CN104434902A - Application of chlorogenic acid in preparing medicines for treating leucoderma - Google Patents
Application of chlorogenic acid in preparing medicines for treating leucoderma Download PDFInfo
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Abstract
The invention provides an application of chlorogenic acid in preparing medicines for treating leucoderma. Tests show that the chlorogenic acid can be used for improving the pathological change of a leucoderma model, reducing the expression of TNF-alpha and IL-4 on a part of skin lesion and promoting the generation of melanocyte, so that a theoretical basis is provided for the clinic treatment of leucoderma.
Description
Technical field
The present invention relates to the purposes of chlorogenic acid in the leukodermic medicine of preparation treatment.
Background technology
Chlorogenic acid (Chlorogenic acid, CGA), have another name called chlorogenic acid, chemistry 3-0-caffeoyl guinic acid (3-0-caffeoylquinic acid) by name, the carboxylic phenolic acid be made up of caffeic acid (Caffeic acid) and quinic acid (Quinic acid).
Chlorogenic acid is the product of plant aerobic respiration metabolism; it is the principle active component in many Chinese crude drugs and fruit and vegetable; there is multiple biological activity, as: cardiovascular protective effect, antioxidation, uvioresistant and radiation resistance, antimutagenic and antitumaous effect, antibacterial action, antivirus action, blood lipid-reducing blood sugar-decreasing effect, immunoregulation effect etc.All have a wide range of applications in the field such as medication chemistry and food.
Summary of the invention
Technical scheme of the present invention there is provided the purposes of chlorogenic acid in the leukodermic medicine of preparation treatment.
The invention provides the purposes of chlorogenic acid in the leukodermic medicine of preparation treatment.
Wherein, described medicine is the medicine of the expression reducing skin lesion place TNF-α and IL-4.
Wherein, described medicine is the medicine promoting that melanocyte generates.
Wherein, described medicine is effective ingredient by chlorogenic acid, adds the preparation that pharmaceutically acceptable adjuvant or complementary composition are prepared from.
Wherein, described preparation is oral formulations, ejection preparation or externally applied transdermal drug-delivery preparation.
Wherein, the using dosage of described preparation is: 1-100mg/kg.Further preferably, the using dosage of described preparation is: 1-30mg/kg.
The present invention's test shows, chlorogenic acid can improve the pathological change of vitiligo model, reduces the expression of skin lesion place TNF-α and IL-4, promotes melanocytic generation, for its clinical treatment vitiligo provides theoretical foundation.
Detailed description of the invention
Example 1: pharmacodynamics test research in leukodermic body treated by chlorogenic acid
1. material
1) animal
Black Cavia porcellus, body weight 300 ~ 350g, male and female half and half.
2) reagent and trial drug
Hydroquinone, methoxsalen sheet, chlorogenic acid; Tryrosinase, acetylcholine esterase (CHE), monoamine oxidase, MAO (MAO), malonaldehyde (MDA) measure test kit
2. experimental technique
1) model preparation and grouping administration
Get black Cavia porcellus, shave with shaver and get back wool area 5cm × 5cm, by body weight random packet, often organize 10, i.e. dosage group (30mgkg), the heavy dose of group (60mg/kg) of chlorogenic acid in blank group (waiting capacity normal saline), model group (waiting capacity normal saline), positive controls (methoxsalen sheet 2.8mg/kg), chlorogenic acid small dose group (15mg/kg), chlorogenic acid.Wherein blank group smears normal saline 0.05ml in depilation district, and all the other are respectively organized and all smear 5% hydroquinone 0.5ml in depilation district, every day 2 times, smear 11 days continuously, prepare Vitiligo guinea-pig model.Within 11st day, start each group of every day of according to dosage administration respectively in 1 hour after smearing hydroquinone respectively, every day 1 time, successive administration 50 days.
2) observation index and mensuration
1. efficacy determination
After experiment terminates, perusal guinea pig back skin pigment distributes.Curative effect judging standard is (with Cavia porcellus agents area center 3cm
2be an observation unit): excellent is tested district (3cm
2) pigment recovers normal substantially; Goodly there is pigment area > 50% for tested district; In there is pigment area < 50% for tested district; Difference for tested district skin be pale or white macula shape, total effective rate in excellent add good.
2. Determination of Blood Rheology
40min after last administration, gets blood in abdominal aortic, centrifugal blood plasma, for the mensuration of CHE, MAO, MDA, TYR activity, surveys hemorheology index (adopting YDA-IU blood rheological instrument to measure).
3. the mensuration of dermal melanin
By each group of Cavia porcellus bark fetching skin 1cm × 1cm, fix with 10% formaldehyde, routine paraffin wax embedded section, carries out melanin dyeing with ferrous sulfate (Lillie method), and observing epidermal basal cell and the distribution of spine cell's melanin and hair follicle has melanic Cavia porcellus number.Each specimen observes 10 high power fields, calculates the par containing the basal cell of melanin granule in every 100 epidermal basal cells.Grading: "-" indicates without melanin; " ± " represents accidental melanin; "+" represents that 30% ~ 50% has melanin; " ++ " represents that 51% ~ 85% has melanin; " +++ " represents that more than 85% has melanin.
3. statistical method
All data all use the process of SPSS 19.0 statistical software.
4. result
1) observation of curative effect
Perusal model group guinea epidermis color is all obviously pale, part hair also turns white, have significance with normal group comparing difference, after each dosage group treatment of chlorogenic acid, curative effect is obvious, perusal guinea pig skin also melanism gradually, chlorogenic acid each administration group color is brownish black, relatively normal group, and has a small amount of pigmentation, compare with positive controls and have significant difference, the results are shown in Table 1.
Table 1 chlorogenic acid is to the observation of curative effect (3cm of Vitiligo guinea-pig model
2, n=10)
Note: compare * P < 0.05, * * P < 0.01 with model group
2) chlorogenic acid is on the impact of each index of model guinea pigs hemorheology
After modeling, model group guinea pig whole blood viscosity and plasma viscosity obviously raise, chlorogenic acid 15mg/kg, and 30mg/kg, 60mg/kg obviously can reduce whole blood viscosity and the plasma viscosity of model guinea pigs.Whole blood viscosity and the plasma viscosity of chlorogenic acid each administration group reduction model guinea pigs are obviously better than positive group, the results are shown in Table 2
The impact of each index of the hemorheology of table 2 chlorogenic acid on Vitiligo guinea-pig model
Note: compare * P < 0.05, * * P < 0.01 with model group
3) chlorogenic acid is on the impact of MAO, CHE, MDA, TYR index in model guinea pigs blood plasma
After modeling, model group guinea pig plasma MAO, CHE and MDA content obviously increases, tryrosinase content obviously reduces, in chlorogenic acid 15mg/kg, 30mg/kg, 60mg/kg guinea pig plasma, the content of MAO, CHE and MDA all obviously reduces, and chlorogenic acid 15mg/kg, 30mg/kg, 60mg/kg obviously can increase the content of tryrosinase in dermal melanin.The results are shown in Table 3.
Table 3 chlorogenic acid is on the impact of MAO, CHE, MDA, TYR level in the blood plasma of Vitiligo guinea-pig model
Note: compare * P < 0.05, * * P < 0.01 with model group
4) chlorogenic acid is on the melanic impact of guinea pig model
After modeling, the black of model group basal layer and spinous layer obviously lacks, chlorogenic acid 15mg/kg, 30mg/kg, 60mg/kg obviously can increase the dermal melanin of model guinea pigs, and obviously increase epiderm skin basal cell and spine cell's melanin distribute and melanin in skin follicle.The results are shown in Table 4.
Table 4 chlorogenic acid is on the melanic impact of guinea pig model
5. conclusion
Above-mentioned experimental result shows, chlorogenic acid obviously can increase the dermal melanin of experimental vitiligo model guinea pigs, obvious increase epiderm skin basal cell and the distribution of spine cell's melanin, melanin in skin follicle can be made significantly to increase, tryrosinase content also significance increase in blood, and significance improves hemorheological indexes; Illustrate that chlorogenic acid can increase experimental vitiligo guinea pig model melanin and generate, reduce dermal melanin and decompose and improve blood circulation, good therapeutical effect is played to vitiligo.
Example 2: the impact that chlorogenic acid is expressed Vitiligo guinea-pig model immune organ organ coefficient and skin lesion district TNF-α, IL-10, IL-4, IFN-γ
1. material
1) animal
Black Cavia porcellus, body weight 300 ~ 350g, male and female half and half.
2) reagent and trial drug
Hydroquinone, methoxsalen sheet, chlorogenic acid
2. experimental technique
1) experimental Vitiligo guinea-pig model preparation
Adopt hydroquinone (hydroquinone) preparation experiment Vitiligo guinea-pig model.Get healthy black Cavia porcellus, male and female half and half, shave get back wool area 5cm × 5cm with shaver, every day smears the hydroquinone 0.5ml of 5% in depilation district, every day 2 times, smears 50 days continuously, prepares Vitiligo guinea-pig model.
2) animal grouping and administration
Above-mentioned model guinea pigs is divided into 5 groups at random, be respectively model group, positive group, chlorogenic acid small dose group (15mg/kg), dosage group (30mg/kg), the heavy dose of group (60mg/kg) of chlorogenic acid in chlorogenic acid, often organize 10; Separately get 10 non-modeling Cavia porcelluss as blank group.Each medicine group Cavia porcellus plays beginning administration on 10th respectively at modeling, and blank group and model group Cavia porcellus such as to give respectively at the normal saline of capacity, successive administration 50 days.
3) animal process
After treatment terminates, put to death Cavia porcellus, get agents area center 1cm × 1cm skin, neutral formalin is fixed, routine paraffin wax embedding, section.And get Thymus of Guinea Pigs, spleen is weighed, and calculates thymus coefficient.
4) Immunohistochemical Method observes IL-4, TNF-α
The conventional dewaxing of section is to water.The multiple antigen of hot repair: 0.01M citrate buffer (pH6.0), electric furnace or microwave-oven-heating are to the rear power-off of boiling, and section is immersed, and after cooling, PBS washs 1-2 time.30%H2O21 part+distilled water 10 parts mixing, room temperature 10 minutes is with inactivating endogenous enzyme.PBS washes 5 minutes × 3 times.Normal goats immune serum is closed, and incubated at room 15 minutes, incline serum deprivation, do not wash.Drip the HMB45 antibody pressing 1:100 dilution, 4 DEG C are spent the night.PBS washes 2 minutes × 3 times.Drip Radix Cochleariae officinalis enzyme labelling strepto-avidin working solution, hatch 10 minutes for 37 DEG C.PBS washes 5 minutes × 3 times.DAB develops the color, and controls the response time, 20s-80s, to the satisfaction that develops the color under mirror.Distilled water wash, color development stopping.Haematoxylin is slightly redyed, and hydrochloride alcohol breaks up, and running water returns blue 1 minute.Dehydration, transparent, mounting.Microscopic examination.
3. statistical method and analysis
All data all use the process of SPSS 19.0 statistical software.
The expression of IL-4, TNF-α:
Percentage ratio according to dye levels and pigmented cells carries out sxemiquantitative scoring.Each section random selecting 5 place high power field (200 ×), mark according to positive cell number, standards of grading are: 0 point, < 25% pigmented cells; 1 point, 25%-50% pigmented cells; 2 points, 50%-75% pigmented cells; 3 points, >75% pigmented cells.Dye levels standards of grading are: 0 point, dye-free; 1 point, light yellow; 2 points, brown color; 3 points of sepias.Dye levels is divided and adds that pigmented cells percentage ratio divides and draw total mark, be divided into 4 grades according to integration: 0 point, negative (-); 2 points, the weak positive (+); 3-4 divides, positive (++); 5-6 divides, strong positive (+++).Statistics adopts rank test method (Kruskal-Wallis method).
4. result
1) chlorogenic acid is on the impact of model guinea pigs immune organ organ coefficient
The organ coefficient of model group Thymus of Guinea Pigs and spleen obviously reduces, and remarkable with blank group comparing difference, after medication, each dosage group of chlorogenic acid can increase model guinea pigs Thymus and spleen and organ coefficient, remarkable with model group comparing difference.Show, chlorogenic acid can strengthen the function of the immunity of model guinea pigs.In table 5.
Table 5 chlorogenic acid is on the impact of model guinea pigs immune organ organ coefficient
Note: compare with Normal group, * P < 0.05, * * P < 0.01; Compare with model group, #P < 0.05, ##P < 0.01
2) chlorogenic acid impact that Vitiligo guinea-pig model skin lesion district TNF-α, IL-10, IL-4, IFN-γ is expressed
TNF-α positive cell is mainly distributed in stratum basale, spinous layer, granular layer, and minority positive cell is distributed in high dermis; IL-4 positive cell is mainly distributed in stratum basale, spinous layer, and matched group is weak positive expression.Model group skin lesion district TNF-α and IL-4 expression are apparently higher than Normal group, and difference has significance (P<0.01); Treatment group skin lesion district TNF-α and IL-4 expression are starkly lower than model group, and difference has significance (P<0.01).
Table 6 chlorogenic acid is on the impact (positive number of cases) (rank test) of skin lesion district TNF-alpha expression
| Group | Number of cases | - | + | ++ | +++ |
| Blank group | 10 | 4 | 5 | 1 | 0 |
| Model group | 10 | 0 | 0 | 2 | 8 |
| Chlorogenic acid group | 10 | 4 | 4 | 2 | 0 |
Note: model group and Normal group comparing difference have significance (P<0.01); Treatment group and model group comparing difference have significance (P<0.01)
The impact (positive number of cases) (rank test) that table 7 chlorogenic acid is expressed skin lesion district IL-4
| Group | Number of cases | - | + | ++ | +++ |
| Blank group | 10 | 4 | 4 | 2 | 0 |
| Model group | 10 | 0 | 0 | 3 | 7 |
| Chlorogenic acid group | 10 | 3 | 5 | 2 | 0 |
Note: model group and Normal group comparing difference have significance (P<0.01); Treatment group and model group comparing difference have significance (P<0.01)
5. conclusion
Chlorogenic acid can improve the pathological change of vitiligo model, reduces the expression of skin lesion place TNF-α and IL-4, promotes melanocytic generation, for its clinical treatment vitiligo provides theoretical foundation.
Claims (7)
1. the purposes of chlorogenic acid in the leukodermic medicine of preparation treatment.
2. purposes according to claim 1, is characterized in that: described medicine is the medicine of the expression reducing skin lesion place TNF-α and IL-4.
3. purposes according to claim 1, is characterized in that: described medicine is the medicine promoting that melanocyte generates.
4. the purposes according to claim 1-3 any one, is characterized in that: described medicine is effective ingredient by chlorogenic acid, adds the preparation that pharmaceutically acceptable adjuvant or complementary composition are prepared from.
5. purposes according to claim 4, is characterized in that: described preparation is oral formulations, ejection preparation or externally applied transdermal drug-delivery preparation.
6. purposes according to claim 5, is characterized in that: the using dosage of described preparation is: 1-100mg/kg.
7. purposes according to claim 6, is characterized in that: the using dosage of described preparation is: 1-30mg/kg.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410778100.7A CN104434902A (en) | 2014-12-15 | 2014-12-15 | Application of chlorogenic acid in preparing medicines for treating leucoderma |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410778100.7A CN104434902A (en) | 2014-12-15 | 2014-12-15 | Application of chlorogenic acid in preparing medicines for treating leucoderma |
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|---|---|
| CN104434902A true CN104434902A (en) | 2015-03-25 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016095087A1 (en) * | 2014-12-15 | 2016-06-23 | 四川九章生物科技有限公司 | Uses of chlorogenic acid in preparation of medicines for treating vitiligo |
| CN109419795A (en) * | 2017-08-28 | 2019-03-05 | 四川九章生物科技有限公司 | A kind of combination medicine for treating tnf family cytokines related disease |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09151130A (en) * | 1995-12-01 | 1997-06-10 | Yakurigaku Chuo Kenkyusho:Kk | Medicine for treating trichopoliosis and leukoplakia |
| CN102985058A (en) * | 2010-06-30 | 2013-03-20 | 雀巢产品技术援助有限公司 | Use of caftaric acid and lactic bacterium in food supplement for regulating skin pigmentation |
-
2014
- 2014-12-15 CN CN201410778100.7A patent/CN104434902A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09151130A (en) * | 1995-12-01 | 1997-06-10 | Yakurigaku Chuo Kenkyusho:Kk | Medicine for treating trichopoliosis and leukoplakia |
| CN102985058A (en) * | 2010-06-30 | 2013-03-20 | 雀巢产品技术援助有限公司 | Use of caftaric acid and lactic bacterium in food supplement for regulating skin pigmentation |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016095087A1 (en) * | 2014-12-15 | 2016-06-23 | 四川九章生物科技有限公司 | Uses of chlorogenic acid in preparation of medicines for treating vitiligo |
| CN109419795A (en) * | 2017-08-28 | 2019-03-05 | 四川九章生物科技有限公司 | A kind of combination medicine for treating tnf family cytokines related disease |
| CN109419795B (en) * | 2017-08-28 | 2021-02-26 | 四川九章生物科技有限公司 | Combined medicine for treating tumor necrosis factor family related diseases |
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Application publication date: 20150325 |
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