CN102836367A - Bacteria-resisting, inflammation-diminishing and itching-relieving traditional Chinese medicine composition and preparation method and application thereof - Google Patents
Bacteria-resisting, inflammation-diminishing and itching-relieving traditional Chinese medicine composition and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine composition, and in particular relates to a bacteria-resisting, inflammation-diminishing and itching-relieving traditional Chinese medicine composition and a preparation method and application thereof. The traditional Chinese medicine composition avoids having toxic and side effects on human bodies and has the functions of resisting bacteria, diminishing inflammation, relieving itching, regulating vaginal flora disturbance, increasing the quantity of vaginal lactobacillus and improving the immune functions of the bodies. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 20-30 parts of fructus cnidii, 20-30 parts of giant knotweed rhizome, 20-30 parts of radix sophorae flavescentis, 10-20 parts of dandelions, 10-15 parts of densefruit pittany root-bark, 10-20 parts of radix stemonae, 5-10 parts of amur corktree bark, 5-10 parts of dahurian angelica root, 5-10 parts of white alum, 3-6 parts of cloves and 30-50 parts of golden larch bark. The traditional Chinese medicine composition has the functions of clearing away heat and toxic materials, dispelling wind and eliminating dampness and killing parasites to relieve itching and can be used on health products such as sanitary towels in aid of gynecological disease treatment.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition, be specifically related to a kind of anti-inflammation itching relieving Chinese medicine composition.
Background technology
Along with all kinds of antibiotic that with the penicillin are representative constantly occur, bacterial infection disease is effectively controlled, thereby makes antibiotic become the powerful mean of human opposing pathogenic bacterial infection.But in recent years, people use antibiotic to cause drug-resistance of bacteria constantly to strengthen for a long time widely, and the drug resistance frequency increases gradually, so that conventional antibiotic can't meet clinical needs.Significant variations along with global spectrum of disease and medical model; Progressively integrating with of China and World Economics, culture and drug level; Compatriots are enhanced to antibiotic understanding; Recognize many untoward reaction and corresponding chemical sproof influence thereof that antibiotic brings, people have been full of expectation to Chinese medicine maintaining healthy and disease preventing and treating.Medical circle also expects to tackle through rational use of drug and exploitation new Chinese medicine the fastbacteria of continuous generation.
Along with people's growth in the living standard at present, health care product be also more and more, especially there is health-care effect in gynecological, and health sanitary napkin is the health care of women articles for use, and pathogenic microorganism is had the wide spectrum killing action.To common gram positive bacteria such as staphylococcus aureus, streptococcus, common gram negative bacteria such as escherichia coli, bacillus pyocyaneus, common fungus such as Candida albicans, acrothesium floccosum etc. all have good antibacterial activity.But it adds the method for Bactericidal medicines such as hibitane, domiphen bromide, metronidazole in various sanitary towels, because user is had pessimal stimulation and toxic and side effects, is substituted by Chinese herbal medicine type sanitary towel gradually.
Summary of the invention
The object of the invention provides a kind of Chinese medicine compound compositions; By multiple natural plants through refining; Pure Chinese medicine elite is processed, and avoids human body is produced toxic and side effects, have antibiotic, antiinflammatory, antipruritic, adjustment vaginal microbial flora is disorderly and increase the effect of vagina lactobacillus quantity, human body immunity improving function.
Technical scheme of the present invention is for providing a kind of anti-inflammation itching relieving Chinese medicine composition, comprises that following parts by weight of Chinese traditional medicine raw material processes: Fructus Cnidii 20-30, Rhizoma Polygoni Cuspidati 20-30, Radix Sophorae Flavescentis 20-30, Herba Taraxaci 10-20, Cortex Dictamni 10-15, Radix Stemonae 10-20, Cortex Phellodendri 5-10, Radix Angelicae Dahuricae 5-10, Alumen 5-10, Flos Caryophylli 3-6, Cortex Pseudolaricis 30-50.
Preferably, above-mentioned anti-inflammation itching relieving Chinese medicine composition comprises that following parts by weight of Chinese traditional medicine raw material processes: Fructus Cnidii 30, Rhizoma Polygoni Cuspidati 30, Radix Sophorae Flavescentis 30, Herba Taraxaci 20, Cortex Dictamni 15, the Radix Stemonae 20, Cortex Phellodendri 10, the Radix Angelicae Dahuricae 10, Alumen 10, Flos Caryophylli 6, Cortex Pseudolaricis 50.
Preferably, above-mentioned anti-inflammation itching relieving Chinese medicine composition water extract that is each raw material of Chinese medicine or ethanol extract are as effective ingredient.
Preferably, above-mentioned anti-inflammation itching relieving Chinese medicine composition is a solution, the ratio of volume and raw material of Chinese medicine quality of solution be 1ml:0.5g to 1ml:1g.
The method for preparing that a kind of anti-inflammation itching relieving Chinese medicine composition is provided of the present invention is with following parts by weight of Chinese traditional medicine raw material: Fructus Cnidii 20-30, Rhizoma Polygoni Cuspidati 20-30, Radix Sophorae Flavescentis 20-30, Herba Taraxaci 10-20, Cortex Dictamni 10-15, Radix Stemonae 10-20, Cortex Phellodendri 5-10, Radix Angelicae Dahuricae 5-10, Flos Caryophylli 3-6, reflux, extract; Filtering, must filtrate, is that to add mass percent be 70% alcohol for the raw material of Chinese medicine of Alumen 5-10 part, Cortex Pseudolaricis 30-50 part with weight fraction again; After twice of the alcohol extraction; The gained alcohol extract joins gained filtrating, slowly adds ethanol while stirring and makes the alcohol amount that contains of filtrating be 70%-80%, airtight cold preservation 48 hours; Filter; Filtrate recycling ethanol, being dissolved in water filters refined liquid, the volume and the raw material of Chinese medicine mass ratio of refined liquid be 1ml:0.5g to 1ml:1g.
In the method for preparing of preferred above-mentioned anti-inflammation itching relieving Chinese medicine composition, each weight part ratio of said raw material of Chinese medicine is specially: Fructus Cnidii 30, Rhizoma Polygoni Cuspidati 30, Radix Sophorae Flavescentis 30, Herba Taraxaci 20, Cortex Dictamni 15, the Radix Stemonae 20, Cortex Phellodendri 10, the Radix Angelicae Dahuricae 10, Alumen 10, Flos Caryophylli 6, Cortex Pseudolaricis 50.
Another technical scheme of the present invention is that a kind of above-mentioned application of anti-inflammation itching relieving Chinese medicine composition aspect the medicated premix of sanitary towel is provided.
Beneficial effect of the present invention is: Chinese medicine compound of the present invention by Fructus Cnidii, Rhizoma Polygoni Cuspidati, etc. Chinese medicine form, be to process by the pure Chinese medicine elite of multiple natural plants after refining, gynaecopathia, dermatosis etc. is had significant curative effect.Can use on health products such as sanitary towel auxiliary gynecopathy therapeutic.Avoid human body is produced toxic and side effects, have antibiotic, antiinflammatory, antipruritic, adjustment vaginal microbial flora is disorderly and increase the effect of vagina lactobacillus quantity, human body immunity improving function.
The specific embodiment
By specifying technology contents of the present invention, structural feature, realized purpose and effect, give explanation below in conjunction with embodiment is detailed.
Chinese medicine composition of the present invention can adopt the conventional method for preparing of Chinese medicine to prepare.
Embodiment 1
Prescription 1: prepare raw material of Chinese medicine Fructus Cnidii 20g, Rhizoma Polygoni Cuspidati 20g, Radix Sophorae Flavescentis 20g, Herba Taraxaci 10g, Cortex Dictamni 10g, Radix Stemonae 10g, Cortex Phellodendri 5g, Radix Angelicae Dahuricae 5g, Alumen 5g, Flos Caryophylli 3g, Cortex Pseudolaricis 30g.By prescription (except that Alumen, Cortex Pseudolaricis) proportioning reflux, extract,, filter, after filtrating adds recipe quantity Alumen and Cortex Pseudolaricis extracting solution (twice of 70% alcohol extraction); Be concentrated into 1ml:0.5g to 1ml:1g, medicinal liquid put cold after, slowly add ethanol while stirring and make and reach regulation and contain alcohol amount (70～80%); Airtight cold preservation 48h; Filter, filtrate recycling ethanol, being dissolved in water filters refined liquid.Have an effect with liquid form at last.1ml:1g is that every 1ml medicinal liquid is equivalent to 1g prescription crude drug.
Prescription 2: the composition of the raw material of Chinese medicine of above-mentioned anti-inflammation itching relieving Chinese medicine composition also can be: Fructus Cnidii 30g, Rhizoma Polygoni Cuspidati 30g, Radix Sophorae Flavescentis 30g, Herba Taraxaci 20g, Cortex Dictamni 15, the Radix Stemonae 20, Cortex Phellodendri 10, the Radix Angelicae Dahuricae 10, Alumen 10, Flos Caryophylli 6, Cortex Pseudolaricis 50.
Prescription 3: the composition of the raw material of Chinese medicine of above-mentioned anti-inflammation itching relieving Chinese medicine composition also can be: Fructus Cnidii 25g, Rhizoma Polygoni Cuspidati 25g, Radix Sophorae Flavescentis 25g, Herba Taraxaci 15g, Cortex Dictamni 12g, Radix Stemonae 15g, Cortex Phellodendri 7g, Radix Angelicae Dahuricae 7g, Alumen 7g, Flos Caryophylli 4g, Cortex Pseudolaricis 40g.
From clinical angle; Each dose can have certain floating in the prescription; This prescriptions of Chinese medicine (external) safety non-toxic; Effect with heat-clearing and toxic substances removing, expelling wind and removing dampness, killing parasites for relieving itching, to common gram positive bacteria such as staphylococcus aureus etc., common gram negative bacteria such as escherichia coli, bacillus pyocyaneus etc., common fungus such as Candida albicans etc. all have good antibacterial activity; Good anti-inflammatory anti-itch is arranged simultaneously, regulate effects such as vaginal microbial flora disorder, human body immunity improving power.
The extraction process for purification of embodiment 2, Chinese medicine compound of the present invention
Chinese medicine composition of the present invention can adopt the conventional method for preparing of Chinese medicine to prepare.The optimum decoction and alcohol sedimentation technique that uses.
1, water extract-alcohol precipitation
Decoction and alcohol sedimentation technique means in the Chinese medicine water extracting liquid, adds ethanol and makes and reach difference and contain the alcohol amount, and some drugs composition dissolubility in alcoholic solution reduces separates out deposition, makes the water extract be able to purified method after the solid-liquid separation.Specific operation process is: Chinese medicine compound water extract of the present invention is concentrated into 1: 1 (g: ml), medicinal liquid put cold after, slowly add ethanol while stirring and make and reach 70% and contain alcohol amount, airtight cold preservation 24～48h filters, filtrate recycling ethanol obtains refined liquid.
Should note following problem during operation: (1) medicinal liquid should suitably concentrate, to reduce amount of ethanol.But (1: 1-1: 0.5), if overrich, effective ingredient is prone to be wrapped in the deposition and causes damage should to control concentrating degree.(2) can add ethanol, volatilization loss after the spissated medicinal liquid cooling in order to avoid ethanol is heated.(3) accelerate slowly to stir.Should stir medicinal liquid fast, slowly add ethanol, local determining alcohol is too high to cause the effective ingredient loss that is wrapped to avoid.(4) airtight cold preservation.Can prevent the ethanol volatilization, promote to separate out sedimentary sedimentation, be convenient to filter operation.
2, the extract of Chinese medicine composition of the present invention also can carry out activated carbon decolorizing
Get the 50ml conical flask; Add compound extract behind the 20.0ml water extract-alcohol precipitation respectively,, add an amount of active carbon (0.5%, 1%, 1.5%, 2.0%) by designing requirement; The concussion decolouring corresponding time (2 hours), the active carbon of adding 1.5% has decolorizing effect preferably.
Embodiment 3
One, Chinese medicine compound of the present invention is carried out the antibacterial action checking:
1, preparation
1.1 culture medium preparation:, prepare various corresponding culture medium according to the difference of strain
1.2 other article
Oxford cup, tweezers, paint daubs, normal saline, clean tube, inoculating loop, liquid-transfering gun head, waste liquid cylinder, positive drug, trial drug, alcohol burner, liquid-transfering gun, different strain (staphylococcus aureus, Staphylococcus albus, bacillus pyocyaneus, escherichia coli, Candida albicans, fat corrected milk(FCM) bacillus).
1.3 actication of culture
Get clean tube, add an amount of MH broth bouillon (in the preparation of 2.7g dry powder preparation 100ml fluid medium ratio) through autoclaving, put cold back at clean bench and get an amount of bacterium with inoculating loop and put into test tube, mixing is cultivated observed result behind 18～24h.(staphylococcus aureus (JP), Staphylococcus albus (BP), bacillus pyocyaneus (LN), escherichia coli (BD), Candida albicans (BN), fat corrected milk(FCM) bacillus are put into 37 ° of C incubators and cultivate; Candida albicans is put into 26.5 ° of C incubators and is cultivated).
2, experimental technique
The Oxford agar diffusion method carries out the antibacterial action primary dcreening operation
The experimental implementation step
(1) Oxford cup, tweezers, paint daubs, normal saline, clean tube, inoculating loop, waste liquid cylinder, positive drug, trial drug autoclaving.
(2) preparation bacteria suspension: get clean tube, add an amount of normal saline, the reuse liquid-transfering gun is got activatory in right amount bacterium liquid, puts into test tube, mixing.
(3) draw the 500ul bacteria suspension to solid medium with liquid-transfering gun, the reuse paint daubs evenly smears to each several part in the culture medium with bacterium liquid.
(4) put to the culture medium of smearing even bacterium liquid with tweezers gripping Oxford cup, the Oxford cup is fully contacted with culture medium, must not leave the slit.Oxford cup placement location is moderate.
(5) in each Oxford cup, behind the adding 150 μ l medicinal liquids, carefully culture dish is placed incubator and cultivate.
3, experimental result
3.1 for the antibacterial circle diameter (cm) that the prescription extracting solution of embodiment 1 and positive control drug are as shown in table 1.
Table 1
Positive control drug is that commercially available JIEERYIN 2 closes 1 washing liquid and does positive control drug.Experimental result shows that the aqueous extract of Chinese medicine compound of the present invention all has bacteriostasis preferably to antibacterials such as staphylococcus aureus, Staphylococcus albus, bacillus pyocyaneus, escherichia coli.
Two, pharmacodynamic study in the body of traditional Chinese medicine compound extract of the present invention
(1) acute toxicity test of Chinese medicine compound of the present invention
Laboratory animal: the healthy KM mice of cleaning level, buy by the Changsha, Hunan Experimental Animal Center.Certificate licence: SCK (Hunan) 2009-0012
Experimental technique:
Acute toxicity testing: 30 of mices are got in trial test, and male and female half and half are divided into 5 groups at random, 6 every group.Being tried thing is maximum concentration (3g/ml crude drug) with the concentrated solution, dilutes successively by 1 times of method, and mice is adopted the oral administration gavage administration, and the administration volume is 0.4ml/10g.5 dose groups institute administered agents dosage are followed successively by 120g/kg, 60g/kg, 30g/kg, 15g/kg, 7.5g/kg body weight (administration concentration is the crude drug amount).Observed 7 days continuously after the administration, do not see tangible toxic reaction, and it is dead not have 1 example.Because of receiving the restriction of drug level and administration volume, can not find the dosage that causes dead mouse, so can't measure the LD50 of Chinese medicine compound mouse stomach administration.
Maximum tolerance determination: get 20 of mices, male and female half and half, the maximum concentration (120g/kg) that can prepare with medicine and the maximum volume (0.4ml/10g body weight) that allows to use once give whole mouse stomach administrations, observe 14 continuously after the administration.The mice phenomenon that appearance in first day is quiet after administration, gathering is motionless, do not have tangible toxic reaction, and activity recovery in the 3rd day normally, do not have 1 routine death in 14 days.The maximum tolerated dose that records the prescription gastric infusion thus is a crude drug 120g/kg body weight.
(2) skin toxicology test of Chinese medicine compound of the present invention
1, sensitization of skin test
(1) experimental technique
1.1 divide into groups: get 50 of healthy guinea pigs (being bought by the Changsha, Hunan Experimental Animal Center), the left side, back cuts off becomes mildewed depilation; The about 9cm2 of depilation scope; At random be divided into 5 group by body constitution amount, sex next day: 70% ethanol normal control group, Drug therapy high concentration group, Drug therapy low concentration group, 2,4-dinitro-chloro-benzene positive controls; The JIEERYIN group, 10 every group.The high and low solubility group of Drug therapy (crude drug amount 1g/mL, 0.25g/mL).
1.2 sensitization contact: get 70% ethanol 0.5mL, Drug therapy high concentration group and low concentration group 0.5mL, 0.1%2 respectively; 4-dinitro-chloro-benzene 0.2mL, 50% JIEERYIN group 0.5ml is applied to each depilation district, treated animal left side; Continue 6 hours, the 7th day and the 14th day be retry 1 time in kind.Excite contact: in last give tried thing sensitization after 14 days, get 70% ethanol 0.5mL, Drug therapy high concentration and low concentration group 0.5mL, O.1%2,4-dinitro-chloro-benzene 0.2mL respectively; 50% JIEERYIN group 0.5ml is applied to each depilation district, treated animal right side, removes after 6 hours and is tried thing; At once observe; Observed the skin allergy situation once more in 24 hours, 48 hours, 72 hours then, the result presses document, skin allergy standards of grading such as table 2.
Table 2
| The order of reaction | Anaphylaxis | |
| 0 | Erythema | No erythema |
| 1 | Slight erythema | |
| 2 | Moderate erythema | |
| 3 | The severe erythema | |
| 4 | Edema and erythema | |
| 0 | Edema | No edema |
| 1 | Mild edema | |
| 2 | Intermediate edema | |
| 3 | The severe edema |
Result's scoring: reaction meansigma methods and the sensitization rate of calculating each group by the last table order of reaction.
(2) experimental result
Experimental result sees that table 3 is the anaphylaxis result (sensitization contact) of Chinese medicine compound of the present invention to guinea pig skin, and table 4 is the anaphylaxis result (excite contact) of herbal mixture of the present invention to guinea pig skin.
Table 3
Table 4
The above results shows that Chinese medicine compound of the present invention can not cause anaphylaxis to guinea pig skin.
(3) antiinflammatory action of Chinese medicine compound of the present invention
Experimental technique: 60 mices (are bought by the Changsha, Hunan Experimental Animal Center.Certificate licence: SCK (Hunan) 2009-0012) is divided into 6 groups at random.Each group is respectively: matched group (distilled water); The JIEERYIN group; Dexamethasone group; The high, medium and low dose groups of prescription (crude drug amount 1g/mL, 0.5g/mL, 0.25g/mL).Mouse right ear front and back two sides coating 1 time (0.1mL) is respectively organized in experiment, and matched group is coated with the equivalent distilled water, administration 7 days.1 hour each administration group is with distilled water flush away medicinal liquid after the administration in the 7th day, and is clean with the dry cotton ball sassafras.After the last administration 1 hour, with liquid-transfering gun on a mice left side ear smear each 10 μ L of xylene tow sides.Behind the 30min, the dislocation of mice cervical vertebra is caused death, cut two ears along the auricle baseline.Lay round auricle at the same position of two ears respectively with diameter 6mm card punch, the electronics Libra is weighed.Calculate inhibitory rate of intumesce.
Inhibitory rate of intumesce (%)=(A ﹣ B)/A * 100%
In the formula, A represents blank control group swelling degree, and the B representative is tried drug group or positive control medicine group swelling degree.
Swelling degree=left ear weight-auris dextra weight
Experimental result:
Table 5
| Group | Ear swelling degree (g) | Suppression ratio (%) |
| The distilled water group | 0.00551±0.00129## | |
| Dexamethasone group | 0.00113±0.00037** | 79.49 |
| The JIEERYIN group | 0.00505±0.00152## | 8.35 |
| High dose group | 0.00305±0.00140**## | 44.65 |
| Middle dose groups | 0.00355±0.00174**## | 35.57 |
| Low dose group | 0.00327±0.00099**## | 40.65 |
Annotate: compare * * p＜0.01 with the distilled water group; Compare ##p＜0.01 with Dexamethasone group
All can suppress mice caused by dimethylbenzene xylene ear swelling degree from the visible administration group of table 5.Compare with blank control group, the medication group all has inhibitory action to the swelling situation of mice, and wherein best with the inhibition effect of dexamethasone, suppression ratio reaches 79.49%, and JIEERYIN group suppression ratio is minimum, and each dose groups of prescription all has good inhibitory effect.
(4) Chinese medicine compound of the present invention causes the influence (itching-relieving action) of guinea pig skin pruritus to histamine phosphate
Experimental technique
Get 50 of Cavia porcelluss (purchase of Changsha, Hunan Experimental Animal Center); Male and female half and half are divided into 5 groups, i.e. the normal saline matched group at random; The dexamethasone positive controls, prescription high dose group (crude drug content 1g/ml), prescription low dose group (crude drug 0.5g/ml).Test right back foot depilation in preceding 24 hours, depilation place administration 100 μ L cause when itching depilation place are abraded with fine sandpaper, and (area 1cm * 1cm), administration is 1 time again with slight oozing of blood degree of being.Every animal 0.5h behind the last coating begins only to drip 0.01% histamine phosphate, 50 μ L/ at the wound surface place; Every separated 3min drips mass concentration 0.01%, 0.02%, 0.03%, 0.04%, 0.05% successively ... Progressive concentration; Each 50 μ L/ only; Right back foot occurs later licking until Cavia porcellus, later to lick the right back histamine phosphate total amount that gives when sufficient be itch-threshold to occur Cavia porcellus at last.
Experimental result
Experimental result is seen table 6.Table 6 is the influence of Chinese medicine compound extract of the present invention to Cavia porcellus pruritus due to the histamine phosphate.
Table 6
| Group | Itch-threshold (μ g) |
| Normal saline | 9±5.16# |
| JIEERYIN | 62.5±39.24* |
| Dexamethasone | 293.00±134.45**## |
| High dose group | 42.00±37.94* |
| Low dose group | 25.63±15.91# |
Annotate: compare * p＜0.01, * * p＜0.05 with normal saline; Compare #p＜0.01 with the JIEERYIN group
After experimental result showed traditional Chinese medicine compound extract coating of the present invention, high dose group itch-threshold and normal saline matched group comparing difference had significance; High dose group and JIEERYIN matched group be there was no significant difference relatively, shows that prescription can obviously improve Cavia porcellus itch-threshold due to the histamine phosphate, and tangible itching-relieving action is arranged, and its effect is suitable with JIEERYIN.
(5) traditional Chinese medicine compound extract of the present invention is to Immune Effects
1, traditional Chinese medicine compound extract of the present invention is to the influence of T, bone-marrow-derived lymphocyte
(1) experiment material
1.1 medical material: herbal mixture, all medical materials are all available from Yao and Sheng Academy of Traditional Chinese Medicine.
1.2 reagent concanavalin A, Con A (Con A), LPS
1.3 the healthy KM mice of animal cleaning level is provided by the Changsha, Hunan Experimental Animal Center.Certificate licence: SCK (Hunan) 2009-0012
1.4 instrument C0-150 CO2 gas incubator, the RT-2100C ELIASA, Suzhou purifies superclean bench, OLYMPUS IX51 inverted microscope.JA1203N type electronic balance.
(2) experimental technique
2.1 the animal modeling according to bibliographical information, is diluted cyclophosphamide dry powder with normal saline, be mixed with 4mg/mL concentration, by the 40mg/kg standard mice is given intraperitoneal injection.Injection is three days continuously, sets up mouse immune and suppresses model.
2.2 animal divides into groups
With 40 of model mices, male and female half and half are divided into model control group, herbal mixture high concentration group (1g/ml) low concentration group (0.25g/ml) at random, 50% JIEERYIN group, and other gets 10 normal mouses is the blank group.Gastric infusion, one day 1 time, continuous 7d, matched group gives the isometric(al) distilled water by the 0.02ml/kg administration.1h after the last administration puts to death animal and under aseptic condition, gets spleen and experimentize: 1. making of mouse boosting cell suspension: the aseptic spleen of getting, preparation individual cells; The centrifugal 5min of 1000rmin-1 removes supernatant, kowtows loose cell precipitation gently; Add the 0.4mL distilled water 30s that vibrates gently, the hypotonic dissolution erythrocyte adds 2 times of dense normal saline of 0.4mL (1.7%) then solution recovery etc. is oozed; The same centrifugal 5min of rotating speed, after serum-free RPMI1640 liquid was washed 2 times again, cell precipitation was resuspended in the RPMI RPMI-1640 that contains 10% calf serum; The microscopically counting transfers to 5 * 10 with cell density
5/ mL, this is subsequent use splenocyte.2. lymphproliferation response: add the splenocyte suspension 100 μ L that make in the every hole of flat culture plate, 96 holes respectively, the whole mass concentration of ConA 100 μ L is 7.5 μ gmL-1, and matched group then adds with 100 μ LRPMI-1640 culture fluid, every mice 3 holes.Culture plate is put into 37 ℃ of 5%CO2 incubators, cultivate 48h.After cultivating end, mtt assay is surveyed cell optical density (OD).
Statistical method: all experimental datas are all represented with
± s.Continuous data adopts SPSS 13.0 statistical softwares to carry out check between variance analysis and group.
Experimental result
Can be known to the influence of T, bone-marrow-derived lymphocyte propagation that by table 7 Chinese medicine compound of the present invention high dose group all can extremely significantly promote immunosuppressed mice T, bone-marrow-derived lymphocyte propagation (P < 0.01), the short cultivation effect of low dose group is not obvious.
Table 7
| Group | T lymphocyte (OD) | Bone-marrow-derived lymphocyte (OD) |
| Normal group | 0.0562±0.0024 △△ | 0.0488±0.0065 △ |
| Model group | 0.0428±0.0008 ** | 0.0390±0.0020* |
| The JIEERYIN group | 0.0486±0.0027 * | 0.0483±0.0086 △ |
| High dose group | 0.0547±0.0027 △△ | 0.0537±0.0063 △△ |
| Low dose group | 0.0495±0.0032 | 0.0430±0.0068 |
Compare with normal group: < 0.05, * * p < 0.01 for * p; With model group than △ p 0.05, △ △ p < 0.01
2, Chinese medicine compound of the present invention is to the influence of hemolytic antibody generation and organ index
(1) experiment material: the same.
(2) experimental technique
2.1 the animal modeling according to bibliographical information, is diluted cyclophosphamide dry powder with normal saline, be mixed with 4mg/mL concentration, by the 40mg/kg standard mice is given intraperitoneal injection.Injection is three days continuously, sets up the immunosuppressed mice model.
2.2 divide into groups and administration
Be divided into normal control group and herbal mixture high and low dose group and JIEERYIN group at random, 10 every group.Normal group is irritated the clothes distilled water, and Chinese medicine compound group of the present invention is irritated clothes high and low dose 0.02ml/g,, the JIEERYIN group is irritated clothes 50% JIEERYIN 0.02ml/g, successive administration 7d.Inject 5% chicken red blood cell suspension 0.2mL in 7th every mouse peritoneal and carry out immunity, continuously 7d.
2.3 serum hemolysin experiment
40min after the last administration wins eyeball and gets blood, the centrifugal 5min of 3000r/min.
Serum is got dilute serum 1mL and is mixed in 0.5mL 5% chicken erythrocyte suspension and 0.5mL 10% complement (GPS) with 100 times of normal saline dilution, in 37 ℃ of calorstats, behind the insulation 30min, puts 0 ℃ of refrigerator 30min stopped reaction.Centrifugal, get supernatant, the colorimetric in spectrophotometer 540nm place, photometry density value.Other establishes normal saline and does blank, returns to zero when getting its supernatant as colorimetric.Calculate and respectively organize mice haemolysis OD value.
2.4 organ index is measured
Take off neck and put to death mice, get spleen immediately, according to formula calculating organ index with thymus is weighed.Organ index=(organ weights/mice body weight) * 10
2.5 statistical method
Data are represented with
± s; Relatively use one factor analysis of variance between each group, carry out statistical analysis with SPSS11.0.
(3) experimental result
3.1 serum hemolysin antibody generates: compare with model group; High concentration group mice serum hemolytic antibody generates all has rising, and the result sees table 8: Chinese medicine compound of the present invention explains that to the influence
that the mice serum hemolytic antibody generates Chinese medicine compound of the present invention can promote the generation of immunosuppressed mice hemolytic antibody.
Table 8
| Group | N (only) | Optical density (OD) |
| The blank group | 10 | 0.60390±0.040959 |
| Model group | 10 | 0.52120±0.050352 ** |
| The JIEERYIN group | 10 | 0.58000±0.021448 △△ |
| The high concentration group | 10 | 0.53620±0.033895 ** |
| Low concentration group | 9 | 0.56622±0.050884 |
Annotate: compare with the normal control group: * * p 0.01, compare with model group: △ △ p < 0.01
3.2 organ index:
Compare with matched group; Like table 9: Chinese medicine compound of the present invention is to the influence (n=10 of mice organ index;
thymus index is at height; Remarkable rising (p < 0.05) is all arranged in the low dose group mice, and the high concentration group raises more obviously (p < 0.01), explains that Chinese medicine compound of the present invention can promote mouse immune organ thymus; The weightening finish of spleen, the prompting compound recipe can improve the immunosuppressed mice immune organ and increase.
Table 9
| Group | Index and spleen index | Thymus index |
| The blank group | 0.08790120±0.009228541 △△ | 0.04117619±0.007334765 △△ |
| Model group | 0.06729571±0.005778540 * | 0.02667668±0.003446494 * |
| The JIEERYIN group | 0.08065789±0.008821144 | 0.02747108±0.004779367 |
| The high concentration group | 07713666±.010022596 △△ | 0.03566412±0.004301745 △△ |
| Low concentration group | 0.08256155±0.010442171 △ | 0.02999935±0.006153046 △ |
Compare with normal group: < 0.05, * * p < 0.01 for * p; With model group than △ p 0.05, △ △ p < 0.01
3, Chinese medicine compound of the present invention influences the mice phagocytic function
(1). experiment material: the same.
(2) experimental technique
2.1 the animal modeling according to bibliographical information, is diluted cyclophosphamide dry powder with normal saline, be mixed with 4mg/mL concentration, by the 40mg/kg standard mice is given intraperitoneal injection.Injection is three days continuously, sets up the immunosuppressed mice animal model.
2.2 divide into groups and administration
Be divided into large and small dose groups of normal control group and Chinese traditional compound medicine and JIEERYIN group at random, 10 every group.Normal group is irritated the clothes distilled water, and Chinese medicine compound group of the present invention is irritated clothes high and low dose 0.02ml/g, and the JIEERYIN group is irritated clothes 50% JIEERYIN 0.02ml/g, successive administration 8d.The last administration after 40 minutes in mouse tail intravenous injection india ink diluent.2min and 5min incorporate in the 2ml0.01% sodium carbonate liquor in mouse orbit venous blood collection 0.02ml behind the injection prepared Chinese ink.Put to death mice and get its liver, spleen is weighed.Get 0.02ml mice blood and be dissolved in the 2ml0.01% sodium carbonate liquor, measure absorbance in the 650nm place as the blank group.Calculate phagocytic index (k) and engulf coefficient (a): k=(logA1-logA2)/(t1-t2) by following formula; A=body weight/(liver weight+spleen heavy) * k1/3. wherein A1, A2 represents the absorbance of twice blood sampling successively respectively, and t1-t2 represents the interval of twice blood sampling.
3 experimental results
By table 10: prescription is to the influence (n=10 of immunosuppressed mice phagocytic function;
is visible, and Chinese medicine compound of the present invention can improve immunosuppressed mice phagocyte phagocytic index and engulf coefficient.
Table 10
| Group | Phagocytic index K | Engulf coefficient a |
| Model group | 0.0367±0.0979 △△ | 3.669±1.102 △△ |
| Normal blank control group | 0.0959±0.0136 ** | 5.217±1.423 ** |
| The JIEERYIN group | 0.0511±0.0110 *△△ | 4.484±0.511 |
| The high concentration group | 0.0495±0.0110 *△△ | 5.036±1.491 ** |
| Low concentration group | 0.0646±0.0169 **△△ | 5.079±0.160 * |
Compare with model group: < 0.05, * * p < 0.01 for * p; With normal group than △ p 0.05, △ △ p < 0.01
(6) Chinese medicine compound of the present invention is to the disorderly regulating action of mouse vagina flora
1 experiment material:
Laboratory animal: 50 of cleaning level Kunming kind female mices, in 8～10 ages in week, body weight 18～22g is provided by the Changsha, Hunan Experimental Animal Center.Certificate licence: SCK (Hunan) 2009-0012
Medicine and reagent: penicillin sodium salt is that east wind Pharmaceutical limited company in group Jiangxi produces in the river, specification: 800,000 units, lot number: 041045-3.The penicillin sodium salt formulations prepared from solutions: existing preparation before each the use, use physiological saline solution to transfer to concentration and be 12.5mg/mL.
The preparation of MRS agar culture medium: the culture medium of dissolving 25g agar in the 1L distilled water adds following each component: peptone, meat extract 10g, yeast extract 5g, glucose 20g, K2HPO42g, dibasic ammonium citrate 2g, sodium acetate 5g, Tween 80 1mL, MgS0
47H
2O 0.58g, MnSO
44H
2O 0.25g transfers pH6.2～6.4,121 ℃ sterilization 15min.
2 experimental techniques
Animal divides into groups: get 50 of Kunming kind female mices, be divided into 10 of normal control groups (C) at random, antibiotic causes 40 of mouse vagina dysbacteriosis model group (A); After the modeling success, model be divided at random high and low dose group, each 10 of JIEERYIN positive controls.
Vaginal dysbacteriosis modelling: except that the normal control group; Other each groups are got 50 μ L penicillin sodium saline solution (concentration is 12.5mg/mL) with micro sample adding appliance and are carried out mouse vagina flushing administration; Every day 1 time, continuous 5d, matched group each normal saline that injects equal volume every day.
Medication: after the mouse vagina dysbacteriosis model modeling success; Inject each 50 μ L of prescription high dose group (crude drug content 1g/ml), low dose group (crude drug content 0.25g/ml) and JIEERYIN group (JIEERYIN medicinal liquid dilution 50%) for the model mice intravaginal; Continuous 7d, every day 1 time.Normal control group and model group each normal saline that injects equal volume every day.
The mensuration of mouse vagina lactobacillus and analysis: behind the last administration 12h, get 50 μ L normal saline with micro sample adding appliance, wash mouse vagina repeatedly 8 times, 10 times of serial dilutions of vaginal washing fluid of getting 30 μ L are to 10-6.Drip kind on the MRS selective medium with micro sample adding appliance, carry out count plate (CFU/mL) (testing repetition 3 times) after cultivating through 37 ℃, 48h.
3 experimental results
Through setting up antibiotic induced mouse vagina dysbacteriosis model; Use the lactobacillus in the mouse vagina secretions of back to measure with the method for plate culture count to prescription, the result sees table 11: respectively organize mouse vagina lactobacillus testing result (viable count CFU/mL) after the medication.
Table 11
| Group | Number of animals | CFU |
| The normal saline group | 10 | 395±200.28* |
| The JIEERYIN group | 10 | 92.58±23.59 |
| High dose group | 10 | 799.99±134.69**# |
| Low dose group | 10 | 359.26±92.01* |
| Model group | 10 | 13.33±5.44# |
Annotate: compare * p＜0.01, * * p＜0.05 with model group; Compare #p＜0.01 with normal group
Experimental result shows: the continuous vaginadouche administration of penicillin sodium saline solution causes the vaginal microbial flora disorder after 5 days, and vagina lactobacillus quantity reduces, and compares with the normal control group, and difference has the significance meaning to show the vaginal dysbacteriosis modelling.Inject prescription extracting solution 50 μ L in the mouse vagina, after continuous 7 days, obviously improve lactobacillus quantity, recovery also surpasses C group, compares with model group, and difference has utmost point significance meaning.Explain that disorder has regulating action and increases vagina lactobacillus quantity this prescription for the mouse vagina flora.
Chinese medicine compound of the present invention has following effect:
1, antibacterial action: adopt conventional in-vitro antibacterial experiment, JIEERYIN XIYE is made positive control, the extract minimum inhibitory concentration and the minimum bactericidal concentration of clear and definite Chinese medicine preparation of the present invention.To common gram positive bacteria such as staphylococcus aureus, streptococcus; Common gram negative bacteria such as escherichia coli, bacillus pyocyaneus, common fungus such as Candida albicans, Aspergillus fumigatus, acrothesium floccosum etc. all have good antibacterial activity.
2, antiinflammatory action: study its antiinflammatory action with auricle edema, subcutaneous injection Oleum Tiglii induced mice ear corridor swelling model due to conventional acute inflammation model such as the xylene; Set up positive control (JIEERYIN is made positive control), the extract of clear and definite Chinese medicine preparation of the present invention has tangible antiinflammatory action.
3, itching-relieving action: cause the rat skin of dorsum of foot with conventional histamine phosphate and cause its itching-relieving action of scale-model investigation of itching; Set up positive control (also available JIEERYIN XIYE is made positive control), the be significantly improved effect of itch-threshold of the extract of clear and definite Chinese medicine preparation of the present invention.
4, adjustment vaginal microbial flora is disorderly and increase vagina lactobacillus quantity
5, promote immunologic function: the immunological experiment that utilization is conventional; Set up positive control (also available JIEERYIN XIYE is made positive control); The extract of clear and definite Chinese medicine preparation of the present invention has the phagocytic function that promotes Turnover of Mouse Peritoneal Macrophages; Promote splenic T, bone-marrow-derived lymphocyte to transform the effect of human body immunity improving function.
The above is merely embodiments of the invention; Be not so limit claim of the present invention; Every equivalent structure or equivalent flow process conversion that utilizes description of the present invention to do; Or directly or indirectly be used in other relevant technical fields, all in like manner be included in the scope of patent protection of the present invention.
Claims (7)
1. anti-inflammation itching relieving Chinese medicine composition; It is characterized in that, comprise that following parts by weight of Chinese traditional medicine raw material processes: Fructus Cnidii 20-30, Rhizoma Polygoni Cuspidati 20-30, Radix Sophorae Flavescentis 20-30, Herba Taraxaci 10-20, Cortex Dictamni 10-15, Radix Stemonae 10-20, Cortex Phellodendri 5-10, Radix Angelicae Dahuricae 5-10, Alumen 5-10, Flos Caryophylli 3-6, Cortex Pseudolaricis 30-50.
2. anti-inflammation itching relieving Chinese medicine composition according to claim 1; It is characterized in that, comprise that following parts by weight of Chinese traditional medicine raw material processes: Fructus Cnidii 30, Rhizoma Polygoni Cuspidati 30, Radix Sophorae Flavescentis 30, Herba Taraxaci 20, Cortex Dictamni 15, the Radix Stemonae 20, Cortex Phellodendri 10, the Radix Angelicae Dahuricae 10, Alumen 10, Flos Caryophylli 6, Cortex Pseudolaricis 50.
3. anti-inflammation itching relieving Chinese medicine composition according to claim 1 is characterized in that, said Chinese medicine composition is that water extract or the ethanol extract of each raw material of Chinese medicine is as effective ingredient.
4. anti-inflammation itching relieving Chinese medicine composition according to claim 1 is characterized in that said Chinese medicine composition is a solution, the ratio of volume and raw material of Chinese medicine quality of solution be 1ml:0.5g to 1ml:1g.
5. the method for preparing of an anti-inflammation itching relieving Chinese medicine composition is characterized in that, with following parts by weight of Chinese traditional medicine raw material: Fructus Cnidii 20-30, Rhizoma Polygoni Cuspidati 20-30, Radix Sophorae Flavescentis 20-30, Herba Taraxaci 10-20, Cortex Dictamni 10-15, Radix Stemonae 10-20, Cortex Phellodendri 5-10, Radix Angelicae Dahuricae 5-10, Flos Caryophylli 3-6; Reflux, extract, is filtered, and must filtrate; Again with weight fraction be the raw material of Chinese medicine of Alumen 5-10 part, Cortex Pseudolaricis 30-50 part to add mass percent be 70% alcohol, after twice of the alcohol extraction, the gained alcohol extract joins gained filtrating; Slowly adding ethanol while stirring makes the alcohol amount that contains of filtrating be 70%-80%; Airtight cold preservation 48 hours filters filtrate recycling ethanol; Being dissolved in water filters refined liquid, the volume and the raw material of Chinese medicine mass ratio of refined liquid be 1ml:0.5g to 1ml:0.5g.
6. the method for preparing of anti-inflammation itching relieving Chinese medicine composition according to claim 5; It is characterized in that each weight part ratio of said raw material of Chinese medicine is specially: Fructus Cnidii 30, Rhizoma Polygoni Cuspidati 30, Radix Sophorae Flavescentis 30, Herba Taraxaci 20, Cortex Dictamni 15, the Radix Stemonae 20, Cortex Phellodendri 10, the Radix Angelicae Dahuricae 10, Alumen 10, Flos Caryophylli 6, Cortex Pseudolaricis 50.
Like each described anti-inflammation itching relieving Chinese medicine composition of claim 1-4 in the application aspect the medicated premix of sanitary towel.
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| CN105079211A (en) * | 2015-08-11 | 2015-11-25 | 金科伟业（中国）有限公司 | Antibacterial and anti-inflammation traditional Chinese medicine extract and application and preparation thereof |
| CN105727353A (en) * | 2016-02-05 | 2016-07-06 | 株洲千金药业股份有限公司 | Liquid medicine having bacteriostatic and sterilizing effects and application thereof |
| CN107158127A (en) * | 2017-05-24 | 2017-09-15 | 广西中医药大学第附属医院 | A kind of multi-effective skin-wash liquid |
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| CN107233606A (en) * | 2017-05-31 | 2017-10-10 | 苏州蔻美新材料有限公司 | A kind of degerming panty liner of skin care |
| CN108618973A (en) * | 2018-06-25 | 2018-10-09 | 李淑华 | Pure Chinese medicine wet sanitary napkins |
| CN111588672A (en) * | 2020-06-30 | 2020-08-28 | 河北农业大学 | Edible itching-relieving emulsion for mosquito bites and preparation method thereof |
| CN112206259A (en) * | 2020-10-30 | 2021-01-12 | 陕西理工大学 | Traditional Chinese medicine composition for treating vaginitis, traditional Chinese medicine lotion and application |
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Denomination of invention: The invention relates to an antibacterial, anti-inflammatory and antipruritic Chinese medicine composition, a preparation method and application thereof Effective date of registration: 20201217 Granted publication date: 20150318 Pledgee: Industrial Bank Limited by Share Ltd. Huian branch Pledgor: BARON (CHINA) Co.,Ltd. Registration number: Y2020350000142 |