CN113975323B - Traditional Chinese medicine composition for treating atopic dermatitis and preparation method and application thereof - Google Patents

Abstract

The invention relates to a traditional Chinese medicine composition for treating atopic dermatitis, a preparation method and application thereof. The application provides a traditional Chinese medicine composition for treating atopic dermatitis, which is prepared from the following traditional Chinese medicines in parts by weight: 3-10 g of ephedra, 5-20 g of forsythia, 5-15 g of cicada slough, 5-20 g of trumpet creeper, 20-40 g of phaseolus calcaratus, 20-35 g of sea buckthorn, 5-20 g of cortex dictamni, 5-20 g of fructus kochiae and 5-12 g of raw liquorice. The application provides an application of a traditional Chinese medicine composition in preparing a medicine for treating atopic dermatitis. The traditional Chinese medicine composition provided by the application can obviously reduce skin erythema, bleeding points, dry skin, scaling or moss-like change of atopic dermatitis, reduce percutaneous water loss, reduce the pH value of the skin and improve the histopathological expression of skin lesions. The traditional Chinese medicine composition provided by the application is superior to a positive control drug mometasone furoate in the aspects of reducing percutaneous water loss and scratching behavior.

Description

Traditional Chinese medicine composition for treating atopic dermatitis and preparation method and application thereof

Technical Field

The invention relates to the field of traditional Chinese medicine compositions, and relates to a traditional Chinese medicine composition for treating atopic dermatitis, and a preparation method and application thereof.

Background

Atopic Dermatitis (AD) is a chronic recurrent, pruritic, inflammatory skin disease with inflammatory exudation, eczematoid lesions, lichenification, and dry skin as its major clinical manifestations. The disease usually occurs from infants, the prevalence rate of AD of children of 1-7 years old in China is 12.94%, and the prevalence rate of AD of adults is 4.6%, and the disease tends to rise year by year. In addition, AD often combines asthma, allergic rhinitis and food allergy, has increased susceptibility to bacterial and viral infections, has a tendency to suffer from anxiety, depression, attention deficit hyperactivity disorder and other diseases, seriously affects the physical and mental health of patients, and has become an increasingly serious public health problem worldwide.

Atopic dermatitis is related to hereditary, allergy and other factors, but the exact etiology and pathogenesis of atopic dermatitis are not completely elucidated, and it is considered that it is closely related to hereditary, environmental, skin barrier damage, neuroimmune disorder and other factors. AD belongs to the categories of diseases such as 'infantile eczema', 'infantile eczema' and 'fetal wound healing' in the traditional Chinese medicine, doctors of all ages discuss the pathogenesis of AD abundantly, and the AD is mostly considered to be attacked by deficient innate endowment, fetal toxin, heat or improper acquired regulation, spleen dysfunction, dampness generation, heat transformation, repeated rheumatism and heat pathogen, skin accumulation and internal and external pathogenic factors. Wind-damp-heat accumulation is one of the most common types of traditional Chinese medicine syndromes in AD clinic.

Ephedra, forsythiae, nelumbo-Phaseolus angularis Tang comes from Shang Han Lun (treatise on Cold-induced diseases by differentiation of Yang brightness disease and pulse-pattern of diseases)' chapter 262; for cold injury, blood stasis and heat in the interior, the body must be yellow, and Ma Huang Lian, lian jiao, chi Dou Tang is the main one. The prescription is prepared from ephedra, forsythia, phaseolus calcaratus, raw catalpa bark, almond, ginger, honey-fried licorice root and Chinese date, and has the functions of dispersing wind-damp heat evil, strengthening spleen and regulating the middle warmer and clearing damp-heat. Lian Yin in the prescription, song Ben (Song Ben's "Shang Yun" Lian Gen of Lian Qiao, nowadays, clinical practice usually involves Lian Qiao, so it is also called as Ma Huang Lian Qiao Xiao Dou Tang. The raw catalpa bungei white-bark is bitter in taste and cold in nature, has the functions of clearing heat and promoting diuresis, calming adverse-rising energy and stopping vomiting, and killing parasites and relieving itching, but is not prepared in the current pharmacy, and is mostly replaced by the white mulberry root-bark, virgate wormwood herb and the like in later doctors.

The Chinese ephedra, forsythia and phaseolus calcaratus decoction (named as Chinese ephedra, forsythia and phaseolus calcaratus decoction) substituted by the fructus forsythiae and the cortex mori has certain effects on the functions of clearing away damp-heat, killing parasites and relieving itching, and the Chinese date, the honey-fried licorice root and the ginger in the formula are sweet, warm or pungent in flavor and are not beneficial to expelling damp-heat pathogenic factors, so that the clinical curative effect of the formula on treating AD is influenced. Atopic dermatitis is characterized by severe itching and eczematous skin lesions, and the condition of atopic dermatitis is easy to recur, and the atopic dermatitis is lingering and difficult to cure after long-term invasion of blood. Therefore, the effect of using the ephedra, forsythia and red bean soup alone on the rheumatic heat accumulation type atopic dermatitis is poor, such as the effects of improving skin damage of atopic dermatitis patients and effectively controlling pruritus are not satisfactory.

Disclosure of Invention

In order to improve the itching relieving and skin damage effects of ephedra, forsythia and red bean soup, the application provides a traditional Chinese medicine composition for treating atopic dermatitis.

In a first aspect, the application provides a traditional Chinese medicine composition for treating atopic dermatitis, which is prepared from the following traditional Chinese medicines in parts by weight: 3-10 g of ephedra, 5-20 g of fructus forsythiae, 5-15 g of periostracum cicada, 5-20 g of trumpet creeper, 20-40 g of phaseolus calcaratus, 20-35 g of sea buckthorn, 5-20 g of cortex dictamni, 5-20 g of fructus kochiae and 5-12 g of raw liquorice.

Preferably, the traditional Chinese medicine composition is prepared from the following traditional Chinese medicines in parts by weight: 3-9 g of ephedra, 10-15 g of fructus forsythiae, 5-15 g of periostracum cicada, 10-15 g of trumpet creeper, 20-40 g of phaseolus calcaratus, 25-35 g of sea buckthorn, 10-15 g of cortex dictamni, 10-15 g of fructus kochiae and 5-15 g of raw liquorice.

Further preferably, the traditional Chinese medicine composition is prepared from the following traditional Chinese medicines in parts by weight: 4-8 g of ephedra, 13-18 g of fructus forsythiae, 6-12 g of periostracum cicada, 13-18 g of trumpet creeper, 25-35 g of phaseolus calcaratus, 25-35 g of sea buckthorn, 13-18 g of cortex dictamni, 13-18 g of fructus kochiae and 6-10 g of raw liquorice.

More preferably, the traditional Chinese medicine composition is prepared from the following traditional Chinese medicines in parts by weight: 6 g of ephedra, 15 g of forsythia, 9 g of cicada slough, 15 g of trumpet creeper, 30 g of phaseolus calcaratus, 30 g of sea buckthorn, 15 g of cortex dictamni, 15 g of fructus kochiae and 9 g of raw liquorice.

In a second aspect, the present application provides a method for preparing a Chinese medicinal composition for treating atopic dermatitis, comprising the steps of: weighing the components according to the proportion, mixing uniformly, extracting by using an organic solvent, filtering, collecting filtrate, concentrating and drying to obtain the product.

The organic solvent is water or lower alcohol such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, etc.

In a third aspect, the present application provides an application of a Chinese medicinal composition in preparing a medicament for treating atopic dermatitis.

The medicine is a preparation consisting of a traditional Chinese medicine composition and a pharmaceutically acceptable carrier.

The preparation is tablet, capsule, granule, oral liquid, sustained release preparation, controlled release preparation, gel, ointment, salve, cream, suppository, injection, powder for injection, patch, dripping pill, suspension, etc.

The pharmaceutically acceptable carrier is one or more of a binder, a filler, a disintegrant, a lubricant, a suspending agent, a solvent, a surfactant, a flavoring agent, and a wetting agent.

The binder is selected from cellulose derivatives, alginates, starch, water, dextrin, gelatin, polyvinylpyrrolidone, or the like.

The filler is selected from starch, sucrose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose or glucose.

The disintegrant is selected from microcrystalline cellulose, sodium carboxymethyl starch, crospolyvinylpyrrolidone, low-substituted hydroxypropyl cellulose or croscarmellose sodium.

The lubricant is selected from stearic acid, polyethylene glycol, calcium carbonate, sodium bicarbonate, superfine silica gel powder, talcum powder or magnesium stearate.

The suspending agent is selected from silica gel micropowder, beeswax, cellulose or solid polyethylene glycol.

The wetting agent is selected from glycerol, tween-80, ethoxylated hydrogenated castor oil or lecithin.

The solvent is selected from ethanol, liquid polyethylene glycol, isopropanol, tween-80, glycerol, propylene glycol or vegetable oil selected from soybean oil, castor oil, peanut oil or blend oil.

The surfactant is selected from sodium dodecyl benzene sulfonate, stearic acid, polyoxyethylene-polyoxypropylene copolymer, sorbitan fatty acid or polysorbate (Tween) and the like.

The correctant is selected from aspartame, sucralose, essence, steviosin, acesulfame potassium, citric acid or saccharin sodium.

The application has the following beneficial effects:

1. the traditional Chinese medicine composition has the advantages of simple prescription and easy preparation

The traditional Chinese medicine composition provided by the invention comprises nine traditional Chinese medicines, namely ephedra, fructus forsythiae, phaseolus calcaratus, campsis grandiflora, cortex dictamni, sea buckthorn, fructus kochiae, periostracum cicadae and raw liquorice.

In the formula, the ephedra herb is pungent and slightly bitter in taste, enters lung and bladder channels, and can induce sweat, relieve exterior syndrome, ventilate lung, relieve asthma, induce diuresis and reduce edema; it can disperse pathogenic dampness, and can also disperse lung qi upwards, regulate water passage, and transport bladder downwards to induce diuresis and relieve swelling, and treat diseases due to pathogenic wind attacking exterior and lung failing to disperse and descend.

Lian Qiao, bitter and pungent in flavor and slightly cold in nature, enters lung, heart and small intestine meridians. The recipe has the effects of clearing heat and removing toxicity, relieving swelling and dissipating stagnation, and dispelling wind-heat, and is called "Chuang Jia Sheng Yao" in the pearl sac, because it can treat heat in the upper energizer, especially remove toxicity and cure abscess and dissipate stagnation.

Cicada slough, sweet in flavor and cool in nature, enters lung and liver meridians; disperse wind-heat, relieve sore throat and promote eruption, improve vision and remove nebula, extinguish wind and relieve spasm. It is good at treating measles without adequate eruption, rubella itching, furuncle and pyogenic infections.

Campsis grandiflora, slightly cold in nature, pungent and sour in taste; it enters liver and pericardium meridians. Has the functions of clearing heat, cooling blood and activating blood circulation, and is mainly used for treating rubella, skin itch, acne and the like.

Red bean, neutral in nature, sweet in taste; it enters heart and small intestine channels, has the functions of inducing diuresis to alleviate edema, removing toxicity and expelling pus, and is used for edema and fullness, beriberi edema, jaundice and dark urine, rheumatism and heat arthralgia, carbuncle swelling and sore toxicity, intestinal abscess and abdominal pain, etc.

Sea buckthorn, sour and astringent in taste and warm in nature; it has the functions of invigorating spleen, promoting digestion, relieving cough, eliminating phlegm, promoting blood circulation to disperse blood clots, etc.

Cortex Dictamni Radicis, cold in nature; is bitter in taste; spleen, stomach and bladder meridians entered; has the functions of clearing away heat, eliminating dampness, dispelling wind and removing toxicity. It is indicated for all kinds of damp-heat accumulation in the skin, such as damp-heat sores, eczema, rubella itching, scabies, etc.

Broom cypress fruit, cold in nature, pungent and bitter in taste; it enters kidney and bladder meridians. Has the functions of clearing heat, promoting diuresis, dispelling wind and relieving itching, and is a common medicine for skin diseases; it is mainly used for treating rubella, eczema, sore, scabies, pudendum itching, etc.

Raw licorice root: sweet in flavor and neutral in nature, enter heart, spleen, stomach and lung meridians. Has the functions of invigorating spleen and replenishing qi, clearing away heat and toxic material, eliminating phlegm and stopping cough, relieving spasm and pain, and harmonizing the medicines.

The nature, taste, channel tropism and efficacy main treatment of the 9 medicines are comprehensively analyzed, and the medicines can be roughly divided into the following four groups according to the characteristics of symptoms and signs of atopic dermatitis, rheumatism and heat evil and skin accumulation:

first, the Chinese ephedra, forsythia and raw licorice root are combined as the medicine for dispersing exterior evil. The three medicines have the effects of dispersing blood stasis and heat, and reaching the exterior to discharge pathogenic factors, and also have the effects of inducing diuresis to alleviate edema, detoxifying and resolving masses, and dispelling wind and relieving itching.

Secondly, a medicine combination for cooling blood and invigorating blood circulation, namely Chinese trumpetcreeper, red bean and sea backthern. The three medicines aim at the characteristics of atopic dermatitis which is lingering and difficult to cure, enters blood for a long time and enters collaterals for a long time, and have the functions of clearing heat and cooling blood, activating blood and reducing swelling, and harmonizing the middle warmer and strengthening the spleen.

Thirdly, the antipruritic medicine composition-broom cypress fruit, dittany bark and cicada slough. The two medicines aim at the characteristics of atopic dermatitis accompanied by severe pruritus, and are used for clearing away damp-heat, dispelling wind and arresting itching, and the curative effects are greatly increased by mutually reinforcing.

Compared with the existing ephedra, forsythia and phaseolus calcaratus decoction original prescription (ephedra, forsythia, phaseolus calcaratus, almond, mulberry bark, ginger, honey-fried licorice root and Chinese date), the traditional Chinese medicine composition provided by the invention eliminates three pungent, sweet and warm medicines of ginger, honey-fried licorice root and Chinese date, so as to prevent the disadvantage of promoting generation of damp heat (the clinical effect is not ideal) caused by the ingredients; the sea-buckthorn is used for replacing the almond, so that the cough relieving effect is achieved, and the blood circulation promoting, the blood stasis removing, the spleen strengthening and the digestion promoting effects are achieved; raw gan Cao is used to replace gan Cao (prepared Licorice root), so it excels in clearing heat and removing toxicity, and can tonify qi and harmonize middle energizer. The cortex dictamni is used for replacing the raw Chinese catalpa bark (which is not prepared in the pharmacy nowadays), and the broom cypress fruit and the cicada slough are added, so that the effects of clearing away damp-heat, dispelling wind and relieving itching are achieved by combining the three medicines; the campsis grandiflora is added to clear heat and cool blood, and the components can be combined to treat atopic dermatitis and rheumatic fever which are the most common syndrome types in the traditional Chinese medicine clinical, have the effects of treating exterior and interior simultaneously, regulating qi and blood, being mild in medicine property, eliminating pathogenic factors without damaging vital qi, and having the effects of dispelling wind and clearing heat, detoxifying and resolving masses, cooling blood and activating blood, and promoting diuresis and relieving itching.

2. The traditional Chinese medicine composition provided by the application has the following overall effects:

1) Can obviously relieve atopic dermatitis erythema, bleeding spot, xerosis cutis, scale or moss-like change, reduce percutaneous water loss, reduce pH value of skin, and improve pathological expression of skin injury tissue.

2) The traditional Chinese medicine composition provided by the application is superior to a positive control drug mometasone furoate in the aspects of reducing the percutaneous water loss and the scratching behavior.

In addition, mometasone furoate, a common medicine for clinically treating atopic dermatitis, is a corticoid medicine, has good effect on relieving symptoms such as inflammation, pruritus and the like, and is mainly used for eczema, neurodermatitis, atopic dermatitis and skin pruritus. However, due to long-term and repeated onset of atopic dermatitis, the product often has side effects of burning skin, itching and stinging, skin atrophy, hirsutism, skin infiltration, secondary infection, skin striation pigmentation, etc. after long-term use. The research shows that the effect of the traditional Chinese medicine composition provided by the application is equivalent to that of the traditional Chinese medicine composition, and no side effect exists, which is a treatment target realized by the conventional traditional Chinese medicine compound.

Description of the drawings:

FIG. 1: the skin lesions of the mice in each group are generally observed on the 27 th day of the experiment, and the mice in each group are a normal group, a model group, an original square group, an example group, a control group, a comparative example 1 group and a comparative example 2 group from left to right in the figure;

FIG. 2 is a schematic diagram: the skin histopathology (HE × 10) of each group of mice was shown in the upper row from left to right in the figure as a normal group, a model group, an original square group, and an example group; the lower row is from left to right, and is a control group, a comparative example 1 group and a comparative example 2 group in turn.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

Examples 1 to 5: traditional Chinese medicine composition for treating atopic dermatitis

1. The composition of the traditional Chinese medicine composition is shown in table 1:

table 1: the composition of each Chinese medicinal composition (unit: mg, g or kg)

2. The preparation method comprises the following steps:

weighing the components according to the proportion, decocting in water for 2 times, the first time for 20 minutes, the second time for 20 minutes, collecting filtrate, concentrating and drying to obtain the traditional Chinese medicine composition.

Comparative examples 1 to 2:

the formulation and preparation process are the same as in the examples, except that:

compared with the examples, the comparative example 1 has no cortex dictamni, fructus kochiae and periostracum cicadae in the medicine composition, has the main effects of dispelling wind and arresting itching and clearing damp and heat, is added aiming at the symptoms of severe pruritus of atopic dermatitis, and the comparative example 1 removes the three medicines and aims to compare and observe the itching relieving curative effect of the examples.

Comparative example 2 no seabuckthorn drug is contained in the drug composition, the main effects of the seabuckthorn drug are clearing heat and cooling blood, nourishing blood and moistening dryness, and the seabuckthorn drug is added mainly aiming at the symptoms of desquamation and lichenification caused by collateral invasion and blood invasion of atopic dermatitis due to long-term illness, and the skin moistening effect of the drug in comparative observation examples is removed in comparative example 2.

Table 2: pharmaceutical composition of comparative example (unit: mg, g or kg)

	Comparative example 1	Comparative example 2
			Chinese ephedra	6	6
Forsythia fruit	15	15
			Cicada slough	0	9
Chinese trumpet creeper	15	15
			Semen Phaseoli	30	30
Sea-buckthorn	30	0
			Cortex Dictamni Radicis	0	15
Kochia scoparia (L.) Schrad	0	15
			Radix Glycyrrhizae	9	9

Experimental example 1: investigating the effects of the Chinese medicinal composition

1 materials and methods

1.1 Experimental animals

SPF grade 6 week old male BALB/c mice, body mass (20. + -.2 g), purchased from Beijing Wintolite laboratory animal technology, inc. (license number: SYXK (Kyoto) 2016-0006). The feed is bred in an animal house of Beijing university of traditional Chinese medicine, the environment temperature is 25 +/-2 ℃, the humidity is 40-60%, and the feed can be freely drunk by drinking water every day.

1.2 drugs and reagents

2, 4-dinitrofluorobenzene (Sigma, USA), acetone (analytical pure homemade reagent), olive oil (Muelilia, spain); xylene (analytically pure reagent), absolute ethanol (analytically pure reagent), hematoxylin staining solution (Zhonghua Jinqiao, lot number 20042002), eosin staining solution (Zhonghua Jinqiao, lot number 20060901), differentiation solution (Yili, lot number 20200730), and bluing staining solution (Yili, lot number 20200528).

All the Chinese medicinal materials were purchased from Beijing Tongrentang drugstore.

Mometasone furoate gel (Specification: 0.1%, huarun Sanjiu (Nanchang) pharmaceutical Co., ltd., batch No. 2006007J)

1.3 instruments and devices

An independent aeration cage IVC rearing cage (Jiangsu. Suhang scientific and technological equipment Co., ltd.), a skin multi-probe tester (Courage + Khazakz, germany); histoCore fully automatic closed tissue hydroextractor (Leica, germany), histoCore Arcadia H paraffin embedding machine (Leica, germany), histoCore Arcadia C tissue section cold stage (Leica, germany), RM2255 fully automatic microtome (Leica, germany), HI1210 slide and HI1220 slide bakeware (Leica, germany), revolve FL inverted integral fluorescence microscope (Echo, USA).

1.4 Molding method

Reference to the literature ^[1] The method for preparing the AD mouse model by repeatedly stimulating the 2, 4-Dinitrofluorobenzene (DNFB) hapten is an animal model which has higher score and is more suitable for the AD research of traditional Chinese medicine on the traditional Chinese medicine syndrome goodness of fit at present ^[2] . The specific method comprises the following steps: the hair on the back of the mice was removed by electric hair clippers and depilatory cream before sensitization, and the area was about 2.0cm × 3.0cm. 200. Mu.L of 1% DNFB solution was dropped onto 1cm × 1cm gauze, and the solution was evenly applied to the back skin 2 times a week (i.e., days 1,4, 8, and 11). After 14d, 200. Mu.L of 0.2% DNFB solution was applied to the dorsal skin evenly for 2 times/week, and the above procedure was repeatedOperation, the model group and each drug treatment group were given the above treatments; normal group mice were smeared with an equal amount of matrix solution (acetone: olive oil = 4.

1.5 grouping and administration

(1) Grouping: 70 male BALB/c mice are divided into a normal group, a model group, a positive control group (mometasone furoate group, which is called as a control group for short), a ephedra, forsythia and phaseolus calcaratus soup original group (called as an original formula group for short), an example group and a comparative example 1-2 group (a formula of the comparative example 1 and the comparative example 2 in the application) by a random number table method, and 10 mice are divided into 7 groups.

(2) Administration: each group of mice was individually drug-intervened on day 20 of the experiment. The original prescription group, the example group and the comparative example 1-2 groups are used for gastric lavage by corresponding traditional Chinese medicine formulas. After the dosage of each group of mice is converted according to the equivalent dose coefficient reduction algorithm of the mice and the human, the dosage of the traditional Chinese medicine formula of the original formula group, the example group, the comparative example 1 group and the comparative example 2 group is unified to be 14.4 g/kg.d, and the gavage is carried out twice in the morning and at the evening for 7 days continuously. Uniformly smearing a proper amount of 0.1% mometasone furoate gel on the skin lesion of the mouse in the control group, and continuously smearing the mometasone furoate gel once a day for 7 days; meanwhile, the normal group and the model group were administered with the same amount of normal temperature distilled water for intragastric administration twice a day for 7 consecutive days.

1.6 animals were sacrificed and the material was taken

On the 28 th day of the experiment, after 4% chloral hydrate was anesthetized, the eyeball was removed and blood was taken to kill each group of mice, and skin tissue at the skin lesion was taken under aseptic conditions and fixed and stored in 4% paraformaldehyde solution.

1.7 index observations

(1) Visual inspection and scoring of skin lesions

Reference to the literature ^[3] And performing skin lesion scoring evaluation. I.e. the lesions included 4 symptoms: (1) erythema/hemorrhage; (2) drying and scabbing; (3) denudation/erosion; (4) edema. Each symptom can be rated as 0-3 according to the skin damage severity: the score 0 is asymptomatic, the score 1 is mild, the score 2 is moderate, and the score 3 is severe. The sum of all symptom scores is the skin severity, the higher the score is, the more serious the skin damage is, the maximum score is not more than 12, and statistics are made on the data. The mice in each group were compared for erythema, edema, dryness, fluid infiltration and scabbing on the backAnd waiting for AD-like skin lesion expression, and photographing and recording.

(2) Percutaneous Water loss (TWEL)

The shaved area of the back of the mouse was flattened, the skin of the mouse was lightly touched using a Tewameter TM300 probe, the percutaneous water loss rate at the skin lesion was measured using a skin meter, the measurement time was set to 20 seconds/time using a standard measurement method, and the average value was read. Maintaining relative consistency in time of each test, indoor environment, test persons and other conditions.

(3) pH value of skin

The pH value of the lesion is measured using a pH 905 probe, and the measurement is repeated 3 times each time, and the results are averaged. Maintaining relative consistency in time of each test, indoor environment, test persons and other conditions.

(4) Scratching behavior

The mice were placed in a single cage and the number of scratching of the mice within 30min was recorded and counted using a video camera. Continuous scratching of the ears and abdomen by hind limbs is considered as 1 scratching activity, and when the hind paw falls to the ground or is licked, this scratching is finished ^[4] . The movements of the forelimbs and scratching of the face and back of the hind paw were not counted.

(5) Determination of serum IgE, IL-6 and IL-10 contents

Mouse eyeball blood is picked, placed at room temperature for 5min, centrifuged to obtain serum (4 deg.C, 5000r min) ^-1 And 10 min), storing in a refrigerator at the temperature of 20 ℃ below zero, and measuring the content of IgE, IL-6 and IL-10 in the serum of the mouse by an ELISA method.

(6) Histopathological observation of skin

Fixing the skin of a sensitization area on the back of a mouse with 4% paraformaldehyde solution for 24 hours, taking a part of the skin, embedding paraffin, conventionally slicing, staining hematoxylin-eosin (HE), and observing histopathological changes under a microscope.

1.8 statistical treatment

Data analysis was performed using SPSS 25.0 software. The measurement data is expressed by mean + -SD, the comparison among groups is performed by one-way analysis of variance (ANOVA), and the comparison between every two groups is performed by t test. P <0.05 is statistically significant for the differences.

2 results of

2.1 visual inspection and assessment of lesion scores

The mice in the normal group have no skin damage in the whole experiment process; the skin lesion of the AD model group mouse presents a pathological process similar to the clinic at different time points in the replication process, namely erythema, bleeding point → skin edema and ulceration → new blood crust → blood crust hardening → blood crust shedding and new scar coexistence → skin is scattered at the erythema bleeding point, scar hyperplasia, scaling or lichenification and the like. The skin lesions of the mice of the control group, the mice of the example group and the mice of the comparative example group are improved in different degrees, wherein the curative effect of the control group and the mice of the example group is more obvious, and the bleeding points of the skin lesions, dry skin scales and lichenification lesions are obviously reduced or improved (see figure 1 in detail).

The skin damage scoring results show that the original formula group, the example group, the control group and the comparative example 1 are all obviously reduced compared with the model group, the example group is more obvious, and the curative effect is better than that of the original formula group and the comparative examples 1-2 (shown in table 3).

TABLE 3 assessment of skin damage scores

Group of	Number of samples (n)	3d	10d	20d	27d
						Normal group	6	--	--	--	--
Model set	6	1.83±0.75	3.50±1.05	5.33±1.03	3.17±1.17
						Original square group	6	1.83±0.75	2.83±0.75	4.33±1.03	1.83±0.75 **#
Group of embodiments	6	1.83±0.75	3.50±1.05	4.33±1.03	1.17±0.75 **
						Control group	6	2.00±0.90	3.33±1.03	4.50±1.05	1.00±0.63 **
Comparative example 1 group	6	2.33±0.52	4.00±1.41	4.83±1.33	2.17±0.75 *#
						Comparative example 2 group	6	2.67±0.52	3.67±1.21	5.00±1.10	2.33±0.52 ##

Note: compared with model group ^* P<0.05， ^** P<0.01; compared with the example group ^# P<0.05， ^## P<0.01

2.2 transdermal Water loss (TWEL)

TEWL is a condition that indicates water loss by measuring the water vapor pressure gradient at the skin surface, reflecting the water permeability barrier of the skin. When the barrier function is damaged, the TEWL value is increased; conversely, TEWL decreases, suggesting that the barrier is tending to repair.

As shown in table 4: the TWEL value of the skin damage area of the model group mice is obviously increased compared with that of the normal group. Compared with the model group, the original square group, the example group, the control group and the comparative examples 1-2 are all obviously reduced, and the example group is most obvious and superior to the original square group, the control group and the comparative examples 1-2.

TABLE 4 analysis of transdermal Water loss (TWEL)

Group of	Number of samples (n)	TWEL
			Normal group	6	7.49±1.55
Model set	6	48.37±6.96 ▲▲
			Original square group	6	27.07±2.95 ▲▲ ** ##
Group of embodiments	6	18.42±3.67 ▲▲ **
			Control group	6	26.76±4.24 ▲▲ ** ##
Comparative example 1 group	6	31.98±2.21 ▲▲ ** ##
			Comparative example 2 group	6	32.61±2.83 ▲▲ ** ##

Note: compared to the normal group: ^▲ P<0.05， ^▲▲ P<0.01; comparison with model group<0.05，**P<0.01; compared with the example group ^# P<0.05， ^## P<0.01

2.3 skin pH

Under physiological conditions, the pH value of the skin is in weak acidity, and the capability of the skin for resisting external erosion, elasticity, luster, moisture and the like are all in the optimal state. When the pH value is increased, the skin is easily damaged by external chemical stimulation, and on the contrary, when the skin is damaged by inflammation, rash and the like, the pH value is also changed.

As shown in table 5: compared with the normal group, the pH value of the skin damage area of the mice in the model group is obviously increased. Compared with the model group, the original square group, the example group, the control group and the comparative example 2 are all obviously reduced, and the example group is superior to the control group and the comparative example 1.

TABLE 5 skin pH analysis

Group of	Number of samples (n)	Skin pH value
			Normal group	6	6.39±0.04
Model set	6	6.94±0.05 ▲▲
			Original square group	6	6.87±0.05 ▲▲*
Group of embodiments	6	6.84±0.03 ▲▲**
			Control group	6	6.77±0.03 ▲▲**##
Comparative example 1 group	6	6.90±0.03 **#
			Comparative example 2 group	6	6.89±0.03 ▲**

Note: compared to the normal group: ^▲ P<0.05， ^▲▲ P<0.01; compared with model group ^* P<0.05，**P<0.01; compared with the example group ^# P<0.05， ^## P<0.01。

2.4 analysis of scratching behavior

Scratching behavior is one of the common indicators for assessment of the degree of itching.

As shown in table 6: the scratching behavior counts in the lesion areas of the model mice were significantly increased compared to the normal group. Compared with the model group, the original square group, the example group, the control group and the comparative examples 1-2 are all obviously reduced, and the itching relieving effect of the example group is obviously better than that of the original square group, the control group and the comparative examples 1-2.

TABLE 6 analysis of scratching behavior

Group of	Number of samples (n)	Number of scratching
			Normal group	6	5.50±1.38
Model set	6	42.67±3.33 ▲▲
			Original square group	6	31.67±4.41 ▲▲**#
Group of embodiments	6	26.83±5.71 ▲▲**
			Control group	6	34.50±4.04 ▲▲**##
Comparative example 1 group	6	36.00±3.63 ▲▲**##
			Comparative example 2 group	6	39.00±2.97 ▲▲**##

Note: compared to the normal group: ^▲ P<0.05， ^▲▲ P<0.01; compared with model group ^* P<0.05， ^** P<0.01; compared with the example group ^# P<0.05， ^## P<0.01

2.5 serum IgE, IL-6, IL-10 content

Elevated serum IgE is the most powerful cue for allergic disease. Interleukin-6 (Interleukin-6, IL-6) regulates the growth and differentiation of a variety of cells, and increased levels play a major role in the assessment of stress response, infection, chronic inflammation, and the like. IL-10 is an anti-inflammatory factor that acts to down-regulate the inflammatory response and antagonize inflammatory mediators.

As shown in table 7: compared with the normal group, the serum IgE and IL-6 of the model group mice are obviously increased, and the IL-10 content is obviously reduced; compared with the model group, the original prescription group, the example group, the control group and the comparative examples 1-2 have the advantages that serum IgE and IL-6 are obviously reduced, the IL-10 content is obviously increased, and the example group is superior to other groups.

TABLE 7 analysis of serum IgE, IL-6 and IL-10 content

Group of	Number of samples (n)	IgE(ng/ml)	IL-6(pg/ml)	IL-10(pg/ml)
					Normal group	6	52.89±5.58	15.70±3.30	205.20±6.22
Model set	6	65.53±4.09 ▲▲	34.50±1.52 ▲▲	171.23±8.72 ▲▲
					Original square group	6	57.71±4.62 ▲**	23.18±2.07 ▲▲**##	180.27±8.80 ▲▲#
Group of embodiments	6	53.38±2.02 **	18.80±1.37 ▲**	191.38±11.16 ▲**
					Control group	6	53.76±4.97 **	19.17±1.78 ▲**	176.01±3.91 ▲▲##
Comparative example 1 group	6	59.76±2.73 ▲▲*##	25.90±1.80 ▲▲**##	172.63±12.19 ▲▲##
					Comparative example 2 group	6	62.49±0.85 ▲▲##	27.98±3.57 ▲▲**##	173.51±7.44 ▲▲##

Note: compared to the normal group: ^▲ P<0.05， ^▲▲ P<0.01; compared with model group ^* P<0.05，**P<0.01; compared with the example group ^# P<0.05， ^## P<0.01。

2.6 Observation of skin histopathology

As shown in fig. 2: the normal group of mice has regular skin structure, normal cell morphology, complete and clear epithelial layer, normal and clear subcutaneous tissue structure, and no obvious inflammation expression such as vasodilatation, inflammatory cell infiltration and the like. The epidermis of the mouse in the model group is thickened, and the thickening is mainly performed on an acanthocyte layer and mild spongiform edema, the thickening and descending of an epithelial layer, the hyperplasia of skin appendages, the edema of a dermal layer, the dilatation and congestion of small blood vessels, the infiltration of lymphocytes, eosinophils and mast cells is obvious, and the infiltration of epidermal hyperkeratosis, inflammatory cells and the like are accompanied. Compared with the model group, the original prescription group, the example group, the control group and the comparative examples 1-2 can improve the pathological changes of the skin of the model mouse in different degrees, such as the thinning of epithelial hyperplasia, the disappearance of edema of the dermis layer, the obvious reduction of small blood vessel dilatation hyperemia and inflammatory infiltration, and the like, and the original prescription group, the example group, the control group and the comparative examples 1-2 are more obvious.

Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that modifications and improvements can be made thereto without departing from the scope of the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Attached: reference to the literature

[1]Kim TH,Kim GD,Ahn HJ,et al.The inhibitory effect of naringenin on atopic dermatitis induced by DNFB in NC/Nga mice[J].Life Sci.2013 Oct 10；93(15):516-524.

[2] Zhang Rubia, shiyanmei, li XiuMin, etc. based on the characteristics of Chinese and Western medicine clinical syndromes, animal model analysis of atopic dermatitis [ J ]. Chinese traditional medicine J, 2021,46 (4): 762-766.

[3]Stalder JF,Taieb A,Atherton DJ,et al.Severity scoring of atopic dermatitis:the SCORAD index.European task force on atopic dermatitis[J].Dermatology,1993,186(1):23-31.

[4]Yasushi Kuraishi,Tetsuro Nagasawa,Kazuko Hayashi,Masamichi Satoh.Scratching behavior induced by pruritogenic but not algesiogenic agents in mice[J].European Journal of Pharmacology,1995,275(3):229-233.

Claims (7)

1. The traditional Chinese medicine composition for treating atopic dermatitis is characterized by being prepared from the following traditional Chinese medicines by weight: 3-10 g of ephedra, 5-20 g of forsythia, 5-15 g of cicada slough, 5-20 g of trumpet creeper, 20-40 g of phaseolus calcaratus, 20-35 g of sea buckthorn, 5-20 g of cortex dictamni, 5-20 g of fructus kochiae and 5-12 g of raw liquorice.

2. The traditional Chinese medicine composition for treating atopic dermatitis is characterized by being prepared from the following traditional Chinese medicines by weight: 3-9 g of ephedra, 10-15 g of fructus forsythiae, 5-15 g of periostracum cicada, 10-15 g of trumpet creeper, 20-40 g of phaseolus calcaratus, 25-35 g of sea buckthorn, 10-15 g of cortex dictamni, 10-15 g of fructus kochiae and 5-15 g of raw liquorice.

3. The traditional Chinese medicine composition of claim 1, wherein the traditional Chinese medicine composition is prepared from the following traditional Chinese medicines by weight: 4-8 g of ephedra, 13-18 g of forsythia, 6-12 g of cicada slough, 13-18 g of trumpet creeper, 25-35 g of phaseolus calcaratus, 25-35 g of sea buckthorn, 13-18 g of cortex dictamni, 13-18 g of fructus kochiae and 6-10 g of raw liquorice.

4. The traditional Chinese medicine composition of claim 1, wherein the traditional Chinese medicine composition is prepared from the following traditional Chinese medicines by weight: 6 g of ephedra, 15 g of forsythia, 9 g of cicada slough, 15 g of trumpet creeper, 30 g of phaseolus calcaratus, 30 g of sea buckthorn, 15 g of cortex dictamni, 15 g of fructus kochiae and 9 g of raw liquorice.

5. A preparation method of a traditional Chinese medicine composition for treating atopic dermatitis is characterized by comprising the following steps: weighing the traditional Chinese medicines according to the proportion of the traditional Chinese medicine composition of any one of claims 1 to 4, uniformly mixing, extracting with water or lower alcohol, filtering, collecting filtrate, concentrating and drying to obtain the traditional Chinese medicine composition.

6. Use of a Chinese medicinal composition according to any one of claims 1 to 4 in the manufacture of a medicament for the treatment of atopic dermatitis.

7. The use of claim 6, wherein the medicament is a formulation comprising a combination of Chinese herbs and a pharmaceutically acceptable carrier.

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