CN104825519B - The Preparation method and use of vernonia anthelmintica phenolic acid part - Google Patents
The Preparation method and use of vernonia anthelmintica phenolic acid part Download PDFInfo
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Abstract
The present invention relates to the preparation methods and purposes of a kind of vernonia anthelmintica phenolic acid part.This method is that the vernonia anthelmintica fruit that will be dried crushes, and is extracted using organic solvent, resin separation purification obtains vernonia anthelmintica willd extract, and main component is phenolic acid compound.The mass percent 20%-99% of phenolic content in position.The content of main active 3,4- two-caffeoyl quinic acid, 3,5- two-caffeoyl quinic acid and 4,5- two-caffeoyl quinic acid is respectively 3-26%, 5-34% and 5-40% in vernonia anthelmintica phenolic acid part.Prove through preliminary experiment: the vernonia anthelmintica phenolic acid part that the method obtains through the invention has the function of increasing melanocyte in hair follicle, promotes melanin genesis, can be used for treating leucoderma.Method and process of the present invention is simple, and production cost is low, stable effective ingredients, is easy to quality control.The present invention provides new medicinal part and compound for therapy of vitiligo.
Description
Technical field
The present invention relates to a kind of preparation method of vernonia anthelmintica phenolic acid part and the purposes of leucoderma medicament, belong to
National medicine technical field.
Background technique
Leucoderma (vitiligo, OMIM:#193200) is a kind of common posteriority depigmentation disease, main special
Sign is depigmentation caused by skin and hair melanocyte selective destruction.Leucoderma has larger between different geographical and race
Difference, the deeper crowd of the colour of skin more easily ill, the illness rate of China's leucoderma is 0.09%-2.7%.
The damage of leucoderma can betide the skin, mucous membrane and hair at any position of whole body, but exposure portion, and easily friction
Position is easier to occur.Skin lesion is symmetric more, or is distributed along nerve segment.The complete depigmentation spot of pigment is milky, boundary
Clear, size and form is different, and the hair for being grown on hickie area can also bleach, and serious person's hickie can general hair whole body.Leucoderma tissue
Pathological characters are the reduction or disappearance of skin lesion area melanocyte, and melanin granule missing, can find in early days on skin lesion periphery in cell
Inflammatory cell infiltration based on lymphocyte.
Leucoderma pathogenesis is unknown, and presently, there are a variety of pathogenesis hypothesis, but any hypothesis has its limitation,
And different type leucoderma pathogenesis may be different.The cause of disease and pathogenesis of leucoderma mainly have autoimmunity hypothesis, lose
Pass hypothesis, melanocyte autoclasia hypothesis etc..
Vernonia anthelmintica, Vernonia anthelmintica (L.) Willd, alias India mountain fennel, is composite family
(Asteraceae) the annual tall and big draft of Vernonia (Vernonia), medicinal part are mellow fruit, and autumn harvesting is dried in the air
It is dry.Its property three-level is xeothermic, and it is successive dynasties tradition Uygur that effect, which mainly removes abnormal phlegm, expelling parasite, detumescence, eliminating cold to stop pain,
Cure the main classical drug for treating leucoderma.
In vernonia anthelmintica fruit containing Sesquiterpene lactones, flavonoids, include epoxy octadecenoic acid and its glyceride type,
The multiple compounds such as triterpenes and sterols and caffeoyl quinic acid class.
Vernonia anthelmintica is with a long history in civil medication, has clinical application steady in a long-term, therefore before its development and application
Scape is wide.In uighur medicine theory, vernonia anthelmintica three-level is xeothermic, and effect mainly removes abnormal phlegm, expelling parasite, disappears
Swollen, eliminating cold to stop pain.Vernonia anthelmintica seed is that the marketed products of main ingredient and preparation have insect-expelling saligna injection liquid, card power cumin honey
White bar of cream, compound Vernonia anthelmintica ball, compound kaliziranding, compound card power cumin piece, drive Buss piece etc..Flavone compound
And flavones site treatment leucoderma is it has been reported that still phenolic acid compound and phenolic acid part treatment leucoderma have not been reported.
The extract of vernonia anthelmintica, flavones position and flavone compound for treating leucoderma have more report at present
Road, but phenolic acid part and phenolic acid compound have not been reported.
Summary of the invention
Present invention aims at provide the preparation method and purposes of a kind of vernonia anthelmintica phenolic acid part.This method be by
Dry vernonia anthelmintica fruit crushes, and is extracted using organic solvent, and resin separation purification obtains vernonia anthelmintica willd extract, main
Wanting ingredient is the phenolic acid compounds such as caffeoyl quinic acid, two-caffeoyl quinic acid.Measure the quality percentage of phenolic content
Compare 20%-99%.Main active 3,4- two-caffeoyl quinic acid, bis- caffeoyl of 3,5- in vernonia anthelmintica phenolic acid part
The content of base quininic acid and 4,5- two-caffeoyl quinic acid is respectively 3-26%, 5-34% and 5-40%.It is demonstrate,proved through preliminary experiment
Bright: the vernonia anthelmintica phenolic acid part that the method obtains through the invention has melanocyte, promotion in increase hair follicle black
The effect of element synthesis, can be used for treating leucoderma.Method and process of the present invention is simple, and production cost is low, and effective component is steady
It is fixed, it is easy to quality control.The present invention provides new medicinal part and compound for therapy of vitiligo.
A kind of preparation method of vernonia anthelmintica phenolic acid part of the present invention, follows these steps to carry out:
It a. is 0-95% methanol, ethyl alcohol or aqueous solution, room temperature-with concentration after dry vernonia anthelmintica fruit being crushed
95 DEG C are extracted 1-5 times, extraction time 0.5-3 hour, are cooled to room temperature, and are filtered, and combined extract is concentrated under reduced pressure into no alcohol taste,
The solid-to-liquid ratio for obtaining alcohol extract, vernonia anthelmintica fruit and methanol or ethanol water is 1:3-30；
B. after being 0-15% methanol, ethyl alcohol or aqueous dissolution dispersion with concentration for the extract that step a is obtained, filtering,
It is separated again with resin, is first washed with water decontamination, then with concentration be 10-95% methanol or ethanol elution, collected methanol or ethyl alcohol is washed
De- liquid, reduced pressure eliminate solvent, dry to get to containing 3,5- two-caffeoyl quinic acid, 4,5-, bis- coffee acyl quinine
The vernonia anthelmintica phenolic acid part of acid and 3,4- two-caffeoyl quinic acid.
Resin described in step b be DM-130, AB-8, HPD-450, NKA-9, HPD-100, D101, HPD-400,
HPD-500, HPD-600, HPD-300, HPD-417, polyamide or octadecylsilane chemically bonded silica.
Containing phenolic acid compound in the vernonia anthelmintica phenolic acid part that the method obtains is mass percent 20%-
99%.
Main active 3,4- two-caffeoyl quinic acid, 3 in the vernonia anthelmintica phenolic acid part that the method obtains,
The content of 5- two-caffeoyl quinic acid and 4,5- two-caffeoyl quinic acid is respectively 3-26%, 5-34% and 5-40%.
Purposes of the vernonia anthelmintica phenolic acid part that the method obtains in the drug for preparing anti-curing vitiligo.
The preparation method and purposes of vernonia anthelmintica phenolic acid part of the present invention, the expelling parasite spot obtained by this method
Turtledove chrysanthemum phenolic acid part pharmacodynamic tests prove that, there is significant treatment leucoderma wind action, and in identical clinical equivalent agent
Under amount, the effect of vernonia anthelmintica phenolic acid part is better than vitiligo capsule.
Detailed description of the invention
Fig. 1 is HE of the present invention dyeing observation skin tissue morphology's variation diagram；
Fig. 2 be phenolic acid part of the present invention to the leucoderma mouse skin number of hair follicle containing melanin influence (melanin dye,
200 times) figure.
Specific embodiment
Embodiment 1: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica fruit medicinal material 10kg is crushed, the aqueous solution of 30 times of volumes, heating and refluxing extraction 3 is added
It is secondary, it 95 DEG C of temperature, the time 3 hours, is cooled to room temperature, filters, combined extract is concentrated under reduced pressure into no alcohol taste, obtains alcohol extract；
Obtained alcohol extract is dispersed to 100g/L containing crude drug with concentration for 5% ethyl alcohol, is filtered, then inhaled with AB-8 type macropore
Attached resin column first with 1 times of column volume water elution, then with 8 times of column volume concentration is 40% ethanol elution, collects ethanol eluate,
It being concentrated under reduced pressure, vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5- two-caffeoyl quinic acid and 3 at dry powder,
The vernonia anthelmintica phenolic acid part 390g of 4- two-caffeoyl quinic acid.
Embodiment 2: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica fruit medicinal material 10kg is crushed, it is 95% ethyl alcohol that 3 times of volumetric concentrations, which are added, is heated to reflux
It extracts 5 times, room temperature the time 3 hours, is cooled to room temperature, and filtering, combined extract is concentrated under reduced pressure into no alcohol taste, obtains alcohol extracting
Object；
Obtained alcohol extract is dispersed to 50g/L containing crude drug with concentration for 5% ethyl alcohol, is centrifuged (4000 revs/min), DM-
130 type large pore resin absorption columns first with 3 times of column volume water elutions, then with 1 times of column volume concentration are 95% ethanol elution, collect
Ethanol eluate is concentrated under reduced pressure, and vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5-, bis- caffeoyl at dry powder
The vernonia anthelmintica phenolic acid part 420g of base quininic acid and 3,4- two-caffeoyl quinic acid.
Embodiment 3: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica fruit medicinal material 10kg is crushed, it is 30% ethyl alcohol that 15 times of volumetric concentrations, which are added, is heated back
Stream extracts 3 times, and 75 DEG C of temperature, time 2 h is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, is obtained alcohol extracting
Object；
Obtained extract is dispersed to 80g/L containing crude drug with concentration for 10% methanol, is centrifuged (4000 revs/min),
HPD-100 type large pore resin absorption column first with 4 times of column volume water elutions, then with 6 times of column volume concentration is 50% ethanol elution,
Ethanol eluate is collected, is concentrated under reduced pressure, vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5-, bis- coffee at dry powder
The vernonia anthelmintica phenolic acid part 440g of coffee acyl group quininic acid and 3,4- two-caffeoyl quinic acid.
Embodiment 4: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica fruit medicinal material 10kg is crushed, the concentration that 18 times of volumes are added is 60% methanol, heating
Refluxing extraction 2 times, temperature 50 C the time 0.5 hour, is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, obtains
Alcohol extract；
Obtained extract is dispersed to 100g/L containing crude drug with concentration for 15% methanol, is centrifuged (4000 revs/min),
NKA-9 type large pore resin absorption column first with 5 times of column volume water elutions, then with 5 times of column volume concentration is that 60% methanol elutes, receives
Collect meoh eluate, be concentrated under reduced pressure, vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5-, bis- coffee at dry powder
The vernonia anthelmintica phenolic acid part 460g of acyl group quininic acid and 3,4- two-caffeoyl quinic acid.
Embodiment 5: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica fruit medicinal material 10kg is crushed, it is that 10% ethyl alcohol is heated to reflux that 12 times of volumetric concentrations, which are added,
It extracts 2 times, 80 DEG C of temperature, time 2 h is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, is obtained alcohol extract；
Obtained extract is dispersed to 30g/L containing crude drug with concentration for 10% ethyl alcohol, is filtered, then inhaled with D101 type macropore
Attached resin column first with 8 times of column volume water elutions, then with 6 times of column volume concentration is 80% ethanol elution, collects ethanol eluate,
It being concentrated under reduced pressure, vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5- two-caffeoyl quinic acid and 3 at dry powder,
The vernonia anthelmintica phenolic acid part 564g of 4- two-caffeoyl quinic acid.
Embodiment 6: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica fruit medicinal material 10kg is crushed, it is 95% ethyl alcohol that 18 times of volumetric concentrations, which are added, is heated back
Stream extracts 2 times, and temperature 60 C the time 1 hour, is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, obtains alcohol extracting
Object；
Obtained extract is water-dispersible to 50g/mL containing crude drug, it is centrifuged (4000 revs/min), with HPD400 type macropore
Adsorption resin column first with 6 times of column volume water elutions, then with 6 times of column volume concentration is that 65% methanol elutes, collects methanol elution
Liquid is concentrated under reduced pressure, and vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5- two-caffeoyl quinic acid at dry powder
With the vernonia anthelmintica phenolic acid part 490g of 3,4- two-caffeoyl quinic acid.
Embodiment 7: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica medicine fruit material 10kg is crushed, it is 95% methanol that 30 times of volumetric concentrations, which are added, is heated back
Stream extracts 2 times, and 65 DEG C of temperature, time 2 h is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, is obtained alcohol extracting
Object；
Obtained extract is dispersed to 60g/mL containing crude drug with concentration for 10% ethyl alcohol, is centrifuged (4000 revs/min),
HPD600 type large pore resin absorption column first with 3 times of column volume water elutions, then with 8 times of column volume concentration is 95% ethanol elution, receives
Collect ethanol eluate, be concentrated under reduced pressure, vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5-, bis- coffee at dry powder
The vernonia anthelmintica phenolic acid part 524g of acyl group quininic acid and 3,4- two-caffeoyl quinic acid.
Embodiment 8: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica medicine fruit material 10kg is crushed, it is 95% methanol that 15 times of volumetric concentrations, which are added, is heated back
Stream extracts 2 times, and 55 DEG C of temperature, time 2 h is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, is obtained alcohol extracting
Object；
Obtained extract is dispersed to 80g/mL containing crude drug with concentration for 10% ethyl alcohol, is centrifuged (4000 revs/min),
HPD500 type large pore resin absorption column first with 3 times of column volume water elutions, then with 6 times of column volume concentration is 75% ethanol elution, receives
Collect ethanol eluate, be concentrated under reduced pressure, vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5-, bis- coffee at dry powder
The vernonia anthelmintica phenolic acid part 498g of acyl group quininic acid and 3,4- two-caffeoyl quinic acid.
Embodiment 9: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica medicine fruit material 10kg is crushed, it is 75% methanol that 20 times of volumetric concentrations, which are added, is heated back
Stream extracts 2 times, and temperature 60 C, time 2 h is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, is obtained alcohol extracting
Object；
Obtained extract is dispersed to 60g/mL containing crude drug with concentration for 15% ethyl alcohol, is centrifuged (4000 revs/min),
HPD450 type large pore resin absorption column first with 1 times of column volume water elution, then with 4 times of column volume concentration is 75% ethanol elution, receives
Collect ethanol eluate, be concentrated under reduced pressure, vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5-, bis- coffee at dry powder
The vernonia anthelmintica phenolic acid part 433g of acyl group quininic acid and 3,4- two-caffeoyl quinic acid.
Embodiment 10: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica medicine fruit material 10kg is crushed, it is 55% methanol that 30 times of volumetric concentrations, which are added, is heated back
Stream extracts 2 times, and temperature 70 C the time 3 hours, is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, obtains alcohol extracting
Object；
Obtained extract is dispersed to 100g/mL containing crude drug with concentration for 10% ethyl alcohol, is centrifuged (4000 revs/min),
HPD417 type large pore resin absorption column first with 1 times of column volume water elution, then with 8 times of column volume concentration is 85% ethanol elution, receives
Collect ethanol eluate, be concentrated under reduced pressure, vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5-, bis- coffee at dry powder
The vernonia anthelmintica phenolic acid part 603g of acyl group quininic acid and 3,4- two-caffeoyl quinic acid.
Embodiment 11: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica medicine fruit material 10kg is crushed, it is 45% methanol that 30 times of volumetric concentrations, which are added, is heated back
Stream extracts 2 times, 55 DEG C of temperature, the time 1 hour, is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, obtains alcohol extracting
Object；
Obtained extract is dispersed to 90g/mL containing crude drug with concentration for 15% ethyl alcohol, is centrifuged (4000 revs/min),
HPD300 type large pore resin absorption column first with 2 times of column volume water elutions, then with 6 times of column volume concentration is 65% ethanol elution, receives
Collect ethanol eluate, be concentrated under reduced pressure, vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5-, bis- coffee at dry powder
The vernonia anthelmintica phenolic acid part 491g of acyl group quininic acid and 3,4- two-caffeoyl quinic acid.
Embodiment 12: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica medicine fruit material 10kg is crushed, it is 95% methanol that 30 times of volumetric concentrations, which are added, is heated back
Stream extracts 2 times, and temperature 45 C, time 2 h is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, is obtained alcohol extracting
Object；
Obtained extract is dispersed to 90g/mL containing crude drug with concentration for 10% ethyl alcohol, is centrifuged (4000 revs/min), gathers
Amide resin column first with 1 times of column volume water elution, then with 8 times of column volume concentration is 85% ethanol elution, collects ethanol elution
Liquid is concentrated under reduced pressure, and vacuum drying contains 3,5- two-caffeoyl quinic acid, 4,5- two-caffeoyl quinic acid at dry powder
With the vernonia anthelmintica phenolic acid part 586g of 3,4- two-caffeoyl quinic acid.
Embodiment 13: the preparation of vernonia anthelmintica phenolic acid part:
Dry vernonia anthelmintica medicine fruit material 10kg is crushed, it is 55% methanol that 30 times of volumetric concentrations, which are added, is heated back
Stream extracts 2 times, and 65 DEG C of temperature, time 2 h is cooled to room temperature, combined extract, is concentrated under reduced pressure into no alcohol taste, is obtained alcohol extracting
Object；
It using octadecylsilane chemically bonded silica, is purified repeatedly by preparing liquid phase, obtaining purity respectively is 99%
3,4- two-caffeoyl quinic acid, 3,5- two-caffeoyl quinic acid, 4,5- two-caffeoyl quinic acid；
2.6 milligrams of 3,4- two-caffeoyl quinic acid, 3.4 milligrams of 3,5- two-caffeoyl quinic acid and 4 are weighed respectively
Milligram 4,5- two-caffeoyl quinic acid is uniformly mixed to get the vernonia anthelmintica phenolic acid part for being 99% to total phenol acid content.
Embodiment 14: the constituent analysis of vernonia anthelmintica phenolic acid part:
Assay is carried out to obtained vernonia anthelmintica phenolic acid part with spectrophotometry, the results showed that expelling parasite spot
Total phenol acid content mass percent is 20%-99% in turtledove chrysanthemum phenolic acid part.Shown using liquid-phase chromatographic analysis: in expelling parasite ringdove
3,4- two-caffeoyl quinic acid, bis- coffee acyl Kui of 3,5- two-caffeoyl quinic acid and 4,5- in chrysanthemum medicine fruit material raw material
Peaceful acid content mass percent is respectively 0.3-3%, 0.5-3%, 0.6-3%, the expelling parasite that the method obtains through the invention
Ringdove chrysanthemum phenolic acid part 3,4- two-caffeoyl quinic acid, bis- coffee acyl quinine of 3,5- two-caffeoyl quinic acid and 4,5-
Content mass percent is respectively 3-26%, 5-34%, 5-40% in acid, the results showed that the method for the invention can be mentioned significantly
High 3,4- two-caffeoyl quinic acid, 3,5- two-caffeoyl quinic acid and 4,5- two-caffeoyl quinic acid content.
Embodiment 15: vernonia anthelmintica phenolic acid part urgency toxicity assessment:
20 cleaning grade kunming mouses, 5 week old, weight 20g, equilibrium are randomly divided into 2 groups (administration group and control groups),
Every group 10, half male and half female.After being deprived of food but not water 16h, administration group intragastric administration on mice gives the phenolic acid part of concentration 0.6g/mL, gives
Medicine volume be 40mL/kg, gastric infusion 1 time.Naive mice stomach-filling equal amount of distilled water.It is observed continuously in 4h after administration, after
Every morning is observed 1 time, is observed continuously 14 days.Mouse weight is measured during experiment daily, administration group and control group weight are administered
The equal no difference of science of statistics in front and back, the results are shown in Table 1；Appearance, diet, behavioral activity, secretion, excreta of animal etc. are observed simultaneously
Without significant abnormal；Dead mouse is had no during test；After the test, mouse distinguishes sacrificed by decapitation, dissection, each internal organs color
Normally, no abnormality seen is organized, the results are shown in Table 2；Calculate the maximum dosage-feeding (MLD) of phenolic acid part are as follows: 24g/kg/d (is equivalent to life
Dose 564.7g crude drug/kg), it is 6776.67 times of clinical adult dosage.Illustrate that phenolic acid part is substantially non-toxic, clinic is used
The safety of medicine is big；
1 phenolic acid part acute toxicity test mouse changes of body mass of table (g,N=10)
2 phenolic acid part of table is to acute toxicity test in mice result
Embodiment 16: effect of the vernonia anthelmintica phenolic acid part to leucoderma model mice:
C57BL/6 mouse 70, weight 18-22g, male and female half-and-half, shave off back hair with shaver, area is about
2cm × 2cm, equilibrium are randomly divided into 6 groups: blank control group, model group, positive drug (vitiligo capsule) control group, extract is low,
Middle and high dosage group, every group 12.The quinhydrones of model group and each administration group in hair removal section smearing 5%, each 0.5mL, 2 times a day,
Blank control group smears equal amount of distilled water in hair removal section；Continuous 50 days, vitiligo mice model is established, during this period, each group is small
Mouse loses hair or feathers 1 time for average every 3 days in tested skin region；After quinhydrones modeling 20 days, there is decoloration phenomenon in mouse skin and hair；From
Start within modeling the 21st day, each group after smearing 5% quinhydrones 1 hour, by setting dosage, starts gastric infusion: blank control respectively
Isometric distilled water is given in group, model group stomach-filling, and test medicine is given in administration group stomach-filling, 1 time a day, successive administration 30 days, real
After testing, mouse is put to death.
1. visually observing the curative effect of drug therapy leucoderma:
Experimental result is shown, during modeling, the visible tested area's skin color of each group modeling mouse of naked eyes is pale, and hair is raw
It is long slowly to extend with the modeling time, there is hickie, new piliation becomes white,；After treating, naked eyes are as it can be seen that vitiligo capsule
Group and each dosage group of phenolic acid part can be different degrees of the tested area's skin color of recovery vitiligo mice, new piliation also shows extensive
It is again black.Under identical clinical equivalent dosage, phenolic acid part large dosage group is better than vitiligo capsule group, the results are shown in Table 3；
Observation of curative effect of 3 phenolic acid part of table to vitiligo mice caused by quinhydrones
Note: compared with model group,*P<0.05。
2. histological observation
The variation of 2.1 HE dyeing observation skin tissue morphology:
Blank control group: mouse skin is methodically arranged, and hair follicle structure is complete, and number is more, and pigment content is more and color is deep；
Leucoderma model group: mouse skin is obviously thinning, and cuticula slightly thickens, the visible oedema of stratum spinosum epidermidis, hair follicle number
Amount is few, and structure is imperfect, sebaceous glands disintegration, hair shaft depigmentation.Subcutaneous tissue has no that hair follicle is distributed；
Vitiligo capsule group: mouse skin caliper recovery is normal, and keratoderma has no and obviously thickens, epidermis and corium without
Oedema, basal layer cell have pigmentation, and hair follicle increases, structural integrity, and pigment content increases, and color is deepened；
Phenolic acid part low dose group: mouse skin is obviously thinning, and part cuticula thickens, epidermal cell Mild edema, spine
The visible oedema of cellular layer, skin corium and the visible a small amount of hair follicle distribution of subcutaneous tissue；
Phenolic acid part middle dose group: mouse skin caliper recovery is normal, and epidermis and corium have no obvious oedema, and basal layer is thin
Born of the same parents have pigmentation, and hair follicle structure is complete, and number and pigment content obviously increase, skin corium and the visible hair follicle point of subcutaneous tissue
Cloth；
Phenolic acid part high dose group: mouse skin caliper recovery is normal, and keratoderma is thinning, and epidermis and corium are without obvious
Oedema, basal layer cell have pigmentation, and hair follicle structure is complete, and number obviously increases, and pigmentary colours are obviously deepened, skin corium and
The visible hair follicle distribution of subcutaneous tissue, is shown in Fig. 1.
2.2 melanin dyeing observation dermal melanin distribution situation:
Model group mouse, compared with blank control group, not only hair follicle is few, and most of hair follicle is practically free of black
Element；Each administration group mouse, compared with model group, hair follicle increases, and wherein the middle and high dosage group melanin of phenolic acid part obviously increases
Add.Under identical clinical equivalent dosage, the effect of vernonia anthelmintica phenolic acid part is better than vitiligo capsule, the results are shown in Table 4, Fig. 2；
4 phenolic acid part of table causes the influence of vitiligo mice skin hair follicle containing melanin number to quinhydrones
Note: compared with blank control group,#P<0.05；Compared with model group,*P<0.05。
3. mice serum and skin histology tyrosinase assay:
The experimental results showed that model group mouse, compared with blank control group, the content of TYR is dramatically increased in serum, skin
The content of TYR significantly reduces in tissue；Each dosage group of phenolic acid part can obviously reduce the content (P of TYR in vitiligo mice serum
< 0.05), and increase content (P < 0.05) effect of TYR in vitiligo mice skin histology better than vitiligo capsule, as a result see
Table 5；
Influence of 5 phenolic acid part of table to vitiligo mice serum and skin TYR caused by quinhydrones
Note: compared with blank control group,#P<0.05；Compared with model group,*P<0.05。
After C57 mouse quinhydrones modeling, there is hickie in model group mouse skin；The visible spongiosis of histological observation,
Hair shaft depigmentation, hair follicle containing melanin are reduced；Hair follicle structure is imperfect in skin histology, sebaceous glands disintegration；TYR contains in serum
Amount dramatically increases, and TYR content significantly reduces in skin histology, shows quinhydrones damage model group mouse hair follicles institutional framework and hair
Melanocyte in capsule.
Model group is compared, phenolic acid part effect group can improve influence of the quinhydrones modeling to C57 mouse, restore leucoderma model
The tested area's skin color of mouse；Histological observation restores normal without obvious oedema, hair shaft depigmentation, the quantity of hair follicle containing melanin
Range；Hair follicle structure is complete in skin histology, and number obviously increases；TYR content restores normal range (NR) in serum, in skin histology
The content of TYR dramatically increases.Phenolic acid part can increase melanogenesis protein content in skin histology, increase the number of hair follicle containing melanin
Amount preferably restores the tested area's skin color of mouse.
In conclusion vernonia anthelmintica phenolic acid part can increase melanocyte quantity in vitiligo mice hair follicle, promote
Melanin genesis can be used for preparing the purposes in the drug for the treatment of leucoderma.
Claims (1)
1. a kind of purposes of vernonia anthelmintica phenolic acid part in the drug for preparing anti-curing vitiligo, it is characterised in that press following step
It is rapid to carry out:
It a. is 0-95% methanol, ethyl alcohol or aqueous solution with concentration after dry vernonia anthelmintica fruit being crushed, -95 DEG C of room temperature
It extracts 1-5 times, extraction time 0.5-3 hour, is cooled to room temperature, filter, combined extract is concentrated under reduced pressure into no alcohol taste, obtains
The solid-to-liquid ratio of alcohol extract, vernonia anthelmintica fruit and methanol or ethanol water is 1: 3-30；
B. after being 0-15% methanol, ethyl alcohol or aqueous dissolution dispersion with concentration for the extract that step a is obtained, filtering, then use
Resin be DM-130, AB-8, HPD-450, NKA-9, HPD-100, D101, HPD-400, HPD-500, HPD-600, HPD-300,
HPD-417, polyamide or octadecylsilane chemically bonded silica separation, are first washed with water decontamination, then with concentration be 10-95% methanol
Or ethanol elution, collect methanol or ethanol eluate, reduced pressure eliminates solvent, dry to get to containing 3,5-, bis- caffeoyl
The expelling parasite spot of base quininic acid 5-34%, 4,5- two-caffeoyl quinic acid 5-40%, 3,4- two-caffeoyl quinic acid 3-26%
Turtledove chrysanthemum phenolic acid part containing phenolic acid compound is mass percent 20%-99% in the vernonia anthelmintica phenolic acid part.
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| CN102526153A (en) * | 2012-02-27 | 2012-07-04 | 新疆维吾尔自治区维吾尔医药研究所 | Vernonia anthelmintica flavone components, preparation method and application thereof |
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