CN106729444A

CN106729444A - A kind of gel breast of repairing skin wound surfaces and preparation method thereof

Info

Publication number: CN106729444A
Application number: CN201710085048.0A
Authority: CN
Inventors: 胡荣; 刘量; 王冬东; 沈新宇; 廖凯; 刘帅
Original assignee: Yangzhou University
Current assignee: Yangzhou University
Priority date: 2017-02-17
Filing date: 2017-02-17
Publication date: 2017-05-31

Abstract

Gel breast of a kind of repairing skin wound surfaces and preparation method thereof, belongs to biomedicine technical field, will be crushed after cucumber seed, curcuma zedoary, the rhizome of davallia, asarum, ramulus mori, cape jasmine, frankincense and myrrh mixed freezing, adds water distillation, obtains volatile oil and lower floor's aqueous；By liquid refrigeration, centrifugation, upper strata liquid is obtained；Chitosan hydrochloride is soaked with upper strata liquid, gel liquid is obtained；Arlacel-80 volatile oil is dissolved, oil phase is obtained；Polyoxyethylene sorbitan monooleate lower floor's aqueous are dissolved, water phase is obtained；By oil phase and water phase homogeneous, emulsion oil-in-water is obtained；By emulsion oil-in-water and gel liquid mixing homogeneous, the gel breast of repairing skin wound surfaces is obtained.Can be used for the treatment of the surface of a wound such as frostbite, burn and scald, external cause wound, radiation injury and scabies and diabetes.

Description

A kind of gel breast of repairing skin wound surfaces and preparation method thereof

Technical field

The invention belongs to biomedicine technical field, the preparation side of the gel breast of more particularly to a kind of repairing skin wound surfaces Method.

Background technology

Frostbite, burn and scald, wound and furuncle, malignant boil, sore subcutaneous ulcer, bedsore, diabetes, varicose ulcer of lower extremity, radiation Property ulcer etc. disease be clinically common skin callus, for the crowd of hypoimmunity, chronic inflammation not more for a long time Easily induce other diseases.To above-mentioned dermatopathy, conventional medicine such as hydrogen peroxide, Iodophor, chlorohexidene and antibiotic, The chemicalses such as hormone are virose to cell, have suppression to make epithelialization and angiogenesis while wound infection is resisted With being unfavorable for the reparation of wound.The rarely seen pure of the Chinese herbal medicine extracts such as aloe, Asian puccoon, pomegranate cuts the medication of trauma model Report, heals extremely limited to the complicated condition of the injury.Growth factor class species is various, but has the danger for causing hyperplastic scar or even canceration Danger, still in the experimental stage.And with the fast development of the industries such as traffic, building, the traumatic wounds of skin are more and more, companion There is the Disease impaired wound healing such as diabetes, varication, treat thorny.

The content of the invention

The present invention seeks to propose a kind of gel breast that can effectively accelerate union of wounded skin.

The technical scheme is that：Mass percent >=0.5% of volatile oil, the quality of aurantiin in gel breast Percentage >=0.005%.

Consulting the compound Chinese patent medicine content quality control index that 2015 editions pharmacopeia include is mostly a composition or does not have, such as external application Preparation：" trauma ointment " is referred to by Asian puccoon, garden burnet, cape jasmine, rheum officinale, the root of large-flowered skullcap, golden cypress, borneol seven flavor medicine thing prescription, content Quality Control Mark is the jamaicin in golden cypress；" Anyang refines cream " is by Radix Aconiti, Radix Aconiti Kusnezoffii, the root of three-nerved spicebush, radix ampelopsis, the root of Dahurain angelica, bletilla, semen momordicae, wood Logical, pawpaw, Rhizoma Sparganii curcuma zedoary, Radix Angelicae Sinensis, the radix paeoniae rubrathe, Chinese cassia tree, rheum officinale, the capsule of weeping forsythia, dragon's blood, asafoetide, frankincense, myrrh, catechu, menthol, water Poplar acid methyl esters, the four traditional Chinese medicine thing prescription of borneol 20, content quality control index is medicinal extract amount；" YANGHE JIENING GAO " is by fresh burdock grass, fresh The fresh lopseed of wind, Radix Aconiti, cassia twig, rheum officinale, Radix Angelicae Sinensis, Radix Aconiti Kusnezoffii, unprocessed Radix Aconiti Lateralis, earthworm, stiff silkworm, the radix paeoniae rubrathe, the root of Dahurain angelica, radix ampelopsis, bletilla, Rhizome of chuanxiong, teasel root, windproof, schizonepeta, excrementum pteropi, the banksia rose, citron, dried orange peel, Chinese cassia tree, frankincense, myrrh, storax, muscone 20 Seven flavor medicine thing prescription, without content quality control index；" awei huapi plaster, awei huapi gao " is by rhizoma cyperi, the bark of official magnolia, Rhizoma Sparganii, curcuma zedoary, Radix Angelicae Sinensis, Radix Aconiti Kusnezoffii, life Monkshood, garlic, the fruit of Rangoon creeper, the root of Dahurain angelica, pangolin, semen momordicae, dung beetle, Radix picrorrhizae, rheum officinale, castor bean, frankincense, myrrh, aloe, blood Exhaust, realgar, Chinese cassia tree, camphor, the four traditional Chinese medicine thing prescription of asafoetide 20, without content quality control index；" Asian puccoon ointment " by Asian puccoon, Radix Angelicae Sinensis, Windproof, glutinous rehmannia, the root of Dahurain angelica, frankincense, myrrh seven flavor medicine thing prescription, without content quality control index.

The present invention --- gel breast mainly considers cucumber with volatile oil content and naringin content as quality control index In seed, curcuma zedoary, the rhizome of davallia, asarum, ramulus mori, cape jasmine, frankincense, myrrh this eight tastes formula herbs,《Chinese book on Chinese herbal medicine》Described in cucumber Seed has reunion of fractured tendons and bones, and wind-dispelling, effect of dissolving phlegm, its active ingredient is mainly volatile oil, also there is flavone compound；Pharmacopeia pair The quality control index of curcuma zedoary, asarum, frankincense, myrrh this few taste medicinal material is all the content of volatile oil；Pharmacopeia is to the rhizome of davallia, cape jasmine It is respectively aurantiin, Gardenoside and ethanol extract with the quality control index of ramulus mori, the active ingredient of ramulus mori is mainly flavonoid Thing.With pharmacopeia and《Chinese book on Chinese herbal medicine》It is foundation, determines that inventive gel milk content quality control index there are two, one is total volatile oil； Two is the flavone compound in terms of aurantiin.Containing flavones ingredient Chinese herbal medicine, flavonoids is polyhydroxy based component and many band phenol more Hydroxyl, aurantiin is flavones ingredient, and there is phenolic hydroxyl group No. 5 carbon connection α hydroxyls, No. 7 carbon connection β hydroxyls, 4 ' positions in molecular structure, Content is higher in gel breast, representative.

The present invention combines traditional Chinese medicine promoting blood circulation to remove blood stasis, the medication principle of relieving rigidity of muscles and knitting bone, using cucumber seed, curcuma zedoary, the rhizome of davallia, thin Pungent, ramulus mori, cape jasmine, frankincense, the plant medicinal material of myrrh eight prepare effect gel breast, through experiment, can be used for frostbite, burn and scald, outer Because of the treatment of the surface of a wound such as wound, radiation injury and scabies and diabetes.

A kind of preparation method it is another object of the present invention to propose above gel breast.

The present invention is comprised the following steps：

1）Cucumber seed, curcuma zedoary, the rhizome of davallia, asarum, ramulus mori, cape jasmine, frankincense and myrrh are mixed, is crushed after freezing, added water and soak After be placed in Sheng medicinal cupping in distilled, respectively obtain volatile oil and lower floor's aqueous；The liquid after medicinal cupping Chinese medicine slag is removed will be contained cold Hide, then through centrifugation, obtain upper strata liquid；

2）By chitosan hydrochloride using being mixed after the immersion of above-mentioned upper strata liquid, gel liquid is obtained；

3）Arlacel-80 is dissolved using above-mentioned volatile oil, oil phase is obtained；By polyethenoxy sorbitan list oil Acid esters is dissolved using lower floor's aqueous, obtains water phase；

4）Add water mutually and homogeneous oil phase, obtain emulsion oil-in-water；

5）Emulsion oil-in-water and the gel liquid are mixed, through homogeneous, the gel breast of repairing skin wound surfaces is obtained.

In inventive gel breast：The effects such as cucumber seed is replenished the calcium, zhuanggu, regulation channels and collaterals, promotion human body cell regeneration；Curcuma zedoary Promoting the circulation of qi blood-breaking, Xiao Ji Zhi Tong are antiviral；Asarum sedation and analgesia, anti-inflammation；Cape jasmine can hemostasia and detumescence；Ramulus mori has work promoting blood circulation and removing blood stasis With；The rhizome of davallia has promoting blood circulation and hemostasis；Frankincense, myrrh are respectively provided with promoting blood circulation and stopping pain, detumescence and promoting granulation effect；Water soluble chitosan is good to be stopped Blood and relieving pain, antibacterial, biodegradation and compatibility can accelerate the high-quality of the surface of a wound to heal.Above-mentioned medicine auxiliary material comprehensive function, Neng Gou Biological property during corium unstability to Skin Cell produces good stable state reparation.

Further, cucumber seed of the present invention, curcuma zedoary, the rhizome of davallia, asarum, ramulus mori, cape jasmine, frankincense, myrrh, dehydration mountain The mixing quality ratio of pears alcohol monoleate, polyoxyethylene sorbitan monooleate and chitosan hydrochloride is 30: 5: 10: 10 ∶10∶5∶5∶5∶2∶5∶13.Wherein cucumber seed, curcuma zedoary, the rhizome of davallia, asarum, ramulus mori, cape jasmine, frankincense, myrrh press monarch Principle of medication metrology recipe, the active ingredient of extraction is distributed in volatile oil and its lower floor's aqueous and liquid；Anhydro sorbitol list Oleate is that oil soluble emulsifying agent can make oil dispersed into oil droplet, and polyoxyethylene sorbitan monooleate is water-emulsifiable Agent, in the presence of external force oil phase being made to be dispersed in water phase well, the total consumption of emulsifying agent is generally 2 ~ 10% in emulsion Scope, and hydrophilic emulsifier and lipophilic emulsifier collocation use preferable, hydrophilic emulsifier when preparing oil-in-water emulsion Agent consumption is more than lipophilic emulsifier, usually its more than 2 times；Chitosan hydrochloride forms gel in being dispersed in liquid, has Certain viscosity, can keep the physical stability of emulsion, and the merging and water-oil phase for preventing oil droplet are layered.Each spice is used in formula Amount is than being determined according to principles above and experiment, it is ensured that the shaping of gel breast and the performance of effect.

Step 1 of the present invention）In, the gross mass of cucumber seed, curcuma zedoary, the rhizome of davallia, asarum, ramulus mori, cape jasmine, frankincense and myrrh with The mixing ratio of water is 1: 15.The amount of water for carrying out steam distillation is 15 times of spice gross mass, 15 times of water 5h of medicinal material amount Can extract the effective components such as volatile oil complete, it is ensured that steam distillation is smoothed out.

Step 1）In, 15 times of water and distillation time of medicinal material amount are to be verified in plant-size equipment for 5h. The water distillation 5h amount of liquid medicine satisfactions of 15 times of medicinal material amounts prepare gel liquid consumption requirement；5h can extract the effective components such as volatile oil Completely, continuing the effective component extracted amounts such as volatile matter distillation oil will not increase, and the short volatile oil extracting of distillation time is incomplete.

The step 1）In, the temperature environment of the liquid refrigeration is 4 DEG C, and cold preservation time is 16h；The speed of the centrifugation It is 8000 r/min, centrifugation time is 0.5h.Removing when the liquid after the dregs of a decoction is heated makes vegetable protein, mucilaginous substance, resin etc. miscellaneous Matter is flocculated, and is precipitated when refrigeration contributes to separate out under 4 DEG C of low temperature environments, and it is complete that refrigeration 16h ensures that impurity is separated out, and in addition 8000 R/min centrifugations improve acceleration of gravity, and it is bottomed that the centrifugation time of 0.5h makes the impurity being suspended in liquid settle completely, supernatant medicine Liquid is easily poured out.

The step 2）In, the time of immersion is 1h.The time for using liquid to soak chitosan hydrochloride is 1h, because of salt Acidified chitosan is macromolecular compound, and the winding of its molecule is agglomerating to need certain hour ability spread apart, the big expansion of the big quality of molecule Laxity, so needing the swelling time of 1h.

The condition of homogeneous is 3000 r/min, homogeneous 5min is further continued for after oil phase is instilled into water phase, because water-oil phase is mutual It is immiscible, it is necessary to mechanical force smashes oil reservoir makes into broken small oil droplet, oil droplet is emulsified agent parcel and is dispersed in water phase, and this emulsification is through testing Verify and can complete emulsification within 5~10 minutes with the shearing force of 3000 r/min.The too high generation whirlpool of speed is stirred and is not easy into air Drive out of；Mixing time is long, easily demulsification, that is, the oil droplet for being emulsified agent parcel is drawn, and oil droplet merges influence emulsification effect Rate.

Brief description of the drawings

Fig. 1 is the chemical structural drawing of aurantiin.

Fig. 2 is the dissolving situation photo for differentiating gel breast with dilution method.

Fig. 3 is to differentiate that gel breast foreign minister dyes blue situation photo with decoration method.

Fig. 4 is the indentification by TLC figure of aurantiin in gel breast（Result is observed under 365nm uviol lamps）.

Fig. 5 is the indentification by TLC figure of aurantiin in gel breast（Result is observed under 254nm uviol lamps）.

Fig. 6 is the liquid chromatogram of aurantiin standard reference material, and abscissa is the time（Point）, for qualitative；Ordinate is Absorbance, for quantitative.

Fig. 7 is that the liquid chromatogram abscissa of gel breast need testing solution is the time（Point）, for qualitative；Ordinate is to inhale Luminosity, for quantitative.

Fig. 8 is that the liquid chromatogram abscissa of negative gel breast need testing solution is the time（Point）, for qualitative；Ordinate It is absorbance, for quantitative.

Fig. 9 is basis of microscopic observation normal rats dermal pathology slice map.

Figure 10 is basis of microscopic observation scalding model group dermal pathology slice map.

Figure 11 is basis of microscopic observation gel breast group scalding healing tissue pathological slice figure.

Figure 12 is basis of microscopic observation positive drug control group scalding healing tissue pathological slice figure.

Specific embodiment

First, the preparation of gel breast：

1st, cucumber seed 1.5kg, the kg of curcuma zedoary 0.25, the kg of the rhizome of davallia 0.5, asarum 0.5kg, ramulus mori 0.5kg, cape jasmine 0.25 are weighed Kg, the kg of frankincense 0.25, the kg mixing coarse crushings of myrrh 0.25, plus 60kg water are distilled in multi-function extractor.Divide after 5h Take volatile oil 0.31kg and lower floor aqueous 1kg in collector.

Extractor Chinese medicine liquid is released, 4 DEG C of refrigeration 16h, 8000 r/min 30 min of centrifugation pour out supernatant 45kg, discard Bottom settles impurity.

2nd, weigh in the kg of chitosan hydrochloride 0.65 addition step 1 gained supernatants, immersion more than 1h stirs evenly to obtain gel again Liquid 45.65kg.

3rd, in Arlacel-80 0.1kg being dissolved in into the volatile oil collected by step 1, oil phase 0.41kg is obtained.

During the kg of polyoxyethylene sorbitan monooleate 0.25 dissolved in into step 1 gained lower floor aqueous, water phase is obtained 1.25kg。

Oil phase is instilled into water phase under 3000 r/min processing conditions again, is continued homogeneous 5min and is obtained emulsion oil-in-water 1.66kg。

The 4th, step 3 gained emulsion oil-in-water is instilled the gel medicine of step 2 gained under 3000 r/min processing conditions In liquid, continue homogeneous 5min and obtain gel breast 47kg.

2nd, the discriminating of gel breast emulsion types：

1st, dilution method：

10mL tool plug test tubes two are taken, taking a little gel breast with spoon point places wherein, then is separately added into 5mL ether and distillation Water, shaking for several times, is as a result shown in Fig. 2.Fig. 2 shows that gel breast can not mix with ether, assembles agglomerating（Such as the left side test tube in Fig. 2 Display）, but can be diluted with water and mix integral（Right side test tube such as Fig. 2 shows）.Above discrimination test illustrates that gel breast is outer It is mutually aqueous medium, as oil-in-water type.

2nd, decoration method：

Enter 1 drop aqueous solution of methylene blue to gel emulsion droplet, draw on a small quantity, drop one is dropped on slide, and covered puts microscope Lower observation, is as a result shown in Fig. 3.Fig. 3 shows that the foreign minister of gel breast is that the decentralized medium for joining together is dyed to blueness, and each is newborn Drop is that dispersed phase volatile oil is not colored, and methylenum careuleum is water-soluble dye, illustrates that gel breast foreign minister is aqueous medium, as water bag Oil type.

3rd, gel breast dewatering ability experiment：

By gel breast with the centrifugation 5hr of 4000r/min, do not occur being layered, flocculate and phenomenon that emulsion droplet merges.The experiment phase When the physically stable implementations that 1 year is placed in gel breast, that is, have good stability.

4th, gel breast volatile oil content testing

By 2015 editions pharmacopeia volatile oil content testing methods, precision weighs gel breast 100g, in putting volatile oil extractor, vapor Oil mass of being volatilized in distillation to collector no longer increases（About 1h）, obtain volatile oil 0.66g.

It is computed, the mass percent of volatile oil is 0.66% in gel breast.

5th, the discriminating of aurantiin and assay in gel breast

Aurantiin is flavones ingredient, and there is phenolic hydroxyl group No. 5 carbon connection α hydroxyls, No. 7 carbon connection β hydroxyls, 4 ' positions in molecular structure, As shown in Figure 1.

The preparation of aurantiin standard reference material solution：Precision weighs aurantiin standard items, plus methyl alcohol excusing from death 10min is made 62 μ g/ mL solution.

The preparation of gel breast need testing solution：Precision weighs solidifying rubber latex sample 10g, plus methyl alcohol 20mL excusing from death 15min, mistake Filter, filtrate is with methanol constant volume to 25mL.

The preparation of negative gel breast need testing solution：Take formula ratio cucumber seed, curcuma zedoary, asarum, ramulus mori, cape jasmine, frankincense, do not have Medicine, by the preparation of implementation method one, gel breast：Similarly hereinafter method operation preparation feminine gender gel is newborn for item, and precision weighs negative gel milk sample Product 10g, plus methyl alcohol 20mL excusing from death 15min, filtering, filtrate is with methanol constant volume to 25mL.

The preparation of rhizome of davallia control medicinal material solution：Coarse crushing rhizome of davallia medicinal material 5g is taken, plus 10min, filter are extracted in methyl alcohol excusing from death Except the dregs of a decoction are standby.

1st, the indentification by TLC of gel breast index components aurantiin

By above-mentioned four kinds of solution, through 0.45 μm of membrane filtration, quantitative point is on the lamellae of same silica G 254 respectively, with toluene, Ethyl acetate, formic acid and water volume ratio are that the mixed system of the ︰ 3 of 1 ︰, 12 ︰ 2.5 is solvent, are dried after expansion, respectively in 365nm Observed with 254nm uviol lamps：Test sample, standard items and control medicinal material show aurantiin spot on same position, see Fig. 4 and Fig. 5.1 is aurantiin standard reference material solution thin-layer chromatography migration situation in Fig. 4,5, and the single spot of aurantiin is presented；2 is cloudy Property gel breast need testing solution thin-layer chromatography migration situation, the spot with the same migration value of aurantiin is not presented；3 is rhizome of davallia medicine Material solution thin-layer chromatography migration situation, is presented the spot with the same migration value of aurantiin；4 is gel breast need testing solution thin layer Analysis migration situation, is presented the spot with the same migration value of aurantiin.

Fig. 4 and Fig. 5 show：Fluorescence spot under 365nm（Fig. 4））Compared with 254nm because fluorescent quenching produce blackening（Figure 5, official method）Easily observe, point out pharmacopeia this thin-layer identification method to be improved as the case may be.

2nd, the measure of gel breast index components naringin content

2.1 chromatographic conditions

Chromatographic column：Symmetry C₁₈（5 μm, 4.6 × 250 mm）；Mobile phase：The mixture of methyl alcohol and 3% aqueous acetic acid Product is than being 40 ︰ 60；Flow velocity：0.8 mL/min；Detection wavelength：283 nm；Column temperature：25℃；Sample size：10 μL.

2.2 linear relationships are investigated

It is accurate respectively to draw the aurantiin reference substance solution 0.25,0.5,1.0,1.5,2.0,2.5 that concentration is 62 μ g/ mL , 3.0 mL, put in 10mL volumetric flasks, with methanol dilution to scale, shake up, respectively by 2.1 chromatographic condition sample introductions determine peak area. With peak area as ordinate（Y）, aurantiin concentration is abscissa（X）Linear regression is carried out, Y=8590.3X+1609, R2=is obtained 0.9991, show that aurantiin is in good linear relation in 1.55 ~ 18.6 μ g/mL concentration ranges and peak area.

2.3 specificities, precision, stability, reappearance, average recovery experiment

By 2.1 lower chromatographic conditions respectively by aurantiin reference substance solution, gel breast need testing solution, negative gel breast test sample As a result solution, injecting chromatograph shows that remaining chromatographic peak does not disturb the measure of aurantiin, sees Fig. 6, Fig. 7, Fig. 8.

Fig. 6 is the liquid chromatogram of aurantiin standard reference material solution, aurantiin appearance after sample introduction 15.806min.

Fig. 7 is the liquid chromatogram of gel breast need testing solution, and the synergy of retention time 15.476 is in gel breast In aurantiin behavior.

Fig. 8 is the liquid chromatogram of negative gel breast need testing solution, compared with Fig. 7, aurantiin chromatogram is not presented simply Peak.

Precision draws aurantiin reference substance solution, repeats sample introduction 5 times, records aurantiin peak area, calculates RSD=0.93%, Show that instrument precision is good；Precision draws same need testing solution, respectively in different time injection in 0,2,4,8,12,24 hours Chromatograph, records aurantiin peak area, calculates RSD=1.30%, shows that need testing solution is good in 24 hours internal stabilities；It is accurate Draw same need testing solution 5 times, order sample introduction determines the aurantiin peak area of each sample introduction, calculates the RSD of naringin content =1.76%, show that the assay method reappearance is good；Accurate equivalent draws same 9 parts of need testing solution, and every three parts accurate respectively 1 mL, 2 mL and 3 mL same concentration aurantiin reference substance solutions are added, is mixed, by 2.4.1 lower chromatographic condition sample introduction, determined Its average recovery rate is 98.92%, RSD=1.47%.

According to methodological study and practical measurement, by 2.2 lower normal equations：

Y=8590.3X+1609, R2=0.9991, the mass percent for calculating aurantiin in gel breast are 0.005319%.

6th, reparative experiment of the gel breast to burned rats：

Experimental animal is SD rats, is provided by Yangzhou University's comparative medicine center, and animal productiong licensing number is SXCXK（Soviet Union） 2012-0004, is SYXK using credit number（Soviet Union）2012-0029）.

Rat is randomly divided into 4 groups, every group 12, respectively with model group, gel breast group, positive controls（Commercially available moistening MEBO）With Normal group sub-cage rearing.

1st, modeling and administration：

Rat adapt to environment after one week fasting overnight can't help water.Back about 12cm is applied with 8% sodium sulfide solution²Depilation, hair removal section Warm water is cleaned, medical 75% ethanol disinfection, after normal raising 1d, observes and determine that hair removal section is abnormal without redness, infection, inflammation etc. Situation, carries out scalding model preparation.Caused to anaesthetize with 10% chloraldurate intraperitoneal injection（0.003 mL/g）, the depilation of 75% ethanol disinfection Area.Modeling：Boiling water heats the min of counterweight 20, and slightly pressure is placed in rat back 15s, and preparing every burned rats gross area is 12cm²Deep II degree of scalding model.The surface of a wound is cleaned after scalding 5 hours with physiological saline, 2d starts administration.Experimental group gives solidifying Latex, positive controls give MEBO, to be dipped with cotton swab and be grouped single cage raising after being applied to site of injury, give sooner or later daily Each 1 time of medicine, each dosage 2g/ only, drink water during experiment by normal feed.

Observed in each time point after administration and record wound healing situation, 4 rat materials of every group of each time point mark, Remaining is used to observe healing time.

Method：With modeling same method anesthetized rat, surface of a wound skin is removed with sterile surgical scissors, continued after wound coating Single cage is raised.Skin histology is fixed with 4% paraformaldehyde, section, HE dyeing, in optical microphotograph Microscopic observation.

2nd, Testing index：

2.1 Wound healing rates

Respectively at administration the 5th, 10,15,20d, describe the rat surface of a wound with pan paper, to weigh paper weight, area linear equation y= 362.71x+0.0102, R²=0.9991 calculates surface of a wound area；Wound healing rate is calculated simultaneously：

2.2 wound healing times

Heal evaluation index completely：All come off with forming a scab, epiderm skin completion and the degree that becomes mildewed are index, during record incrustation Between and healing time, carry out statistical test.

2.3 new capillary vessels are counted

Respectively at the 5th, 10,15d modeling same method anesthetized rat is administered, materials, HE dyeing, new capillary vessel is counted.

2.4 expression, rat plasma TNF-α and the vWF for determining rat skin NF- κ B

After administration 15d, eyeball takes blood, determines rat plasma TNF-α and vWF, and modeling same method anesthetized rat removes skin, Determine the expression of rat skin NF- κ B.

2.5 statistical procedures

Observation by light microscope HE coloration results, experiment the data obtained with(average) ± s（Standard deviation）Represent, application The softwares of SPSS 17.0 carry out statistical analysis, and measurement data is using one-way analysis of variance and does Multiple range test.WithP<0.05,P <0.01 is that difference is statistically significant.

3rd, result of the test：

Influence of the 3.1 gels breast to wound tissue healing rate

There are body shakes in burned rats, are trembled after administration and substantially weaken, and non-goodbye jitter two days later are administered, just Step shows that gel breast has analgesic effect.

Whole experiment process, compared with model group, gel breast is organized and positive controls scab forming time and wound healing time It is respectively provided with statistical significance（P<0.01）；Gel breast group surface of a wound scab forming time, healing time are earlier than positive controls（P< 0.05）, with statistical significance.5 after administration, each group Rat Wound Healing rate rises with the increase of time after 10,15,20d, Each time period gel breast group and positive controls Wound healing rate are above model group（P<0.01）, with statistical significance；With Positive controls compare, and gel breast group is in 5d（P<0.05）With 10d, 15d, 20d（P<0.01）With statistical significance, see Table 1, table 2.

The each group scab forming time of table 1 and healing time comparing ( , n ＞ 4)

Note：A is compared with model group in table： p<0.01；B is compared with positive controls：p<0.01。

The each group different time points wound tissue healing rate of table 2 comparing ( , n ＞ 4)

The expression of 3.2 each group rat skins administration 15dNF- κ B

NF- κ B positive expressions in model group epidermis epithelial cell, nucleus shows brown color, its integral optical density value (16.1 ± 2.0) compared with normal group (6.4 ± 1.6), positive controls (11.1 ± 1.7), gel breast group (6.6 ± 3.5), it is respectively provided with Significant difference (P<0.01)；NF- κ B positive cell rates in model group epidermis epithelial cell（63±1.4）With normal group（24± 0.8）, positive controls (38 ± 1.3), gel breast group (27 ± 4.1) compare, be respectively provided with significant difference (P<0.01).Sun Property control group epidermis epithelial cell in NF- κ B positive expressions significantly reduce, part nucleus and endochylema are in brown color, gel breast group Only see a few cell core and endochylema expression NF- κ B, its positive cell rate（27±4.1）With positive controls（38±1.3）Compare, With significant difference (P<0.01).Gel breast group rat skin tissue NF- κ B integral optical density values (6.6 ± 3.5）With sun Property control group (11.1 ± 1.7）Compare, with significant difference (P<0.01).Show the therapeutic intervention of gel breast, effectively suppression The activation of NF- κ B has been made, so as to inhibit inflammatory reaction, 3 has been shown in Table.

The positive expression of the rat skin tissue of table 3 administration 15d NF- κ B（, n=4）

Note:A is that model group compares with normal group, positive controls, gel breast group in tableP< 0.01；B is gel breast group and sun Property control group comparesP< 0.01；C is that gel breast group compares with normal group, P > 0.05。

3.3 each group rat plasmas are administered the measurement result of 15d TNF-αs and vWF

Model group Plasma TNF-α and vWF expression have higher than Normal group, positive controls and gel breast treatment group Statistical significance (P<0.01)；Gel breast treatment group Plasma TNF-α and vWF expression are significantly lower than positive controls, have Statistical significance (P< 0.01)；Show the therapeutic intervention of gel breast, the expression of TNF-α and vWF is effectively inhibited, so as to suppress Inflammatory reaction and endothelial injuries.Model group Plasma TNF-α and vWF expression is proportionate (r=0.01), is shown in Table 4.

The testing result of each group rat plasma 15d vWF of table 4 and TNF-α（, n=4）

Note:A is that model group compares with Normal group, positive controls, gel breast group in table, P<0.01；B is gel breast group Compare with positive controlsP< 0.01；C is that gel breast group compares with normal group, P > 0.05；Model group vWF and TNF-α Expression is proportionate（r=0.01）.

3.4 scalding healing tissues are administered 15d pathology section examination results

Scalding healing tissue pathological slice observation result is shown in accompanying drawing 3 ~ 6（× 100, HE are dyeed）.

Normal skin tissue is complete as seen from Figure 9, clear layer, and fat is more ruddy, flexible, dermal collagen fiber arrangement Neatly, epidermis rule, without inflammatory cell.

Model group rats wound tissue lymphocyte still largely infiltrates as seen from Figure 10, epidermis ulceration, with a large amount of inflammation Disease cellular infiltration, arrangement of collagen fibers is disorderly, and color and luster is dark.

The wound tissue healing of gel breast group is preferable as seen from Figure 11, has no inflammatory cell infiltration, and epidermis rule, color and luster is approached Normal skin, arrangement of collagen fibers is neat.

The healing of positive controls rat wound tissue is preferable as seen from Figure 12, has no inflammatory cell infiltration, and epidermis is relatively regular, Arrangement of collagen fibers is more neat.

3.5 new capillary vessels are counted

Each group new capillary vessel is in the trend fallen after rising, and each group new capillary vessel number reached maximum at the 10th day, with After decline, new capillary vessel is not further added by 15 days, partly starts in closure state, and gel breast group and positive controls are new Angiogenic number compared with model group, with obvious otherness（p<0.01、p<0.05）, it is shown in Table 5.

The new capillary vessel of table 5 is counted（, n=4）

Note：A is compared with model group in tablep<0.01；B is compared with positive controlsp<0.05。

Claims

1. a kind of gel of repairing skin wound surfaces is newborn, it is characterised in that：The gel breast in volatile oil mass percent >= 0.5%, mass percent >=0.005% of aurantiin.

2. repairing skin wound surfaces as claimed in claim 1 gel breast preparation method, it is characterised in that comprise the following steps：

3. preparation method according to claim 2, it is characterised in that the cucumber seed, curcuma zedoary, the rhizome of davallia, asarum, ramulus mori, Cape jasmine, frankincense, myrrh, Arlacel-80, polyoxyethylene sorbitan monooleate and chitosan hydrochloride Mixing quality ratio is 30: 5: 10: 10: 10: 5: 5: 5: 2: 5: 13.

4. preparation method according to claim 2, it is characterised in that the step 1）In, cucumber seed, curcuma zedoary, the rhizome of davallia, The gross mass of asarum, ramulus mori, cape jasmine, frankincense and myrrh and the mixing ratio of water are 1: 15.

5. the preparation method according to Claims 2 or 3 or 4, it is characterised in that the step 1）In, the distillation time is 5h。

6. preparation method according to claim 2, it is characterised in that the step 1）In, the temperature of the liquid refrigeration Environment is 4 DEG C, and cold preservation time is 16h；The speed of the centrifugation is 8000 r/min, and centrifugation time is 0.5h.

7. preparation method according to claim 2, it is characterised in that the step 2）In, the time of immersion is 1h.

8. preparation method according to claim 2, it is characterised in that the step 4）The condition of middle homogeneous is 3000 r/ min。

9. the preparation method according to claim 2 or 8, it is characterised in that after oil phase is instilled into water phase under processing condition again Continue homogeneous 5min.

10. preparation method according to claim 2, it is characterised in that the step 5）The condition of middle homogeneous is 3000 r/ min。