CN102836367B - 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 antipruritic Chinese medicine composition of a kind of anti-inflammation and its preparation method and application.
Background technology
Along with all kinds of antibiotic taking penicillin as representative constantly occur, bacterial infection disease is effectively controlled, thus antibiotic is become powerful mean that the mankind resist pathogenic bacterial infection.But in recent years, people use antibiotic to cause the drug resistance of antibacterial constantly to strengthen for a long time widely, and Resistant frequency increases gradually, so that conventional antibiotic cannot meet clinical needs.Along with global disease spectrum and the important change of medical model, progressively integrating with of China and World Economics, culture and drug level, compatriots are enhanced to antibiotic understanding, recognize the impact of many untoward reaction that antibiotic brings and corresponding drug resistance thereof, people are filled with expectation to Chinese medicine maintaining healthy and disease preventing and treating.Medical circle is also expected to tackle by rational use of drug and exploitation new Chinese medicine the fastbacteria constantly produced.
At present along with the raising of people's living standard, health care product also gets more and more, and especially have health-care effect to gynecological, health sanitary napkin is health care of women articles for use, has broad spectrum kill effect to pathogenic microorganism.To common gram positive bacteria as staphylococcus aureus, streptococcus, common gram negative bacteria is as escherichia coli, bacillus pyocyaneus, and common fungus such as Candida albicans, acrothesium floccosum etc. all have good antibacterial activity.But it adds the method for the Bactericidal medicines such as hibitane, domiphen bromide, metronidazole in various sanitary towel, owing to having pessimal stimulation and toxic and side effects to user, gradually by Chinese herbal medicine type sanitary towel is substituted.
Summary of the invention
Object of the present invention provides a kind of Traditional Chinese medicine compound composition, by multiple natural plants through refining, pure Chinese medicine elite is made, and avoids producing toxic and side effects to human body, has antibacterial, antiinflammatory, antipruritic, adjustment disturbed vaginal flora increase Lactobacillus vaginalis quantity, improve the effect of body's immunity.
The Chinese medicine composition of technical scheme of the present invention for providing a kind of anti-inflammation antipruritic, the raw material of Chinese medicine comprising following weight portion is made: 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, the raw material of Chinese medicine that the Chinese medicine composition that above-mentioned anti-inflammation is antipruritic comprises following weight portion is made: 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, the Chinese medicine composition that above-mentioned anti-inflammation is antipruritic is that the water extract of each raw material of Chinese medicine or ethanol extract are as effective ingredient.
Preferably, the antipruritic Chinese medicine composition of above-mentioned anti-inflammation is solution, the volume of solution and the ratio of raw material of Chinese medicine quality be 1ml:0.5g to 1ml:1g.
The preparation method of the Chinese medicine composition providing a kind of anti-inflammation antipruritic of the present invention, raw material of Chinese medicine by following weight portion: 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, filter, obtain filtrate, be Alumen 5-10 part again by weight fraction, the raw material of Chinese medicine of Cortex Pseudolaricis 30-50 part adds the alcohol that mass percent is 70%, after alcohol extraction twice, gained alcohol extract joins gained filtrate, slowly adding ethanol while stirring makes the alcohol content of filtrate be 70%-80%, airtight cold preservation 48 hours, filter, filtrate recycling ethanol, be dissolved in water and filter to obtain refined liquid, the volume of refined liquid and raw material of Chinese medicine mass ratio be 1ml:0.5g to 1ml:1g.
In the preparation method of the Chinese medicine composition that preferred above-mentioned anti-inflammation is antipruritic, each weight part ratio of described 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.
The Chinese medicine composition application the medicated premix of sanitary towel in of another technical scheme of the present invention for providing a kind of above-mentioned anti-inflammation antipruritic.
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 by multiple natural plants through refine after pure Chinese medicine elite make, to gynaecopathia, dermatosis etc., there is significant curative effect.Can be used on the health products such as sanitary towel, auxiliary gynecopathy therapeutic.Avoid producing toxic and side effects to human body, there is antibacterial, antiinflammatory, antipruritic, adjustment disturbed vaginal flora increase Lactobacillus vaginalis quantity, improve the effect of body's immunity.
Detailed description of the invention
By describing technology contents of the present invention, structural feature in detail, being realized object and effect, be explained in detail below in conjunction with embodiment.
Chinese medicine composition of the present invention can adopt the customary preparation methods 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 Alumen, Cortex Pseudolaricis) proportioning reflux, extract, filter, after filtrate adds recipe quantity Alumen and Cortex Pseudolaricis extracting solution (70% alcohol extraction twice), be concentrated into 1ml:0.5g to 1ml:1g, after medicinal liquid lets cool, slowly add ethanol while stirring and make to reach regulation alcohol content (70 ~ 80%), airtight cold preservation 48h, filter, filtrate recycling ethanol, is dissolved in water and filters to obtain refined liquid.Finally have an effect in liquid form.1ml:1g and every 1ml medicinal liquid are equivalent to 1g prescription crude drug.
Prescription 2: the composition of the raw material of Chinese medicine of the Chinese medicine composition that above-mentioned anti-inflammation is antipruritic 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 the Chinese medicine composition that above-mentioned anti-inflammation is antipruritic 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 Point of View of Clinical, in prescription, each dose can have certain floating, this prescriptions of Chinese medicine (external) safety non-toxic, there is effect of heat-clearing and toxic substances removing, expelling wind and removing dampness, killing parasites for relieving itching, to common gram positive bacteria if staphylococcus aureus etc., common gram negative bacteria are as escherichia coli, bacillus pyocyaneus etc., common fungus such as Candida albicans etc. all has good antibacterial activity; There is good anti-inflammatory anti-itch simultaneously, regulate disturbed vaginal flora, improve the effects such as immunity of organisms.
The Hydrolysis kinetics method of embodiment 2, Chinese medicine compound of the present invention
Chinese medicine composition of the present invention can adopt the customary preparation methods of Chinese medicine to prepare.Optimum use decoction and alcohol sedimentation technique.
1, water extract-alcohol precipitation
Decoction and alcohol sedimentation technique means in Chinese medicine water extracting liquid, adds ethanol and makes to reach different alcohol content, and some drugs composition dissolubility in alcoholic solution reduces separates out precipitation, makes the method that Aqueous extracts is refined after solid-liquid separation.Specific operation process is: Chinese medicine compound Aqueous extracts of the present invention is concentrated into 1: 1(g: ml), after medicinal liquid lets cool, slowly add ethanol while stirring and make to reach 70% alcohol content, airtight cold preservation 24 ~ 48h, filter, filtrate recycling ethanol, obtains refined liquid.
Following problem should be noted: (1) medicinal liquid should suitably concentrate, to reduce ethanol consumption during operation.But should control concentrating degree (1: 1-1: 0.5), if overrich, effective ingredient is easily wrapped in precipitation and causes damage.(2) ethanol can be added, the volatilization loss in order to avoid ethanol is heated after concentrated medicinal liquid cooling.(3) slow quickening stirs.Medicinal liquid should be stirred fast, slowly add ethanol, be wrapped loss to avoid the too high effective ingredient that causes of local determining alcohol.(4) airtight cold preservation.Can prevent ethanol from volatilizing, promote the sedimentation of separating out precipitation, be convenient to filtration operation.
2, the extract of Chinese medicine composition of the present invention also can carry out activated carbon decolorizing
Get 50ml conical flask, add compound extract after 20.0ml water extract-alcohol precipitation respectively, by designing requirement, add appropriate active carbon (0.5%, 1%, 1.5%, 2.0%), concussion decolouring corresponding time (2 hours), the active carbon adding 1.5% has good decolorizing effect.
Embodiment 3
One, antibacterial action checking is carried out to Chinese medicine compound of the present invention:
1, preparation
1.1 culture medium preparations: according to the difference of strain, prepare various corresponding culture medium
1.2 other article
Oxford cup, tweezers, paint daubs, normal saline, clean tube, inoculating loop, liquid transfer 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 appropriate MH broth bouillon (by 2.7g dry composition 100ml fluid medium proportions) and, through autoclaving, after clean bench lets cool, get appropriate bacterium with inoculating loop put into test tube, mixing, observed result after cultivation 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
Odontothrips loti carries out antibacterial action primary dcreening operation
Laboratory operating procedures
(1) Oxford cup, tweezers, paint daubs, normal saline, clean tube, inoculating loop, waste liquid cylinder, positive drug, trial drug autoclaving.
(2) prepare bacteria suspension: get clean tube, add appropriate normal saline, then get appropriate activated bacterium liquid with liquid-transfering gun, put into test tube, mixing.
(3) 500ul bacteria suspension is drawn in solid medium with liquid-transfering gun, then with paint daubs by each several part in bacterium liquid uniform application to culture medium.
(4) put in the culture medium of smearing even bacterium liquid with tweezers gripping Oxford cup, Oxford cup is fully contacted with culture medium, must not gap be left.Oxford cup placement location is moderate.
(5), after adding 150 μ l medicinal liquids in the cup of each Oxford, carefully culture dish is placed incubator and cultivate.
3, experimental result
3.1 antibacterial circle diameters (cm) for the prescription extracting solution of embodiment 1 and positive control drug being are as shown in table 1.
Table 1
Positive control drug is that commercially available 2-in-1 1 washing liquid of JIEERYIN does positive control drug.Experimental result shows that the aqueous extract of Chinese medicine compound of the present invention has good bacteriostasis to antibacterials such as staphylococcus aureus, Staphylococcus albus, bacillus pyocyaneus, escherichia coli.
Pharmacodynamic study in the body of two, 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 grade, is bought by Changsha, Hunan Experimental Animal Center.Certificate licence: SCK (Hunan) 2009-0012
Experimental technique:
Acute toxicity testing: mice 30 is got in trial test, male and female half and half are divided into 5 groups at random, often organize 6.Tested material take concentrated solution as maximum concentration (3g/ml crude drug), dilutes successively by 1 times of method, and mice adopts oral administration gavage administration, and administration volume is 0.4ml/10g.The drug dose that 5 dosage groups give is followed successively by 120g/kg, 60g/kg, 30g/kg, 15g/kg, 7.5g/kg body weight (administration concentration is crude drug amount).After administration, Continuous Observation 7 days, has no obvious toxic reaction, and dead without 1 example.Because of the restriction by drug level and administration volume, can not find the dosage causing dead mouse, therefore the LD50 of Chinese medicine compound mouse stomach administration cannot be measured.
Maximum tolerance determination: get mice 20, male and female half and half, the maximum volume (0.4ml/10g body weight) that the maximum concentration (120g/kg) can prepared with medicine and permission use once gives whole mouse stomach administration, Continuous Observation 14 days after administration.Mice is except first day appearance after administration is quiet, assemble except motionless phenomenon, and without obvious toxic reaction, and movable recovery in the 3rd day is normal, and in 14 days, nothing 1 example is dead.The maximum tolerated dose recording prescription gastric infusion is thus crude drug 120g/kg body weight.
(2) skin toxicology test of Chinese medicine compound of the present invention
1, Skin sensitization
(1) experimental technique
1.1 groupings: get healthy guinea pig (being bought by Changsha, Hunan Experimental Animal Center) 50, left dorsal cuts off and becomes mildewed, depilation, depilation scope is about 9cm2, next day is divided into 5 groups: 70% ethanol Normal group, Drug therapy high concentration group, Drug therapy low concentration group at random by weight, sex, 2,4-dinitro-chloro-benzene positive controls, JIEERYIN group, often organizes 10.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, be applied to district of losing hair or feathers on the left of each treated animal, continue 6 hours, the 7th day and the 14th day in kind retry 1 time.Excite contact: after last is to tested material sensitization 14 days, get 70% ethanol 0.5mL, Drug therapy high concentration and low concentration group 0.5mL, O.1%2 respectively, 4-dinitro-chloro-benzene 0.2mL, 50% JIEERYIN group 0.5ml, is applied to district of losing hair or feathers on the right side of each treated animal, removes tested material after 6 hours, at once observe, then again observed skin allergy situation in 24 hours, 48 hours, 72 hours, result presses document, and skin allergy standards of grading are as table 2.
Table 2
| The order of reaction | Anaphylaxis | |
| 0 | Erythema | Without erythema |
| 1 | Slight erythema | |
| 2 | Moderate erythema | |
| 3 | Severe erythema | |
| 4 | Edema erythema | |
| 0 | Edema | Without edema |
| 1 | Mild edema | |
| 2 | Intermediate edema | |
| 3 | Severe edema |
Result is marked: the reaction meansigma methods and the sensitization rate that calculate each group by the upper table order of reaction.
(2) experimental result
Experimental result is the anaphylaxis result (sensitization contact) of Chinese medicine compound of the present invention to guinea pig skin in table 3, 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, 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 Changsha, Hunan Experimental Animal Center.Certificate licence: SCK (Hunan) 2009-0012) be divided into 6 groups at random.Each group is respectively: matched group (distilled water); JIEERYIN group; Dexamethasone group; The high, medium and low dosage group of prescription (crude drug amount 1g/mL, 0.5g/mL, 0.25g/mL).Test two sides coating 1 time (0.1mL) before and after each group of mouse right ear, matched group is coated with equivalent distilled water, administration 7 days.After administration in 7th day, 1 hour each administration group washes away medicinal liquid with distilled water, clean with dry cotton ball sassafras.After last administration 1 hour, smear each 10 μ L of dimethylbenzene tow sides with liquid-transfering gun at the left ear of mice.After 30min, by lethal for mice cervical dislocation, cut two ears along auricle baseline.Lay round auricle at the two same positions of ear respectively with diameter 6mm card punch, electronics Libra is weighed.Calculate inhibitory rate of intumesce.
Inhibitory rate of intumesce (%)=(A ﹣ B)/A × 100%
In formula, A represents blank group swelling, and B represents test medicine group or positive control medicine group swelling.
Swelling=left ear weight-auris dextra weight
Experimental result:
Table 5
| Group | Ear swelling degree (g) | Suppression ratio (%) |
| Distilled water group | 0.00551±0.00129## | |
| Dexamethasone group | 0.00113±0.00037** | 79.49 |
| JIEERYIN group | 0.00505±0.00152## | 8.35 |
| High dose group | 0.00305±0.00140**## | 44.65 |
| Middle dosage group | 0.00355±0.00174**## | 35.57 |
| Low dose group | 0.00327±0.00099**## | 40.65 |
Note: compare * * p < 0.01 with distilled water group; ##p < 0.01 is compared with Dexamethasone group
Administration group as seen from Table 5 all can suppress mice caused by dimethylbenzene xylene ear swelling degree.Compare with blank group, the swelling situation of medication group to mice has inhibitory action, and wherein best with the inhibition of dexamethasone, suppression ratio reaches 79.49%, and JIEERYIN group suppression ratio is minimum, and prescription each dosage group all has good inhibitory action.
(4) Chinese medicine compound of the present invention causes the impact (itching-relieving action) of guinea pig skin pruritus on histamine phosphate
Experimental technique
Get Cavia porcellus (purchase of Changsha, Hunan Experimental Animal Center) 50, male and female half and half, are divided into 5 groups at random, i.e. saline control group, 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 first 24 hours, depilation place administration 100 μ L, causes when itching and is abraded by depilation place fine sandpaper, with slight oozing of blood for degree (area 1cm × 1cm), then administration 1 time.Every animal 0.5h after last coating starts to drip 0.01% histamine phosphate 50 μ L/ at wound surface place only, mass concentration 0.01%, 0.02%, 0.03%, 0.04%, 0.05% is dripped successively every 3min ... progressive concentration, each 50 μ L/ only, until Cavia porcellus occurs later licking right back foot, with finally occur Cavia porcellus later lick right back sufficient time the histamine phosphate total amount that gives for itch-threshold.
Experimental result
Experimental result is in table 6.Table 6 is for Chinese medicinal compound extract of the present invention is on the impact of Cavia porcellus pruritus caused by 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# |
Note: compare * p < 0.01, * * p < 0.05 with normal saline; #p < 0.01 is compared with JIEERYIN group
After experimental result shows traditional Chinese medicine compound extract coating of the present invention, high dose group itch-threshold and saline control group comparing difference have significance; High dose group compares there was no significant difference with JIEERYIN matched group, shows that prescription can significantly improve Cavia porcellus itch-threshold caused by histamine phosphate, has obvious itching-relieving action, and its effect is suitable with JIEERYIN.
(5) traditional Chinese medicine compound extract of the present invention is on the impact of immunologic function
1, traditional Chinese medicine compound extract of the present invention is on the impact of T, bone-marrow-derived lymphocyte
(1) experiment material
1.1 medical materials: herbal mixture, all medical materials are all purchased from Yao and Sheng Academy of Traditional Chinese Medicine.
1.2 reagent concanavalin A, Con As (Con A), LPS
The healthy KM mice of 1.3 animal cleaning grade, is provided by Changsha, Hunan Experimental Animal Center.Certificate licence: SCK (Hunan) 2009-0012
1.4 instrument C0-150 CO2 gas incubator, RT-2100C microplate reader, Suzhou purification superclean bench, OLYMPUS IX51 inverted microscope.JA1203N type electronic balance.
(2) experimental technique
2.1 animal models, according to bibliographical information, by cyclophosphamide dry powder normal saline dilution, are mixed with 4mg/mL concentration, give intraperitoneal injection by 40mg/kg standard to mice.Continuous injection three days, sets up mouse immune inhibition.
2.2 animal groupings
By model mice 40, 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), 50% JIEERYIN group at random, and separately getting 10 normal mouses is blank group.Gastric infusion, one day 1 time, continuous 7d, matched group gives isometric(al) distilled water by 0.02ml/kg administration.1h after last administration, put to death animal and aseptically get spleen and test: 1. mouse boosting cell suspension is obtained: asepticly getting spleen, prepare individual cells, the centrifugal 5min of 1000rmin-1, remove supernatant, kowtow loose cell precipitation gently, add 0.4mL distilled water to vibrate gently 30s, hypotonic dissolution erythrocyte, then adding 0.4mL 2 times of dense normal saline (1.7%) makes solution recover isotonic, the centrifugal 5min of same rotating speed, after serum-free RPMI1640 liquid washes 2 times again, cell precipitation is resuspended in the RPMI RPMI-1640 containing 10% calf serum, counted under microscope, cell density is adjusted to 5 × 10
5/ mL, this is splenocyte for subsequent use.2. lymphproliferation response: add obtained splenocyte suspension 100 μ L in the 96 every holes of hole flat-bottomed plates 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 hole.Culture plate is put into 37 DEG C of 5%CO2 incubators, cultivate 48h.After cultivation terminates, mtt assay surveys cell optical density (OD).
Statistical method: all experimental datas all with
± s represents.Continuous data adopts SPSS 13.0 statistical software to carry out checking between variance analysis and group.
Experimental result
From table 7 Chinese medicine compound of the present invention on the impact of T, B lymphocyte proliferation, high dose group all extremely significantly can suppress mice T, B lymphocyte proliferation (P<0.01) by Promote immunity, and it is not obvious that low dose group urgees cultivation effect.
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* |
| 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: * p<0.05, * * p<0.01; With model group than △ p<0.05, △ △ p<0.01
2, Chinese medicine compound of the present invention generates and the impact of organ index hemolytic antibody
(1) experiment material: the same.
(2) experimental technique
2.1 animal models, according to bibliographical information, by cyclophosphamide dry powder normal saline dilution, are mixed with 4mg/mL concentration, give intraperitoneal injection by 40mg/kg standard to mice.Continuous injection three days, sets up immunosuppressed mice model.
2.2 grouping and administrations
Be divided into Normal group and herbal mixture high and low dose group and JIEERYIN group at random, often organize 10.Normal group gavages distilled water, and Chinese medicine compound group of the present invention gavages high and low dose 0.02ml/g, JIEERYIN group gavages 50% JIEERYIN 0.02ml/g, successive administration 7d.Inject 5% chicken red blood cell suspension 0.2mL to for 7th every mouse peritoneal and carry out immunity, continuous 7d.
2.3 serum hemolysin experiments
40min after last administration, wins eyeball and gets blood, the centrifugal 5min of 3000r/min.
Serum normal saline dilution 100 times, gets dilute serum 1mL in 0.5mL 5% chicken erythrocyte suspension and 0.5mL 10% complement (guinea pig serum) mixing, after being incubated 30min, sets to 0 a DEG C refrigerator 30min stopped reaction in 37 DEG C of calorstats.Centrifugal, get supernatant, in spectrophotometer 540nm place colorimetric, densitometric value.Separately establish normal saline to do blank, get its supernatant and return to zero as during colorimetric.Calculate each group of mice haemolysis optical density value.
2.4 organ indexs measure
De-neck execution mice, gets spleen immediately and thymus is weighed, and calculates organ index according to the following formula.Organ index=(organ weights/Mice Body weight) × 10
2.5 statistical method
Data are used
± s represents, compares with one factor analysis of variance, carry out statistical analysis with SPSS11.0 between each group.
(3) experimental result
3.1 Hemolysins generate: compare with model group, and high concentration group mice serum hemolytic antibody generates all has rising, the results are shown in Table 8: the impact that Chinese medicine compound of the present invention generates mice serum hemolytic antibody
illustrate that Chinese medicine compound of the present invention can suppress the generation of mice hemolytic antibody by Promote immunity.
Table 8
| Group | N(only) | Optical density (OD) |
| Blank group | 10 | 0.60390±0.040959 |
| Model group | 10 | 0.52120±0.050352 ** |
| JIEERYIN group | 10 | 0.58000±0.021448 △△ |
| High concentration group | 10 | 0.53620±0.033895 ** |
| Low concentration group | 9 | 0.56622±0.050884 |
Note: compare with Normal group: * * p<0.01, compares with model group: △ △ p<0.01
3.2 organ indexs:
Compare with matched group, as table 9: Chinese medicine compound of the present invention on the impact of mice organs index (n=10,
thymus index is at height, remarkable rising (p<0.05) is all had in low dose group mice, and high concentration group raises more obviously (p<0.01), illustrate that Chinese medicine compound of the present invention can promote mouse immune organ thymus, the weightening finish of spleen, prompting compound recipe can improve immunosuppressed mice immune organ and increase.
Table 9
| Group | Index and spleen index | Thymus index |
| Blank group | 0.08790120±0.009228541 △△ | 0.04117619±0.007334765 △△ |
| Model group | 0.06729571±0.005778540 * | 0.02667668±0.003446494 * |
| JIEERYIN group | 0.08065789±0.008821144 | 0.02747108±0.004779367 |
| High concentration group | 07713666±.010022596 △△ | 0.03566412±0.004301745 △△ |
| Low concentration group | 0.08256155±0.010442171 △ | 0.02999935±0.006153046 △ |
Compare with normal group: * p<0.05, * * p<0.01; With model group than △ p<0.05, △ △ p<0.01
3, Chinese medicine compound of the present invention affects mice phagocytic function
(1). experiment material: the same.
(2) experimental technique
2.1 animal models, according to bibliographical information, by cyclophosphamide dry powder normal saline dilution, are mixed with 4mg/mL concentration, give intraperitoneal injection by 40mg/kg standard to mice.Continuous injection three days, sets up immunosuppressed mice animal model.
2.2 grouping and administrations
Be divided into Normal group and the large and small dosage group of Chinese traditional compound medicine and JIEERYIN group at random, often organize 10.Normal group gavages distilled water, and Chinese medicine compound group of the present invention gavages high and low dose 0.02ml/g, and JIEERYIN group gavages 50% JIEERYIN 0.02ml/g, successive administration 8d.Last administration after 40 minutes in mouse tail intravenous injection india ink diluent.After injecting prepared Chinese ink, 2min and 5min is in mouse orbit venous blood collection 0.02ml, incorporates in 2ml0.01% sodium carbonate liquor.Put to death mice and get its liver, spleen is weighed.Get 0.02ml Mouse Blood and be dissolved in 2ml0.01% sodium carbonate liquor as blank group, measure absorbance in 650nm place.By following formulae discovery phagocytic index (k) and engulf coefficient (a): k=(logA1-logA2)/(t1-t2), a=body weight/(liver weight+spleen weight) × k1/3. wherein A1, A2 represents the absorbance of successively twice blood sampling respectively, and t1-t2 represents the interval of twice blood sampling.
3 experimental results
By table 10: prescription on the impact of immunosuppressed mice phagocytic function (n=10,
visible, 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 group | 0.0959±0.0136 ** | 5.217±1.423 ** |
| JIEERYIN group | 0.0511±0.0110 *△△ | 4.484±0.511 |
| 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: * p<0.05, * * p<0.01; With normal group than △ p<0.05, △ △ p<0.01
(6) Chinese medicine compound of the present invention is to the regulating action of mouse vagina Flora Disturbance
1 experiment material:
Laboratory animal: cleaning grade Kunming kind female mice 50, in 8 ~ 10 week age, body weight 18 ~ 22g, is provided by Changsha, Hunan Experimental Animal Center.Certificate licence: SCK (Hunan) 2009-0012
Medicine and reagent: penicillin sodium salt is that Jiangxi east wind Pharmaceutical limited company of Jiang Zhong group produces, specification: 800,000 units, lot number: 041045-3.Prepared by penicillin sodium saline solution: existing preparation before each use, being adjusted to concentration with physiological saline solution is 12.5mg/mL.
Prepared by MRS agar culture medium: the culture medium of dissolving 25g agar in 1L distilled water, adds following ingredients: 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, adjusts pH6.2 ~ 6.4,121 DEG C of sterilizing 15min.
2 experimental techniques
Animal divides into groups: get Kunming kind female mice 50, be divided into Normal group (C) 10 at random, antibiotic causes mouse vagina dysbacteriosis model group (A) 40; Modeling success after, model be divided at random each 10 of high and low dose group, JIEERYIN positive controls.
Vaginal dysbacteriosis model is set up: except Normal group, get 50 μ L penicillin sodium saline solution (concentration is 12.5mg/mL) with micro sample adding appliance for other each group and carry out mouse vagina flushing administration, every day 1 time, continuous 5d, matched group injects the normal saline of same volume every day at every turn.
Medication: after mouse vagina dysbacteriosis model modeling success, 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%) each 50 μ L are injected to model mice intravaginal, continuous 7d, every day 1 time.Normal group and model group inject the normal saline of same volume every day at every turn.
The Sequencing and analysis of mouse vagina lactobacillus: after last administration 12h, gets 50 μ L normal saline with micro sample adding appliance, repeatedly rinses mouse vagina 8 times, gets vaginal washing fluid 10 times of serial dilutions of 30 μ L to 10-6.Drip kind on MRS selective medium with micro sample adding appliance, through 37 DEG C, 48h carries out count plate (CFU/mL) (experiment repetition 3 times) after cultivating.
3 experimental results
Mouse vagina dysbacteriosis model is caused by setting up antibiotic, lactobacillus after using prescription with the method for plate culture count in mouse vagina secretions measures, and the results are shown in Table 11: each group mouse vagina lactobacillus testing result (viable count CFU/mL) after medication.
Table 11
| Group | Number of animals | CFU |
| Normal saline group | 10 | 395±200.28* |
| 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# |
Note: compare * p < 0.01, * * p < 0.05 with model group; #p < 0.01 is compared with normal group
Experimental result shows: the continuous vaginadouche administration of penicillin sodium saline solution caused disturbed vaginal flora after 5 days, Lactobacillus vaginalis quantity reduce, compare with Normal group, difference have significant to show vaginal dysbacteriosis model is set up.Inject prescription extracting solution 50 μ L in mouse vagina, after continuous 7 days, significantly improve lactobacillus quantity, recover and exceed levels found in normal controls, comparing with model group, difference has pole significant.Illustrate that this prescription has regulating action for mouse vagina Flora Disturbance and increases Lactobacillus vaginalis quantity.
Chinese medicine compound of the present invention has following effect:
1, antibacterial action: adopt conventional Antimicrobial test, JIEERYIN XIYE makes positive control, specifies extract minimum inhibitory concentration and the minimum bactericidal concentration of Chinese medicine formula of the present invention.To common gram positive bacteria as staphylococcus aureus, streptococcus; Common gram negative bacteria is as escherichia coli, bacillus pyocyaneus, and common fungus such as Candida albicans, Aspergillus fumigatus, acrothesium floccosum etc. all have good antibacterial activity.
2, antiinflammatory action: study its antiinflammatory action with conventional acute inflammation model auricle edema, subcutaneous injection Oleum Tiglii induced mice ear corridor swelling model caused by dimethylbenzene, set up positive control (JIEERYIN makes positive control), the extract of specifying Chinese medicine formula of the present invention has obvious antiinflammatory action.
3, itching-relieving action: cause 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 can make positive control by JIEERYIN XIYE), the extract of specifying Chinese medicine formula of the present invention is significantly improved the effect of itch-threshold.
4, adjust disturbed vaginal flora and increase Lactobacillus vaginalis quantity
5, Promote immunity function: use conventional immunological experiment, set up positive control (also can make positive control by JIEERYIN XIYE), the extract of specifying Chinese medicine formula of the present invention has the phagocytic function promoting Turnover of Mouse Peritoneal Macrophages, promote that splenic T, bone-marrow-derived lymphocyte transform, improve the effect of body's immunity.
The foregoing is only embodiments of the invention; not thereby the scope of the claims of the present invention is limited; every utilize description of the present invention to do equivalent structure or equivalent flow process conversion; or be directly or indirectly used in other relevant technical fields, be all in like manner included in scope of patent protection of the present invention.
Claims (6)
1. the Chinese medicine composition that an anti-inflammation is antipruritic, it is characterized in that, be made up of the raw material of Chinese medicine of following weight portion: 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. the Chinese medicine composition that anti-inflammation according to claim 1 is antipruritic, it is characterized in that, be made up of the raw material of Chinese medicine of following weight portion: 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. the Chinese medicine composition that anti-inflammation according to claim 1 is antipruritic, is characterized in that, described Chinese medicine composition is made up of as effective ingredient the water extract of each raw material of Chinese medicine or ethanol extract.
4. the Chinese medicine composition that anti-inflammation according to claim 1 is antipruritic, is characterized in that, described Chinese medicine composition is refined liquid, and the volume of described refined liquid and the ratio of raw material of Chinese medicine quality are 1ml:0.5g to 1ml:1g.
5. the preparation method of the Chinese medicine composition that an anti-inflammation is antipruritic, it is characterized in that, raw material of Chinese medicine by following weight portion: 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, filter, obtain filtrate, be Alumen 5-10 part again by weight fraction, the raw material of Chinese medicine of Cortex Pseudolaricis 30-50 part adds the alcohol that mass percent is 70%, after alcohol extraction twice, gained alcohol extract joins gained filtrate, slowly adding ethanol while stirring makes the alcohol content of filtrate be 70%-80%, airtight cold preservation 48 hours, filter, filtrate recycling ethanol, be dissolved in water and filter to obtain refined liquid, volume and the raw material of Chinese medicine mass ratio of described refined liquid are 1ml:0.5g to 1ml:1g.
6. the preparation method of the Chinese medicine composition that anti-inflammation according to claim 5 is antipruritic, it is characterized in that, each weight part ratio of described 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.
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| CN103611185B (en) * | 2013-12-03 | 2016-02-24 | 安徽缘远博爱生物技术有限公司 | A kind of Radix Sophorae Flavescentis sterilization soluble stanching gauze |
| 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 |
| 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 |
| CN112206259B (en) * | 2020-10-30 | 2022-09-16 | 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 |