CN104116957B - Pharmaceutical composition for treating organ fibrosis and preparation method and application thereof - Google Patents

Abstract

The invention provides a pharmaceutical composition for treating organ fibrosis. The pharmaceutical composition is an agent prepared from the following raw material medicines in parts by weight: 7-13 parts of ephedra, 7-13 parts of almond, 15-20 parts of caulis bambusae in taeniam, 10-15 parts of scutellaria baicalensis, 10-15 parts of the root bark of white mulberry, 6-10 parts of ginseng, 15-20 parts of astragalus, 15-20 parts of the root of red-rooted salvia, 10-15 parts of Chinese angelica, 2-4 parts of crocus sativus, 6-10 parts of ligusticum wallichii, 15-20 parts of radix ophiopogonis, 15-20 parts of asparagus fern, 15-20 parts of the root of straight ladybell and 4-6 parts of liquorice. The invention also provides a preparation method and application of the pharmaceutical composition. According to monarch, ministerial and adjuvant drugs, the various medicines are clear in functions and supplement with one another, the essence of embattled speculative scientific research methodology in a compound compatibility medicine unique in traditional Chinese medicine is fully exerted, the effects of stopping asthma, dredging collaterals and strengthening healthy energy can be achieved, the composition has a good treatment effect on'excessive superficial and deficient root' special syndrome types of pulmonary fibrosis in the formula, and a novel choice is provided for clinical medication.

Description

Fibrotic pharmaceutical composition of a kind for the treatment of organs and its production and use

Technical field

The present invention relates to Fibrotic pharmaceutical composition of a kind for the treatment of organs and its production and use.

Background technology

Pulmonary fibrosiss (Pulmonary fibrosis/PF) disease is that many reasons cause pneumonia and alveolar persistence Damage and the destruction repeatedly of extracellular matrix (Extracelluluar matrix/ECM), reparations, reconstruction and over-deposit so as to Cause the class spectrum of disease that normal lung tissue structure changes and function is lost, be that many reasons cause the common of chronic lung disease Final result, is the main pathological basis of respiratory failure, and its cause of disease and pathogenesis are complicated, although its definite pathogenesis is at present still Thoroughly do not illustrate, but the regulation and control of cytokine profiles and its network are the factors of the utmost importance that it occurs development pathomechanism.

Pulmonary fibrosiss belong to the intractable great disease of the high class of international concern degree, and research both at home and abroad is thought：Fibrosiss The cureless basic reason of chronic disease, be also its disable, lethal main cause, have nearly 50% death relevant with fibrosiss, And the cureless basic reason of various chronic lung diseases is that it has fibrosis lesion.Pulmonary fibrosiss (PF) are organ fibrosis In most representative, most typically most Chang Fazhe, its cause of disease is complicated, in addition to atypical pneumonia can cause PF, many lung pattern diseases Cause PF, at least more than the 150 kinds causes of disease are associated.In treatment, non-specific antiinflammatory, immunosuppressant are still only limitted at present And the method such as glucocorticoid, its common feature is the absence of specific treatment, and in a large number using antibiotic and hormone, this is inherently It is to cause fibrosiss and increase Fibrotic major reason.With its sickness rate, the continuous rising of mortality rate, PF researchs become World Focusing focus, therefore seek other therapies such as Chinese medicine into the task of top priority.

It is considered that the fibrosiss of organ cannot be reversed once being formed.However, since last century the nineties, in a large number Experimental studies results show, can be reverse including the organ fibrosis including pulmonary fibrosiss.The institute of various organ fibrosis The reverse similar with conditioning generating process may pass through correct treatment, is because on cellular level and molecular level each The process that kind of fibrosiss are formed is just closely similar, the characteristics of with reaching the same goal by different routes.These similar generating process promptings, Yi Zhongqi The result of study of official's fibrosiss mechanism, is also applied for other various organ fibrosis；To a kind of the effective of organ fibrosis Therapeutic Method, is also beneficial (Border WA.Evidence that to preventing, mitigating other various histoorgan fibrosiss TGF-βshould be a therapeutic target in diabetic nephropathy.Kidney Int,1998, 54:1390-1391.).In view of this, both at home and abroad many scholars concentrate strength on from the beginning of a kind of reverse of organ fibrosis, gradually The research that various organ fibrosis are reversed is expanded to, a large amount of evidences that organ fibrosis can be reversed is obtained, including lung Fibrosiss.

Although pulmonary fibrosiss have treatment reversibility, still lack satisfied western modern medicine means of prevention at present, this may be with The correlations such as multifactor, pathological changes Mutiple Targets, the multiformity of pathogenesis of pulmonary fibrosiss morbidity.As a kind of typical complexity Disease, the doctor trained in Western medicine chemicalses Jing based on single target spot often seems helpless, and the traditional medicine such as Chinese medicine is in organ fibrosis Treatment aspect possess very big potentiality and advantage, the Chinese medicine based on determined curative effect develops the medicine of anti-organ fibrosis, special It is not exploitation pulmonary fibrosis resistant, hepatic fibrosis, the medicine of renal fibrosiss, prospect is increasingly wide.

The traditional Chinese medical science thinks that pulmonary fibrosiss belong to typically " deficiency in origin and excess in superficiality " disease, its pathogenesis is " disease for a long time → this empty ", " empty, resistance fold ", can belong to traditional Chinese medical science network disease category (Li Rong. treat the theory of Chinese medical science base of pulmonary fibrosiss by moxa-moxibustion fei shu region below the heart Yu Plinth with treat foundation. Liaoning Journal of Traditional Chinese Medicine, 2004,31 (4):291-293.).Yang Ke etc. reports a kind of Fuzheng Huayu Recipe, main Composition is wanted to be Herb Gynostemmae Pentaphylli, Radix Salviae Miltiorrhizae, Cordyceps, for anti-Pulmonary Fibrosis in Rats, effect is obvious.The patent No.：CN 201110162621.6, there is provided a kind of pharmaceutical composition for treating pulmonary fibrosiss, its raw material is consisted of：Radix Ginseng 5-15 parts, pellet Ginseng 5-15 parts, Radix Angelicae Sinensis 1-11 parts, Rhizoma Polygonati 5-15 parts, Fructus Perillae 5-15 parts, Semen Sinapis Albae 1-11 parts, Fructus Aurantii Immaturus 1-11 parts, Herba Ardisiae Japonicae 5- 15 parts, effect that human embryo lung (HEL) diploid fibroblast 2BS cell strains can be suppressed to breed treats pulmonary fibrosiss.The patent No.： 201110162761.3, there is provided a kind of pharmaceutical composition for treating pulmonary fibrosiss, its raw material is consisted of：Radix Ginseng 5-15 parts, pellet Ginseng 7.5-22.5 parts, Folium Ginkgo 7.5-22.5 parts, Radix Asparagi 7.5-22.5 parts, Rhizoma Polygonati 7.5-22.5 parts, Pericarpium Trichosanthiss 7.5-22.5 Part, Radix Angelicae Sinensis 2.5-7.5 parts, Radix Notoginseng 2.5-7.5 parts, can improve organ fibrosis patient clinical symptom and life quality.

For current treatment, Chinese medicine in the treatment to organ fibrosis (especially pulmonary fibrosiss), Great progress is achieved, therefore, from the Fibrotic new drug of Chinese medicine angle research and development treating organs, it has also become current Study hotspot.

The content of the invention

It is an object of the invention to provide Fibrotic new pharmaceutical compositions of a kind for the treatment of organs and preparation method thereof and using On the way.

The invention provides a kind of Fibrotic pharmaceutical composition for the treatment of organs, it is by the crude drug of following weight proportion The preparation being prepared from：

Herba Ephedrae 7-13 parts, Semen Armeniacae Amarum 7-13 parts, Caulis Bambusae In Taenia 15-20 parts, Radix Scutellariae 10-15 parts, Cortex Mori 10-15 parts, Radix Ginseng 6-10 Part, Radix Astragali 15-20 parts, Radix Salviae Miltiorrhizae 15-20 parts, Radix Angelicae Sinensis 10-15 parts, Stigma Croci 2-4 part, Rhizoma Chuanxiong 6-10 parts, Radix Ophiopogonis 15-20 parts, day Winter 15-20 part, Radix Adenophorae (Radix Glehniae) 15-20 parts, Radix Glycyrrhizae 4-6 parts.

Further, it is the preparation being prepared from by the crude drug of following weight proportion：

10 parts of Herba Ephedrae, 10 parts of Semen Armeniacae Amarum, 18 parts of Caulis Bambusae In Taenia, 12 parts of Radix Scutellariae, 12 parts of Cortex Mori, 8 parts of Radix Ginseng, 18 parts of the Radix Astragali, Radix Salviae Miltiorrhizae 18 parts, 12 parts of Radix Angelicae Sinensis, 3 parts of Stigma Croci, 8 parts of Rhizoma Chuanxiong, 18 parts of Radix Ophiopogonis, 18 parts of Radix Asparagi, 18 parts of Radix Adenophorae (Radix Glehniae), 5 parts of Radix Glycyrrhizae.

Wherein, the Herba Ephedrae is Herba Ephedrae (processed)；The Radix Astragali is Radix Astragali Preparata；The Radix Glycyrrhizae is Radix Glycyrrhizae Preparata.

Wherein, described pharmaceutical composition be by the crude drug of the weight proportion water or/and ethanol extraction for activity Composition, adds the preparation that pharmaceutically conventional adjuvant or complementary composition is prepared from.

Wherein, the preparation is Jing gastrointestinal absorption preparations.

Present invention also offers the preparation method of aforementioned pharmaceutical compositions, it comprises the steps：

(1) crude drug is weighed by the weight proportion；

(2) each medicine is added water to cook, merges decocting liquid, added pharmaceutically available adjuvant or complementary composition is prepared into often Regulation agent.

Present invention also offers aforementioned pharmaceutical compositions are in treatment or the Fibrotic medicine of auxiliary treatment organ is prepared Purposes.

Present invention research finds that aforementioned pharmaceutical compositions coordinate moxibustion treating organs fibrosiss, and its therapeutic effect is more preferably.

Further, the medicine is the medicine for treating pulmonary fibrosiss.

Further, the medicine is the medicine for reducing platelet derived growth factor content in lung tissue.

It is the traditional Chinese medical science to have been accepted as " organic conception ", " determination for the treatment of based on pathogenesis obtained through differentiation of symptoms and signs ", " compound recipe is used ", " compound compatibility medication is such as used military forces " at present Most scientific maximally effective several big advantages, wherein " compound compatibility medication is such as used military forces " is the original theory of Chinese medicine of very science.In Each medicine tight compatibility all linked with one another as arraying troops for battle in prescription is cured, is effective handss that it is better than doctor trained in Western medicine chemistry medicine composition Section.TCM prescription theory thinks that each prescription not only needs to select the appropriate compatibility of suitable medicine according to etiology and pathogenesis, while Also the basic structure of prescription should be met, i.e. the prescription compatibility theory of " monarch, minister, help, make ", the prescription of so-called " monarch, minister, help, make " Compatibility, is exactly that it sets up the scientific matching on the comprehensive judgement basis to disease pathogenesis.Middle medical square is by too many levels, many Pulmonary fibrosiss are so had fall ill multifactor, pathological changes Mutiple Targets, pathogenesis various by the biological mechanism of target spot Integrate adjustment Property the difficult disease of the complicated feature such as (" more than three "), treatment or its symptom regulated and controled, can obtain than doctor trained in Western medicine chemical drugs more preferably, More longlasting therapeutic effect, and the toxic and side effects of doctor trained in Western medicine chemical drugs are avoided, and this curative effect is built upon above-mentioned traditional Chinese medical science tradition On original theoretical accurate instruction, pharmaceutical composition of the present invention just have followed this principle.

Be to prevent that it causes too quickly anxiety to weak patient with Herba Ephedrae as monarch drug in side, be by its processed with honey, to delay its property, With for Herba Ephedrae (processed).Semen Armeniacae Amarum, Cortex Mori, Caulis Bambusae In Taenia, Radix Glycyrrhizae Preparata have an antiasthmatic effect, therefore are allowed to for ministerial drug, strengthen Dingchuan effect of Herba Ephedrae, It is " helping its advantage to press down its shortcoming " and Cortex Mori, Caulis Bambusae In Taenia, Radix Glycyrrhizae Preparata can also slow down the dry strong of Herba Ephedrae.Pulmonary fibrosiss are network disease, Pathogen usually intruding into collateral in protracted disease, it is necessary to activating blood circulation to dissipate blood stasis and dredge the collateral, therefore the Radix Salviae Miltiorrhizae for having this effect, Radix Angelicae Sinensis, Stigma Croci, Rhizoma Chuanxiong are adjusted the assistant for making left Medicine, assists a ruler in governing a country the monarch and his subjects and makes contributions from a side.And righting qi-restoratives aspect：Radix Ginseng, Radix Astragali QI invigorating are consolidated；Chinese angelica blood supplementing；" Radix Salviae Miltiorrhizae simply When four things ", it is that pole says that Radix Salviae Miltiorrhizae possesses " SIWU TANG " effectiveness of replenishing and activating blood；Radix Ophiopogonis, Radix Asparagi, Radix Adenophorae (Radix Glehniae) nourish lung yin；Radix Scutellariae lung heat clearing Heat, and the fire of the product of the property delayed heat.Therefore ginseng stilbene returns red Radix Asparagi and Radix Ophiopogonis Radix Adenophorae (Radix Glehniae) Radix Scutellariae same adjuvant drug sent as right wing of totally eight tastes by more than, from Assist the monarch and his subjects' potentiation in another side.Radix Glycyrrhizae has heat clearing away emergency to act on, and coordinating the actions of various ingredients in a prescription is good at again, therefore with it as medicine is made, coordinates full side.

Above monarch each group flavour of a drug post is clearly demarcated, complements each other, and has given full play to the exclusive compound recipe of Chinese medicine and has matched somebody with somebody Analyze mentally sexology research methodology marrow of 5 medications as arranged army and structuring the formation, the effect of can altogether play Dingchuan dredging collateral righting, for The targeted pulmonary fibrosiss of invention prescription " the real deficiency in origin of mark " special pattern of syndrome has good therapeutic effect, provides for clinical application A kind of new selection.

Specific embodiment

The preparation of the pharmaceutical composition of the present invention of embodiment 1

Take 10 grams of Herba Ephedrae (processed), 10 grams of Semen Armeniacae Amarum, 18 grams of Caulis Bambusae In Taenia, 12 grams of Radix Scutellariae, 12 grams of Cortex Mori, 8 grams of Radix Ginseng, Radix Astragali Preparata 18 Gram, 18 grams of Radix Salviae Miltiorrhizae, 12 grams of Radix Angelicae Sinensis, 3 grams of Stigma Croci, 8 grams of Rhizoma Chuanxiong, 18 grams of Radix Ophiopogonis, 18 grams of Radix Asparagi, 18 grams of Radix Adenophorae (Radix Glehniae), 5 grams of Radix Glycyrrhizae Preparata. Each flavour of a drug are weighed according to quantity, are doped with enough water purification, and intense fire changes slow fire and decocts again 10 minutes to 15 minutes after being fried boiling, and collect decocting liquid Afterwards, decoction 2 times is added water, merges each decocting liquid, obtain final product decoction.

The preparation of the pharmaceutical composition of the present invention of embodiment 2

Take 13 grams of Herba Ephedrae (processed), 13 grams of Semen Armeniacae Amarum, 15 grams of Caulis Bambusae In Taenia, 10 grams of Radix Scutellariae, 10 grams of Cortex Mori, 6 grams of Radix Ginseng, Radix Astragali Preparata 15 Gram, 15 grams of Radix Salviae Miltiorrhizae, 10 grams of Radix Angelicae Sinensis, 2 grams of Stigma Croci, 6 grams of Rhizoma Chuanxiong, 15 grams of Radix Ophiopogonis, 15 grams of Radix Asparagi, 15 grams of Radix Adenophorae (Radix Glehniae), 4 grams of Radix Glycyrrhizae Preparata. After first extracting 2 times with 70%v/v ethanol, merge alcohol extract, medicinal residues are boiled 2 times again with decocting, merge decocting liquid.Respectively by alcohol extraction After liquid, decocting liquid concentration, mixing adds appropriate dextrin, soluble starch, pelletizes, and obtains final product granule.

The preparation of the pharmaceutical composition of the present invention of embodiment 3

Take 7 grams of Herba Ephedrae (processed), 7 grams of Semen Armeniacae Amarum, 20 grams of Caulis Bambusae In Taenia, 15 grams of Radix Scutellariae, 15 grams of Cortex Mori, 10 grams of Radix Ginseng, 20 grams of Radix Astragali Preparata, 20 grams of Radix Salviae Miltiorrhizae, 15 grams of Radix Angelicae Sinensis, 4 grams of Stigma Croci, 10 grams of Rhizoma Chuanxiong, 20 grams of Radix Ophiopogonis, 20 grams of Radix Asparagi, 20 grams of Radix Adenophorae (Radix Glehniae), 6 grams of Radix Glycyrrhizae Preparata.Respectively Flavour of a drug are weighed according to quantity, and Radix Ginseng is ground as superfine end, are doped with enough water purification, with high heat remaining each medicine are decocted to boiling, change text Fire, then decoct 10 minutes to 15 minutes or so, decocting liquid is then collected, Radix Ginseng powder is blended into, mix thoroughly；Medicinal residues add water decoction 2 times, convert Powder is as before, merging filtrate, after concentration, drying, addition appropriate Microcrystalline Cellulose is encapsulated after mixing, obtains final product capsule.

The preparation of the pharmaceutical composition of the present invention of embodiment 4

Decoction prepared by Example 1, stands, and takes supernatant, after concentration, dry, pulverize, standby；Take appropriate poly- second two Alcohol 4000, after melting, above-mentioned medicated powder is added in melting substrate, is incubated at 75-85 DEG C；Using various pill dripping machine, drip inside/outside Footpath and drop speed appropriateness, in being instilled dimethicone, collect drop pill, obtain final product drop pill.

The preparation of the pharmaceutical composition of the present invention of embodiment 5

Decoction prepared by Example 1, after concentration, plus proper starch, dextrin or/and Microcrystalline Cellulose, granulation.

Beneficial effects of the present invention are illustrated below by way of test example.

Therapeutical effect of the medicine of the present invention of test example 1 to Rat Experimental pulmonary fibrosiss

1st, materials and methods

1.1 material

1.1.1 animal

Experiment uses SPF level healthy SDs rat 160, male and female half and half, 200 ± 20g of body weight, adaptability to feed one week, room temperature 18～25 DEG C, relative humidity 55～70%, drinking water is tap water, rat free water and feed during experiment.

1.1.2 primary drug and reagent

Medicine of the present invention：Prepare by embodiment 1, water-bath after decocting is concentrated into finite concentration, then needed for being made into distilled water Solution, by middle dosage concentration, i.e. 1.5g crude drugs/ml.

Stock solution zeocin bleomycin 125mg/, the used time is diluted to the solution of 4mg/ml with aseptic 0.9%NaCl solution. Folium Artemisiae Argyi.The normal saline of 250ml/ bottles 0.9%.RNAstore Sample preservation liquid.Total tissue RNA extracts kit.AccuPower GreenStar qPCR PreMix (PCR kit for fluorescence quantitative).RocketScript^TM RT PreMix (template Reverse Transcriptase kit).

1.1.3 key instrument

DF110 type electronic analytical balance scales, 20 DEG C of ultra cold storage freezers of ﹣, real-time fluorescence quantitative PCR instrument.

1.2 animal packets and modeling

Previous experiments are randomly divided into into 5 groups with SD rats：Blank group, model group, medicine middle dosage treatment group of the present invention (save Claim our group), moxibustion treatment matched group, moxa-moxibustion coordinates we's treatment matched group (save and claim moxibustion medicine group), wherein blank group 10, its Remaining per group 30.Pulmonary Fibrosis in Rats is made using classical modeling method current in the world, i.e. tracheal strips injection bleomycin Model：Under etherization, rat is fixed on operating-table, skin of neck is cut after sterilization, successively separated, fully expose gas Pipe, with 1ml syringes quantitative bleomycin diluent (5mg/kg) is extracted, and tracheal strips is disposably injected, immediately by upright The means such as rotation, make that medicine is full and uniform to intersperse among its lung tissue, sterilization, suture, and rat can wake up and freely look for after a few minutes Food.

1.3 method

Start within the 7th day after modeling treatment, medicine middle dose group of the present invention presses 10 times of people's dosage, i.e. 5g crude drugs/kg is to big Mus gavage；Blank group and model group by etc. capacity normal saline gavage as control；Moxibustion group treatment takes its " fei shu ", " region below the heart Yu " cave, Shu Points orientation reference《Subject of experimental acupuncture and moxibustion》And《Chinese Veterinary Acupuncture》Method, using moxibustion with seed-sized moxa cone, left and right replaces moxibustion, Strengthen per cave 3, often strengthen 5mg；When moxa-moxibustion coordinates our treatment of control group, ibid, but while also with our gavage, method is ibid for moxibustion.With On, 10 days are for a course for the treatment of, continuously 3 courses for the treatment of of treatment, take a day off between the course for the treatment of.In addition to blank group, each group animal is in reality There is different degrees of death during testing.After treatment end, animal is put to death using vertebra dislocation method, quickly open breast and take lung group Knit, after weighing rapidly, 150mg tissues are taken respectively and is put into RNAstore reagent (Sample preservation liquid) in proportion, use Parafim (sealed membrane) is sealed rapidly, is immediately placed in Refrigerator store.

Evaluate lung morphology to change, it is as shown in the table according to the sorting technique of Szapiel：

The alveolitises of table 1 and pulmonary fibrosiss grade scale

Gene expression detection method：Laboratory animal lung tissue sample rna is extracted, reverse transcription is carried out, Platelet is carried out The in good time quantitative fluorescence analysis of mRNA of derivative growth factors/PDGF (platelet derived growth factor).

Statistical method：Operation is wrapped in SPSS13.0 software statistics, measurement data is usedRepresent, two are adopted to sample The t inspections of independent sample.Represent that difference is statistically significant with P ＜ 0.05.

Main concrete steps：

1.3.1 pathologic state colony observation and graphical analyses：

Laboratory animal lung tissue embedding step is as follows:(1) lung tissue of rats is fixed：Induced lung evacuation, in 10% Property formalin invade at ambient temperature bubble 24h；(2) serial dehydration is as follows：50% ethanol 4h；70% ethanol 4h；80% ethanol Overnight (time is not too long)；90% ethanol 2h；The 2.5h of 95% ethanol I；The 2.5h of 95% ethanol II；The 1.5h of 100% ethanol I； The 1.5h of 100% ethanol II；(3) it is transparent：The 14min of dimethylbenzene I (depending on circumstances)；15min (depending on circumstances) (4) the waxdip bag of dimethylbenzene II Bury：The 1.5h of 52-54 degree wax I (after 1.5 hours overnight)；The 3h of 52-54 degree wax II；52-54 degree waxes；Embedding (wax stone is stored in 4 DEG C) (5) Section：Microscope slide is cleaned, dried, steeping acid, deionized water rinsing is clean, dry, 100% ethanol pad, 0.125% poly of painting rely Propylhomoserin, 37 DEG C of collected overnights are standby.By slide pencil label before section.By lung tissue wax stone, thick 5 μm section is continuously cut. Open up piece, drag for piece, 37 DEG C of incubators overnight, are cut into slices 4 DEG C and preserved.Tissue slice is carried out by a conventional method HE, micro- sem observation is simultaneously Photographic analysis.

1.3.2 HE colouring methods：

Concrete operation step：(1) embedded section is put into into room temperature from 4 DEG C；(2) conventional dewaxing：Before dimethylbenzene dewaxing, Tissue is placed in 60min or 60 DEG C of calorstat at ambient temperature toasts 20min；Tissue be individually placed to dimethylbenzene (I, 11) immersion 20min in；(3) conventional rehydration：100% ethanol I, II each 5min；95% ethanol I, II each 5min；80% ethanol 3min；70% ethanol 3min；60% ethanol 3min；50% ethanol 3min；Ddw3min；(4) haematoxylin dyeing 3-5min；(5) certainly Water rushes 5-15min (Microscopic observation after 5min)；(6) 1% hydrochloric acid, 70% ethanol color separation (quick to survey twice)；(7) tap water punching Wash 10min；(8) 1% ammonia return blue 30s, and tap water is rinsed；(9) eosin stains 10min；(10) gradient alcohol dehydration：95% second Alcohol I (fast), 95% ethanol II (fast), the slow 3min of 95% ethanol III：100% ethanol I, 100% ethanol II, 100% ethanol III are each 2min；(11) dimethylbenzene is transparent：Dimethylbenzene I, II each 10min；(12) neutral gum mounting；(13) micro- sem observation is taken pictures point Analysis.

1.3.3 RT-PCR detections：

(1) total serum IgE is extracted：1st, take out appropriate lung tissue to weigh, in being put into the mortar for filling liquid nitrogen, liquid nitrogen is more than lung group Knit, then grind lung tissue, it is degree to be ground to without clearly visible granule.The instruction homogenate of by specification, after repeatedly homogenate terminates Centrifugation.It is dried 2 to 5 minutes under room temperature condition, retains precipitation, the RNase-fress for instilling about 20-50 microlitre or so dissolves heavy Form sediment, to be allowed to be completely dissolved, -80 DEG C of refrigerators are stored in afterwards.

(2) purity and quantitative (using nucleic acid-protein quantitative instrument measure, RNA prep pure Tissue Kit description is entered Row RNA purity and Concentration Testing)

(3) reverse transcription (RocketScriptTMRT PreMix description carries out RNA reverse transcription analyses)

(4) RT-RCP reactions (according to the method operation that TIANGEN companies provide)

(5) cDNA synthesis (analysis gene expression)

2nd, result

2.1 overview

2.1.1 perusal：Blank group Rat Development is normal, activity flexibly, hair color moisten, receive food can, body weight increase just Often.Lethargy, emaciated physique, mobility are poor after modeling for model group, have withered tachypnea phenomenon, hair color, poor appetite, body weight increase Slowly.We's group is about the same with moxibustion group, body is slightly thin, it is movable can, tachypnea obvious, fur owe to moisten, receive food can, body weight Increases slowly.Moxibustion medicine group body is slightly thin, mobility can, without obvious tachypnea situation, by hair gloss, receive can, body weight have growth.

2.1.2 body weight change：From the 7th, 14,30 days body weight analysis (the results are shown in Table 2) of rat.Can be observed by form When the 7th day, each group rat body weight compares with blank group, and body weight is on a declining curve, with statistical significance (P ＜ 0.01 or P ＜ 0.05)；After modeling 14 days, still in decline or stagnant long trend, model group compares most obvious (P ＜ with blank group to the body weight of each group 0.01)；After 30 days, the body weight of rat all has different degrees of ascendant trend, but the rat body of blank group after decline or stagnant length Other four groups are remained above again.

The each rat body weight of table 2 compare (N=8, unit：g)

Note：Compare with blank group, ▲ P ＜ 0.05, ▲ ▲ P ＜ 0.01；Compare with model group, * P ＜ 0.05

2.1.3 perusal lung tissue：Blank group bilateral pulmonary is in pale red, Smooth and moist, soft texture, high resilience. After 30 days, substantially, hardness increases the Lung neoplasm of model group, and congestion is serious.In remaining each group, the pathologic of moxibustion medicine group changes Minimum, prescription group of the present invention is taken second place.

2.2 pathologic state colonies are observed

2.2.1 HE dyeing：Blank group lung tissue structure changes without alveolitises, edema and fibrosiss.Model group at 30 days, Alveolar structure is destroyed, expression of collagen in lung deposition, changes in fibrosiss；And our group, moxibustion group, moxibustion medicine group are then compared with model group Gently, wherein, moxibustion medicine group is relatively most light.

HE is dyeed and the change of light Microscopic observation pulmonary fibrosiss degree see the table below shown in (table 3)：

The each group lung fibrosis in rats alveolitises degree of table 3 impact (N=8, unit：Point)

Note：Compare with blank group, ▲ P ＜ 0.05, ▲ ▲ P ＜ 0.01；Compare with model group, * P ＜ 0.05

2.2.2 in good time quantitative fluorescence analysis results (table 4) of the mRNA of PDGF (platelet derived growth factor)：

Each group lung tissue of rats PDGF of table 4 mRNA result tables ()

Note：Compare with blank group, ▲ P ＜ 0.05, ▲ ▲ P ＜ 0.01；Compare with model group, * P ＜ 0.05, * * P ＜ 0.01

Therefore, to compare with blank group, each group PDGF value substantially increases (P ＜ 0.05, P ＜ 0.01).And and model Group is compared, and the PDGF of each treatment group then has substantially reduction, and prompting moxibustion medicine group therapy effect is most obvious, and its PDGF value is minimum, its It is secondary for our group.

3. discuss

The cause of disease and pathogenesis of organ fibrosis is more complicated, but the pathogenesis and pathology of various organ fibrosis turn Return it is essentially identical, therefore using more multiple pulmonary fibrosiss as object of study, you can deduce other various Fibrotic pathology substantially Lapse to.With deepening continuously for studying fibrosiss, it has been recognized that the cytokine of various cell releases is at Fibrotic Have very important significance in hair growth promoting exhibition.In these cytokines, transforming growth factor-β (transforming Growth factor- β, TGF-β) and platelet derived growth factor (Platelet derivative growth factors, PDGF it is considered as) to participate in histoorgan fibrosis progression and the stronger cytokine of effect.

Occur in evolution in fibrosiss, PDGF is very important regulatory factor, it is by adjusting extracellular matrix Metabolism and mediated cell synthesis collagen, stimulate secreted by alveolar macrophages fibronectin.PDGF has to fibroblast There are strong chemotaxiss and mitogenic activities, be allowed to carry out DNA replication dna, so that hypertrophy.PDGF can also be in transcriptional level Upper enhancing procollagen mRNA expression, so that the synthesis of III procollagen type increases in fibroblast, and also makes glue before its I, V-type Original synthesis increases, etc..PDGF especially plays a significant role in early, the mid-term of pulmonary fibrosiss.

After injury of lung, the PDGF of pulmonary begins to increase.Also, substantial amounts of result of study is proved, in pulmonary fibrosiss disease In change, the gene expression of PDGF increases；Early stage can then mitigate vascular remodeling and pulmonary fibrosiss degree by antagonism PDGF；Work as PDGF When expression declines, pulmonary fibrosiss degree also accordingly mitigates.

As can be seen here, the expression that present invention test passes through intervention PDGF, with sufficient scientific basis；Medicine group of the present invention Compound and its coordinate moxa-moxibustion therapy from affect PDGF expression visual angle incision, can fully point out pharmaceutical composition of the present invention and its Coordinate the curative effect effect of moxibustion treatment；And can deduce, pharmaceutical composition of the present invention and its cooperation moxibustion treatment are to other organs Fibrosiss also have therapeutical effect.

To sum up, pharmaceutical composition energy effectively treatment of the present invention or auxiliary treatment organ are particularly the fibrosiss of lung, improve it Index of correlation, its symptom is alleviated to improve the therapeutic effect of its life quality so as to reach, and avoids doctor trained in Western medicine chemicalses Serious toxic and side effects, for clinical application a kind of new selection is provided.

Claims (8)

1. the Fibrotic pharmaceutical composition of a kind for the treatment of organs, it is characterised in that：It is by the crude drug system of following weight proportion Standby preparation：

10 parts of Herba Ephedrae, 10 parts of Semen Armeniacae Amarum, 18 parts of Caulis Bambusae In Taenia, 12 parts of Radix Scutellariae, 12 parts of Cortex Mori, 8 parts of Radix Ginseng, 18 parts of the Radix Astragali, 18 parts of Radix Salviae Miltiorrhizae, 12 parts of Radix Angelicae Sinensis, 3 parts of Stigma Croci, 8 parts of Rhizoma Chuanxiong, 18 parts of Radix Ophiopogonis, 18 parts of Radix Asparagi, 18 parts of Radix Adenophorae (Radix Glehniae), 5 parts of Radix Glycyrrhizae.

2. pharmaceutical composition according to claim 1, it is characterised in that：The Herba Ephedrae is Herba Ephedrae (processed)；The Radix Astragali is to process The Radix Astragali；The Radix Glycyrrhizae is Radix Glycyrrhizae Preparata.

3. pharmaceutical composition according to claim 1 and 2, it is characterised in that：Described pharmaceutical composition is by the weight The water or/and ethanol extraction of the crude drug of proportioning is active component, adds pharmaceutically conventional adjuvant or complementary composition The preparation being prepared from.

4. the pharmaceutical composition according to claims 1 to 3 any one, it is characterised in that：The preparation is Jing gastrointestinal tract Absorbable preparation.

5. the preparation method of Claims 1 to 4 any one described pharmaceutical composition, it is characterised in that：It comprises the steps：

(1) crude drug is weighed by the weight proportion；

(2) each medicine is added water to cook, merges decocting liquid, added pharmaceutically available adjuvant or complementary composition is prepared into conventional system Agent.

6. Claims 1 to 4 any one described pharmaceutical composition is preparing treatment or the Fibrotic medicine of auxiliary treatment organ In purposes.

7. purposes according to claim 6, it is characterised in that：The medicine is the medicine for treating pulmonary fibrosiss.

8. purposes according to claim 7, it is characterised in that：The medicine is to reduce platelet derived growth factor in lung tissue The medicine of content.