CN104546992A - Folium cortex eucommiae extract as well as preparation method and application thereof - Google Patents
Folium cortex eucommiae extract as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a folium cortex eucommiae extract which is a water extract or an ethanol extract of folium cortex eucommiae. The folium cortex eucommiae extract contains no less than 9% of chlorogenic acid, no less than 1% of geniposidic acid and no less than 1% of aucubin. Furthermore, the invention also provides a preparation method and an application of the folium cortex eucommiae extract. The folium cortex eucommiae extract, as a traditional Chinese medicine extract, is low in side effect, capable of being used for a long term, free from toxic and side effects to organisms, safe and effective, capable of improving the life quality of patients and having a significant curative effect on psoriasis; and therefore, the folium cortex eucommiae extract has a broad application prospect in treating psoriasis.
Description
Technical field
The present invention relates to Folium Eucommiae extract and its production and use.
Background technology
Psoriasis is commonly called as " psoriasis ", is that a kind of chronic inflammatory skin of common also easily recurrence is sick, with micaceous scales, red film disease, petechial hemorrhage for Clinical symptoms, is about 0.123% at the sickness rate of China.In recent years, onset of psoriasis is in rising trend, and there is obvious seasonality at its initial stage, and in the winter, the heavy summer is light, and no matter men and women, old and young all can fall ill, and the current cause of disease is not bright, still lacks effective Therapeutic Method.
Psoriasis is divided into 4 types at present, and psoriasis vulgaris, arthropathic psoriasis, erythrodermic psoriasis and psoriasis pustulosa, wherein psoriasis vulgaris accounts for more than 99%.Psoriasis vulgaris is plain edition psoriasis, and other three types are called special type psoriasis.According to successive dynasties famous doctor of Chinese Medicine summary of experience, psoriasis vulgaris is generally divided into 4 types: i.e. blood-heat type (blood heat wind-dryness type), blood-deficiency type (blood-deficiency and wind-dry type), blood-dryness type, blood stasis type (the stagnant skin type of the stasis of blood); In special type psoriasis, pustule type is generally dialectical is septicopyemia type (damp and hot accumulate malicious type), and erythrodermic is scorchingly hot type (heat in blood poison Sheng type), and joint type is cold-dampness type or wind-damp numbness type.
The main clinical manifestation of psoriasis vulgaris is: white unicorn bits, shinny thin film and petechial hemorrhage.Main pathology shows as: parakeratosis, and in angleplied laminate or under angleplied laminate, the microabscess (Munro microabscess) be made up of neutrophilic granulocyte as seen, granular layer is thinning or disappear.Acanthosis, trochanterellus extends.The buckling of vessel expansion of dermal papilla portion, tube wall slightly thickens.The light to moderate inflammatory cell infiltration of upper part of dermis.Psoriasis vulgaris is comparatively common, accounts for more than 90% greatly.
Psoriasis pustulosa is the type that in psoriasis, the state of an illness is comparatively serious, point general property psoriasis and Local Psoriasis.Clinically with the aseptic pustule of foxtail millet grain size on general rubescent speckle basis, the whole body for feature, often raise and hypoproteinemia with Gao Re and leukocyte, even can threat to life.This type is more rare clinically, accounts for psoriasis people's 0.77%.
Psoriatic tissue pathology is that epidermal keratinocyte can not full maturity, is full of air between micelle, and refracted ray becomes seen silvery white micaceous scales clinically; Intradermal papilledema swells and charges into epidermis close to horny layer, scratches clinically to scrape squama and can damage nipple blood vessel and occur petechial hemorrhage.Doctor trained in Western medicine is thought, psoriatic generation is main relevant with autoimmune, metabolism and endocrine dysfunction, and climate change, anxious state of mind, infection, wound etc. are risk factor.
Psoriatic pathogenesis is complicated, illustrated not yet completely at present, its pathogenesis roughly may be summarized to be: on the basis of certain genetic background, multiple virulence factor stimulation body immune system causes the immune system dysfunction based on T cell, inflammatory cell migrates to epidermis and at local infiltration, cause local inflammation and keratinocyte paraplasm, finally cause the generation that psoriasis pathology changes.
Psoriasis is a kind of common chronic inflammatory skin disease being formed as main pathological change with keratinocyte hyperplasia, inflammatory cell infiltration and new vessels, is dermatology common disease, frequently-occurring disease.Primary disease etiology unknown, treatment difficulty, the method that there is no is eradicated completely, current Western medicine many uses tretinoin and this disease of immunosuppressant treatment, and it has advantage in control psoriasis activity, but have that untoward reaction is many, the problem such as relapse rate height after drug withdrawal.
Folium Eucommiae is the dried leaves of Eucommiaceae plant Cortex Eucommiae Eucommia ulmoides Oliv., micro-pungent, temperature.Return liver, kidney channel.Invigorating the liver and kidney, bone and muscle strengthening.For deficiency of the liver and kindey, have a dizzy spell, soreness of waist and knee joint, muscles and bones flaccidity is soft.Have no Folium Eucommiae extract and treat psoriatic report.
Summary of the invention
Technical scheme of the present invention there is provided a kind of new Folium Eucommiae extract and its production and use.
A kind of Folium Eucommiae extract, it is water or the ethanol extraction of Folium Eucommiae, and in described Folium Eucommiae extract, chlorogenic acid content is not less than 9%, and Geniposidic acid is not less than 1%, and aucubin is not less than 1%.
Preferably, in described Folium Eucommiae extract, chlorogenic acid content is 9.38% ~ 39.11%, and geniposide acid content is 1.03% ~ 10.33%, and aucubin is 1.17% ~ 10.68%.
Preferred further, in described Folium Eucommiae extract, chlorogenic acid content is 10.21% ~ 32.37%, and geniposide acid content is 1.03% ~ 9.38%, and aucubin is 1.17% ~ 9.56%.
Preferred further again, in described Folium Eucommiae extract, chlorogenic acid content is 10.21% or 32.37%, and geniposide acid content is 1.03% or 2.24%, and aucubin is 1.24% or 2.07%.
Wherein, described Folium Eucommiae extract is prepared by the following method:
Get Folium Eucommiae, with water or 30-80% ethanol extraction 1 ~ 2 time, each 1 ~ 2 hour, by extracting solution or its concentrated solution after phenylethylene absorption with macroporous adsorbent resin, use 10-90% ethanol gradient elution, collect eluent, merge, concentrate drying, to obtain final product.
Preferably, described Folium Eucommiae extract is prepared by the following method:
Get Folium Eucommiae, with 60% ethanol in 70 DEG C of lixiviates twice, at every turn with 15 times amount volumes, each 1 hour, merge extractive liquid, concentration and recovery ethanol, concentrated solution, with after D-101 macroporous adsorbent resin resin saturated adsorption, uses 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collects eluent, merge 10% and 30% eluent, or merge 70% and 90% eluent, concentrate drying, to obtain final product;
Or, get Folium Eucommiae, with 30% ethanol in 60 DEG C of lixiviates twice, each 2 hours, each use 12 times amount volume, merge extractive liquid, concentration and recovery ethanol, concentrated solution is with after HPD-100 macroporous adsorbent resin resin saturated adsorption, use 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collect eluent, merge 70% and 90% eluent, or get 30% eluent, concentrate drying, to obtain final product;
Or, get Folium Eucommiae, with 20 times of water in 80 DEG C of lixiviates once, each 1.5 hours, extracting solution, with after XAD-16 macroporous adsorbent resin resin saturated adsorption, used 20% and 60% ethanol gradient concentration ethanol eluting successively, collect eluent and 60% eluent of 20% respectively, concentrate drying, to obtain final product;
Or, get Folium Eucommiae, with 12 times of water in 60 DEG C of lixiviates twice, each 2 hours, extracting solution, with after AB-8 macroporous adsorbent resin resin saturated adsorption, used 10%, 90% ethanol gradient concentration ethanol eluting successively, collect eluent, collect 10% eluent and 90% eluent respectively, concentrate drying, to obtain final product.
The present invention prepares the method for aforementioned Folium Eucommiae extract, it comprises the steps: to get Folium Eucommiae, with water or 30-80% ethanol extraction 1 ~ 2 time, by extracting solution or its concentrated solution after phenylethylene absorption with macroporous adsorbent resin, use 10-90% ethanol gradient elution, collect eluent, merge, concentrate drying, to obtain final product.
Preferably, described Folium Eucommiae extract preparation method is as follows:
Get Folium Eucommiae, with 60% ethanol in 70 DEG C of lixiviates twice, at every turn with 15 times amount volumes, each 1 hour, merge extractive liquid, concentration and recovery ethanol, concentrated solution, with after D-101 macroporous adsorbent resin resin saturated adsorption, uses 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collects eluent, merge 10% and 30% eluent, or merge 70% and 90% eluent, concentrate drying;
Or, get Folium Eucommiae, with 30% ethanol in 60 DEG C of lixiviates twice, each 2 hours, each use 12 times amount volume, merge extractive liquid, concentration and recovery ethanol, concentrated solution is with after HPD-100 macroporous adsorbent resin resin saturated adsorption, use 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collect eluent, merge 70% and 90% eluent, or get 30% eluent, concentrate drying;
Or, get Folium Eucommiae, with 20 times of water in 80 DEG C of lixiviates once, each 1.5 hours, extracting solution, with after XAD-16 macroporous adsorbent resin resin saturated adsorption, used 20% and 60% ethanol gradient concentration ethanol eluting successively, collect eluent and 60% eluent of 20% respectively, concentrate drying;
Or, get Folium Eucommiae, with 12 times of water in 60 DEG C of lixiviates twice, each 2 hours, extracting solution, with after AB-8 macroporous adsorbent resin resin saturated adsorption, used 10%, 90% ethanol gradient concentration ethanol eluting successively, collect eluent, collect 10% eluent and 90% eluent respectively, concentrate drying.
Present invention also offers the purposes of aforesaid Folium Eucommiae extract in the psoriatic medicine of preparation treatment.
Wherein, described medicine is the medicine for the treatment of psoriasis vulgaris or psoriasis pustulosa.
Present invention also offers one and treat psoriatic medicine, it be with aforesaid Folium Eucommiae extract for active component, add the preparation that pharmaceutically acceptable adjuvant or complementary composition are prepared from.
Wherein, described preparation is oral formulations, external preparation.
Wherein, described oral formulations is tablet, granule, powder, pill, oral liquid, capsule.
Psoriasis be a kind ofly to be determined by polygenic inheritance, dysimmune chronic inflammation proliferative skin disorders that multi-environment factor stimulates induction, worldwide general population prevalence is 0.1%-3%, China's prevalence is 0.123%, and annual morbidity is 0.1%.Although this disease does not jeopardize the life of people, but its course of disease is grown and obstinate refractory, can be there is crippling psoriasis arthropathica in severe patient, erythrodermic silver shoulder is sick, even involves the organs such as eye, liver, kidney, cardiovascular, lung, thus have a strong impact on the quality of life of patient.Some patient can occur that mental maladjustment is as ashamed, nervous, anxiety, melancholy etc., is reluctant to participate in doings, affects daily life, brings great misery to patient body and psychology.Due to this sick recurrent exerbation, often repeatedly need treat, bring heavy financial burden thus to patient and family thereof.Current psoriasis cannot be cured, and traditional treatment mostly is immunosuppressant and tretinoin medicines except Chinese medicine, and these medicines of prolonged application easily produce the serious side effect such as dependency and hepatorenal damage.
Folium Eucommiae extract is Chinese medicine extract, and side effect is little, can life-time service, does not produce toxic and side effects to body, safe and effective, improves the quality of life of patient, has significant curative effect to psoriasis, therefore Folium Eucommiae extract treatment psoriasis has broad application prospects.
Obviously, according to foregoing of the present invention, according to ordinary technical knowledge and the customary means of this area, not departing under the present invention's above-mentioned basic fundamental thought prerequisite, the amendment of other various ways, replacement or change can also be made.
The detailed description of the invention of form by the following examples, is described in further detail foregoing of the present invention again.But this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example.All technology realized based on foregoing of the present invention all belong to scope of the present invention.
Detailed description of the invention
The preparation of embodiment 1 Folium Eucommiae extract of the present invention
Folium Eucommiae with 60% ethanol in 70 DEG C of lixiviates twice, each use 15 times amount volume, each 1 hour, merge extractive liquid, concentration and recovery ethanol, concentrated solution is with after D-101 macroporous adsorbent resin resin saturated adsorption, use 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collect eluent, merge 10% and 30% eluent, concentrate drying obtains extract sample 1 (content: chlorogenic acid=11.27%, Geniposidic acid=1.87%, aucubin=1.17%); Merge 70% and 90% eluent, concentrate drying obtains extract sample 2 (content: chlorogenic acid=32.37%, Geniposidic acid=2.24%, aucubin=2.07%)
The preparation of embodiment 2 Folium Eucommiae extract of the present invention
Folium Eucommiae with 30% ethanol in 60 DEG C of lixiviates twice, each use 12 times amount volume, each 2 hours, merge extractive liquid, concentration and recovery ethanol, concentrated solution is with after HPD-100 macroporous adsorbent resin resin saturated adsorption, use 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collect eluent, merge 70% and 90% eluent, concentrate drying obtains extract sample 1 (content: chlorogenic acid=11.24%, Geniposidic acid=9.38%, aucubin=9.56%); 30% eluent concentrate drying obtains extract sample 2 (content: chlorogenic acid=38.55%, Geniposidic acid=1.18%, aucubin=1.36%)
The preparation of embodiment 3 Folium Eucommiae extract of the present invention
By Folium Eucommiae with 20 times of water in 80 DEG C of lixiviates once, each 1.5 hours, extracting solution is with after XAD-16 macroporous adsorbent resin resin saturated adsorption, use 20% and 60% ethanol gradient concentration ethanol eluting successively, collect eluent, 20% eluent concentrate drying obtains extract sample 1 (content: chlorogenic acid=23.04%, Geniposidic acid=1.29%, aucubin=3.54%); 60% eluent concentrate drying obtains extract sample 2 (content: chlorogenic acid=10.21%, Geniposidic acid=1.03%, aucubin=1.24%)
The preparation of embodiment 4 Folium Eucommiae extract of the present invention
By Folium Eucommiae with 12 times of water in 60 DEG C of lixiviates twice, each 2 hours, extracting solution is with after AB-8 macroporous adsorbent resin resin saturated adsorption, use 10%, 90% ethanol gradient concentration ethanol eluting successively, collect eluent, 10% eluent concentrate drying obtains extract sample 1 (content: chlorogenic acid=39.11%, Geniposidic acid=10.33%, aucubin=10.68%); 90% eluent concentrate drying obtains extract sample 2 (content: chlorogenic acid=9.38%, Geniposidic acid=1.11%, aucubin=2.31%)
The preparation (tablet) of embodiment 5 medicine of the present invention
Prescription
Take the extract sample 2 of embodiment 1 preparation by recipe quantity, add starch, hyprolose sieves mixing, add polyvidone ethanol wet granulation, dry, granulate, adds Pulvis Talci, tabletting.
The preparation (granule) of embodiment 6 medicine of the present invention
Prescription
Take the extract sample 1 of embodiment 1 preparation by prescription, add starch, hyprolose sieves mixing, ethanol wet granulation, dry, granulate, adds magnesium stearate, pack.
The preparation (powder) of embodiment 7 medicine of the present invention
Prescription
Take extract sample 1 prepared by embodiment 2 by prescription, add mannitol and to sieve mixing, pack.
The preparation (pill) of embodiment 8 medicine of the present invention
Prescription
Take the extract sample 2 of embodiment 2 preparation by prescription, add starch, hyprolose sieves mixing, add carboxymethylstach sodium water liquid pill, dry, granulate, adds magnesium stearate, pack.
The preparation (oral agents) of embodiment 9 medicine of the present invention
Take the extract sample 1 of embodiment 3 preparation by prescription, adjust volume with water for injection, fill.
The preparation (capsule) of embodiment 10 medicine of the present invention
Prescription
Take the extract sample 2 of embodiment 3 preparation by prescription, add starch slurry and granulate, dry, granulate, loads capsule.
Beneficial effect of the present invention is further illustrated below with effect experiment:
Test example 1 Folium Eucommiae extract of the present invention treats the psoriatic test of pesticide effectiveness (psoriasis vulgaris model)
1. material
1) animal
6-8 week age SCID mice 50, weight 18-22g, Sichuan University's experimental animal center provides.Animal rank: one-level, credit number: No. 10.
2) medicine
Medicine test sample 1 (embodiment 1 extract sample 1);
Medicine test sample 2 (embodiment 1 extract sample 2);
Medicine test sample 3 (embodiment 2 extract sample 1);
Acitretin Capsules, purchased from Huabang Pharmaceutical Co., Ltd., Chongqing, lot number: 2013010
2. experimental technique
1) experiment grouping
Healthy guinea pig, adopts random digits table to be divided into 5 groups at random: (1) model control group; (2) positive controls; (3) medicine test sample 1 group; (4) medicine test sample 2 groups; (5) medicine test sample 3 groups.
2) animal model preparation
By SCID mice isolating female from male culture, raise 1 week, mice all can Adaptable growth environment.Choose homeliness type or psoriasis in plaques patient (signing Informed Consent Form).Every patient chooses cutis laxa position and comprises 2cm × 2cm psoriatic lesion and do fusiform incision, cuts skin holostrome, is trimmed to 0.6cm × 0.6cm skin graft of suitable depth, with the aseptic packaging of normal saline gauze parcel in order to transplanting.3% chloral hydrate anesthesia (0.1ml/10g) is given according to SCID Mouse Weight, preserved skin unhairing, cut off epidermis and be separated to fascia layer, by the skin graft 3-0 silk suture pruned, and with being soaked with the cotton balls pressure dressing of normal saline, mice sub-cage rearing after skin grafts, whole process completed in 2 hours.At the PBMC of skin lesions circumferece injection through the graft supplier of ConA process after 10 days, cell number is 1 × 10
6/ only.Remove outside wrapping after 2 weeks, observe skin graft and survive situation, after determining to cause merit, carry out next step experiment.
3) administration
Positive drug control group gives Acitretin Capsules suspensoid 2.25mg/kg, and gavage amount is each 1mL/100g, 2 times/d; Medicine test sample 1 group, test sample 2 groups and test sample 3 groups all by 1mL/100g gastric infusion, dosage 100mg/kg, 1 time/d; Model control group normal saline every day gavage.Continue medication 10 days.
4) detect
After medication terminates, cervical dislocation puts to death mice, and get its back and transplant skin lesion, 10% formaldehyde (formalin solution) fixes 24h.Skin lesion is trimmed to suitable size, dehydration, embedding, section, HE dyes.Optical microphotograph Microscopic observation is respectively organized and is transplanted the change such as skin lesion epidermis and intradermal inflammatory cell, and at 200 times of visual field inside counting epidermis cell numbers of plies and epidermal thickness, counting high dermis blood vessel peripheral lymphocyte and mononuclear cell number, calculate Backer scoring, respectively organize diversity between data.
Above-mentioned Cavia porcellus gets blood before not putting to death, and centrifugal 2000r/rain, gets serum, adopts enzyme linked immunological kit, carries out detection optical density by the requirement of test kit description, according to standard curve result of calculation, calculates cAMP/cGMP ratio according to result.
5) statistical method
All data all use the process of SPSS 13.0 statistical software, and experimental result measurement data is used
representing, is that difference has statistical significance with P<0.05.Multisample mean compares employing variance analysis.
6) Backer scoring
Laboratory animal improvement Ba ke r point system method is as follows: horny layer parakeratosis 0.5-1.5 divides (light-heavy), hyperkeratosis 1.5 points, it is that bar-shaped 0.5-1.5 divides (light-heavy) that trochanterellus extends, acanthosis 1.0 points, granular layer is thinning or disappear 1.0 points, skin corium inflammatory cell infiltration 0.5-1.5 divides (light-heavy), nipple is stretched pestle shape and becomes 1.0 points, telangiectasis 1.0 points
3. result
(1) each group Baker scoring, Streptococcus counting, spine cell's number
Each group of light microscopic changes as follows: positive controls, moderate parakeratosis, the about thick 24-32 layer of spinous layer, epidermis is moderate wooden club sample hypertrophy, partial visible point shape granular layer and hyperkeratosis, a small amount of mononuclear cell infiltration around intradermal blood vessel.Medicine test sample 1 group: slight parakeratosis, the about thick 21-29 layer of spinous layer, epidermis is slight wooden club sample hypertrophy, substantially without granular layer, a small amount of mononuclear cell infiltration around intradermal blood vessel.Medicine test sample 2 groups: slight parakeratosis, the about thick 20-28 layer of spinous layer, epidermis is slight wooden club sample hypertrophy, substantially without granular layer, a small amount of mononuclear cell infiltration around intradermal blood vessel.Medicine test sample 3 groups: slight parakeratosis, the about thick 20-29 layer of spinous layer, epidermis is slight wooden club sample hypertrophy, substantially without granular layer, a small amount of mononuclear cell infiltration around intradermal blood vessel.Negative control group: severe parakeratosis, the about thick 26-38 layer of spinous layer, epidermis is severe wooden club sample hypertrophy, substantially without granular layer, moderate mononuclear cell infiltration around intradermal blood vessel.
Table 1 respectively group Baker scoring, Streptococcus counting, spine cell's number (
n=10)
Note: compare with model control group,
ap<0.01; Compare with positive controls
bp<0.05;
Result shows, and medicine test sample 1, test sample 2 and test sample 3 groups and model control group have significant difference (P<0.01); Compared with positive controls, there were significant differences (P<0.05) for medicine test sample 2 and test sample 3 groups.
Can find out, Folium Eucommiae extract can improve Baker scoring, alleviates the Streptococcus counting that intradermal in Animal Models of Psoriasis infiltrates, illustrates that Folium Eucommiae extract of the present invention has good therapeutical effect to psoriasis vulgaris, wherein, best with test sample 2.
(2) on the content of serum cAMP, cGMP and the impact of ratio
Table 2 on the impact of the content of serum cAMP, cGMP and ratio (
n=10)
Note: compare with blank group,
ap<0.01; Compare with model group
bp<0.01; Compare with positive group
cp<0.05
Psoriasis guinea pig model serum cAMP content all significantly reduces, and cGMP content all significantly raises (P<0.01).Compare with model group, medicine test sample is respectively organized and is all significantly raised (P<0.01) with positive controls cAMP content, cGMP content all reduces (P<0.01), and cAMP/cGMP ratio all raises (P<0.01); Medicine test sample 1, test sample 2 and test sample 3 respectively group all have statistical significance (P<0.05) with positive controls cGMP content and cAMP comparision contents, wherein, best with test sample 2.
(3) on the impact of Serum VEGF
Table 3 on the impact of Serum VEGF (
n=10)
Note: compare with blank group,
ap<0.01; Compare with model group
bp<0.01; Compare with positive group
cp<0.01;
Psoriasis guinea pig model serum VEGF is significantly higher than normal group, modeling success.Through treatment, each test sample group of medicine and positive controls serum VEGF are all lower than model group (P<0.01), and there were significant differences; Each test sample group of medicine is lower than positive controls (P<0.01), and there were significant differences, wherein, best with test sample 2 (embodiment 1 extract sample 2).
Folium Eucommiae extract of the present invention all effectively can improve the Baker scoring of psoriasis model, improve the content of serum cAMP, cGMP, reduce Serum VEGF, illustrate that Folium Eucommiae extract of the present invention can effectively treat psoriasis vulgaris (Liu Yuanlin etc., " change of patients with psoriasis vulgaris treatment front and back serum E-cad and VEGF level ", Chinese experimental diagnostics JIUYUE the 12nd volume the 9th phase in 2008 the 1162nd page of left hurdle inverse 1 ~ 3 row; Li Fei, " the Calcium Dibutyryladenosine Cyclophosph-ate associating acitretin treatment curative effect of psoriasis vulgaris and the impact on blood cAMP and cGMP ", Dermatology diagnostics magazine volume the 5th phase October the 17th in 2010), and optimum with the curative effect of medicine test sample 2.
Test example 2 Folium Eucommiae extract of the present invention treats the psoriatic test of pesticide effectiveness (psoriasis pustulosa)
Experimentation:
1. material
1) animal
Kunming mouse, weight 18-22g, Sichuan University's experimental animal center provides.Animal rank: one-level, credit number: No. 10.
2) medicine
Medicine test sample 1 (embodiment 3 extract sample 1);
Medicine test sample 2 (embodiment 3 extract sample 2);
2. experimental technique
1) animal grouping
Kunming mouse is divided into 3 groups at random.The each Mus of medicine test sample group presses 1mL/100g gastric infusion, dosage 100mg/kg; Model control group gives normal saline gavage.Observing time point: during non-administration (0 day), after administration the 1st, 2,4,6 day.
2) experimental technique
By one piece of 1cm
2sterile gelatin sponge in 0.75% zymosan A, soak 30min after, operation is inserted in the peritoneal cavity of mice, and gives normal feed.Postoperative 24 hours, 5 mices of phase point when disconnected neck is put to death same, take out gelfoam, the cell eluent 37 DEG C being placed in 0.5ml respectively hatches 30min, with 0.01mol/LPBS0.5ml respectively rinsing gelfoam 3 times, collect eluent, centrifugally after crossing 40 μm of nylon screens be settled to 0.5ml, with automatic blood cell instrument, classified counting of leucocyte is carried out to above-mentioned eluent, read the quantity of polymorphonuclear leukocyte (PMN).
3. result
Table 4 medicine test sample group moves quantity (× 10 at the PMN of different time phases
9/ L)
Note: compare with blank group, * P<0.05; △ P<0.01;
Result shows, after medication, medicine test sample 1 group and test sample 2 groups PMN from second day obviously reduce, significant difference is had compared with blank group, illustrate that Folium Eucommiae extract has the effect (Chen Shengping, the clinical observation of arotinoid ethylester for curing psoriasis pustulosa and experimentation, Third Military Medical University's master thesis 2002) for the treatment of psoriasis pustulosa, wherein, the effect of medicine test sample 2 groups (embodiment 3 extract sample 2) is relatively excellent.
To sum up, Folium Eucommiae extract of the present invention effectively can treat psoriasis, particularly to the determined curative effect of psoriasis vulgaris or psoriasis pustulosa, for psoriatic clinical treatment provides a kind of new medicine.
Claims (13)
1. a Folium Eucommiae extract, is characterized in that: it is water or the ethanol extraction of Folium Eucommiae, and in described Folium Eucommiae extract, chlorogenic acid content is not less than 9%, and Geniposidic acid is not less than 1%, and aucubin is not less than 1%.
2. Folium Eucommiae extract according to claim 1, is characterized in that: in described Folium Eucommiae extract, and chlorogenic acid content is 9.38% ~ 39.11%, and geniposide acid content is 1.03% ~ 10.33%, and aucubin is 1.17% ~ 10.68%.
3. Folium Eucommiae extract according to claim 2, is characterized in that: in described Folium Eucommiae extract, and chlorogenic acid content is 10.21% ~ 32.37%, and geniposide acid content is 1.03% ~ 9.38%, and aucubin is 1.17% ~ 9.56%.
4. Folium Eucommiae extract according to claim 3, is characterized in that: in described Folium Eucommiae extract, and chlorogenic acid content is 10.21% or 32.37%, and geniposide acid content is 1.03% or 2.24%, and aucubin is 1.24% or 2.07%.
5. the Folium Eucommiae extract according to Claims 1 to 4 any one, is characterized in that: described Folium Eucommiae extract is prepared by the following method:
Get Folium Eucommiae, with water or 30-80% ethanol extraction 1 ~ 2 time, each 1 ~ 2 hour, by extracting solution or its concentrated solution after phenylethylene absorption with macroporous adsorbent resin, use 10-90% ethanol gradient elution, collect eluent, merge, concentrate drying, to obtain final product.
6. Cortex Eucommiae extract according to claim 5, is characterized in that: described Folium Eucommiae extract is prepared by the following method:
Get Folium Eucommiae, with 60% ethanol in 70 DEG C of lixiviates twice, at every turn with 15 times amount volumes, each 1 hour, merge extractive liquid, concentration and recovery ethanol, concentrated solution, with after D-101 macroporous adsorbent resin resin saturated adsorption, uses 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collects eluent, merge 10% and 30% eluent, or merge 70% and 90% eluent, concentrate drying, to obtain final product;
Or, get Folium Eucommiae, with 30% ethanol in 60 DEG C of lixiviates twice, each 2 hours, each use 12 times amount volume, merge extractive liquid, concentration and recovery ethanol, concentrated solution is with after HPD-100 macroporous adsorbent resin resin saturated adsorption, use 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collect eluent, merge 70% and 90% eluent, or get 30% eluent, concentrate drying, to obtain final product;
Or, get Folium Eucommiae, with 20 times of water in 80 DEG C of lixiviates once, each 1.5 hours, extracting solution, with after XAD-16 macroporous adsorbent resin resin saturated adsorption, used 20% and 60% ethanol gradient concentration ethanol eluting successively, collect eluent and 60% eluent of 20% respectively, concentrate drying, to obtain final product;
Or, get Folium Eucommiae, with 12 times of water in 60 DEG C of lixiviates twice, each 2 hours, extracting solution, with after AB-8 macroporous adsorbent resin resin saturated adsorption, used 10%, 90% ethanol gradient concentration ethanol eluting successively, collect eluent, collect 10% eluent and 90% eluent respectively, concentrate drying, to obtain final product.
7. prepare the method for Folium Eucommiae extract described in claim 1 ~ 6 any one for one kind, it is characterized in that: it comprises the steps: to get Folium Eucommiae, with water or 30-80% ethanol extraction 1 ~ 2 time, each 1 ~ 2 hour, by extracting solution or its concentrated solution after phenylethylene absorption with macroporous adsorbent resin, use 10-90% ethanol gradient elution, collect eluent, merge, concentrate drying, to obtain final product.
8. method according to claim 7, is characterized in that: it comprises the steps:
Get Folium Eucommiae, with 60% ethanol in 70 DEG C of lixiviates twice, at every turn with 15 times amount volumes, each 1 hour, merge extractive liquid, concentration and recovery ethanol, concentrated solution, with after D-101 macroporous adsorbent resin resin saturated adsorption, uses 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collects eluent, merge 10% and 30% eluent, or merge 70% and 90% eluent, concentrate drying;
Or, get Folium Eucommiae, with 30% ethanol in 60 DEG C of lixiviates twice, each 2 hours, each use 12 times amount volume, merge extractive liquid, concentration and recovery ethanol, concentrated solution is with after HPD-100 macroporous adsorbent resin resin saturated adsorption, use 10%, 30%, 70% and 90% ethanol gradient concentration ethanol eluting successively, collect eluent, merge 70% and 90% eluent, or get 30% eluent, concentrate drying;
Or, get Folium Eucommiae, with 20 times of water in 80 DEG C of lixiviates once, each 1.5 hours, extracting solution, with after XAD-16 macroporous adsorbent resin resin saturated adsorption, used 20% and 60% ethanol gradient concentration ethanol eluting successively, collect eluent and 60% eluent of 20% respectively, concentrate drying;
Or, get Folium Eucommiae, with 12 times of water in 60 DEG C of lixiviates twice, each 2 hours, extracting solution, with after AB-8 macroporous adsorbent resin resin saturated adsorption, used 10%, 90% ethanol gradient concentration ethanol eluting successively, collect eluent, collect 10% eluent and 90% eluent respectively, concentrate drying.
9. the purposes of the Folium Eucommiae extract described in claim 1 ~ 6 any one in the psoriatic medicine of preparation treatment.
10. purposes according to claim 9, is characterized in that: described medicine is the medicine for the treatment of psoriasis vulgaris or psoriasis pustulosa.
11. 1 kinds treatment psoriatic medicines, it is characterized in that: it be with the Folium Eucommiae extract described in claim 1 ~ 6 any one for active component, add the preparation that pharmaceutically acceptable adjuvant or complementary composition are prepared from.
12. medicines according to claim 11, is characterized in that: described preparation is oral formulations, external preparation.
13. medicines according to claim 12, is characterized in that: described oral formulations is tablet, granule, powder, pill, oral liquid, capsule.
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