CN102885878B

CN102885878B - Application of effective parts of astragalus mongholicus and hedysarum polybotrys

Info

Publication number: CN102885878B
Application number: CN201210374823.1A
Authority: CN
Inventors: 李金田; 李娟�; 刘永琦; 景明; 张毅; 蔺兴遥; 李雪燕; 苏蕴; 孙少伯
Original assignee: GANSU CHINESE OF TRADITIONAL CHINESE MEDICINE
Current assignee: GANSU CHINESE OF TRADITIONAL CHINESE MEDICINE
Priority date: 2012-09-29
Filing date: 2012-09-29
Publication date: 2014-10-15
Anticipated expiration: 2032-09-29
Also published as: CN102885878A

Abstract

The invention belongs to the traditional Chinese medicine field and relates to the preparation of effective parts of astragalus mongholicus and hedysarum polybotrys and selection thereof for pulmonary fibrosis model intervention so as to obtain further study on the effective parts having the optimal curative effect. An interstitial pulmonary fibrosis model of rats is established through modeling; and compared with the normal rats, the rats having interstitial pulmonary fibrosis have obvious difference and pathologic change typical cases in various related detection indexes and pathologic changes. The various effective parts of astragalus mongholicus and hedysarum polybotrys are capable of suppressing various related indexes of the interstitial pulmonary fibrosis model of rats induced by bleomycin and preventing the development of interstitial pulmonary fibrosis, wherein low and medium dose groups of hedysarum polybotrys flavone have the optimal effect. The degree of the effect of the effective parts of astragalus mongholicus and hedysarum polybotrys on stopping the progress of interstitial pulmonary fibrosis is related to appropriate concentrations of equivalent dugs. Through comprehensive assessment, the hedysarum polybotrys flavone is better in influence on the lung function, HA, LN, HYP and the like of the rats having interstitial pulmonary fibrosis than other effective parts.

Description

The application of the Radix Astragali, Radix Hedysari effective site

Technical field

The present invention relates to a kind of application of Chinese medicine extract, particularly, relate to the application of the Radix Astragali, Radix Hedysari effective site.

Background technology

Pulmonary fibrosis, lungs stroma is made up of collagen protein, elastoidin and albumen carbohydrate, in the time that fibroblast is subject to chemical or physical property injury, can secretes collagen protein and carry out the repairing of interstitial tissue of lung, and then cause lungs fibrosis; Be after lungs come to harm, the result that human body reparation produces.

Pulmonary fibrosis is many in the morbidity of 40-50 year, and male is mainly in women.Dyspnea is the common sympton of pulmonary fibrosis.When slight pulmonary fibrosis, dyspnea only occurs in the time of aggravating activities, and therefore usually out in the cold or mistaken diagnosis is other diseases.In the time that pulmonary fibrosis makes progress, in the time of tranquillization, also there is dyspnea, can there is carrying out property dyspnea in serious pulmonary fibrosis patients.Other symptoms have dry cough, weak.50% patient has drumstick finger and cyanosis, splits sound in the tiny race of base of lung portion audible and air-breathing end.Though have dyspnea in early days, x-ray rabat possibility normal; The middle and late stage netted or nodular shadow of dispersivity that occurs marching off into political wilderness in two lungs, accidental pleural effusion, thickens or calcification.The serious consequence of lung fibrosis, causes normal lung tissue's structural change, afunction.Be exactly not have in a large number the fibrosis tissue of gas exchange function to replace alveolar, cause oxygen can not enter blood.Patient respiratory is not smooth, anoxia, acidosis, disability, by respirator existence, last exhaustion, death.

At present, the method for the treatment of pulmonary fibrosis is generally treatment with western, as, glucocorticoid (prednisone claims again strong Buddhist nun pine), immunosuppressant or cell toxicity medicament (cyclophosphamide, azathioprine), antioxidant (Ambroxol, taurine), anti-fibrosis medicine (pirfenidone, IT_ receptor antagonist, monoclonal antibody, ), acceptor inhibitor (LTRA, inhibitors of tumor necrosis factor-alpha, the shape flower growth factor-beta inhibitor), immunomodulator (bcg-polysaccharides nucleic acid, levamisole), other drug (colchicine, macrolide antibiotics, interferon-γ, angiotensin-convertion enzyme inhibitor) and gene therapy method etc., treatment by Chinese herbs is also more rare.

Summary of the invention

The technical problem to be solved in the present invention is to overcome existing defect, and a kind for the treatment of by Chinese herbs method of applying the Radix Astragali, Radix Hedysari effective site treatment pulmonary fibrosis is provided.

In order to solve the problems of the technologies described above, the invention provides following technical scheme:

The Radix Astragali, the application of Radix Hedysari effective site in preparation treatment pulmonary fibrosis medicine.

Particularly, the described Radix Astragali, Radix Hedysari effective site are one or more compositionss in astragalus polysaccharides, Radix Astragali flavone, Radix Astragali saponin, Hedysamn polysaccharide, Radix Hedysari flavone, Radix Hedysari saponin.

Preferably, the described Radix Astragali, Radix Hedysari effective site are Radix Hedysari flavone.

The present invention has following beneficial effect:

(1) lung coefficient results shows: the lung coefficient of model group obviously raises; The lung coefficient of the heavy dose of group of Radix Astragali flavone, the heavy dose of group of Radix Astragali saponin, little, the middle dosage group of Radix Hedysari flavone, Radix Hedysari saponin small dose group obviously reduces.Point out most of effective site groups improving in varying degrees bleomycin and cause the lung coefficient of Pulmonary Fibrosis in Rats with Bleomycin-induced, to make lung tissue weight saving, or can improve its morbid state body weight is obviously increased.(2) pulmonary function experimental result shows: Radix Hedysari flavone is little, in, heavy dose of group, little, the middle dosage group of Radix Astragali flavone be obvious in the improvement effect of most indexs, these group therapeutic effect are more excellent.(3) HA serum test experience result shows; Prednisone group, astragalus polysaccharides be little, in heavy dose of group, Radix Astragali flavone, in heavy dose of group, Radix Astragali saponin, heavy dose of group, Radix Hedysari flavone is little, in, the heavy dose of group of value that can significantly reduce HA reduce the local ECM level of lung, pulmonary fibrosis alleviates to some extent.(4) LN serum test experience result shows: prednisone group, Radix Astragali flavone be little, in, heavy dose of group, little, the middle dosage group of Radix Astragali saponin, the value that little, the middle dosage group of Radix Hedysari flavone can significantly reduce LN slows down the collagen aggregate velocity in ECM, and pulmonary fibrosis alleviates to some extent.(5) HYP tissue homogenate test experience result shows: the heavy dose of group of astragalus polysaccharides, Radix Astragali flavone be little, in, heavy dose of group, Radix Hedysari flavone be little, in, heavy dose of group can significantly reduce collagen fiber deposition pulmonary fibrosis is alleviated to some extent.(6) Comprehensive Assessment, Radix Hedysari flavone is excellent compared with other effective site to the influence of lung fibrosis in rats pulmonary function, HA, LN, HYP.

Wherein, hyaluronic acid (HA), laminin，LN (LN) and hydroxyproline (HyP) are in the time that pulmonary fibrosis occurs, and serum and liver organization all raise, and each effective site all has reducing effect to them.

Compared with model group, each effective site group all has some improvement on each index content, illustrates that each effective site group all has certain inhibitory action to the process of pulmonary fibrosis.Treatment difference between different effective site groups, illustrates that drug level is very large to the function influence of disease treatment, thereby makes the grouping of this therapeutic scheme on dosage more meaningful.Illustrate the Radix Astragali, Radix Hedysari effective site little, in, heavy dose of relevant with its drug dose to suppressing the process of pulmonary fibrosis, Comprehensive Assessment, Radix Hedysari flavone is excellent compared with other effective site to the influence of lung fibrosis in rats pulmonary function, HA, LN, HYP.

Brief description of the drawings

Accompanying drawing is used to provide a further understanding of the present invention, and forms a part for description, for explaining the present invention, is not construed as limiting the invention together with embodiments of the present invention.In the accompanying drawings:

Fig. 1 is Technology Roadmap of the present invention;

Fig. 2 is the canonical plotting of HA;

Fig. 3 is the canonical plotting of LN.

Detailed description of the invention

Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described, should be appreciated that preferred embodiment described herein, only for description and interpretation the present invention, is not intended to limit the present invention.

Embodiment 1

The extraction of red, Radix Astragali effective site

1. experiment material

1.1 medicines and reagent

Astragaloside reference substance, control substance of Rutin, glucose reference substance are all purchased from Chinese pharmaceutical biological product inspection institute.95% ethanol, n-butyl alcohol, ethyl acetate, ether are analytical pure.Radix Hedysari, the Radix Astragali are purchased from medical material market, the west of Gansu Province, Dingxi City, through Affiliated Hospital of Gansu University of Traditi's poplar stannum storehouse professor of pharmacy, Gansu Province Drug Testing Institute Zhu Jun scholar professor of pharmacy qualification.

1.2 instrument and equipment

UV-3600 type ultraviolet-uisible spectrophotometer (Japanese Shimadzu), AE-240 electronic balance (prunus mume (sieb.) sieb.et zucc. Teller-Tuo benefit instrument Shanghai branch company), KQ-100 ultrasonic cleaner (ultrasonic technique company limited of city of Kunshan), R-200 Rotary Evaporators (Switzerland's cloth is strange), DZF6050 vacuum drying oven (Shanghai new talent medical apparatus and instruments Manufacturing Co., Ltd).

1.2 experimental technique

1.2.1 the extraction of Radix Hedysari effective site separates

1.2.1.1 Radix Hedysari total polysaccharides

After being volatilized to ethanol, Radix Hedysari medicinal residues after ethanol extraction decoct with water 3 times, each 1h, add for the first time the water of 8 times of amounts, add 6 times of water gagings for twice afterwards, merge three times decocting liquid, filter, it is 1.15(20 DEG C that filtrate decompression is concentrated into relative density) extractum, centrifugalize, discard precipitation, supernatant slowly adds 95% ethanol under stirring, what make solution reaches 70% containing alcohol amount, fully stir evenly, airtight, leave standstill 24 h, filter, precipitation precipitates 2 times by appropriate 70% washing with alcohol, precipitation is dissolved with distilled water, centrifugalize (2500r/min, 15min), discard precipitation, supernatant slowly adds 95% ethanol under stirring, what make solution reaches 70% containing alcohol amount, fully stir evenly, airtight, leave standstill 24 h, filter centrifugal, precipitation precipitates 2 times by 70% washing with alcohol, 50 DEG C following dry, after dry, with the dry thing of water dissolution, obtain solution, chloroform-n-butyl alcohol (5:1) is added in solution by the volume of 0.2 times, extraction (repeated multiple times), removes the albumen in polysaccharide, except after Deproteinization, polysaccharide decolouring, adds heating activated carbon backflow 0.5h(active carbon to add by 3 ﹪ the aqueous solution of polysaccharide), filtered while hot, obtains Radix Hedysari total polysaccharides after filtrate evaporate to dryness.

1.2.1.2 Radix Hedysari total flavones

Get Radix Hedysari medical material, after moistening, cut into the decoction pieces that thickness is 2～3 mm, dry, take 6.5 kg, add people's 70% ethanol, reflux, extract, 3 times, each 1h, add for the first time the ethanol of 8 times of amounts, rear add for twice 6 times amount ethanol, merge three times extracting solution, filter, filtrate leaves standstill 24 h, elimination precipitation, decompression filtrate recycling ethanol is extremely without alcohol taste, let cool, with distilled water diluting to 1:2, then go up polyamide column, upper column liquid concentration: 1g/ml, upper prop speed: 2BV/h, then washing, then wash with 70% ethanol, finally wash with 95% ethanol, eluent reclaims after ethanol, with the water saturated ethyl acetate extraction of equal-volume 5 times, combined ethyl acetate extract, reclaim ethyl acetate, dry in residue water-bath, obtain Radix Hedysari total flavones.

1.2.1.3 Radix Hedysari total saponins

The effluent of upper complete polyamide column is continued to upper D-101 macroporous resin, upper column liquid concentration: 1g/ml, upper prop speed: 2BV/h, then washing, then wash with 70% ethanol, finally wash with 95% ethanol, eluent reclaims after ethanol, uses the water saturated n-butanol extraction of equal-volume 5 times, merge butanol extraction liquid, reclaim n-butyl alcohol, dry in residue water-bath, obtain Radix Hedysari total saponins.

1.2.2 the assay of Radix Hedysari effective site

1.2.2.1 the preparation of reference substance solution

The preparation of glucose reference substance solution: precision takes the glucose reference substance 10.68mg that is dried to constant weight at 105 DEG C, is placed in 100 mL volumetric flasks, dissolves and is diluted to scale with distilled water, shakes up, for subsequent use.Obtaining concentration of glucose is 0.1068mg/ml.

The preparation of control substance of Rutin solution: precision takes control substance of Rutin 50 mg that are dried to constant weight at 105 DEG C, be placed in 25 dry mL volumetric flasks, with dissolve with methanol and be diluted to scale, shake up, precision measures 10ml, is placed in 100ml volumetric flask, adds water to scale, shake up, obtain (every 1ml is containing anhydrous rutin 0.2mg) for subsequent use.Joining concentration is in fact 0.2198mg/ml.

The preparation of astragaloside reference substance solution: precision takes astragaloside reference substance 12.5 mg that are dried to constant weight at 105 DEG C, is placed in 25 dry mL volumetric flasks, with anhydrous alcohol solution and be diluted to scale, shakes up, for subsequent use.Joining concentration is in fact 0.532mg/ml.

1.2.2.2 the preparation of sample solution

Precision takes Radix Hedysari total polysaccharides 0.5002g,, in 50mL volumetric flask, shakes up with distilled water standardize solution, and precision pipettes 10ml and is settled in 100ml volumetric flask, shakes up, then draws 5ml and be settled in 25ml volumetric flask, gets 2ml and measures; Precision takes Radix Hedysari total flavones 1.521g, is settled in 100ml volumetric flask with 70% ethanol, and the accurate 2ml that draws, in 25ml volumetric flask, then draws in 1ml to 25ml volumetric flask, the method under sighting target directrix curve, and from " adding water to 8.0ml ", mensuration in accordance with the law; Precision takes Radix Hedysari total saponins 1.536g,, in 25mL volumetric flask, filters with dehydrated alcohol standardize solution, and precision is got filtrate 0.5ml, adds the vanillin reagent of 0.5ml, to ice-water bath, slowly adds 5ml72% sulphuric acid, shakes up placement, measures.

1.2.2.3 the drafting of standard curve

The drafting of glucose standard curve: above-mentioned dextrose standard sample solution 0.1,0.2,0.4,0.6,0.8,1.0 mL of accurate absorption, be placed in 10mL test tube, it is 1.0ml that each adding distil water makes cumulative volume, shakes up.Add respectively 5% phenol solution 1.5mL, shake up, add rapidly 5 mL concentrated sulphuric acids, jolting 5 minutes, puts in boiling water bath and heats 15 minutes, then puts in psychrolusia cooling 30 minutes, and in wavelength, 490 nm places measure absorbances, and retinue is blank.Data obtain regression equation through regression treatment: A=57.2482C+0.0695, r=0.9984.

The drafting of rutin standard curve: precision measures above-mentioned control substance of Rutin solution 0,1,2,3,4,6,8mL, to 25mL volumetric flask, respectively add water to 8ml respectively, add respectively people 5% sodium nitrite solution 1mL, shake up and place 6 minutes, add people 10% aluminum nitrate solution 1mL, place 6 minutes, add people 1N(4%) sodium hydroxide solution 10mL, add water to again scale, place 15 minutes, put in cuvette, at wavelength, 500 nm places measure absorbance.Data obtain regression equation through regression treatment: A=11.66035C+0.0062, r ²=0.99839.

The drafting of astragaloside standard curve: above-mentioned astragaloside reference substance solution 0.15,0.30,0.45,0.60,0.75 mL of accurate absorption, be placed in respectively tool plug test tube, add respectively dehydrated alcohol 0.75 mL, add respectively again 0.75 mL8% vanillin-concentrated sulphuric acid reagent, being placed in ice bath, to add 7.5ml volume fraction be that 72% sulphuric acid shakes up, and the water bath with thermostatic control of putting into 62 DEG C is incubated 20 minutes, takes out, cooling 5 min of flowing water, shake up.In 30 minutes, in wavelength, 544 nm places measure absorbance, and retinue is blank.Data obtain regression equation through regression treatment: A=21.102C+0.0298, r=0.9993.

1.2.2.4 precision test

Accurate each 5 parts of Radix Hedysari total flavones sample solution, Radix Hedysari total saponins sample solution, Radix Hedysari total polysaccharides sample solution, every part of 0.5 mL of drawing; Measure absorbance, calculate with absorbance, Radix Hedysari total polysaccharides RSD is 2.92%, and Radix Hedysari total flavones RSD is 2.52%, and Radix Hedysari total saponins RSD is 3.08%.

1.2.2.5 stability test

Get total flavones sample solution, within every 15 minutes, survey an absorbance, 2h investigates its stability continuously, and result shows that in 2h, colour developing is all stable, can carry out assay.Get total saponins sample solution, within every 10 minutes, survey an absorbance, 1h investigates its stability continuously, and result shows that in 1h, colour developing is all stable, can carry out assay.Get total polysaccharides sample solution, within every 10 minutes, survey an absorbance, 1h investigates its stability continuously, and result 1h shows that interior colour developing is all stable, can carry out assay.

1.2.2.6 determination of recovery rates

Precision takes Radix Hedysari total polysaccharides 125mg, adds a certain amount of glucose reference substance,, in 100mL volumetric flask, shakes up with distilled water standardize solution; Precision takes Radix Hedysari total flavones 40mg, adds a certain amount of control substance of Rutin,, in 25mL volumetric flask, shakes up by methanol constant volume; Precision takes Radix Hedysari total saponins 25mg, adds a certain amount of astragaloside reference substance,, in 25mL volumetric flask, shakes up with dehydrated alcohol standardize solution.Press the method for assay, measure calculate recovery rate.Measurement result: glucose average recovery rate is that 98.3%, RSD is 2.19%(n=5); Rutin average recovery rate is that 95.8%, RSD is 2.96%(n=5); Astragaloside average recovery rate is that 92.5%, RSD is 3.21%(n=5).

1.2.2.7 sample size is measured

The assay of Radix Hedysari total polysaccharides: the accurate Radix Hedysari total polysaccharides sample solution 2ml that draws measures, and by standard curve method operation, surveys its content according to regression equation, obtains 0.718g/g.

The assay of Radix Hedysari total flavones: the accurate Radix Hedysari total flavones sample solution of drawing, by standard curve method operation, survey its content according to regression equation, obtain 0.526g/g.

The assay of Radix Hedysari total saponins: the accurate Radix Hedysari total saponins sample solution of drawing, by standard curve method operation, survey its content according to regression equation, obtain 0.517g/g.

1.2.2 the extraction of Radix Astragali effective site separates

1.2.2.1 Radix Astragali Mongolici total polysaccharide

After being volatilized to ethanol, astragalus root dregs after ethanol extraction decocts with water 3 times, each 1h, add for the first time the water of 8 times of amounts, add 6 times of water gagings for twice afterwards, merge three times decocting liquid, filter, it is 1.15(20 DEG C that filtrate decompression is concentrated into relative density) extractum, centrifugalize, discard precipitation, supernatant slowly adds 95% ethanol under stirring, what make solution reaches 70% containing alcohol amount, fully stir evenly, airtight, leave standstill 24 h, filter, precipitation precipitates 2 times by appropriate 70% washing with alcohol, precipitation is dissolved with distilled water, centrifugalize (2500r/min, 15min), discard precipitation, supernatant slowly adds 95% ethanol under stirring, what make solution reaches 70% containing alcohol amount, fully stir evenly, airtight, leave standstill 24 h, filter centrifugal, precipitation precipitates 2 times by 70% washing with alcohol, 50 DEG C following dry, after dry, with the dry thing of water dissolution, obtain solution, chloroform-n-butyl alcohol (5:1) is added in solution by the volume of 0.2 times, extraction (repeated multiple times), removes the albumen in polysaccharide, except after Deproteinization, polysaccharide decolouring, adds heating activated carbon backflow 0.5h(active carbon to add by 3 ﹪ the aqueous solution of polysaccharide), filtered while hot, obtains Radix Astragali Mongolici total polysaccharide after filtrate evaporate to dryness.

1.2.2.2 Radix Astragali total flavones

Get Milkvetch Root, after moistening, cut into the decoction pieces that thickness is 2～3 mm, dry, take 6.5 kg, add people's 70% ethanol, reflux, extract, 3 times, each 1h, add for the first time the ethanol of 8 times of amounts, rear add for twice 6 times amount ethanol, merge three times extracting solution, filter, filtrate leaves standstill 24 h, elimination precipitation, decompression filtrate recycling ethanol is extremely without alcohol taste, let cool, with distilled water diluting to 1:1, then go up polyamide column, upper column liquid concentration: 1g/ml, upper prop speed: 2BV/h, then washing, then wash with 70% ethanol, finally wash with 95% ethanol, eluent reclaims after ethanol, with the water saturated ethyl acetate extraction of equal-volume 5 times, combined ethyl acetate extract, reclaim ethyl acetate, dry in residue water-bath, obtain Radix Astragali total flavones.

1.2.2.3 Radix Astragali total saponins

The effluent of upper complete polyamide column is continued to upper D-101 macroporous resin, upper column liquid concentration: 1g/ml, upper prop speed: 2BV/h, then washing, then wash with 70% ethanol, finally wash with 95% ethanol, eluent reclaims after ethanol, uses the water saturated n-butanol extraction of equal-volume 5 times, merge butanol extraction liquid, reclaim n-butyl alcohol, dry in residue water-bath, obtain Radix Astragali total saponins.

2.2.3 the assay of Radix Astragali effective site

2.2.3.1 the preparation of reference substance solution

2.2.3.2 the preparation of sample solution

Precision takes Radix Astragali Mongolici total polysaccharide 0.5005g,, in 50mL volumetric flask, shakes up with distilled water standardize solution, and precision pipettes 10ml and is settled in 100ml volumetric flask, shakes up, then draws 5ml and be settled in 25ml volumetric flask, gets 2ml and measures; Precision takes Radix Astragali total flavones 1.501g, is settled in 100ml volumetric flask with 70% ethanol, and the accurate 2ml that draws, in 25ml volumetric flask, then draws in 1ml to 25ml volumetric flask, the method under sighting target directrix curve, and from " adding water to 8.0ml ", mensuration in accordance with the law; Precision takes Radix Astragali total saponins 1.582g,, in 25mL volumetric flask, filters with dehydrated alcohol standardize solution, and precision is got filtrate 0.5ml, adds the vanillin reagent of 0.5ml, to ice-water bath, slowly adds 5ml72% sulphuric acid, shakes up placement, measures.

2.2.3.3 the drafting of standard curve

2.2.3.4 precision test

Accurate each 5 parts of Radix Astragali total flavones sample solution, Radix Astragali total saponins sample solution, Radix Astragali Mongolici total polysaccharide sample solution, every part of 0.5 mL of drawing; Measure absorbance, calculate with absorbance, Radix Astragali Mongolici total polysaccharide RSD is 3.05%, and Radix Astragali total flavones RSD is 2.63%, and Radix Astragali total saponins RSD is 3.16%.

2.2.3.5 stability test

2.2.3.6 determination of recovery rates

Precision takes Radix Astragali Mongolici total polysaccharide 125mg, adds a certain amount of glucose reference substance,, in 100mL volumetric flask, shakes up with distilled water standardize solution; Precision takes Radix Astragali total flavones 40mg, adds a certain amount of control substance of Rutin,, in 25mL volumetric flask, shakes up by methanol constant volume; Precision takes Radix Astragali total saponins 25mg, adds a certain amount of astragaloside reference substance,, in 25mL volumetric flask, shakes up with dehydrated alcohol standardize solution.Press the method for assay, measure calculate recovery rate.Measurement result: glucose average recovery rate is that 98.3%, RSD is 2.18%(n=5); Rutin average recovery rate is that 96.8%, RSD is 2.85%(n=5); Astragaloside average recovery rate is that 92.8%, RSD is 3.12%(n=5).

2.2.3.7 sample size is measured

The assay of Radix Astragali Mongolici total polysaccharide: the accurate Radix Hedysari total polysaccharides sample solution 2ml that draws measures, and by standard curve method operation, surveys its content according to regression equation, obtains 0.721g/g.

The assay of Radix Astragali total flavones: the accurate Radix Hedysari total flavones sample solution of drawing, by standard curve method operation, survey its content according to regression equation, obtain 0.558g/g.

The assay of Radix Astragali total saponins: the accurate Radix Hedysari total saponins sample solution of drawing, by standard curve method operation, survey its content according to regression equation, obtain 0.531g/g.

Embodiment 2

One, experiment

1 materials and methods

1.1 material

1.1.1 laboratory animal and raising condition

The healthy Wistar rat of SPF level, male and female half and half, body weight (200 ± 10) g, is provided by Gansu Chinese of Traditional Chinese Medicine's scientific experiment center.Quality of Experimental Animals quality certification numbering: SCXK(is sweet) 2011-0001-0001059(is shown in annex 1) SPF level feedstuff (being supplied with by the feed corporation,Ltd that pulls together of Beijing Austria of section) feeds, and freely drink water, prepare raising and test after 3 days.Animal facility use certificate numbering: SCSY(is sweet) 20011-0001-0002753(is shown in annex 2).

1.1.2 main agents

Hydrochloride for injection Bleomycin A5	Be purchased from Harbin bleomycin company limited	Lot number: 110424
			Pentobarbital sodium	Be purchased from Beijing chemical reagents corporation	Lot number: 101021
HA immune radiating immunoassay medicine box	Be purchased from Beijing North Institute of Biological Technology	Lot number: 111220
			LN radioimmunoassay, RIA medicine box	Be purchased from Beijing North Institute of Biological Technology	Lot number: 111220
Hydroxyl dried meat is by sour testing cassete	Be purchased from Nanjing and build up Bioengineering Research Institute	Lot number: 20111210

1.1.3 Experimental agents

Tested medicine: the polysaccharide of the Radix Astragali, Radix Hedysari, flavone, saponin are provided by Gansu Chinese of Traditional Chinese Medicine's scientific experiment center pharmaceutical preparation laboratory, inspection reaches more than 50%.Control drug: prednisone acetate tablets, 5mg/ sheet, Zhejiang Province XianJu Pharmacy stock Co., Ltd, the accurate word H33021207 of traditional Chinese medicines, lot number 110810.

1.1.4 key instrument

GC-9111 type radioimmunity r enumerator	Zhong Jia branch company of Keda Innovation Co., Ltd
		KDC-2044 low speed refrigerated centrifuge	Zhong Jia company of Keda Innovation Co., Ltd
The full-automatic fluorescence microscope of OLYMPUS AX80	Japan
		AE200 electronic analytical balance	Prunus mume (sieb.) sieb.et zucc. Teller---holder benefit instrument Shanghai company limited
SFC-182 type biological microscope	Maike Aodi Industry Group Co Ltd
		EMKA animal lung function detection system	France
Ultraviolet spectrometry Density Measuring Instrument UV-2601	Beijing North divides Rayleigh analytical tool (group) company

1.2 experimental technique

1.2.1 laboratory animal grouping

252 of SPF level Wistar rats, body weight 200 ± 20g, male and female half and half, adaptability was fed after 3 days, according to table of random number method first separate blank group, model group, prednisone group, the Radix Astragali, Hedysamn polysaccharide little, in, heavy dose of group, the Radix Astragali, Radix Hedysari flavone be little, in, heavy dose of group, the Radix Astragali; Radix Hedysari saponin is little, in, heavy dose of group, 12 every group, divide cage to place by sex, 6, every cage, all rats are unified modeling, number by pre-configured 3% the picric acid solution dyeing row labels of going forward side by side.

1.2.2 experimental animal model preparation

Conventional letting animals feed, after 3 days, copies pulmonary fibrosis model, with 3% pentobarbital sodium (40～50mgkg ^-1) intraperitoneal injection anesthesia, wait press from both sides tail without pain reaction after, rat dorsal position is fixed on Mus plate, fixing limbs, fixing head, makes head slightly to layback.Assistant strangles tooth on rat with a rubber band, to front lower place tractive, fasten lower tooth to back upper place tractive (angle is advisable with 60o) with another root rubber band, patient's left hand is held the special laryngoscope of toy, stretch in rat oral cavity, opening is to bottom right, dorsal segment is lifted the jack-up root of the tongue gently on upwards omiting, expose glottis, take advantage of air-breathing moment of animal, homemade tracheal casing pipe (is cut off disposable use venous detaining needle (22G × 25mm/Y-G) to the part of syringe needle, the length of remaining plastics sebific duct is advisable with 6-7cm, with thin wire in tail end is made circular insertion sebific duct as nook closing member, length equates with plastics sebific duct, can not exceed sebific duct, in order to avoid stab rat throat.) take advantage of a situation and insert in rat trachea, in sleeve pipe, inject in advance 2-3 and save little water column, if the interior water column of sleeve pipe is motionless or fiercely ejection or suction, illustrate that intubate enters esophagus, need intubate again; If rhythmical the moving up and down of water column follower respiratory frequency in sleeve pipe, illustrates and inserts trachea.Now, extract rapidly nook closing member, will take out in advance bleomycin (5mgkg ^-1, 5mgml ^-1) lml syringe be connected in tracheal casing pipe, the disposable trachea that pushes.After injection, extract immediately sleeve pipe, animal is upright, and then left rotation and right rotation approximately 1 minute is evenly distributed medicine in lung.In blank group trachea, inject isopyknic normal saline.After animal is naturally clear-headed, conventional SPF raises.

1.2.3 dosage regimen

Within after modeling the 2nd day, start gastric infusion.Every day 1 time, gavage treatment 28 days continuously, fetching mark.The theory that each treatment group dosage converts according to clinical kg body weight dosage multiple, determine that in conjunction with document and principle of comparability the dosage of this experiment is as follows: Radix Astragali 6500g extracts and isolates polysaccharide: 53.6g, saponin: 32.36g, flavone: 32.4g, converts every gram of Radix Astragali and contains polysaccharide: 8.25mg, saponin: 4.98mg, flavone: 4.98mg simultaneously; Radix Hedysari 6500g extracts and isolates polysaccharide: 53.6g, saponin: 32.36g, flavone: 32.4g, converts every gram of Radix Hedysari and contains polysaccharide: 5.72mg, saponin: 4.23mg, flavone: 2.92mg.Adult's the Radix Astragali, the dosage of Radix Hedysari are by 30g(pharmacopeia regulation adult one consumption per day 9～30g) calculate, a consumption per day is 0.5gkg ^-1crude drug.Pressing the direct scaling method of body surface area or man and animal conversion dosage multiple calculates.The rat dosage of a day behave 6 times, the rat crude drug consumption of a day is 3.0gkg ^-1.The dosage that the amount that goes out effective site according to every gram of crude drug actual extracting converses each effective site is as following table: (in table 1,2)

The actual dosage of table 1 Radix Astragali, Hedysamn polysaccharide, saponin, flavone

The actual dosage of table 2 Radix Astragali, Hedysamn polysaccharide, saponin, flavone

The Radix Astragali, Hedysamn polysaccharide, flavone, saponin are mixed with respectively the solution of respective concentration before use with cosolvent 0.5% carboxymethylcellulose sodium solution, every day gastric infusion, gavage volume 1mL100g ^-1body weight.Prednisone group gives prednisone acetate tablets, and reference literature determines that dosage is 3mgKg ^-1d ^-1, with 0.5% carboxymethylcellulose sodium solution preparation; Model group and blank group give equal-volume 0.5% carboxymethylcellulose sodium solution gavage.

1.2.4 laboratory animal collection of specimens and detection

1.2.4.1 the assay method of pulmonary function and drawing materials

Start computer, operation IOX Data Collection Software based, commissioning device, guarantees that system normally works.Open animal respirator, signal conditioner.Set up a new experiment, log-on data collection.Rat is put into animal plethysmography box in batches, trace that case is each puts one for four, each four, do pulmonary function test (pft) under noinvasive and waking state (rf).Test index has: Ti(inspiratory duration), Te(expiratory duration), EIP (air-breathing latter stage pause), EEP (EEP pause), RT (persistent period), PIF(inspiratory airflow peak), PEF(expiratory airflow peak), the flow velocity of EF50(while breathing out 50% tolerance), MV (ventilation per minute), TV(tidal volume), EV(forced volume,expiratory), AV (alveolar ventilation), Penh (bronchoconstriction degree), F(frequency) etc., tested rear saving result.After the pulmonary function completing under noinvasive state, just anaesthetize and have the pulmonary function test (pft) of (rcv) under wound state.Adopt isoflurane inhalation anesthesia, under small animal laryngoscope guiding, carry out tracheal intubation, then rat clinostatism is placed in to the airtight plethysmography box of toy respirator.First trace one section of eupnea, then survey Ti(inspiratory duration), Te(expiratory duration), EIP (pause air-breathing latter stage), EEP (EEP pause), RT (persistent period), PIF(inspiratory airflow peak), PEF(expiratory airflow peak), flow velocity when EF50(breathes out 50% tolerance), MV (ventilation per minute), TV(tidal volume), EV(forced volume,expiratory), cdyn(Cdgn dyanamic compliance), Penh (bronchoconstriction degree), LR(lung resistance), dppl(pleura is pressed and is changed), EEW (EEP merit), pau(pauses), F(frequency) etc.After lung function tests, carry out femoral artery and get blood.Then cut off rat thoracic cavity, careful separation lung tissue, the complete pair lungs of isolating, carefully remove connective tissue around, blot electronic balance weighing lung weight in wet base, calculating lungs coefficient after normal saline washing with filter paper.Win inferior lobe of right lung, do the detection of tissue homogenate in order to lung tissue HYP.

1.2.4.2 HA detects

0,25,50,100,200,400,800ng/mL adopt radio immunoassay, carry out according to test kit description, determination step is as follows: 7 bottles of concentration of (1) HA standard substance are followed successively by:.(2) HA quality controlled serum: dissolve with 0.5mL distilled water before using, after dissolving, at least 15 minutes ability is used for application of sample.(3) BALF, with 0.2mLPBS dilution lung tissue homogenate dried frozen aquatic products, gets 100 μ L and measures.(4) get polystyrene test tube some, after numbering, carry out application of sample by application of sample sequence list, before application of sample, all reagent comprises that testing sample is careful and shakes up, and avoids producing foam as far as possible.For application of sample is accurate, various reagent and sample should first equilibrate to room temperature before application of sample.As table 3, Fig. 1.

Table 3 application of sample sequence list (unit: μ L)

1.2.4.3 LN detects

Adopt radio immunoassay, carry out according to test kit description, determination step is as follows: (1) LN standard substance: with 2.0mL distilled water dissolving S0, with 1.0ml distilled water dissolving S1～S5,0,40,80,160,320,640ng/mL after fully dissolving, its accurate concentration is respectively:.(2) dissolve 125I-LN with 10ml distilled water.(3) dissolve LN antibody with l1mL distilled water.(4) BALF, with 0.2mLPBS dilution lung tissue homogenate dried frozen aquatic products, respectively gets 100 μ L and measures.(5) get round bottom polystyrene test tube some, with marking pen or special pencil numbering NSB, S0-S5 and testing sample pipe etc., then use micro sample-adding according to the form below application of sample.Before application of sample, all reagent (especially separating medium) comprises that testing sample will shake up, and avoids producing foam as far as possible, and preferably equilibrates to room temperature.(as table 4, Fig. 2)

Table 4LN application of sample sequence list (unit: μ L)

1.2.4.4HYP detection

The oxidation product that hydroxyproline produces under the effect of oxidant and dimethylaminobenzaldehyde effect present aubergine, can extrapolate its content according to the depth of its colour generation.Carry out according to test kit description, determination step is as follows: (1) accurately takes tissue wet 30-100ml and puts into test tube, adds hydrolyzed solution 1ml.Mix.Add a cover rear 95 degrees Celsius or boiling water bath hydrolysis 20 minutes.(while being hydrolyzed 10 minutes, mix once, object be make hydrolysis more abundant).(2) adjust pH value to 6.0-6.8 left and right; 1. cooling each test tube flowing water rear each pipe is added to one of indicator, shake up; 2. each pipe adds tune PH first liquid 1.0ml, mixes (now solution should be red); 3. draw with the sample injector of 200 μ L the second liquid of adjusting pH value, after being often added dropwise to, will mix, until the color of indicator becomes yellow green in liquid.Now pH value (approximately adds the second liquid 100 μ L-500 μ L left and right of adjusting pH value) in 6.0～6.8 left and right; 4. then adding distil water, to 10ml, mixes; 5. the appropriate active carbon (about 20-30mg left and right, with colourless being as the criterion of the centrifugal rear clarification of supernatant) of hydrolyzed solution man of getting 34ml dilution, mixes, and 3500 turn ∕ divides centrifugal 10 minutes, gets supernatant 1ml and checks.(3) mix after adding reagent, 60 DEG C of water-baths 15 minutes, cooling after, 3500 turn ∕ divides centrifugal 10 minutes, gets supernatant at 550nm place, 1cm optical path, distilled water zeroing, surveys each pipe absorbance.

Accurately draw normal rat serum 0.5ml, detect by operating procedure.

Surveying blank tube absorbance is 0.075, standard pipe absorbance 0.61.

Result of calculation formula is:

Hydroxyproline content (μ g ∕ ml)=mensuration pipe absorbance-blank tube Xi Guang Du ∕ standard pipe absorbance-blank tube absorbance × standard pipe content (5 μ g ∕ ml) × hydrolyzed solution cumulative volume (ml) ∕ sampling amount (ml) (in table 5)

Table 5HYP operation table

2 experimental results

The variation of 2.1 each group rat general status and lung coefficient

In blank group rat experiment process, without dead, feed and the mental status are good, are quick on the draw, and fur gloss is shinny, and body weight increases gradually, breathes steadily.The model group rat mental status is poor, appetite and movable minimizing, fur is sparse, its colour changed into yellow tarnishes, common fall perpendicular phenomenon, hogback is rolled up, bradykinesia, rapid breathing, health are become thin etc.The situation of all the other each treated animals after administration falls between.

According to formula lung coefficient=lung weight in wet base, (mg) ∕ body weight (g) × 100% is calculated lung coefficient, and as can be seen from Table 6, in model group, astragalus polysaccharides, dosage group induced lung coefficient significantly raises and has notable difference (P ﹤ 0.01) compared with blank group; Dosage group, little, the middle dosage group of Hedysamn polysaccharide induced lung coefficient rising variant (P ﹤ 0.05) in Radix Astragali flavone; Compared with model group, the heavy dose of group of Radix Astragali flavone, the heavy dose of group of Radix Astragali saponin, little, the middle dosage group of Radix Hedysari flavone, Radix Hedysari saponin small dose group induced lung coefficient significantly reduce and have notable difference (P ﹤ 0.01); Astragalus polysaccharides is little, in heavy dose of group, Radix Hedysari saponin, heavy dose of group induced lung coefficient reduces variant (P ﹤ 0.05); (in table 6)

2.2 each group pulmonary function testing results

The lung function index of model group rat compared with blank group: under noinvasive (rf) state, TV, EV, MV, PIF, PEF, EF50 significantly reduce, difference has statistical significance (P ﹤ 0.01), and AV also reduces, and difference has statistical significance (P ﹤ 0.05).Have under wound (rcv) state, TV, EV, cdyn significantly reduce, and difference has statistical significance (P ﹤ 0.01), and EF50 reduces, and difference has statistical significance (P ﹤ 0.05).All risings in various degree of the numerical value of index TV, EV, MV, AV, PIF, PEF under each effective site treatment group pulmonary function rf state compared with model group.Have under wound rcv state, the numerical value of EF50, TV, EV, cdyn also has rising in various degree.(in table 7,8,9,10,11,12,13,14)

Wherein (rf) TV of astragalus polysaccharides small dose group variant compared with model group (P ﹤ 0.05).(rf) MV of dosage variant compared with model group (P ﹤ 0.05) in astragalus polysaccharides.(rf) MV, (rcv) cdyn, the EV of the heavy dose of group of astragalus polysaccharides have significant difference (P ﹤ 0.01) compared with model group.(rf) AV of Radix Astragali flavone small dose group, (rcv) TV, cdyn have significant difference (P ﹤ 0.01) compared with model group, (rf) EV, MV variant compared with model group (P ﹤ 0.05).In Radix Astragali flavone, (rf) AV of dosage group, (rcv) TV, cdyn have significant difference (P ﹤ 0.01) compared with model group, (rf) EV variant compared with model group (P ﹤ 0.05).(rcv) EV, the cdyn of the heavy dose of group of Radix Astragali flavone have significant difference (P ﹤ 0.01) compared with model group.(rcv) EV of Radix Astragali saponin small dose group has significant difference (P ﹤ 0.01) compared with model group, (rf) TV, AV variant compared with model group (P ﹤ 0.05).

The affect result of table 6 on each group of induced lung coefficient

Note: with the comparison of blank group: ^△p < 0.05, ^{△ △}p < 0.01; With model group comparison: ^▲p < 0.05, ^{▲ ▲}p < 0.01;

The result of table 7 to each group of pulmonary function Index Influence

Note: with the comparison of blank group: ^△p < 0.05, ^{△ △}p < 0.01; With model group comparison: ^▲p < 0.05, ^{▲ ▲}p < 0.01

(rcv) EV of Hedysamn polysaccharide small dose group has significant difference (P ﹤ 0.01) compared with model group.(rcv) EV, the cdyn of dosage group variant compared with model group (P ﹤ 0.05) in Hedysamn polysaccharide.(rcv) cdyn of the heavy dose of group of Hedysamn polysaccharide has significant difference (P ﹤ 0.01), (rf) TV variant compared with model group (P ﹤ 0.05) compared with model group.(rf) TV of Radix Hedysari flavone small dose group, MV, AV, (rcv) TV, EV, cdyn have significant difference (P ﹤ 0.01) compared with model group.(rf) EV variant compared with model group (P ﹤ 0.05).In Radix Hedysari flavone, (rf) TV, the MV of dosage group, (rcv) EV, cdyn have significant difference (P ﹤ 0.01) compared with model group, (rf) EV, AV, (rcv) TV variant compared with model group (P ﹤ 0.05).(rcv) TV, EV, the cdyn of the heavy dose of group of Radix Hedysari flavone have significant difference (P ﹤ 0.01), (rf) EV variant compared with model group (P ﹤ 0.05) compared with model group.(rcv) TV variant compared with model group (P ﹤ 0.05) of Radix Hedysari saponin small dose group.(rcv) TV of dosage group variant compared with model group (P ﹤ 0.05) in Radix Hedysari saponin.In Radix Hedysari saponin, the LR of dosage group has significant difference (P ﹤ 0.01) compared with model group.(rf) MV, (rcv) TV variant compared with model group (P ﹤ 0.05) of the heavy dose of group of Radix Hedysari saponin.

Little, the middle dosage group of Radix Hedysari flavone and model group comparison numerical value in the impact of (rf) TV (aspirated volume) are obviously raise, difference has statistical significance (P ﹤ 0.01), Radix Astragali saponin small dose group, the heavy dose of group of Hedysamn polysaccharide and relatively numerical value rising of model group, difference has statistical significance (P ﹤ 0.05).

The heavy dose of group of Hedysamn polysaccharide in the impact of (rf) EV (forced volume,expiratory) is obviously raise with model group comparison numerical value, difference has statistical significance (P ﹤ 0.01), little, the middle dosage group of Radix Astragali flavone, Radix Hedysari flavone be little, in, heavy dose of group and model group relatively numerical value raise, difference has statistical significance (P ﹤ 0.05).

Prednisone group, the heavy dose of group of astragalus polysaccharides, little, the middle dosage group of Radix Hedysari flavone and model group comparison numerical value in the impact of (rf) MV (ventilation per minute) are obviously raise, difference has statistical significance (P ﹤ 0.01), relatively numerical value rising of dosage group, Radix Astragali flavone small dose group and model group in astragalus polysaccharides, difference has statistical significance (P ﹤ 0.05).

The result of table 8 to each group of pulmonary function Index Influence

The result of table 9 to each group of pulmonary function Index Influence

On (rf) AV(alveolar ventilation) impact in little, the middle dosage group of Radix Astragali flavone, Radix Hedysari flavone small dose group and model group comparison numerical value obviously raise, difference has statistical significance (P ﹤ 0.01), relatively numerical value rising of dosage group and model group in Radix Astragali saponin small dose group, Radix Hedysari flavone, difference has statistical significance (P ﹤ 0.05).

Little on Radix Astragali flavone in the impact of (rcv) TV (tidal volume), in, little, the heavy dose of group of heavy dose of group, Radix Hedysari flavone obviously raises with model group comparison numerical value, difference has statistical significance (P ﹤ 0.01), dosage group in Radix Hedysari flavone, Radix Hedysari saponin be little, in, heavy dose of group and model group relatively numerical value raise, difference has statistical significance (P ﹤ 0.05).

On in dosage group, Radix Hedysari flavone in Radix Astragali flavone small dose group, Radix Astragali saponin small dose group, Hedysamn polysaccharide in the impact of (rcv) EV (forced volume,expiratory), heavy dose of group, Radix Hedysari saponin small dose group and model group comparison numerical value obviously raises, difference has statistical significance (P ﹤ 0.01), dosage group, the heavy dose of group of Hedysamn polysaccharide and relatively numerical value rising of model group in astragalus polysaccharides, difference has statistical significance (P ﹤ 0.05).

On (rcv) cdyn(lung compliance) impact in the heavy dose of group of astragalus polysaccharides, Radix Astragali flavone little, in, heavy dose of group, the heavy dose of group of Hedysamn polysaccharide, Radix Hedysari flavone be little, in, heavy dose of group obviously raises with model group comparison numerical value, difference has statistical significance (P ﹤ 0.01).Relatively numerical value rising of dosage group and model group in prednisone group, Hedysamn polysaccharide, difference has statistical significance (P ﹤ 0.05).

In addition, it is little that interstitial pulmonary fibrosis changes dependency to the impact of other lung function index, just no longer carries out statistics and analysis.

The result of table 10 to each group of pulmonary function Index Influence

The result of table 11 to each group of pulmonary function Index Influence

The result of table 12 to each group of pulmonary function Index Influence

The result of table 13 to each group of pulmonary function Index Influence

The result of table 14 to each group of pulmonary function Index Influence

The impact of HA, LN content in 2.3 each group rat blood serums

2.3.1 HA testing result in serum

Compared with blank group, dosage group in model group, astragalus polysaccharides, Radix Astragali saponin small dose group, Hedysamn polysaccharide be little, in heavy dose of group, Radix Hedysari saponin, the value of heavy dose of group rat HA significantly raises, difference has statistical significance (P ﹤ 0.01), astragalus polysaccharides is little, heavy dose of group, the value of Radix Astragali flavone small dose group, little, the heavy dose of group rat of Radix Hedysari flavone HA raise, and difference has statistical significance (P ﹤ 0.05);

Compared with model group, prednisone group, astragalus polysaccharides be little, dosage group in heavy dose of group, Radix Astragali flavone, in heavy dose of group, Radix Astragali saponin, in heavy dose of group, Hedysamn polysaccharide, Radix Hedysari flavone is little, in, heavy dose of group, the value of rat HA significantly reduce, difference has statistical significance (P ﹤ 0.01), in astragalus polysaccharides, the value of dosage group, Radix Astragali flavone small dose group, the heavy dose of group of Hedysamn polysaccharide rat HA reduces, and difference has statistical significance (P ﹤ 0.05); (in table 15)

2.3.2 LN testing result in serum

Compared with blank group, model group, astragalus polysaccharides be little, in, heavy dose of group, Radix Astragali saponin be little, in, heavy dose of group, Hedysamn polysaccharide be little, in, heavy dose of group, the heavy dose of group of Radix Hedysari flavone, Radix Hedysari saponin be little, in, the value of heavy dose of group rat LN significantly raises, difference has statistical significance (P ﹤ 0.01), the value of the heavy dose of group of Radix Astragali flavone rat LN obviously raises, and difference has statistical significance (P ﹤ 0.05).

With model group ratio, prednisone group, Radix Astragali flavone be little, in, heavy dose of group, little, the middle dosage group of Radix Astragali saponin, the value of little, the middle dosage group of Radix Hedysari flavone rat LN significantly reduces, difference has statistical significance (P ﹤ 0.01); In astragalus polysaccharides, the value of dosage group rat LN reduces, and difference has statistical significance (P ﹤ 0.05); (in table 15)

The result of table 15 on the HA in serum, LN impact

HYP content detection result in 2.4 each group lung tissue of rats

Compared with blank group, model group, prednisone group, astragalus polysaccharides be little, in, heavy dose of group, Radix Astragali saponin be little, in, heavy dose of group, Hedysamn polysaccharide be little, in, heavy dose of group, the value of little, the middle dosage group of Radix Hedysari saponin rat HYP significantly raise, difference has statistical significance (P ﹤ 0.01); The value of little, the heavy dose of group rat of Radix Hedysari flavone HYP raises, and difference has statistical significance (P ﹤ 0.05);

With model group ratio, the heavy dose of group of astragalus polysaccharides, Radix Astragali flavone be little, in, heavy dose of group, Radix Hedysari flavone be little, in, the value of heavy dose of group rat HYP significantly reduces, difference has statistical significance (P ﹤ 0.01); The value of the heavy dose of group of Hedysamn polysaccharide rat HYP reduces, and difference has statistical significance (P ﹤ 0.05).(in table 16)

The result of table 16 to the HYP content influence in lung tissue

Discuss

The research of 1 model

Bleomycin (BLM) is a peptide species class antineoplastic agent, and because it has the side effect that obviously causes pulmonary fibrosis, thereby development has worked the animal model that utilizes BLM to induce and set up pulmonary fibrosis.Zoopery proved in trachea disposable injection BLM reproducible go out the animal model similar to people's interstitial pulmonary fibrosis pathological process.At present, the method has become the classical animal model that carries out in the world pulmonary fibrosis research.It is easy and simple to handle because of it that bleomycin copies interstitial pulmonary fibrosis model, and modeling is stable, and the feature that success rate is high, is widely used.This experiment adopts the method for small animal laryngoscope guidance tracheal intubation to copy pulmonary fibrosis model, this method has avoided the skin of neck of incision rat successively to separate and expose the method for trachea, has eliminated the postoperative local infection of incision skin of rat and the inducement of some complication.In modeling process, apply small animal laryngoscope guiding and can reduce the sewing process in classical modeling process, and can reduce the consumption of anaesthetic, in the process of a large amount of animal models, can greatly improve efficiency.Having avoided secondary insult is the improvement to classical modeling method.Modeling method after improvement, have easy, without wound, speed are fast, modeling success rate is high feature.Have by the rising of lung coefficient, pulmonary function (rf) TV, EV, MV, PIF, PEF, EF50, (rcv) TV, EV, cdyn, HA, LN, blank group of HYP and the relatively demonstration of model group the difference that significantly increases the weight of trend, difference has statistical significance (P ﹤ 0.01).It is more successful that result can show that this tests improved modeling method.

The selection of 2 control drug

Glucocorticoid is the medicine that a class has very strong antiinflammatory action and immunosuppressive action, is once once being used as the choice drug for the treatment of pulmonary fibrosis.At present, the animal experiment study of most IPF all selects glucocorticoid to do positive control drug.There are some researches prove, dexamethasone can promote the pulmonary macrophage apoptosis of lung fibrosis in rats; Reduce the generation of inflammatory cytokine, suppress gathering, the infiltration of pulmonary's inflammatory cell in lung tissue; Inhibition apoptosis of vascular endothelial cell, has protective effect to pulmonary's capillary endothelial cell; Reduce fibrocyte and breed the collagen secretion increase that causes etc.Because durative action preparation is more obvious to immune inhibitory action, life-time service can disturb fat and protein metabolism, affect growth promoter, although have report to find that early application Low-dose dexamethasone can alleviate the generation of experimental rat pulmonary fibrosis, its body weight on rat has obvious impact.In the experiment in early stage, choose the positive control drug of dexamethasone, occurred that rat body weight obviously reduces, and had the phenomenon of rats death to occur.And prednisone is middle effect preparation, its half-life and half effect phase are all short than dexamethasone.Therefore in a large amount of long-term application processes, prednisone, compared with dexamethasone, brings out or increases the weight of in prevention and infect, and alleviates the side effect successful of the complication of digestive system.

Based on above reason, therefore this experiment is intended selecting prednisone as positive control drug.Experimental result shows, prednisone group is less on the general status impact of experimental rat, and lung coefficient is close with other medicines treatment group, do not occur the body weight phenomenon such as obviously become thin.And the lung function index of pulmonary fibrosis model rat to BLM induction makes moderate progress, also to HA, LN content in serum, the content of the HYP in lung tissue has certain reducing effect (P ﹤ 0.01) compared with model group.

The selection foundation of 3 indexs

3.1 integral status

Observe from traditional Chinese medical science angle, reactivity, fur color and luster, diet etc., be the important sign of the spirited prosperity and decline of reflection.Be quick on the draw, fur is moist, honey stomach, body contain strong, muscle enriches god; Otherwise bradykinesia, fur are withered, diet difficult enter, emaciated physique, withered bone with deficient medulla, extreme emaciation be without god.Lung governing qi, department breathes, and main Xuan Fasu falls, kidney governing storage, main improving inspiration by invigorating kidney-QI, therefore respiratory movement is the direct sign of reflection lung renal function.Lung qi is abundant, and kidney qi is filled Sheng, breathes normal; Otherwise lung qi virtual loss, declares respectful mistake department, kidney loses envelope and hides, and takes the photograph to receive and haves no right, and Function of Respiratory Movement is not normal.Observe from modern medicine angle: general state Observable, judge the variation of Organism of Rats integral status interstitial pulmonary fibrosis disease;

3.2 lung coefficients

In Pulmonary Fibrosis in Rats with Bleomycin-induced pathogenic process, due to body respiratory dysfunction, secondary causes digestion and the absorption of body nutrient substance, occurs serious hindrance, makes catabolism in body be greater than anabolism, makes body occur poky symptom.Interstitial pulmonary fibrosis patient's lung tissue volume reduces, and the collagen deposition that inflammation, fibrosis cause and hardening, weight increases, lung coefficient increases relatively, in addition lung tissue inflammatory exudation, edema, hyperemia, hemorrhage etc. be also the reason that the weight of lung increases, lung coefficient is one of index of reaction interstitial pulmonary fibrosis degree.Therefore bleomycin cause Pulmonary Fibrosis in Rats with Bleomycin-induced lung tissue weight increase reason comprise above-mentioned two aspects.

In 3.3 respiratory functions, detect

The indexs such as PIF, PEF, EF50, TV, EV, MV, AV, cdyn, LR, penh can judge airway patency in interstitial pulmonary fibrosis disease, breathe the state of muscle strength, lung tissue elasticity, ventilatory function and storage capacity.

3.4 impacts on relevant cell epimatrix

HA content can reacting cells epimatrix collagen fiber synthetic and precipitation situation.LN content can react the attraction of pulmonary epithelial cells, fibroblast and inflammatory cell and the situation of adhesion and T lymphocyte and macrophage secretion lymphokine and be suppressed to the situation of fibrocyte and the synthetic collagen of epithelial cell.HYP is the representative index of collagen fiber deposition.

4 Radixs Astragali, the impact of Radix Hedysari effective site on Pulmonary Fibrosis in Rats with Bleomycin-induced lung coefficient

In this experiment, the mensuration of lung coefficient contributes to assess inflammation and fibrosis.Lung coefficient results shows: the lung coefficient of model group obviously raises compared with blank group; The lung coefficient of the heavy dose of group of Radix Astragali flavone, the heavy dose of group of Radix Astragali saponin, little, the middle dosage group of Radix Hedysari flavone, Radix Hedysari saponin small dose group obviously reduces compared with model group, astragalus polysaccharides is little, in heavy dose of group, Radix Hedysari saponin, heavy dose of group induced lung coefficient reduces.Point out most of effective site groups improving in varying degrees bleomycin and cause the lung coefficient of Pulmonary Fibrosis in Rats with Bleomycin-induced, to make lung tissue weight saving, or can improve its morbid state body weight is obviously increased.

5 Radixs Astragali, the impact of Radix Hedysari effective site on Pulmonary Fibrosis in Rats with Bleomycin-induced pulmonary function

Pulmonary function method of testing has a variety of, and pulmonary function test (pft) method in the past, mainly with tracheotomy, is carried out the mode of tracheal intubation, connects toy respiratory function test macro, test each side index.Leading indicator comprises that FVC, FEV0.2, Cydn, MVV, LR, Re etc. react the variation of pulmonary function.

In order to promote quality of experiments, transformation experiment condition, the French Biotech EMKA of the company pulmonary function equipment of the special introduction of my special interest group pulmonary function laboratory, IOX Data Collection Software based and equipment, can do the noinvasive animal lung function under waking state, have a wound animal lung function by tracheal intubation under aerosol narcotism.This method is surveyed pulmonary function under waking state, and animal freely breathes, and by the pressure receptor sensing on plethysmography box, each index is unaffected.Anesthesia method adopts isoflurane inhalation anesthesia, and anesthesia is fast, clear-headed also fast, only needs the time in several seconds; By tracheo-laryngoscope mode intubate, animal is without wound.Consider, this method has been avoided anesthetics and the impact of operation on animal lung function index to greatest extent.Main test index is: TI (inspiratory duration), TE (expiratory duration), the length of reaction breathing time; PIF (inspiratory airflow peak), PEF (expiratory airflow peak), judges the intensity that reaction is breathed by crest (trough) height of respiratory curve waveform; TV (aspirated volume), EV (forced volume,expiratory), the cubical content that reaction is breathed; RT (intermission) is the middle down-time period of twice breathing; MV (ventilation per minute), the efficiency that reaction is breathed; The amount of gas exchange is effectively carried out in AV (alveolar ventilation) reaction; F (respiratory frequency), the speed that reaction is breathed; EIP (pause air-breathing latter stage), EEP (EEP pause), Penh (bronchoconstriction degree).After the pulmonary function completing under noinvasive state, just anaesthetize and have the pulmonary function test (pft) of (rcv) under wound state.Adopt isoflurane inhalation anesthesia, insert tracheal intubation, then rat clinostatism is placed in to the airtight plethysmography box of toy respirator.First trace one section of eupnea, then survey TI (inspiratory duration), TE (expiratory duration), PIF (inspiratory airflow peak), PEF (expiratory airflow peak), TV (aspirated volume), EV (forced volume,expiratory), RT (intermission), MV (ventilation per minute); F (respiratory frequency), EIP (air-breathing latter stage pause) EEP (EEP pauses), Penh (bronchoconstriction degree) cdyn(Cdgn dyanamic compliance), Penh (bronchoconstriction degree), LR(lung resistance), dppl(pleura presses and changes), EEW (EEP merit), pau(pause) etc.By having wound pulmonary function and noinvasive pulmonary function test, carry out lung capacity, the mensuration of ventilatory function not only can find that there is without extremely distinguishing restricted or obstructive dysfunction of pulmonary ventilation.

Little, the middle dosage group of Radix Hedysari flavone and model group comparison numerical value in the impact of (rf) TV (aspirated volume) are obviously raise, Radix Astragali saponin small dose group, the heavy dose of group of Hedysamn polysaccharide and relatively numerical value rising of model group.The heavy dose of group of Hedysamn polysaccharide in the impact of (rf) EV (forced volume,expiratory) is obviously raise with model group comparison numerical value, little, the middle dosage group of Radix Astragali flavone, Radix Hedysari flavone be little, in, heavy dose of group and model group relatively numerical value raise.Little on Radix Astragali flavone in the impact of (rcv) TV (tidal volume), in, little, the heavy dose of group of heavy dose of group, Radix Hedysari flavone obviously raises with model group comparison numerical value, dosage group in Radix Hedysari flavone, Radix Hedysari saponin be little, in, heavy dose of group and model group relatively numerical value raise.On in dosage group, Radix Hedysari flavone in Radix Astragali flavone small dose group, Radix Astragali saponin small dose group, Hedysamn polysaccharide in the impact of (rcv) EV (forced volume,expiratory), heavy dose of group, Radix Hedysari saponin small dose group and model group comparison numerical value obviously raises, dosage group in astragalus polysaccharides, the heavy dose of group of Hedysamn polysaccharide and model group relatively numerical value raise.These two indexs have reacted above-mentioned treatment group aspirated volume and expiration volume all increases to some extent than model group, have met restrictive ventilatory disorder and have made moderate progress through treatment, and pulmonary fibrosis alleviates to some extent.

Prednisone group, the heavy dose of group of astragalus polysaccharides, little, the middle dosage group of Radix Hedysari flavone and model group comparison numerical value in the impact of (rf) MV (ventilation per minute) are obviously raise, and Radix Astragali flavone small dose group and model group relatively numerical value raise.This index has reflected elasticity and the airway resistance of breathing muscle strength, thorax and lung tissue, is the reliability index of an overall merit pulmonary ventilation function storage level.This met restrictive ventilatory disorder through treatment after make moderate progress, pulmonary fibrosis alleviates to some extent.

On (rf) AV(alveolar ventilation) impact in little, the middle dosage group of Radix Astragali flavone, Radix Hedysari flavone small dose group and model group comparison numerical value obviously raise, in Radix Astragali saponin small dose group, Radix Hedysari flavone dosage group and model group relatively numerical value raise.This index refers to that the amount of fresh air of suction alveolar per minute or per minute and blood carry out the amount of gas exchange.The ventilation that only enters alveolar just has the chance of the ventilation of participating in, and this is the important indicator of reaction pulmonary ventilation function.This met restrictive ventilatory disorder through treatment after make moderate progress, pulmonary fibrosis alleviates to some extent.

On (rcv) cdyn(lung compliance) impact in the heavy dose of group of astragalus polysaccharides, Radix Astragali flavone little, in, heavy dose of group, the heavy dose of group of astragalus polysaccharides, Radix Hedysari flavone be little, in, heavy dose of group obviously raises with model group comparison numerical value, prednisone group, the heavy dose of group of Hedysamn polysaccharide and model group relatively numerical value raise.Lung compliance refers to caused volume when unit pressure changes, in order to reflect the elasticity of lung tissue, generally include lung compliance, chest wall compliance and total compliance, and Cdgn dyanamic compliance is one wherein, it is the compliance of the lung that records when air-flow is not blocked in the breathing cycle, it is subject to the impact of airway resistance, indirectly can reflect the volume-variation of lung; When pulmonary fibrosis, the elastic resistance of lung increases, and the compliance of lung reduces, and expiratory dyspnea appears in patient.Cdyn(Cdgn dyanamic compliance) be a sensitive indicator of reflection pulmonary fibrosis degree, in the time that its value reduces, instruction lung compliance is poor, and now pulmonary fibrosis degree increases the weight of; In the time that its value raises, instruction pulmonary fibrosis degree alleviates.Lung compliance attenuating is mainly seen in Fei Xian Weiization ﹑ Fei Shui Zhong ﹑ pulmonary atelectasis and pneumonia etc. makes lung expand limited Pulmonary Diseases.When pulmonary fibrosis, due to interstitial lung inflammation, fibrosis or honeycomb sample change mutually mixed with normal structure, elastance of lung lowers, and therefore lung compliance reduces.This met restrictive ventilatory disorder through treatment after make moderate progress, pulmonary fibrosis alleviates to some extent.

This experiment is applied in the evaluation procedure of rats with pulmonary interstitial fibrosis model and curative effect of medication measuring pulmonary function technology.The main pathological change of IPF is interstitial lung fibroblast proliferation, and can form honeycomb lung late period.Because interstitial lung exists a large amount of fibroblasts, there is restrictive ventilatory functional disturbance, the compliance of lung capacity and lung reduces.In the dosage group contrast that there were significant differences from each index, just can find several effective site dosage groups of curative effect optimum in this test.Experimental result shows: compared with model group Radix Hedysari flavone little, in, heavy dose of group, little, the middle dosage group of Radix Astragali flavone improve obviously most indexs, these group therapeutic effect are more excellent.

6 Radixs Astragali, the impact of Radix Hedysari effective site on Pulmonary Fibrosis in Rats with Bleomycin-induced serum HA, LN

Someone finds that Serum Laminin (Laminin, LN) and hyaluronic acid (Hyaluronic, HA) can be used as a kind of mark in research Interstitial Lung Disease.The interstitial lung that the discovery HA such as Li accumulate in damage suppresses the gas exchange between air and blood.Lung tissue cell injury, the release cells factor, cytokine activation fibroblast synthetic a large amount of HA, also increase the synthetic of HA after Interstitial cell irriate.HA is a kind of polysaccharide, and it is also connected with other matrix components as a kind of intermediary being connected with cell surface, structure that can stable matrix, and have the effect of modified cells at cell surface.HA is the simplest a kind of glycosaminoglycan of structure in extracellular matrix, and someone thinks that matrix components mainly contains collagen protein, proteoglycan and adhesion glycoprotein, these compositions mix interweave make to be organized into as a whole, to complete certain physiological function.Under pathological state, its content, distribution and serum-concentration can change.In the research of pulmonary fibrosis in the past, just there is scholar to find that the variation of lung local cells epimatrix can cause that in serum, the level of extracellular matrix increases, and think that these compositions that increase derive from lung.

A lot of research points out that HA is relevant with the generation of a lot of lungs illness.Especially in the generation of pulmonary fibrosis, play an important role.HA also participates in the anabolic effect of collagen fiber, can make static fibrocyte activation, forms and has fibroblast synthetic and secretion collagen protein function.On fibroblast film, have HA receptor, HA can be connected with covalency and non-covalent two kinds of modes with fibroblast by membrane receptor.After HA is combined with cell-membrane receptor, activated protein kinase, promotes the synthetic of DNA and protein.Therefore the HA increased content in serum prompting lung local organization Extracellular Matrix Levels increases, and HA content in serum reduces prompting lung local cells epimatrix level and reduces.

HA serum test experience result shows; Compared with model group in little, heavy dose of group, Radix Astragali flavone of prednisone group, astragalus polysaccharides, in heavy dose of group, Radix Astragali saponin, heavy dose of group, Radix Hedysari flavone is little, in, heavy dose of group is by above-mentioned mechanism, the value that significantly reduces HA reduces the local ECM level of lung, and pulmonary fibrosis alleviates (P ﹤ 0.01) to some extent.

ECM comprises collagen, laminin，LN, fibronectin, proteoglycan and elastin laminin etc.And ECM reconstruction disorder is the main manifestations of pulmonary fibrosis.LN is produced by various kinds of cell, comprise I type alveolar epithelial cells and vascular endothelial cell etc., LN has various biological effect, comprise inducing cell adhesion, Growth and Differentiation, also attract, adhere to chrotoplast, inflammatory cell, and stimulate T lymphocyte and macrophage secretion lymphokine, promote the synthetic of collagen, someone thinks that matrix components mainly contains collagen protein, proteoglycan and adhesion glycoprotein, and these compositions mix to interweave to be made to be organized into as a whole, and nationality is to complete certain physiological function.Under pathological state, its content, distribution and serum-concentration can change.LN belongs to the macromole glycoprotein in cellular matrix, is distributed widely in the clear layer of basement membrane, forms basement membrane skeleton together with collagen, and LN is mainly synthetic by the endotheliocyte and the epithelial cell that are positioned at basement membrane under normal circumstances, and in serum, content is less.Many results of study prove, LN has participated in endothelial cell damage process, has brought into play important function especially in pulmonary fibrosis process, become one of monitoring pulmonary fibrosis pathological changes compared with sensitive indicator.The numerical value increase of LN shows epithelial cell damage, constantly has new Growth of Cells reparation and hypertrophy to cause epithelium to thicken.LN increases and also can cause inflammatory cells increased and accumulate in basement membrane, and damage lung tissue, causes fibrosis.

LN serum test experience result shows: compared with model group prednisone group, Radix Astragali flavone little, in, heavy dose of group, little, the middle dosage group of Radix Astragali saponin, little, the middle dosage group of Radix Hedysari flavone is by above-mentioned mechanism, the value that significantly reduces LN slows down the collagen aggregate velocity in ECM, and pulmonary fibrosis alleviates (P ﹤ 0.01) to some extent.

7 Radixs Astragali, the impact of Radix Hedysari effective site on Pulmonary Fibrosis in Rats with Bleomycin-induced lung tissue homogenate HYP

Pulmonary fibrosis is due to due to the over-deposit of ECM, and the speed that in lung, collagen protein forms is greater than the speed of degraded, and collagen fiber increase gradually, finally cause collagen to accumulate gradually.HYP is the speed that the speed of collagen fiber collagen protein formation is greater than degraded, and collagen fiber increase gradually, finally cause collagen to accumulate gradually.HYP is the representative index of collagen fiber deposition.HYP is distinctive class of amino acid in collagen fiber, and in collagen protein, content accounts for 12%～14%, and HYP is mainly present in collagen protein, and its hetero-organization contains HYP hardly.A synthetic significant process of collagen is exactly, proline is converted into HYP through hydroxylase effect, in each tropocollagen molecule, in every a peptide chain, at least to there be 100 HYP residues just can make the triple-helix structure of collagen stable, this stable proportionate relationship of peptide chain in HYP and tropocollagen molecule, makes HYP assay become the obvious index of indirect reflection collagen content and is widely used in pulmonary fibrosis diagnosis.HYP is the peculiar aminoacid of one of composition body collagen protein, except elastin laminin contains a small amount of hydroxyproline (approximately 1%), nearly all hydroxyproline is all present in collagen, the main component of ECM when collagen is organ fibrosis, the Fibrotic degree of hydroxyproline in tissue energy reaction member, organizes the mensuration of hydroxyproline content to become the important method of research organization's collagenic supersession.

HYP tissue homogenate test experience result shows: compared with model group the heavy dose of group of astragalus polysaccharides, Radix Astragali flavone little, in, heavy dose of group, Radix Hedysari flavone be little, in, heavy dose of group is by above-mentioned mechanism, significantly reduce collagen fiber and deposit and make pulmonary fibrosis alleviate to some extent (P ﹤ 0.01).

This experiment is from the ordinary circumstance of rat, in lung coefficient, pulmonary function, serum, the content of the content of HA, LN and lung tissue HYP is observed, experimental result shows, compared with model group, each effective site group is all improved effect in majority parameters, illustrate that each effective site group all has certain inhibitory action to the process of pulmonary fibrosis, all can suppress the progress of pulmonary fibrosis, this explanation drug level is very large to the function influence of disease treatment, thus make this therapeutic scheme little, in, grouping in heavy dose is more meaningful.Illustrate the Radix Astragali, Radix Hedysari effective site little, in, heavy dose of relevant with its drug dose to suppressing the process of pulmonary fibrosis.

The innovative point of 8 tests

The improvement of 8.1 pulmonary fibrosis in rats preparation methoies

In the preparation process of pulmonary fibrosis model in the past, there is the modeling method that splashes into bleomycin and intratracheal instillation bleomycin through nostril.But both have its shortcoming, the latter is classical modeling method.The former modeling success rate is not high, and classical modeling method wound is difficult for more greatly recovering.Therefore this subject group has been improved classical modeling method.

This test copies pulmonary fibrosis model, under small animal laryngoscope guiding, carries out, and this is further Improvement and perfection in classical pulmonary fibrosis model preparation.Adopt small animal laryngoscope guidance tracheal intubation can avoid the skin of neck of incision rat successively to separate and expose the method for trachea, eliminated the postoperative local infection of incision skin of rat and the inducement of some complication.In modeling process, apply small animal laryngoscope guiding and can reduce the sewing process in classical modeling process, and can reduce the consumption of anaesthetic, in the process of a large amount of animal models, can greatly improve efficiency.Having avoided secondary insult is the improvement to classical modeling method.Modeling method after improvement, have easy, without wound, speed are fast, modeling success rate is high feature.This test is obtained a result: the induced lung coefficient of model group significantly raises compared with blank group, and lung function index TV, the EV of rat, MV, AV, cdyn significantly reduce.The value of HA, LN significantly raises this, and just to point out this improved pulmonary fibrosis in rats preparation method be more successful.

The application of 8.2 novel animal lung function systems

My subject is for the accuracy of the lung function index that promotes lung pattern disease experiment and detect and comprehensive, and improve experiment equipment and introduced the animal lung function detection system of French emka company, and application AniRes animal lung function analytical system.Applying Scientific Articles that this system experimentation achievement delivers has had many sections to be published on the magazine that external SCI etc. includes.In the world, the method for animal lung function analysis now is roughly divided into has wound body to retouch case method; Noinvasive body is retouched case method.

8.2.1 there is wound body to retouch case method

Animal is put into airtight body and retouches in case, implement tracheal intubation, animal breath need to have accurate animal respirator Aided Machine ventilation, detects the airway pressure of animal and the variation of lung capacity simultaneously, and then calculates airway resistance and the lung compliance of toy.This method can very accurately detect the minor variations of toy tidal volume, and data result is also the most accurate and the most stable, is mainly used in requiring very high scientific research.Due to its higher precision and sensitivity, well stability and repeatability, apply also the most extensively, be the first-selected system of toy pulmonary function scientific research.Because it can obtain extraordinary experimental result, more than 95% scientific research institution adopts this type systematic in the world.

8.2.2 noinvasive body is retouched case method

Animal is put into body and retouch case, external air source is retouched in case and is inputted a highly stable air to body, ensures that animal has normal oxygen supply, is unlikely to be choked to death, and measures animal turnover body simultaneously and retouch the gas flow of case.The index detecting comprises tidal volume, inspiratory duration and the expiratory duration etc. of animal, and then calculates penh value, for the pulmonary function change of assessment animal indirectly.

Conclusion

This experiment modeling method is slight to integral animal damage, and every detection index and pathological change all have significant difference with normal group, and pathological change typical case.

The Radix Astragali, the each effective site group of Radix Hedysari can, by effectively suppressing the process of Pulmonary Fibrosis in Rats with Bleomycin-induced model of bleomycin induction, stop the development of interstitial pulmonary fibrosis, and wherein the Radix Astragali, little, the middle dosage group of Radix Hedysari flavone effect are more excellent.

The Radix Astragali, Radix Hedysari effective site stop the action intensity of interstitial pulmonary fibrosis process relevant with suitable equivalent drug level.

Comprehensive Assessment, Radix Hedysari flavone is excellent compared with other effective site to the influence of lung fibrosis in rats pulmonary function, HA, LN, HYP.

Finally it should be noted that: the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although the present invention is had been described in detail with reference to previous embodiment, for a person skilled in the art, its technical scheme that still can record aforementioned each embodiment is modified, or part technical characterictic is wherein equal to replacement.Within the spirit and principles in the present invention all, any amendment of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.

Claims

1. the Radix Astragali, the application of Radix Hedysari effective site in preparation treatment pulmonary fibrosis medicine; Wherein, the described Radix Astragali, Radix Hedysari effective site are one or both in Radix Astragali saponin and Radix Hedysari saponin.