CN1308019C - Chinese medicinal composition for treating chronic pelvic inflammation and preparation method thereof - Google Patents
Chinese medicinal composition for treating chronic pelvic inflammation and preparation method thereof Download PDFInfo
- Publication number
- CN1308019C CN1308019C CNB2004100541672A CN200410054167A CN1308019C CN 1308019 C CN1308019 C CN 1308019C CN B2004100541672 A CNB2004100541672 A CN B2004100541672A CN 200410054167 A CN200410054167 A CN 200410054167A CN 1308019 C CN1308019 C CN 1308019C
- Authority
- CN
- China
- Prior art keywords
- radix
- group
- ancient books
- reed mentioned
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to a Chinese medicinal composition for treating acute and chronic pelvic inflammation and a preparation method thereof. The Chinese medicinal composition is an oral preparation prepared from medicinal auxiliary materials and active ingredients extracted by using gentian root, stir-baked baical skullcap root, cape jasmine fruit, sargentgloryvine, oldenlandia, beautiful sweetgum fruit, raw rehmannia, bupleurum root, cyathula root, stir-baked nutgrass galingale rhizome, earthworm and Chinese angelica as raw materials. The Chinese medicine composition of the present invention provides a new Chinese medicinal preparation which has the efficiency of clearing heat, removing toxicity, inducing diuresis, reducing edema, coursing the liver, rectifying qi and stimulating circulation to end stasis and is particularly suitable for treating acute and chronic pelvic inflammation according to the Chinese medicine theory.
Description
Technical field
The present invention relates to field of traditional Chinese medicine pharmacy, be specifically related to a kind of Chinese medicine composition and preparation method for the treatment of urgent chronic pelvic inflammatory disease.
Background technology
Pelvic inflammatory disease is a common gynecological disease, and sickness rate is high.Acute pelvic inflammatory disease is if malpractice or delay often cause serious consequence, but the severe patient threat to life is the main paathogenic factor of infertility, intractable pelvic pain, so be one of most important sick kind in the gynecological clinic diagnosis and treatment practice.
The modern medicine treatment of acute pelvic inflammatory disease and chronic pelvic inflammatory disease acute stage, main means are to use antibiotic, but use antibiotic western medical treatment confirms to form the extensive adhesion of pelvic cavity, serious bilateral adnexa pathological changes, tangible inflammatory mass, abscess etc. to laparoscopy the most unsatisfactory curative effect of case merely, transfer to chronic or the repeatability pelvic inflammatory disease more, finally cause severe complications such as infertility and intractable pelvic pain.Pelvic tissue is very fragile during actute infection, and tear easily or bore a hole, and operation may increase the weight of to infect during actute infection, so acute pelvic inflammatory disease and be not suitable for operative treatment, even if the conservative operation also may cause infertility in young patient.
Chinese medicine acute pelvic inflammatory disease and chronic pelvic inflammatory disease acute stage have instant effect, pain relief significantly, promote inflammatory exudation, abscess, enclosed mass absorption, prevent and dissolve adhesion, shorten the course of disease, complication is few and unique curative effect such as light, and toxic and side effects is little and lack, and can obviously shorten the course of disease, alleviate and reduce sequela with the Western medicine use in conjunction.But the Chinese patent medicine that present TCM Gynecology is treated this type of disease is deficient relatively, and it is very satisfied that the clinical application curative effect is not still made us, and can not satisfy the clinical needs of this type of disease fully.
Summary of the invention
Technical problem to be solved by this invention is according to Chinese medical theory, and a kind of clearing away heat-damp and promoting diuresis that has is provided, and the dispersing liver and promoting blood circulation function is used for acute pelvic inflammatory disease and the definite Chinese medicine composition of chronic pelvic inflammatory disease therapeutic effect.
The Chinese medicine composition that is used for the treatment of acute and chronic pelvic inflammatory disease disclosed by the invention is with Radix Gentianae, Radix Scutellariae (stir-fry), Fructus Gardeniae, Caulis Sargentodoxae, Herba Hedyotidis Diffusae, Fructus Liquidambaris, Radix Rehmanniae, Radix Bupleuri, Radix Cyathulae, Rhizoma Cyperi (stir-fry), Pheretima and ought be classified as the active ingredient of raw material extraction acquisition and the oral formulations that pharmaceutic adjuvant is made.
Wherein each crude drug percentage by weight preferred group becomes: Radix Gentianae 5-10%, Radix Scutellariae 5-10%, Fructus Gardeniae 7-15%, Caulis Sargentodoxae 10-18%, Herba Hedyotidis Diffusae 7-15%, Fructus Liquidambaris 7-15%, Radix Rehmanniae 5-10%, Radix Bupleuri 2-7%, Radix Cyathulae 5-10%, Rhizoma Cyperi 5-10%, Pheretima 10-18% and Radix Angelicae Sinensis 5-10%.
Oral formulations of the present invention is pharmaceutically acceptable various dosage form, comprising: granule, tablet, capsule and oral liquid etc.
This aspect another technical problem to be solved is to provide the preparation method of above-mentioned Chinese medicine composition.
The preparation method of the Chinese medicine composition of the acute and chronic pelvic inflammatory disease of treatment disclosed by the invention comprises the following steps:
1, get Radix Gentianae, Radix Scutellariae, Rhizoma Cyperi, Radix Angelicae Sinensis and add 5-7 and doubly measure 70% alcohol reflux, merge extractive liquid, concentrates and reclaims ethanol to relative density and be the extractum of 1.10～1.15 (60 ℃～70 ℃), and is standby;
2, get after Fructus Gardeniae, Caulis Sargentodoxae, Herba Hedyotidis Diffusae, Fructus Liquidambaris, Radix Rehmanniae, Radix Bupleuri, Radix Cyathulae, Pheretima medical material add 6-8 times of water gaging and soak, decoct and extract, filter, merging filtrate also is concentrated into the extractum that relative density is 1.10～1.14 (60 ℃～70 ℃), adding ethanol makes the alcohol amount of containing reach 60-80%, leave standstill, filter, filtrate recycling ethanol also is evaporated to the extractum that relative density is 1.10～1.15 (60 ℃～70 ℃);
3, the extractum that makes of combining step 1 and 2 is made oral formulations according to a conventional method with pharmaceutic adjuvant.
Ancient medicine generally believes that the heresy damage of turbid damp is appointed and is with two arteries and veins, and two arteries and veins lose in constraint, and " body fluid gushes and overflows, sends out to be leukorrhagia " is the key factor that pelvic inflammatory disease is fallen ill.Theory of Chinese medical science thinks that leukorrheal diseases are that damp is a trouble, and the saying of " all bands are not from wet " is arranged, and "Fu Qingzhu works on obstetric" is cloud also: " husband's leukorrhagia all is a damp syndrome "." element asks lunar sun bright opinion " said: " hinder, be subjected to it earlier down " in wet person, and pelvic cavity occupies the part of the body cavity below the umbilicus, housing the bladder, kidneys and bowels, subjects to the damp invasion and attack most and is disease; As for cold, the hot heresy of sensation, also holding under the arm wet to suffering from more; Or deficiency of spleen-YANG and kidneyYANG, or stagnation of liver-QI with deficiency of the spleen, retention of water-damp in the body, flowing in the part of the body cavity below the umbilicus, housing the bladder, kidneys and bowels is disease." the typhoid fever six scripts is declared opinion " meaning: " bottom conception vessel, wet very hot, body fluid gushes and overflows; be leukorrhagia ", and for example " record is easily known by gynecological " sayed: " conception vessel is long-pending wet, wet Sheng Shengre; because of not giving birth to through changing blood; so corruption and SHENGHUANG band, as gas vacuole arteries and veins inanition or the plain body weakness of puerperal through row, and insufficiency of vital energy and blood; pathogenic factor is invaded in an opponent's defence; objective in the part of the body cavity below the umbilicus, housing the bladder, kidneys and bowels, with the QI and blood knot of fighting mutually, disharmony of nutrient QI with defensive QI is consequently fallen ill ".
The pelvic inflammatory disease pathogenesis is blamed in dampness-heat of the liver and is made a bet, and has the blood stasis retardance concurrently, and damp and hot blood stasis is tied mutually in Liver Channel and is its pathogenesis key, holds concurrently with blood circulation promoting and blood stasis dispelling so control to let out dampness-heat of the liver clearly, must go in order to do dampness-heat of the liver, and blood stasis must remove, and then all diseases are explained by oneself.The present invention prescription is in line with strengthening vital QI to eliminate pathogenic factors, and the Therapeutic Principle for the treatment of both the principal and secondary aspects of a disease and guiding theory are according to letting out dampness-heat in the liver and gallbladder thirty years of age of the method for blood circulation promoting and blood stasis dispelling of holding concurrently clearly.
Be monarch with Radix Gentianae, Radix Scutellariae in the prescription of the present invention.Radix Gentianae is the dry root and rhizome of gentianaceae plant bar leaf Radix Gentianae Gentiana manshurica Kitag., Radix Gentianae Gentiana scabra Bge., three flower Radix Gentianae Gentiana triflora pall. or Gentiana rigescens Franch Gentiana rigescens Franch., bitter cold, go into the liver and gall warp, heat clearing and damp drying, kind especially clear damp-heat in lower-JIAO.It " controls the inferior portion's rheumatism and damp and hot " " curing mainly secret formula " cloud.Compendium of Material Medica meaning " fire is posted in liver and gall mutually, have rush down of no help, so the gas of the liver benefiting gallbladder of Radix Gentianae, just also with the pathogenic heat of its energy eliminating pathogen in the liver gallbladder.The opinion of " medicine justice " is outstanding smart: " fire of the special eliminating pathogen in the liver gallbladder of Radix Gentianae ..., all genus Liver Channel pathogenic heats are for suffering from, and are wonderful with it.Its abnormal smells from the patient is thick and heavy and avale, kind clear damp-heat in lower-JIAO ".
Radix Scutellariae is the dry root of labiate Radix Scutellariae Scutellaria baicalensis Georgi, and bitter in the mouth is cold in nature, pathogenic fire purging dampness.Radix Scutellariae can directly be rolled over to let out fire with its bitter cold, turbid damp that again can dryization liver and gall warp." Bencao Jingshu " cloud: " its property of Radix Scutellariae is respectful clearly, so remove evil; So bitter in the mouth dampness; So YIN-cold wins heat, thus main all heat, pathogenic heat and also damp and hot ... roll over it, then all sick autotherapys.”
" element ask the most pure virginity will discuss greatly " said: " wet excessive in interior, control with bitter heat, help light with acid, with hardship dry it, let out it with light ".Fructus Gardeniae is the dry mature fruit of Maguireothamnus speciosus Fructus Gardeniae Gardenia jasminoidesEllis, and bitter in the mouth is cold in nature, and the property of medicine is fallen clearly, the pathogenic fire reducing heat of dispelling, main clear three warmers pathogenic fire had both been gone into gas, went into blood again, Jiao who removes heat by catharsis, damp and hot heresy is gone from urine, and in the damp and hot card that has blood stasis concurrently, that uses is wonderful especially." explaining abstruseness of the canon of materia medica ": " Zhong Jing uses Fructus Gardeniae, Herba Artemisiae Scopariae more, get its diuresis and exempt from also damp and hot ".Herba Hedyotidis Diffusae is the dry herb of Maguireothamnus speciosus Herba Hedyotidis Diffusae Oldenlandiadiffusa (Willd.) Roxb., and the clearing up internal heat by using drugs of bitter in taste and cold in nature detoxifcation is inducing diuresis and reducing edema, and " Quanzhou book on Chinese herbal medicine " says its " heat clearing away dissipating blood stasis, the pain that disappears detoxifcation ".Big Caulis Sargentodoxae is the dry rattan of Lardizabalaceae plant Caulis Sargentodoxae Sargentodoxa cuneata (Oliv.) Rehd.et Wils., and bitter in the mouth is flat, goes into Liver Channel, but the merit heat-clearing and toxic substances removing, promoting blood circulation to remove obstruction in the collateral.One helping the merit of the monarch and his subjects' clearing away heat-fire, but the blood circulation promoting and blood stasis dispelling of holding concurrently.Fructus Liquidambaris is the drying and ripening fruit ear of Hamamelidaceae plant Chinese sweet gum Liquidambar formosana Hance, function collateral dredging diuretic, too late with amended copy side's all medicine dampness removings function.More than four medicines, clearing away liver-fire, dampness removing heat is to establish for damp and hot excess pathogen, altogether the power of principal drug assistance Radix Gentianae, the dehumidifying of Radix Scutellariae purging liver-fire is share and is thought minister.
Right liver storing blood, body is cloudy and, contain then easy impairment of YIN blood in the liver-fire with sun; Bitter cold is oozed the product of profit, but also injuring YIN due to production of dryness, and so help again with the Radix Rehmanniae yin nourishing, the body of tonifying liver makes heresy go and just do not hindering; Liver controlling conveyance and dispersion rises clockwork spring and reaches, and strongly fragrant in the dampness-heat of the liver, the product of letting out fall in depression of liver-QI, bitter cold, also can depressed liver property, so, help the usefulness of liver again with Radix Bupleuri, the thin smooth liver and gall of Rhizoma Cyperi.Because of assertive evidence has blood stasis concurrently, Gu Zuo is with Radix Cyathulae, Pheretima, Radix Angelicae Sinensis three flavors, the meridian dredging, blood stasis dispelling blood again.Radix Cyathulae " Chinese medicinal herbal " is called it: " removing blood stasis decline ", outside invigorating blood circulation, this product still can dampness removing, and here damp and hot heresy from going down.More than six medicines shared for the assistant.
Modern medicine study proves, main effective ingredient gentiopicrin in the present invention prescription in the Radix Gentianae can transform through the intestinal microbial population effect in vivo and generate erythricine, gentisaldehyde and produce the antiinflammatory drug action, and the main effective ingredient baicalin in the Radix Scutellariae has stronger antibiotic, antiinflammation.The Fructus Gardeniae main chemical compositions is iridoid glycosides and organic acid etc., and iridoid glycosides has jasminoidin, geniposide, Geniposidic acid etc.; Organic acid is the ester that forms of phenylpropionic acid derivative and quininic acid such as chlorogenic acid etc.Jasminoidin can suppress the acetic acid twisting reaction of mice, and xylol and Oleum Tiglii cause mice ear obvious effect is all arranged.That Herba Hedyotidis Diffusae has is antibiotic, antiinflammatory, anti-allergy action.The chemical constituent of Radix Rehmanniae is mainly iridoid glycosides, cupreol, mannitol etc., its water decoction and pure preserved material have significant detumescence effect to rat experiment formaldehyde arthritis, and the test tube experiment shows that Radix Rehmanniae water logging agent has inhibitory action to multiple fungi growth such as trichophyton mentagrophytes, Gypsum Fibrosum sample microspore tinea bacterium and ulotrichy microspore tinea bacterium.The Rhizoma Cyperi master contains flavonoid glycoside, volatilization wet goods.Main effective ingredient is inhibited to maincenter and intestinal smooth muscle in the Rhizoma Cyperi, and lumbar injection also has antipyretic effect simultaneously.The Radix Cyathulae master contains the sterols composition, and the total Saponin of Radix Achyranthis Bidentatae all has excitation to the isolated rat uterus under the different physiological statuss; Experiment is proof also, and Radix Cyathulae has pharmacological actions such as antiinflammatory, analgesia.When being classified as gynecological's key medicine, can enrich blood and can invigorate blood circulation, its main chemical compositions is volatile oil and organic acid two parts.Modern study proves, Radix Angelicae Sinensis has the effect that promotes phagocytic function, antiallergic action, enhancing human body immunity function and hemopoietic function, and ferulic acid in the Radix Angelicae Sinensis and Radix Angelicae Sinensis polysaccharide can obviously promote mononuclear phagocyte that the Cl and the enhancing peritoneal macrophage of Congo red are engulfed erythrocytic ability etc.
In sum, the full side of Chinese medicine composition of the present invention is harmonious, and is hot clearly, wet getting profit, and the stasis of blood also disappears, and then all cards are from removing; And both bodies of tonifying liver helps the usefulness of liver again, and is eliminating evil and just do not hinder, and has heat-clearing and toxic substances removing, inducing diuresis and reducing edema, depressed liver-energy dispersing and QI regulating, blood circulation invigorating efficacies.
The present invention is according to the characteristic of each crude drug in the prescription, appropriate design the extracting method of extractum, the active ingredient of having extracted each component to greatest extent.Because the character instability of main effective ingredient gentiopicrin in the Radix Gentianae in the prescription, heated time destroys long easily the decomposition; Baicalin in the Radix Scutellariae is dissolved in hot water, ethanol, be insoluble to warm water and cold water, after frying in shallow oil with other medicine is mixed, easily enter filter cake during filtration and be used as impurity and remove, lose greatlyyer, and the main effective ingredient in the Rhizoma Cyperi, Radix Angelicae Sinensis mostly is pure soluble components, therefore, Radix Gentianae, Radix Scutellariae, Rhizoma Cyperi, Radix Angelicae Sinensis adopt 70% alcohol reflux, extracting solution after reclaiming ethanol, precipitation, filtration, filtrate for later use.And the effective ingredient in Fructus Gardeniae, Caulis Sargentodoxae, Radix Rehmanniae, Radix Cyathulae, Herba Hedyotidis Diffusae, Pheretima, Radix Bupleuri and the Fructus Liquidambaris is all water-soluble, thereby adopts decocting to boil; But because the rate of extract of water extraction is higher, the taking dose of making behind the preparation is big, and melting is also undesirable, therefore, water extraction is partly carried out precipitate with ethanol, to remove non-medicinal ingredients such as wherein starch, polysaccharide.Respectively alcohol extraction, water are mentioned that alcohol precipitation process is optimized experiment through the employing orthogonal experiment, and moulding process is carried out screening study, drawn above-mentioned preparation technology at last.Adopt thin layer chromatography that the jasminoidin in the preparation, gentiopicrin, Radix Rehmanniae, Radix Cyathulae, Radix Angelicae Sinensis and Rhizoma Cyperi are carried out qualitative analysis simultaneously, adopt high-efficient liquid phase technique that content of baicalin is measured, to guarantee the quality of preparation.
Carry out pharmacodynamic study with Chinese medicine composition of the present invention (imperial a kind of reed mentioned in ancient books pelvic cavity relaxation grain), the result shows: (13.0g crude drug/kg) can obviously reduce the chronic pelvic inflammatory disease rat inflammation pathological changes due to the escherichia coli to imperial a kind of reed mentioned in ancient books pelvic cavity relaxation grain high dose, ((13.0g crude drug/kg) all can obviously reduce the whole blood viscosity that the pelvic inflammatory disease rat model raises, and especially effect is obviously under high shear rate for 6.5g crude drug/kg), high dose in the dragon a kind of reed mentioned in ancient books pelvic cavity relaxation grain; Dragon a kind of reed mentioned in ancient books pelvic cavity relaxation grain low (2.5g crude drug/kg), in (5.0g crude drug/kg), (10.0g crude drug/kg) all can obviously reduce mouse writhing reaction and number of times illustrates that it has certain analgesic effect to high dose group, does not have obviously influence but cause pain for hot plate; Dragon a kind of reed mentioned in ancient books basic, normal, high three dosage groups of pelvic cavity relaxation grain and prednisone group xylol induced mice auricle inflammation have the obvious suppression effect; Middle and high dosage is wet to the mice granuloma induced by implantation of cotton pellets, dry weight all has and obviously alleviates effect; Dragon a kind of reed mentioned in ancient books pelvic cavity relaxation grain has the good in-vitro inhibitory action in Candida albicans, yeast, beta hemolytic streptococcus, golden Portugal bacterium, escherichia coli, Bacillus proteus, bacillus pyocyaneus, streptococcus anaerobius, bacteroides fragilis.
Carrying out the toxicologic study result with Chinese medicine composition of the present invention (imperial a kind of reed mentioned in ancient books pelvic cavity relaxation grain) shows:
Acute toxicity testing is not measured LD in the administration of imperial a kind of reed mentioned in ancient books pelvic cavity relaxation grain mouse stomach
50Situation under, irritate stomach three times in 24 hours, recording the mouse stomach maximum dosage-feeding is 378g/kg, is equivalent to intend 371.80 times of clinical people's consumption.
Long term toxicity test, imperial a kind of reed mentioned in ancient books pelvic cavity relaxation grain people intends clinical consumption and is about 1g crude drug/kg/ days, and be two weeks the course of treatment.With reference to Ministry of Public Health " study of tcm new drug guide (pharmacy, pharmacology, toxicology) ", establish high, medium and low three dosage groups and blank group, carry out the rat long term toxicity test, gastric infusion, administration capacity are 10ml/Kg, 2 times/day, continuous three months.The high dose group dosage is 63g/kg, is equivalent to intend 63 times of clinical people's consumption; In dosage 31.5g/Kg, be equivalent to intend 31.5 times of clinical people's consumption; Low dose group is 15.75g/Kg, is equivalent to intend 15.75 times of clinical people's consumption.2/3 rat is killed in work in 24 hours after the last administration, and two weeks lived and kill remaining rat after the drug withdrawal, measures indexs such as the hematology that respectively organizes rat, blood biochemical respectively, measures the main organs coefficient simultaneously, system's postmortem and microscopy.Found that: the high, medium and low dosage group of imperial a kind of reed mentioned in ancient books pelvic cavity relaxation grain compares with blank group, and three months only middle and high dosage group rats of successive administration have blood Cr obviously to raise, and renal function injury to a certain degree is described, but can recovers normal after two weeks of drug withdrawal.In the administration process, animal diet followed, activity, drainage show no obvious abnormalities, none animal dead.Each treated animal there is no notable difference at aspects such as diet, activity, drainage, the mental status after the drug withdrawal; Though organ coefficient, hematology's testing result change all in normal range; Histopathologic slide does not have specificity and changes.No slowness toxicity takes place after the drug withdrawal.
More than experiment shows that Chinese medicine composition of the present invention has the obvious treatment effect to the rat chronic pelvic inflammatory disease, and has tangible antiinflammatory, analgesia, bacteriostatic improvement effect, and acute toxicity and long term toxicity test show that this product do not have the overt toxicity effect, oral safety.
Description of drawings
Fig. 1: imperial a kind of reed mentioned in ancient books granule improves percentile influence to the mice hot plate method threshold of pain
The specific embodiment
Embodiment 1
Prescription:
Radix Gentianae 90g Radix Scutellariae (stir-fry) 90g Fructus Gardeniae 100g Caulis Sargentodoxae 150g
Herba Hedyotidis Diffusae 120g Fructus Liquidambaris 100g Radix Rehmanniae 90g Radix Bupleuri 60g
Radix Cyathulae 90g Rhizoma Cyperi (stir-fry) 90g Pheretima 150g Radix Angelicae Sinensis 90g
Preparation:
Getting Radix Gentianae, Radix Scutellariae, Rhizoma Cyperi, Radix Angelicae Sinensis is the reflux, extract, secondary with 70% ethanol, each 2 hours, add quintuple ethanol for the first time, and add four times of amounts for the second time; Merge ethanol extract, reclaim ethanol, and be concentrated into relative density 1.05 (60 ℃ of surveys), filter while hot, filtrate decompression is concentrated into relative density 1.32～1.34 (80 ℃ of surveys), and is standby.Other gets other drugs such as Fructus Gardeniae, Caulis Sargentodoxae, Herba Hedyotidis Diffusae, Fructus Liquidambaris and decocts with water secondary, add for the first time 8 times of water gagings, add for the second time 6 times of water gagings, each 1.5 hours, filter, merge medicinal liquid and be concentrated into about 1.10 (the 80 ℃ of surveys) of relative density, adding 95% ethanol makes and contains alcohol amount and reach 70%, left standstill 48 hours, and filtered, filtrate recycling ethanol also is evaporated to relative density 1.32～1.34 (80 ℃ of surveys); Merge with alcohol-extracted extract.Get 1 part of extractum, 2 parts of Icing Sugar, 0.5 part in dextrin, 0.2 part of stevioside, 80% ethanol is an amount of, mixed pelletization, and drying, granulate, packing, promptly.
Embodiment 2 quality testings
1, jasminoidin, gentiopicrin qualitative identification
Get embodiment 1 method and make preparation 10g, fine ground, add methanol 30ml supersound extraction 30 minutes, filter, filtrate evaporate to dryness, residue add water 30ml dissolving and are transferred in the separatory funnel, with the extracted with diethyl ether secondary, each 20ml, ether solution merges standby; Water liquid adds water-saturated n-butanol extraction secondary, and each 30ml merges n-butyl alcohol and washes secondary with 1% ammonia, and each 30ml discards ammoniacal liquor, and n-butyl alcohol evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution.Other gets jasminoidin, gentiopicrin reference substance, adds methanol respectively and makes every 1ml and contain 1.0mg solution, in contrast product solution.According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000) test, draw jasminoidin, gentiopicrin reference substance solution, each 5 μ l of need testing solution respectively, put respectively in same be the silica gel G F of adhesive with the sodium carboxymethyl cellulose
254On the lamellae, upper solution with n-butyl alcohol-ethyl acetate-water (4: 1: 5) is developing solvent, launch more than the 14cm, take out, dry, put under the ultra-violet lamp (254nm) and inspect, in the test sample chromatograph, with jasminoidin and the corresponding position of gentiopicrin reference substance chromatograph on, all show the speckle of same color.
2, Radix Rehmanniae is differentiated
Get the ether solution in above-mentioned 1, volatilize, residue adds methanol 2ml makes dissolving, as need testing solution.Other gets Radix Rehmanniae control medicinal material 1g, decocts with water twice, filters, and merges medicinal liquid and also is concentrated into about 10ml, adds ethanol 30ml precipitation, filter, the filtrate evaporate to dryness, from " residue adds water 20ml " with the test sample preparation method, prepare control medicinal material solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw control medicinal material solution 10 μ l, need testing solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene-ethyl acetate-methanol-formic acid (6: 3: 0.5: 0.2) be developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid.Put under the ultra-violet lamp (365nm) and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
3, Radix Cyathulae is differentiated
Get Radix Cyathulae control medicinal material 1g, decoct with water twice, filter, filtrate merges, and is concentrated into about 20ml, and the extracting twice that adds diethyl ether merges ether and evaporate to dryness, and residue adds methanol 1ml makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw each the 5 μ l of need testing solution in control medicinal material solution and above-mentioned 2 respectively, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be developing solvent with benzene-methanol (9: 0.5), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
4, Radix Angelicae Sinensis, Rhizoma Cyperi are differentiated
Get Radix Angelicae Sinensis control medicinal material 0.5g, Rhizoma Cyperi control medicinal material 1g, add 70% ethanol 20ml supersound extraction 30 minutes respectively, filter, the filtrate evaporate to dryness, residue adds water 20ml dissolving respectively, and the extracting twice that adds diethyl ether merges ether and evaporate to dryness, residue adds methanol 1ml makes dissolving, makes Radix Angelicae Sinensis control medicinal material solution and Rhizoma Cyperi control medicinal material solution respectively.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw Radix Angelicae Sinensis control medicinal material solution 2 μ l, Rhizoma Cyperi control medicinal material solution 5 μ l, need testing solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with cyclohexane extraction-ethyl acetate (9: 1) is developing solvent, launch, take out, dry, spray 10% sulphuric acid ethanol, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with Radix Angelicae Sinensis control medicinal material and the corresponding position of Rhizoma Cyperi control medicinal material chromatograph on, show the fluorescence speckle of same color respectively after.
5, content of baicalin is measured
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica, are mobile phase with methanol-glacial acetic acid-water (48: 1: 52), and the detection wavelength is 274nm.Number of theoretical plate calculates by the baicalin peak should be not less than 2500.
The preparation of reference substance solution: precision takes by weighing at 4 hours baicalin reference substance of 60 ℃ of vacuum dryings an amount of, adds 50% methanol and makes the solution that every 1ml contains 0.1mg, promptly.
The preparation of need testing solution: get the about 10g of embodiment 1 preparation, porphyrize, precision takes by weighing 1.0g, puts in the 100ml measuring bottle, and it is an amount of to add 50% methanol, supersound process 15 minutes is put to room temperature, adds 50% methanol and is diluted to scale, shakes up, filter, discard filtrate just, collect subsequent filtrate, promptly.
Algoscopy: accurate reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
Radix Scutellariae is pressed baicalin (C
21H
18O
11) meter, must not be less than 4.0mg/g.
Embodiment 3 pharmacodynamic studies
I. test material:
1. experimental animal:
Mice: the Kunming kind, 18-22g, ♀, Jiangxi Medical College's Experimental Animal Center provides.
Rat: SD is female, and body weight is the 180-250 gram, and animal housing of Jiangxi Medical College provides.
2. medicine:
Dragon a kind of reed mentioned in ancient books granule extractum: 2.5g crude drug/g extractum (proportion is 1.26), the clinical plan of people is about 1 gram crude drug/Kg/d with dosage.
II. test method and result:
One, to the influence of the rat bacterial vaginitis due to the escherichia coli
Test objective: inject rat uterus with a certain amount of escherichia coli and cause the chronic pelvic inflammatory disease model, take out the uterus after the administration and carry out perusal and histopathology observation, estimate the anti-pelvic inflammatory disease effect of medicine in conjunction with both results.
1. animal grouping: be divided into 6 groups at random, every group 10, be imperial a kind of reed mentioned in ancient books granule height (1.30g crude drug/ml), in (0.65g crude drug/ml), low dose group (0.325g crude drug/ml), FUYANKANGFU KELI group (1.50g crude drug/ml), model group and sham operated rats (distilled water).
2. get 60 of the female not copulation of healthy regular grade SD rats, body weight 180-220 gram, the anesthesia of ip30mg/kg pentobarbital sodium, behind the abdominal part routine disinfection, median incision of lower abdomen touches out internal organs, expose the uterus, the clinical new separating Escherichia coli bacterium liquid (10 of syringe pump 0.2ml
7Individual/as ml), to avoid the careful inserting needle of blood vessel with No. four syringe needles in the uterus crotch and go into uterine cavity, slowly inject 0.2ml/ only after, close the abdominal cavity, the layer-by-layer suture tissue.10 rat operation techniques are the same in addition, only do not inject the escherichia coli suspension as sham operated rats.Operation back animal is refused any disposal, normal nursing, the into water of freely ingesting.After the modeling 10 days, animal is divided into 6 groups at random, every group of 10 animals begin to carry out oral drug therapy, irritate stomach every day once, each 10ml/kg, successive administration seven days.Model group and sham operated rats such as gavage at the capacity distilled water.24h after the last administration behind the 30mg/kg pentobarbital sodium ip anesthetized animal, cuts open the belly, and abdominal aortic blood carries out whole blood viscosity to be measured, the variation substantially in perusal uterus then, and specimen taken carries out PATHOMORPHOLOGICAL OBSERVATION OF PULLORUM.
3. the pathological grading standard sees Table 1:
Table 1 pathological grading standard
| Lesion degree | Intrauterine adhesion, obturation or expansion | The chamber wall construction changes | The epithelial cell degeneration necrosis | The inner membrance congestion and edema | Fibroblast proliferation |
| Normal slight moderate severe | - 0～1/3 1/3～2/3 2/3～1 | - + ++ +++ | - + ++ +++ | - + ++ +++ | - + ++ +++ |
According to the scorching range degree of pathology, by light to heavily being divided into level Four:
Abdominal wall structure: (-) each layer is clear; Unclear or the disappearance of the disorderly systematicness of (+) proper mucous membrane gland structure; (++) flesh layer and mucous layer boundary are unclear; (+++) holostrome structure is unclear.
Epithelial cell: (-) monolayer column; (+) cell is flat or epithelial cell is downright bad comes off＜and 1/3; (++) cell is flat or exuviation is downright bad＞and 1/3; (+++) downright bad fully the disappearance.
The chronic inflammation cellular infiltration: in 10 * 20 times of following several four unit are medium-sized lymphocyte of micrometer grid and anatomy cell number, get its average: (-) each layer do not seen inflammatory cell; (+) only has a large amount of inflammatory cells at proper mucous membrane; (++) flesh layer is visible inflammatory cell (20～30) also; (+++) holostrome all has inflammatory cell, and number＞30.
Fibroblast: (-) no hypertrophy; (+) only has a spot of fibroblast at proper mucous membrane; The also visible fibroblast of (++) flesh layer, and measure more; (+++) holostrome all has fibroblast, and number＞30.
According to above-mentioned standard, the overall pathological changes of rat uterus is divided into level Four.0 group: do not have (+); I group: the inaccessible or expansion of intrauterine adhesion, and chamber wall construction no change.All the other four have below 2 (+); II group: intrauterine adhesion obturation or expansion and chamber wall construction have 2 (+) or 1 (++) in changing, and all the other four have one (++) or more than 3 (+); III group: 2 (++) or 1 (+++) are arranged during intrauterine adhesion obturation or expansion and chamber wall construction change, and all the other four have one (+++) or more than 2 (++).
4. the results are shown in Table 2 and table 3:
The scorching range degree of table 2 rat uterus is (sample number n=10) relatively
| Group | Dosage (g/kg) | The classification of scorching range degree | The Ridit value | 95% fiducial limit | |||
| - | I | II | III | ||||
| Sham operated rats | 8 | 2 | 0 | 0 | 0.178 | 0-0.325 | |
| Model control group | 0 | 1 | 6 | 3 | 0.833 | 0.6505-1.0158 △ | |
| Dragon a kind of reed mentioned in ancient books granule | 3.25 | 0 | 5 | 4 | 1 | 0.619 | 0.4365-0.8015 |
| Dragon a kind of reed mentioned in ancient books granule | 6.5 | 0 | 6 | 4 | 0 | 0.565 | 0.3825-0.7475 |
| Dragon a kind of reed mentioned in ancient books granule | 13 | 2 | 5 | 3 | 0 | 0.403 | 0.2205-0.5855 ▲ |
| The FUYANKANGFU KELI group | 2 | 3 | 5 | 2 | 0 | 0.386 | 0.2305-0.5685 ▲ |
Compare with the blank group:
△P＜0.05; Compare with model control group:
▲P＜0.05.
Can find out that from last table imperial a kind of reed mentioned in ancient books granule high dose and FUYANKANGFU KELI have clearly improvement effect to the rat chronic pelvic inflammatory disease due to the escherichia coli.
The imperial a kind of reed mentioned in ancient books granule of table 3 is to the influence (mPa) of chronic pelvic inflammatory disease rat model whole blood viscosity
| Group | Dosage (g/kg) | Number of animals (n) | 200S -1 | 100S -1 | 40S -1 | 20S -1 | 10S -1 |
| The blank group | 10 | 5.46±0.46 | 6.43±0.69 | 8.30±0.96 | 9.93±1.36 | 11.87±1.93 | |
| Model control group | 10 | 6.35±0.81 △△△ | 7.70±1.11 △△ | 10.12±1.55 △△ | 12.85±2.06 △△ | 16.01±2.55 △△ | |
| The low agent group of dragon a kind of reed mentioned in ancient books | 3.25 | 10 | 6.32±0.62 | 7.71±0.75 | 10.31±1.04 | 12.73±1.12 | 15.19±1.29 |
| Agent group in dragon a kind of reed mentioned in ancient books | 6.5 | 10 | 5.71±0.46 ▲ | 6.88±0.72 ▲ | 9.23±1.16 | 11.47±1.45 ▲ | 13.90±1.70 ▲ |
| The high agent group of dragon a kind of reed mentioned in ancient books | 13 | 10 | 5.64±0.31 ▲▲ | 7.22±0.74 | 9.41±0.88 | 11.60±1.31 | 14.43±1.96 |
| The FUYANKANGFU KELI group | 2 | 10 | 5.59±0.31 ▲▲ | 7.11±0.48 | 9.27±0.80 | 11.25±0.80 ▲▲ | 13.69±1.12 ▲▲ |
Compare with the blank group:
△P＜0.05,
△ △P＜0.01; Compare with model control group:
▲P＜0.05,
▲ ▲P＜0.01.
As can be seen from the above table, middle and high dosage of imperial a kind of reed mentioned in ancient books granule and FUYANKANGFU KELI all can obviously reduce the whole blood viscosity that the pelvic inflammatory disease rat model raises, and especially effect is obvious under high shear rate.
Two, to the influence of mice analgesic activity
(-) writhing method:
Test objective: adopt mouse peritoneal injected chemical medicine, calculate the Mus number and the number of times of mouse writhing reaction, observe the analgesic activity of medicine.
1. animal grouping: animal is divided into 6 groups at random, and 10 every group, the basic, normal, high dosage group of promptly imperial a kind of reed mentioned in ancient books granule, dolantin group, model group and matched group.
2. observation index: model group and matched group give normal saline, other each groups are all irritated stomach and are given relative medicine 20ml/kg, after the administration 60 minutes, remove the matched group intraperitoneal injection of saline, the equal lumbar injection 0.6% acetic acid 0.2ml of each Mus, observe in 10 minutes each group and writhing response mice number of elements (abdominal part indent, stretch hind leg, buttocks is raised) and number of times occur, the results are shown in following table.
The imperial a kind of reed mentioned in ancient books granule of table 4 is to the influence of mice analgesic activity
| Group | Number of animals | Dosage (the g crude drug/kg) | Body Mus number (only) appears turning round | Turn round body number of times (inferior) |
| The blank group | 10 | NS | 0 | 0 |
| The high agent group of agent group dragon a kind of reed mentioned in ancient books dolantin in the low agent group dragon of model group dragon a kind of reed mentioned in ancient books a kind of reed mentioned in ancient books | 10 10 10 10 10 | NS 5 10 20 20mg | 10 △△△ 10 △△△ 10 △△△ 10 △△△ 2 ▲▲▲ | 24.20±6.88 △△△ 18.60±6.63 ▲ 17.20±2.85 ▲▲ 14.80±2.14 ▲▲▲ 0.30±0.67 ▲▲▲ |
Compare with the blank group,
△ △ △P＜0.001; Compare with model group,
▲P＜0.05,
▲ ▲P＜0.01,
▲ ▲ ▲P＜0.001.
As can be seen from the above table, lumbar injection 0.6% glacial acetic acid can cause mice and pain reaction occurs.Model group mouse writhing reaction times showed increased, illustrate that modeling is successful, remove the dolantin group and can obviously reduce mouse writhing reaction times and number of elements, the basic, normal, high dosage group of imperial a kind of reed mentioned in ancient books granule all can obviously reduce mouse writhing reaction and number of times, illustrates that it has certain analgesic effect.
(2) hot plate method:
Test objective: adopt thermostimulation, observe mice and give time of licking metapedes behind the different pharmaceutical, as the index of the analgesic effect of weighing medicine.
1. animal grouping: animal is divided into 5 groups at random, and 14 every group, the basic, normal, high dosage group of promptly imperial a kind of reed mentioned in ancient books granule, dolantin group, blank group (normal saline).
2. observation index: 90 of female mices, water bath with thermostatic control is adjusted to 55 ± 0.5 ℃, the contact water surface at the bottom of the metal dish, the heating back is as thermostimulation, certainly drop into hot plate to the pain threshold of the time of metapedes occurring licking with stopwatch record mice as this Mus, is qualified with pain threshold in second at 5-30, sieve totally 70 of mices, carry out random packet by its pain threshold, it is the same to divide into groups, matched group gives normal saline, and other each groups are all irritated stomach and given relative medicine 20ml/kg, and be administered once every day, continuous 7 days, after the last administration 30,60,120, surveyed the mice pain threshold respectively in 240 minutes, and calculated its threshold of pain and improve percentage rate, the formula calculating down of threshold of pain raising rate.The results are shown in Table 5, table 6 and Fig. 1.
The imperial a kind of reed mentioned in ancient books granule of table 5 causes the influence of pain effect to the mice hot plate method
| Group | Number of animals | Dosage (the g crude drug/kg) | Before giving approximately (s) | Different time licks sufficient incubation period (s) after the administration | |||
| 30min | 60min | 120min | 240min | ||||
| The high agent group of agent group dragon a kind of reed mentioned in ancient books dolantin in the low agent group dragon of blank group dragon a kind of reed mentioned in ancient books a kind of reed mentioned in ancient books | 10 10 10 10 10 | NS 5 10 20 20mg | 16.38±2.22 15.96±3.57 16.35±3.71 16.39±2.58 16.42±2.70 | 18.35±4.92 18.64±5.34 17.93±3.75 16.71±3.17 25.42±4.61 △ | 18.57±5.09 19.42±6.72 19.35±5.51 17.35±3.00 28.71±5.34 △△ | 18.28±4.35 20.64±5.89 19.50±4.92 18.14±4.11 25.35±3.99 △ | 18.35±3.8 21.07±7.33 19.92±7.62 18.28±5.49 26.57±5.69 △ |
Compare with the normal saline group,
△P＜0.05,
△ △P＜0.01.
6 imperial a kind of reed mentioned in ancient books granules are proposed the influence of rate to the mice hot plate method threshold of pain
| Group | Number of animals | 30 minutes (%) | 60 minutes (%) | 120 minutes (%) | 240 minutes (%) |
| Dosage group high dose group dolantin group in the matched group low dose group | 14 14 14 14 14 | 12.02 16.79 9.66 1.95 54.81 | 11.67 21.68 18.35 6.12 74.84 | 13.37 29.32 19.27 10.95 54.38 | 14.41 32.02 21.83 11.80 61.81 |
Compare with matched group,
△P＜0.05,
△ △P＜0.01.
Fig. 1 shows that imperial a kind of reed mentioned in ancient books granule improves percentile influence to the mice hot plate method threshold of pain.
From table 5,6 and Fig. 1 can find out, the dolantin group just can obviously improve the mice pain threshold in 30 minutes after administration, in the time of 60,90,120 minutes, also act on obviously, and imperial a kind of reed mentioned in ancient books granule various dose group does not have tangible raising effect to the mice pain threshold, illustrates that imperial a kind of reed mentioned in ancient books granule is not obvious to the analgesic activity of hot plate method.
Three, to the influence of mouse corrosion disease effect
(1) auricle edema method:
Test objective: give medicine by being coated with again after dimethylbenzene causes inflammatory model, weigh the antiinflammatory action of medicine by the weight difference that compares two ears to Mice Auricle.
1. animal grouping: animal is divided into 6 groups at random, and 10 every group, the basic, normal, high dosage group of promptly imperial a kind of reed mentioned in ancient books granule, prednisone group, FUYANKANGFU KELI group and matched group.
2. observation index: matched group gives normal saline, other each groups are all irritated stomach and are given relative medicine 20ml/kg, successive administration 7 days, the last administration after 60 minutes left auricle be coated with 100% dimethylbenzene 0.02ml/ only, after 4 hours, mice is pulled cervical vertebra put to death, cut two ears along the auricle baseline, lay round auricle at same position respectively with the straight card punch of 8mm, torsion balance is weighed; Every Mus left side auricle deducts the auris dextra sheet and heavily is the swelling degree.The results are shown in following table.
The imperial a kind of reed mentioned in ancient books granule of table 7 is to the influence of mice auricle swelling
| Group | Number of animals | Dosage (the g crude drug/kg) | Left side ear heavy (mg) | Auris dextra heavy (mg) | The swelling degree |
| The blank group | 10 | NS | 21.72±3.94 | 12.38±2.10 | 9.34±2.68 |
| The high agent group of agent group dragon a kind of reed mentioned in ancient books prednisone in the low agent group dragon of FUYANKANGFU KELI group dragon a kind of reed mentioned in ancient books a kind of reed mentioned in ancient books | 10 10 10 10 10 | 3 5 10 20 20mg | 19.75±2.23 19.64±2.17 ▲ 19.70±1.82 ▲ 19.23±2.10 ▲ 18.26±2.07 ▲▲ | 12.21±1.15 12.33±1.52 12.20±1.14 11.59±1.77 11.90±1.82 | 7.54±2.40 7.32±1.48 ▲ 7.50±1.41 ▲ 7.63±1.49 ▲ 6.36±2.12 ▲▲ |
Compare with the normal saline group,
▲P＜0.05,
▲ ▲P＜0.01,
▲ ▲ ▲P＜0.001.
As can be seen from the above table, obviously alleviate effect, illustrate outside basic, normal, high three the agent particle complex groups of imperial a kind of reed mentioned in ancient books granule that other each groups have the obvious suppression effect to mouse ear amount group and prednisone group xylol induced mice auricle inflammation except that the wide swelling degree of FUYANKANG has.
(2) cotton ball granular swelling:
Test objective: give medicine by implanting again after cotton balls causes inflammatory model, weigh the antiinflammatory action of medicine by the weight difference that compares cotton balls to mice.
1. animal grouping: animal is divided into 6 groups at random, and 10 every group, the basic, normal, high dosage group of promptly imperial a kind of reed mentioned in ancient books granule, prednisone group, FUYANKANGFU KELI group and matched group.
2. observation index: under aseptic condition, the heavy aseptic cotton balls of 5mg is implanted each one of oxter, both sides behind the etherization, sew up wound, matched group gives normal saline, and other each groups are all irritated stomach and are given relative medicine 20ml/kg, once a day, successive administration 7 days; Put to death mice after 60 minutes after the last administration, peel off the ball granulation and claim its weight in wet base, baking was weighed after 2 hours in 80 ℃ of baking boxs, was dry weight, the results are shown in following table.
The imperial a kind of reed mentioned in ancient books granule of table 8 is to the influence of mice granuloma induced by implantation of cotton pellets
| Group | Number of animals | Dosage (the g crude drug/kg) | Weight in wet base (mg) | Suppression ratio | Dry weight | Suppression ratio |
| The blank group | 10 | NS | 98.59±12.41 | 22.60±3.81 | ||
| The high agent group of agent group dragon a kind of reed mentioned in ancient books prednisone in the low agent group dragon of FUYANKANGFU KELI group dragon a kind of reed mentioned in ancient books a kind of reed mentioned in ancient books | 10 10 10 10 10 | 3 5 10 20 20mg | 84.99±8.96 ▲▲▲ 97.36±8.01 85.09±8.57 ▲▲▲ 82.24±6.72 ▲▲▲ 78.92±7.91 ▲▲▲ | 13.79 1.25 13.69 16.58 19.95 | 18.05±1.57 ▲▲▲ 20.86±2.02 17.59±1.64 ▲▲▲ 17.47±1.44 ▲▲▲ 16.85±1.87 ▲▲▲ | 20.13 7.70 22.17 22.70 25.44 |
Compare with the normal saline group,
▲P＜0.05,
▲ ▲P＜0.01,
▲ ▲ ▲P＜0.001.
As can be seen from the above table, except that imperial a kind of reed mentioned in ancient books granule low dose group, other each group is wet to the mice granuloma induced by implantation of cotton pellets, dry weight all has and obviously alleviates effect, illustrates that middle and high two the dosage groups of imperial a kind of reed mentioned in ancient books granule, FUYANKANGFU KELI group and prednisone group have the obvious suppression effect to inflammation.
Four, extracorporeal bacteria inhibitor test
Test objective: adopt the certain culture base to isolated bacterial or mycete are cultivated the external bacteriostasis of observation medicine recently.
(1) to pelvic inflammatory disease common pathogen inhibitory action
Dragon a kind of reed mentioned in ancient books granule is mixed with the yellowish-brown supernatant liquid that concentration is 3.15g crude drug/ml with distilled water, and adjusting PH is about 7.2.Boil the 30min sterilization, 4 ℃ of refrigerators are preserved stand-by.FUYANKANGFU KELI is mixed with the yellowish-brown supernatant liquid that concentration is 3.89g crude drug/ml with distilled water, and adjusting PH is about 7.2.Boil the 30min sterilization, 4 ℃ of refrigerators are preserved stand-by.Use earlier strain: escherichia coli, staphylococcus aureus, Bacillus proteus, beta hemolytic streptococcus, each 7 strain of bacillus pyocyaneus provide (clinical separation strain recently) by Jiangxi Province healthcare hospital for women ﹠ children.Escherichia coli, staphylococcus aureus, bacillus pyocyaneus are adopted dextrose bouillon buffering culture fluid, and Bacillus proteus, beta hemolytic streptococcus are adopted serum broth.The contrast document, adopt test tube doubling dilution [6]: 10 of small test tubes getting sterilization place on the test tube rack, add phenol red broth bouillon of glucose or the phenol red broth bouillon 1ml of 1% serum glucose with the every pipe in sterile working again: the 1ml of sterile working, put into the 1st pipe, draw 1ml behind the mixing and put into the 2nd pipe, draw 1ml behind the mixing and put into second pipe, draw 1ml behind the mixing and put into the 3rd pipe, so doing doubly, amount is diluted to the 9th pipe, draw 1ml behind the mixing and discard, the 10th pipe does not add antibacterials and compares; Every pipe (removing the 9th pipe) adds tested antibacterial bacterium liquid 0.1ml, cultivates behind the mixing, and the 9th pipe does not add antibacterial bacterium liquid and does the medicine contrast; Put in 37 ℃ of incubators, cultivated 18-24 hour, observed result during according to tested bacterial growth decomposition glucose produce bacterium and make phenol red indicator discoloration (red yellowing) judge every pipe bacterial growth situation, there is not the test tube antibacterial of variable color not grow, the test tube bacterial growth of variable color, that concentration of managing contained antibacterials that the maximum dilution that does not have variable color is doubly measured is as minimal inhibitory concentration (MIC) then.
The result: imperial a kind of reed mentioned in ancient books granule is crude drug/mJ O.79g to beta hemolytic streptococcus MIC: Bacillus proteus, escherichia coli MIC are crude drug/ml O.20g: to bacillus pyocyaneus, staphylococcus aureus MIC is 0.395g crude drug/ml; FUYANKANGFU KELI is 0.486g crude drug/ml to the MIC of golden Portugal bacterium, escherichia coli, bacillus pyocyaneus; Distortion Du bacterium is 0.243g crude drug/ml, and the MIC of beta hemolytic streptococcus is crude drug/ml O.972g.
The imperial a kind of reed mentioned in ancient books granule of table 9 is to the result of 5 kinds of external bacteriostasis of bacterial strain
| Bacterial strain | Dragon a kind of reed mentioned in ancient books medicinal liquid dilution factor | MIC | |||||||||
| 1∶2 | 1∶4 | 1∶8 | 1∶16 | 1∶32 | 1∶64 | 1∶128 | 1∶256 | 1∶512 ※ | Contrast ★ | ||
| Beta hemolytic streptococcus | - | - | + | + | + | + | + | + | - | + | 1∶4 |
| Staphylococcus aureus | - | - | - | + | + | + | + | + | - | + | 1∶8 |
| Escherichia coli | - | - | - | - | + | + | + | + | - | + | 1∶16 |
| Bacillus proteus | - | - | - | - | + | + | + | + | - | + | 1∶16 |
| Bacillus pyocyaneus | - | - | - | + | + | + | + | + | - | + | 1∶8 |
※Expression has the aseptic medicine contrast of medicine
★Expression has bacterium not have the antibacterial contrast of medicine
Table 1O FUYANKANGFU KELI is to the result of 5 kinds of external bacteriostasis of bacterial strain
| Bacterial strain | FUYANKANGFU medicinal liquid dilution factor | MIC | |||||||||
| 1∶2 | 1∶4 | 1∶8 | 1∶16 | 1∶32 | 1∶64 | 1∶128 | 1∶256 | 1∶512 ※ | Contrast ★ | ||
| Beta hemolytic streptococcus | - | - | + | + | + | + | + | + | - | - | 1∶4 |
| Staphylococcus aureus | - | - | - | + | + | + | + | + | - | + | 1∶8 |
| Bacillus proteus | - | - | - | - | + | + | + | + | - | + | 1∶16 |
| Escherichia coli | - | - | - | + | + | + | + | + | - | + | 1∶8 |
| Bacillus pyocyaneus | - | - | - | + | + | + | + | + | - | + | 1∶8 |
※Expression has the aseptic medicine contrast of medicine
★Expression has bacterium not have the antibacterial contrast of medicine
(2) imperial a kind of reed mentioned in ancient books granule is to the inhibitory action of fungus
Be subjected to reagent thing compound method the same, Candida albicans, each 7 strain of yeast adopt the husky Ruo Shi of guarantor culture medium to cultivate by (clinical separation strain recently) is provided by Jiangxi Province healthcare hospital for women ﹠ children.Test operation, cultural method the results are shown in Table 11,12 all with above-mentioned test.
The imperial a kind of reed mentioned in ancient books granule of table 11 is to the vitro inhibition effect of fungus
| Bacterial strain | Dragon a kind of reed mentioned in ancient books medicinal liquid dilution factor | MIC | |||||||||
| 1∶2 | 1∶4 | 1∶8 | 1∶16 | 1∶32 | 1∶64 | 1∶128 | 1∶256 | 1∶512 ※ | Contrast ★ | ||
| Candida albicans | - | - | - | - | + | + | + | + | - | + | 1∶8 |
| Yeast | - | - | - | + | + | + | + | + | - | + | 1∶8 |
※Expression has the aseptic medicine contrast of medicine
★Expression has bacterium not have the antibacterial contrast of medicine
Table 12 FUYANKANGFU KELI is to the vitro inhibition effect of fungus
| Bacterial strain | FUYANKANGFU medicinal liquid dilution factor | MIC | |||||||||
| 1∶2 | 1∶4 | 1∶8 | 1∶16 | 1∶32 | 1∶64 | 1∶128 | 1∶256 | 1∶512 ※ | Contrast ★ | ||
| Candida albicans | - | - | - | + | + | + | - | + | - | + | 1∶8 |
| Yeast | - | - | - | + | + | + | + | + | - | + | 1∶4 |
※Expression has the aseptic medicine contrast of medicine
★Expression has bacterium not have the antibacterial contrast of medicine
The result: imperial a kind of reed mentioned in ancient books granule is 0.20g crude drug/ml to Candida albicans MIC; To yeast MIC is 0.395g crude drug/ml; The FUYANKANGFU KELI dialogue is read and saccharomycetic MIC is 0.486g crude drug/ml.
(3) imperial a kind of reed mentioned in ancient books granule is to the inhibitory action of anaerobe
Be subjected to reagent thing compound method the same, bacteroides fragilis, streptococcus anaerobius provide (clinical separation strain recently) by Jiangxi Province healthcare hospital for women ﹠ children.By the sterile working streptococcus anaerobius, bacteroides fragilis are diluted with the glucose broth of 10% calf serum at 1: 1000 earlier, be mixed with the required bacterium liquid of test; After dosing finishes with the dosing test tube in vacuum desiccator, extract with vacuum pump that air is positioned over to cultivate in 35 ℃ of incubators again and observed in 48 hours to vacuum state (negative pressure reaches 0.1Torr) in the exsiccator, the results are shown in Table 13,14.
The imperial a kind of reed mentioned in ancient books granule of table 13 is to the vitro inhibition effect of anaerobe
| Bacterial strain | The about liquid dilution factor of dragon a kind of reed mentioned in ancient books | MIC |
| 1∶2 | 1∶4 | 1∶8 | 1∶16 | 1∶32 | 1∶64 | 1∶128 | 1∶256 | 1∶512 ※ | Contrast ★ | ||
| Streptococcus anaerobius | - | - | - | + | + | + | + | + | - | + | 1∶8 |
| Bacteroides fragilis | - | - | + | + | - | + | + | + | - | + | 1∶8 |
※Expression has the aseptic medicine contrast of medicine
★Expression has bacterium not have the antibacterial contrast of medicine
Table 14 FUYANKANGFU KELI is to the vitro inhibition effect of anaerobe
| Bacterial strain | FUYANKANGFU medicinal liquid dilution factor | MIC | |||||||||
| 1∶2 | 1∶4 | 1∶8 | 1∶16 | 1∶32 | 1∶64 | 1∶128 | 1∶256 | 1∶512 ※ | Contrast ★ | ||
| Streptococcus anaerobius | - | - | - | + | + | + | + | + | - | + | 1∶8 |
| Bacteroides fragilis | - | - | - | + | + | + | + | - | - | + | 1∶4 |
※Expression has the aseptic medicine contrast of medicine
★Expression has bacterium not have the antibacterial contrast of medicine
The result: imperial a kind of reed mentioned in ancient books granule is 0.79g crude drug/ml to bacteroides fragilis MIC; To streptococcus anaerobius MIC is 0.395g crude drug/ml; FUYANKANGFU KELI is 0.486g crude drug/ml to the MIC of bacteroides fragilis and streptococcus anaerobius.
Conclusion
1, in imperial a kind of reed mentioned in ancient books granule ((13.0g crude drug/kg) and FUYANKANGFU KELI all can obviously reduce the whole blood viscosity that the pelvic inflammatory disease rat model raises, and especially effect is obviously under high shear rate for 6.5g crude drug/kg), high dose.
2, imperial a kind of reed mentioned in ancient books granule low (2.5g crude drug/kg), in (5.0g crude drug/kg), (10.0g crude drug/kg) all can obviously reduce mouse writhing reaction and number of times illustrates that it has certain analgesic effect to high dose group, does not have obviously influence but cause pain for hot plate.
3, basic, normal, high three dosage groups of imperial a kind of reed mentioned in ancient books granule and prednisone group xylol induced mice auricle inflammation have the obvious suppression effect; Middle and high dosage is wet to the mice granuloma induced by implantation of cotton pellets, dry weight all has and obviously alleviates effect.
4, imperial a kind of reed mentioned in ancient books granule has the good in-vitro inhibitory action in Candida albicans, yeast, beta hemolytic streptococcus, golden Portugal bacterium, escherichia coli, Bacillus proteus, bacillus pyocyaneus, streptococcus anaerobius, bacteroides fragilis.
Embodiment 4
Prescription:
Radix Gentianae 120g Radix Scutellariae (stir-fry) 100g Fructus Gardeniae 140g Caulis Sargentodoxae 200g
Herba Hedyotidis Diffusae 180g Fructus Liquidambaris 160g Radix Rehmanniae 160g Radix Bupleuri 45g
Radix Cyathulae 180g Rhizoma Cyperi (stir-fry) 180g Pheretima 330g Radix Angelicae Sinensis 100g
Preparation:
Getting Radix Gentianae, Radix Scutellariae, Rhizoma Cyperi, Radix Angelicae Sinensis is the reflux, extract, secondary with 70% ethanol, each 2 hours, add quintuple ethanol for the first time, and add six times of amounts for the second time; Merge ethanol extract, reclaim ethanol, and be concentrated into relative density 1.05 (60 ℃ of surveys), filter while hot, filtrate decompression is concentrated into relative density 1.32～1.34 (80 ℃ of surveys), and is standby.Other gets other drugs such as Fructus Gardeniae, Caulis Sargentodoxae, Herba Hedyotidis Diffusae, Fructus Liquidambaris and decocts with water secondary, add for the first time six times of water gagings, add for the second time 6 times of water gagings, each 1.5 hours, filter, merge medicinal liquid and be concentrated into about 1.10 (the 80 ℃ of surveys) of relative density, adding 95% ethanol makes and contains alcohol amount and reach 80%, left standstill 48 hours, and filtered, filtrate recycling ethanol also is evaporated to relative density 1.32～1.34 (80 ℃ of surveys); Merge with alcohol-extracted extract.Get the part extract dry, the powder that gets dry extract is granulated with residue extractum, and drying with the appropriate amount of auxiliary materials mixing, is pressed into 1000, the bag film-coat, and packing, promptly.
Embodiment 5
Prescription:
Radix Gentianae 120g Radix Scutellariae (stir-fry) 90g Fructus Gardeniae 150g Caulis Sargentodoxae 225g
Herba Hedyotidis Diffusae 105g Fructus Liquidambaris 180g Radix Rehmanniae 75g Radix Bupleuri 75g
Radix Cyathulae 75g Rhizoma Cyperi (stir-fry) 75g Pheretima 150g Radix Angelicae Sinensis 120g
Preparation:
Getting Radix Gentianae, Radix Scutellariae, Rhizoma Cyperi, Radix Angelicae Sinensis is the reflux, extract, secondary with 70% ethanol, each 2 hours, add seven times of amount ethanol for the first time, and add quintuple for the second time; Merge ethanol extract, reclaim ethanol, and be concentrated into relative density 1.05 (60 ℃ of surveys), filter while hot, filtrate decompression is concentrated into relative density 1.32～1.34 (80 ℃ of surveys), and is standby.Other gets other drugs such as Fructus Gardeniae, Caulis Sargentodoxae, Herba Hedyotidis Diffusae, Fructus Liquidambaris and decocts with water secondary, add for the first time 7 times of water gagings, add for the second time 8 times of water gagings, each 1.5 hours, filter, merge medicinal liquid and be concentrated into about 1.10 (the 80 ℃ of surveys) of relative density, adding 95% ethanol makes and contains alcohol amount and reach 70%, left standstill 48 hours, and filtered, filtrate recycling ethanol also is evaporated to relative density 1.32～1.34 (80 ℃ of surveys); Merge with alcohol-extracted extract, add an amount of stevioside, boil dissolving, put coldly, add essence, antiseptic is an amount of, adds water to 1000ml, stirs evenly, and leaves standstill, filter, embedding, sterilization, promptly.
Claims (2)
1, a kind of Chinese medicine composition for the treatment of urgent chronic pelvic inflammatory disease is characterized in that said composition is to consist of with percentage by weight: Radix Gentianae 5-10%, Radix Scutellariae (parched) 5-10%, Fructus Gardeniae 7-15%, Caulis Sargentodoxae 10-18%, Herba Hedyotidis Diffusae 7-15%, Fructus Liquidambaris 7-15%, Radix Rehmanniae 5-10%, Radix Bupleuri 2-7%, Radix Cyathulae 5-10%, stir-fry Rhizoma Cyperi 5-10%, Pheretima 10-18% and Radix Angelicae Sinensis 5-10% are that raw material extracts the active ingredient of acquisition and the oral formulations that pharmaceutic adjuvant is made.
2, a kind of preparation method of Chinese medicine composition of treatment urgent chronic pelvic inflammatory disease as claimed in claim 1 is characterized in that this method comprises the following steps:
1) get Radix Gentianae, Radix Scutellariae, Rhizoma Cyperi, Radix Angelicae Sinensis and add 5-7 and doubly measure 70% alcohol reflux, merge extractive liquid,, concentrating and reclaiming ethanol to relative density is 1.10～1.15 extractum, standby;
2) get after Fructus Gardeniae, Caulis Sargentodoxae, Herba Hedyotidis Diffusae, Fructus Liquidambaris, Radix Rehmanniae, Radix Bupleuri, Radix Cyathulae, Pheretima medical material add 6-8 times of water gaging and soak, decoct and extract, filter, merging filtrate and to be concentrated into relative density be 1.10～1.14 extractum, adding ethanol makes the alcohol amount of containing reach 60-70%, leave standstill, filter, filtrate recycling ethanol and to be evaporated to relative density be 1.10～1.15 extractum;
3) combining step 1) and 2) extractum that makes, oral formulations made according to a conventional method with pharmaceutic adjuvant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100541672A CN1308019C (en) | 2004-08-31 | 2004-08-31 | Chinese medicinal composition for treating chronic pelvic inflammation and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100541672A CN1308019C (en) | 2004-08-31 | 2004-08-31 | Chinese medicinal composition for treating chronic pelvic inflammation and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1742918A CN1742918A (en) | 2006-03-08 |
| CN1308019C true CN1308019C (en) | 2007-04-04 |
Family
ID=36138543
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004100541672A Active CN1308019C (en) | 2004-08-31 | 2004-08-31 | Chinese medicinal composition for treating chronic pelvic inflammation and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1308019C (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102600429A (en) * | 2012-03-29 | 2012-07-25 | 常熟市常福有机复合肥有限公司 | Traditional Chinese medicine composition for treating qi and blood stagnation chronic pelvic inflammation and preparation method thereof |
| CN104367727A (en) * | 2014-11-19 | 2015-02-25 | 青岛祥翔生物医药科技有限公司 | Traditional Chinese medicine for treating acute pelvic inflammatory disease caused by damp-heat syndrome |
| CN106353446B (en) * | 2016-08-12 | 2020-06-23 | 上海黄海制药有限责任公司 | Annian granule identification and content determination method |
| CN108267529A (en) * | 2018-03-22 | 2018-07-10 | 佛山市中医院 | The method of quality control of Trauma Yellow-water preparation |
-
2004
- 2004-08-31 CN CNB2004100541672A patent/CN1308019C/en active Active
Non-Patent Citations (3)
| Title |
|---|
| 芍药蒺藜煎在妇科病中的应用 吴葆卿，陕西中医，第12卷第5期 1991 * |
| 龙胆泻肝汤古今应用 赵秀荣，中国史学杂志，第26卷第1期 1996 * |
| 龙胆泻肝汤古今应用 赵秀荣，中国史学杂志，第26卷第1期 1996;芍药蒺藜煎在妇科病中的应用 吴葆卿，陕西中医，第12卷第5期 1991 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1742918A (en) | 2006-03-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102145111B (en) | Traditional Chinese medicine composition for treating colpitis mycotica and preparation method thereof | |
| CN109674958A (en) | A kind of Chinese medicine composition and its preparation method and application with anti-trioxypurine | |
| CN105535505A (en) | Method for preparing medicine composition for treating hepatic calculus | |
| CN103800772A (en) | Gynecological external lotion for preventing and treating vagina inflammatory diseases and preparation method thereof | |
| CN101461926B (en) | Medicament composition for treating gland hyperplasia and preparation method thereof | |
| CN102085248B (en) | Traditional Chinese medicinal composition for treating cervix diseases, method for preparing same and method for detecting same | |
| CN1686185A (en) | Traditional Chinese medicina preparation for treating oral cavity, throat disease and its preparation method | |
| CN1709302A (en) | Chinese medicine formulation for treating chronic pelvic inflammation and its preparing method | |
| CN1308019C (en) | Chinese medicinal composition for treating chronic pelvic inflammation and preparation method thereof | |
| WO2005074952A1 (en) | Chinese medicine for treatment of irritable bowel sysndrome and the preparation thereof | |
| CN1270742C (en) | Antiseptic Chinese medicine composition and its prepn process | |
| CN109568430B (en) | Preparation method and application of immunopotentiator | |
| CN1850202A (en) | Vagina external-use medicine composition and its preparing method and use | |
| CN100389796C (en) | Chinese medicinal preparation for treating chronic prostatitis and its manufacturing method | |
| CN1748779A (en) | Medicinal composition for treating chronic prostatitis and its preparation method and use | |
| CN1186052C (en) | Medicine for treatment of pelvic inflammation, its preparation and preparing method | |
| CN1298351C (en) | Chinese medicine oral preparaton for treating urinary system infestation and its preparation method | |
| CN100408055C (en) | Chinese medicinal composition for treating traumatic diseases, its preparation method and quality control method | |
| CN115607586A (en) | Application of dandelion and monomer compound thereof in mite killing | |
| CN101721437B (en) | Preparation method of medicine composition used for treating chronic pharyngitis | |
| CN110613712A (en) | Chinese medicinal composition and its application in treating vasculitis and endothelial injury | |
| WO2004084924A1 (en) | Pharmaceutical composition for treatment of bph and preparation thereof | |
| CN1220515C (en) | Medicine for treating throat disease | |
| CN114796419B (en) | Traditional Chinese medicine composition for treating acute pancreatitis and application thereof | |
| CN116850236B (en) | Externally-applied traditional Chinese medicine composition for treating psoriasis as well as preparation and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20221107 Address after: 201499 5th floor, building 1, 1888 Wangyuan Road, Fengxian District, Shanghai Patentee after: Shanghai Shuofang Pharmaceutical Technology Co.,Ltd. Address before: 200023 Shanghai city Luwan District Ruijin road Haixing Plaza No. 1 16A Patentee before: Shanghai Haitian Medical Technology Development Co.,Ltd. |