, Semen Lablab Album
It is more significant with caspase-8 protein activations, thus infer that radix polygonati officinalis may be mainly by inhibiting in mitochondria pathway with lotus leaf
Caspase-9 protein activations, Semen Lablab Album may be by inhibiting in mitochondria pathway in caspase-9 albumen and TNF approach
Caspase-8 protein activations, and then inhibit the activation of caspase-3 and play and alleviate the effect that PM causes A549 Apoptosis.
The screening of embodiment 5 accelerates the Chinese medical extract of macrophage phagocytosis PM
RAW macrophages to be tested are divided into 632 groups, respectively blank control group (Control), model group (Model)
And the 0.3125mg/ml of 105 Chinese medical extracts, 0.625mg/ml, 1.25mg/ml, 2.5mg/ml, 5mg/ml, 10mg/ml's
6 concentration groups, after the Chinese medical extracts of 630 Chinese medical extract group respective concentrations intervenes 1h, model group and Chinese medical extract
Group acts on 48h with the PM of 200 μ g/ml, and the neutral red solution that a concentration of 0.075mol/ml is then added per hole is incubated 3h, uses PBS
It washes 2 times, measures absorbance, go out to improve the traditional Chinese medicine extraction that RAW macrophages swallow PM best results by index screening of absorbance
Object.
As a result show that it is radix polygonati officinalis, mulberry leaf to improve the Chinese medical extract of RAW macrophages phagocytosis PM best results.According to some
The mass ratio of embodiment, Rhizoma Polygonati Odorati extract and mulberry-leaf extract is 8:1.
6 traditional Chinese medicine compatibility theory of embodiment instructs prescription to design
The optimal Chinese medical extract tested according to above-mentioned in-vitro screening, we are set up using Tsinghua University Lee tip project
Vertical drug matching network Herb network carry out prescription design, in further determining that above-mentioned optimal Chinese medical extract is corresponding
Whether the combination between any two that medicine is formed had application in successive dynasties Chinese medicinal formulae, if using, just it is believed that two
The combination of the extract of person has enough rationality of combination prescription and safety.See Fig. 5 .1 (figure in Fig. 5 .1 is selected from Li S*,
Zhang B,Jiang D,Wei YY,Zhang NB.Herb network construction and co-module
analysis for uncovering the combination rule of traditional Chinese herbal
formulae.BMC Bioinformatics 2010；11(S11):S6), the node in the network is Chinese medicine, the color of node
Herbal nature is indicated, while being the combination of two between Chinese medicine, the thicker combination for indicating two taste Chinese medicines in successive dynasties Chinese medicinal formulae in side
The possibility of compatible use is higher.By the corresponding Chinese medicine of Chinese medical extract screened respectively in aforementioned four model (such as
Prevent the best mulberry leaf of oxidative stress effect, Exocarpium Citri Rubrum；The best Rhizoma Atractylodis Macrocephalae of prevention of inflammation effect, mulberry leaf, Semen Lablab Album；Prevent cell to wither
Die the best radix polygonati officinalis of effect, Semen Lablab Album；Promoting the best radix polygonati officinalis of macrophage phagocytosis effect, mulberry leaf) co-map is to Herb
In network, the combination of Exocarpium Citri Rubrum, Rhizoma Atractylodis Macrocephalae, mulberry leaf, Semen Lablab Album, radix polygonati officinalis this traditional Chinese medicine of the five flavours is obtained, has as a result shown them in the successive dynasties
Usually match between any two in Chinese medicinal formulae and occurs together.In addition, the optimal dosage according to experiment in vitro determines Chinese medicine
The compatibility dosage of composition is Exocarpium Citri Rubrum extract, Rhizoma Atractylodis Macrocephalae extract, mulberry-leaf extract, Semen Lablab Album extract and Rhizoma Polygonati Odorati extract matter
Measure ratio 1:2:4:8:32.
Influence of 7 Chinese medicine composition of embodiment to PM drainage rates in zebra fish body
The preparation of Chinese medicine composition：Exocarpium Citri Rubrum, Rhizoma Atractylodis Macrocephalae, mulberry leaf, Semen Lablab Album, radix polygonati officinalis are heated back with 10 times of amount distilled water respectively
Stream extraction 2 times, each 2h, filtering, merging filtrate is concentrated under reduced pressure at 50-60 DEG C, then freeze-dried, obtains this 5 taste Chinese medicine
Extract；By Exocarpium Citri Rubrum extract, Rhizoma Atractylodis Macrocephalae extract, mulberry-leaf extract, Semen Lablab Album extract and Rhizoma Polygonati Odorati extract in mass ratio 1:
2:4:8:32 weigh simultaneously mixing, obtain powdered Chinese medicine composition.
Experiment is divided into two big groups with zebra fish-egg：I.e. A groups are not added with 1- phenyl -2- thiocarbamides (PTU) and B groups addition PTU
(solvent is fertilized eggs culture solution, 0.0045%) concentration is that wherein 3 groups of component A, that is, blank control group is not (as mass fraction
Add PM), model group (only addition PM), Chinese medicine composition group (addition PM and Chinese medicine composition), 3 group of B component (with A groups),
Per 20 pieces of fertilized eggs of group.It is fertilized after 8h, by grouping situation, each group is not added with or is added accordingly PTU and Chinese medicine composition (administration
Mode is directly to replace with original fertilized eggs culture solution containing PTU or the culture solution of Chinese medicine composition (and)), after fertilization for 24 hours,
By grouping situation, each group is not added with or is added accordingly PM, i.e., to yolk bag microinjection magnetic ferric oxide nano ball, (solvent is to steam
Distilled water, a concentration of 5mg/mL) 2 μ l.The zebra fish of 48h (i.e. after fertilization 72h) after injection is placed in stereomicroscopy microscopic observation, is clapped
According to and preserve.
As a result see Fig. 6 .1, show compared with A groups, the zebrafish embryo relative transparent of B groups, and embryonism is without significance difference
It is different, show that injection PTU has played the effect for inhibiting melanin secretion, eliminates zebra fish dermal melanin to observation PM excretions
The interference of process；In B groups, compared with blank group, observes black deposit in model group zebrafish embryo yolk bag, show magnetic
Property ferric oxide nano ball is successfully injected；In B groups, compared with model group, in Chinese medicine composition group zebrafish embryo yolk bag
Black deposit relatively disperses, and shows excretion facilitation of the Chinese medicine composition to PM in zebra fish body.
8 Chinese medicine composition of embodiment causes PM the influence of mouse injury of lungs
The preparation of Chinese medicine composition：Exocarpium Citri Rubrum, Rhizoma Atractylodis Macrocephalae, mulberry leaf, Semen Lablab Album, radix polygonati officinalis are heated back with 10 times of amount distilled water respectively
Stream extraction 2 times, each 2h, filtering, merging filtrate is concentrated under reduced pressure at 50-60 DEG C, then freeze-dried, obtains this 5 taste Chinese medicine
Extract；By Exocarpium Citri Rubrum extract, Rhizoma Atractylodis Macrocephalae extract, mulberry-leaf extract, Semen Lablab Album extract and Rhizoma Polygonati Odorati extract in mass ratio 1:
2:4:8:32 weigh simultaneously mixing, obtain powdered Chinese medicine composition.
Mouse is randomly divided into 5 groups, respectively blank control group (Control), and model group (Model) and Chinese medicine composition are high
(2g/ml), in (1g/ml), low (0.5g/ml) dosage group, every group 5.First week, blank control group, model group mouse were daily
Gavage 0.2ml distilled water, the daily gavage 0.2ml corresponding concentration Chinese medicine compositions of Chinese medicine composition group mouse；After a week, blank pair
According to daily 0.9% physiological saline of tracheal instillation 0.4ml of group mouse while gavage 0.2ml distilled water, the daily tracheae of model group mouse
Instillation 0.4ml PM2.5 suspensions (25mg/ml is configured with 0.9% physiological saline) while gavage 0.2ml distilled water, Chinese traditional medicine composition
The Chinese medicine composition of the daily tracheal instillation 0.4ml PM2.5 suspensions of object group mouse while gavage 0.2ml corresponding dosages, until experiment
Terminate, entire experiment process continues 30 days.
1. Chinese medicine composition causes PM the influence of mouse injury of lungs
The experiment mice neck that breaks is put to death dissection and obtains fresh lung tissue and observes after experiment.
As a result see Fig. 7 .1, display control group lung form rule is complete, without apparent lesion；The blackening of model group lung tissue,
There is atrophy；There is part blackening in Chinese medicine composition group, and lung's state is better than model group, and middle and high concentration group effect be better than it is low
Concentration group.
2. Chinese medicine composition causes PM the influence of mouse oxidative stress status
Mice serum and lung homogenate carry out SOD, ROS, GSH-Px and MDA assay after to experiment.
As a result see that Fig. 7 .2, display Chinese medicine composition low, middle and high dose groups can reduce ROS and MDA contents, increased SOD
With GSH-Px contents, it was demonstrated that the Chinese medicine composition can improve the oxidative stress status of mouse, and middle and high dosage group and model really
Group is compared, ROS and MDA contents significantly reduce, SOD and GSH-Px contents significantly increase, this illustrates Chinese medicine composition to injury of lungs
There is good remission effect.
3. Chinese medicine composition causes PM the influence of mouse inflammatory conditions
Mouse bronchial irrigating solution, lung homogenate and serum progress IL-6, IL-8, IL-1 β and TNF-α contain after to experiment
It is fixed to measure.
As a result see that Fig. 7 .3, display Chinese medicine composition low, middle and high dose groups can reduce IL-6, IL-8, IL-1 β and TNF-
The content of α, it was demonstrated that the Chinese medicine composition can improve the inflammatory conditions of mouse, and low, middle and high dose groups and model group phase really
Than significantly above four indexs decline.
The above is merely preferred embodiments of the present invention, be not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of Chinese medicine composition, which is characterized in that the bulk pharmaceutical chemicals of the Chinese medicine composition include Chinese yam, arillus longan, mulberry leaf, tangerine
Two kinds or more of combination in red, radix polygonati officinalis, bulbus fritillariae cirrhosae, Rhizoma Atractylodis Macrocephalae, Semen Lablab Album, lotus leaf.
2. Chinese medicine composition according to claim 1, which is characterized in that the bulk pharmaceutical chemicals of the Chinese medicine composition by Chinese yam,
Two kinds or more of group in arillus longan, mulberry leaf, Exocarpium Citri Rubrum, radix polygonati officinalis, bulbus fritillariae cirrhosae, Rhizoma Atractylodis Macrocephalae, Semen Lablab Album, lotus leaf is combined into.
3. Chinese medicine composition according to claim 1, which is characterized in that the bulk pharmaceutical chemicals of the Chinese medicine composition by Chinese yam,
Two kinds or more of group in arillus longan, mulberry leaf, Exocarpium Citri Rubrum, radix polygonati officinalis, bulbus fritillariae cirrhosae is combined into.
4. Chinese medicine composition according to claim 1, which is characterized in that the bulk pharmaceutical chemicals of the Chinese medicine composition by Rhizoma Atractylodis Macrocephalae,
Two or three of group in Semen Lablab Album, mulberry leaf is combined into.
5. Chinese medicine composition according to claim 1, which is characterized in that the bulk pharmaceutical chemicals of the Chinese medicine composition by radix polygonati officinalis,
Two or three of group in Semen Lablab Album, lotus leaf is combined into.
6. Chinese medicine composition according to claim 1, which is characterized in that the bulk pharmaceutical chemicals of the Chinese medicine composition by radix polygonati officinalis and
Mulberry leaf form.
7. Chinese medicine composition according to claim 1, which is characterized in that the bulk pharmaceutical chemicals of the Chinese medicine composition by Exocarpium Citri Rubrum,
Two kinds or more of group in Rhizoma Atractylodis Macrocephalae, mulberry leaf, Semen Lablab Album, radix polygonati officinalis is combined into.
8. Chinese medicine composition according to claim 7, which is characterized in that in the Chinese medicine composition, Exocarpium Citri Rubrum extract,
Rhizoma Atractylodis Macrocephalae extract, mulberry-leaf extract, Semen Lablab Album extract, Rhizoma Polygonati Odorati extract mass ratio be 1-2:1-4:2-6:6-10:30-
34。
9. the pharmaceutical preparation of injury of lungs caused by a kind of prevention or alleviation pellet, which is characterized in that the drug system
Agent includes claim 1-8 any one of them Chinese medicine compositions.
10. claim 1-8 any one of them Chinese medicine composition lung in preparation prevention or caused by alleviating pellet
Application in the pharmaceutical preparation of damage.
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