A kind of external anti-atopic dermatitis essential oil, preparation method and application

Abstract

The invention discloses a kind of external anti-atopic dermatitis three grass antipruritic formula essential oil, preparation method and application, the formula essential oil comprises the following raw materials: 2-5mL of lithospermum erythrorhizon essential oil, 3-5mL of Ecliptae essential oil, 2-5mL of madder essential oil, 2-5mL of schisandra chinensis essential oil, 2-4mL of ebony essential oil, 2-4g of borneol, all medicines are used together, and the whole party plays the effects of cooling blood and activating blood circulation, removing rash and removing spots, nourishing yin and nourishing body fluid, and anti-allergic. The essential oil of the present invention is extracted by pure Chinese medicine, treats both the symptoms and the root cause, has remarkable drug effect, few side effects, low recurrence rate, high use compliance, and the raw materials of Chinese medicine of the present invention are abundant, the preparation process is simple, the cost is low and the effect is good, it is an innovation in the treatment of atopic dermatitis medicine, and has a good market prospect.

Description

Technical Field

[0001] The invention relates to a formula essential oil for external use against atopic dermatitis, a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine. Background Art

[0002] Atopic dermatitis (AD) is a chronic, recurrent, pruritic skin disease caused by an imbalanced immune response. It is mainly characterized by severe itching and eczematous rashes, often accompanied by symptoms such as redness, dryness, flaking, and erosion of the skin. AD occurs in all age groups, and 30% of children and 8% of adult AD patients are often accompanied by symptoms such as anxiety, sleep disorders, and depression, which seriously affect the physical and mental health and quality of life of patients. At present, Western medicine treatment of AD mostly uses local glucocorticoid hormones, immunomodulators and antihistamines, which can temporarily relieve the clinical symptoms of patients, but long-term use is ineffective and has large side effects and is not easy to tolerate. Traditional Chinese medicine believes that the etiology and pathology of AD are mostly spleen and lung qi deficiency, hyperactivity of heart fire, and exogenous rheumatism evil that accumulates in the body and turns into heat. The traditional Chinese medicine thinking based on pathological dialectical treatment has a definite effect in the treatment of AD, with few side effects, and has significant advantages. Under the guidance of traditional Chinese medicine theory, oral Chinese medicine can improve the physical constitution of AD patients, improve sleep quality, relieve clinical symptoms such as nervous anxiety, and has achieved good results. However, oral Chinese medicine is inconvenient to use and has a poor taste, and long-term use is a huge challenge for patients. External Chinese medicine prescriptions are easy to carry, have definite efficacy, and can directly reach the lesion, which has obvious clinical advantages. Therefore, it is of great significance to explore innovative Chinese medicine external treatment drugs that are convenient and efficient for the treatment of atopic dermatitis. Summary of the invention

[0003] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a three-herb antipruritic formula essential oil for external use against atopic dermatitis, a preparation method and application thereof, which can effectively treat atopic dermatitis.

[0004] In order to achieve the above object, one of the technical solutions of the present invention is:

[0005] A three-herb antipruritic essential oil for external use against atopic dermatitis comprises the following raw materials: 2-5 mL of lithospermum erythrorhizon essential oil, 3-5 mL of Eclipta prostrata essential oil, 2-5 mL of madder essential oil, 2-5 mL of schisandra chinensis essential oil, 2-4 mL of ebony essential oil and 2-4 g of borneol.

[0006] Preferably, the essential oil comprises the following raw materials: 3 mL of Lithospermum officinale essential oil, 3 mL of Eclipta prostrata essential oil, 3 mL of Rubia cordifolia essential oil, 3 mL of Schisandra chinensis essential oil, 2 mL of Prunus mume essential oil, and 3 g of borneol.

[0007] One of the technical solutions of the present invention is: a method for preparing the essential oil, comprising the following steps:

[0008] (1) Preparation of essential oils

[0009] a. Lithospermum essential oil, Eclipta essential oil, Schisandra essential oil, Plum essential oil: weigh a certain amount of Lithospermum, Eclipta, Schisandra or Plum, soak in ultrapure water, add zeolite and reflux extraction to collect Lithospermum essential oil, Eclipta essential oil, Schisandra essential oil or Plum essential oil;

[0010] b. Rubia essential oil: crush the dried madder, weigh a certain amount of madder, soak it in ultrapure water, add zeolite and reflux to extract, to obtain a yellow flaky solid, collect the yellow flaky solid and dry it; dissolve the dried flaky solid in sweet almond base oil to prepare madder essential oil;

[0011] (2) Take lithospermum erythrorhizon essential oil, Eclipta prostrata essential oil, Rubia cordifolia essential oil, Schisandra chinensis essential oil, and Prunus mume essential oil, mix them, add an equal volume of sweet almond base oil to dilute the mixed essential oil, add borneol to dissolve and mix well.

[0012] Furthermore, the lithospermum officinale powder or Eclipta prostrata powder is first crushed to 4-6 mm, and then soaked in ultrapure water.

[0013] Furthermore, the amount of ultrapure water added is 8-12 times by weight.

[0014] Further, the soaking time is 2-3 h.

[0015] Furthermore, the reflux extraction temperature is 80-90°C.

[0016] Furthermore, in step b, the ratio of the dried flaky solid to the sweet almond base oil is 20-40 mg: 4-8 mL.

[0017] Preferably, in step b, the ratio of the dried flaky solid to the sweet almond base oil is 30 mg:6 mL.

[0018] One of the technical solutions of the present invention is: an application of the essential oil in preparing an anti-atopic dermatitis drug.

[0019] Beneficial Effects

[0020] The famous prescriptions for treating wind-related skin diseases in successive dynasties all follow the method of "treating wind first and treating blood, and blood circulation will eliminate wind by itself". The three-herb antipruritic formula essential oil of the present invention is composed of lithospermum, madder, Ecliptae, Schisandra, Plum, and Borneol. Among them, sweet and cold salty lithospermum is the monarch, and the smell is fragrant, and there is the power of clearing and dispersing, clearing heat and cooling blood, promoting blood circulation and detoxifying, and removing rash and spots. With madder and Ecliptae as ministers, the effect of lithospermum cooling blood and dredging menstruation is enhanced, which can not only promote blood circulation and remove blood stasis, but also nourish the liver and kidney. With Plum and Schisandra as assistants, Plum is sour and astringent, produces body fluid, and astringes the lungs. Pharmacological experiments have shown that it has an anti-allergic effect. Schisandra is sour, sweet and warm, invigorates qi and produces body fluid, nourishes the kidney and calms the heart, astringes and consolidates, and the medicinal property is sweet and warm, which can help all medicines to reach the whole body, to help lithospermum nourish yin and cool blood. With bitter and slightly cold Borneol as a messenger, its qi is scattered, and there is the power of clearing and dispersing. The combination of these medicines has the effects of cooling blood and activating blood circulation, clearing rashes and removing spots, nourishing yin and body fluid, and resisting allergies. It is effectively used to treat atopic dermatitis.

[0021] The essential oil of the invention is extracted from pure Chinese medicine, has a therapeutic effect on both the symptoms and the root cause, has significant efficacy, few side effects, low recurrence rate, and high use compliance. In addition, the Chinese medicine of the invention has abundant raw materials, a simple preparation process, low cost and good effect, is an innovation in drugs for treating atopic dermatitis, and has a good market prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The effects of different concentrations of essential oils on cell activity (n=5).

[0023] Figure 2 The effect of three-herb antipruritic formula essential oil on organ indexes of AD mice (n=6). Compared with the blank group, ## P <0.01; compared with the model group, ** P <0.01.

[0024] Figure 3 The effect of three-herb antipruritic formula essential oil on the scratching frequency of AD mice after enhanced stimulation (n=6). Compared with the blank group, ## P <0.01; compared with the model group, ** P <0.01.

[0025] Figure 4 The effect of three grass antipruritic formula essential oil on ear swelling of AD mice (n=6). Compared with the blank group, ## P <0.01; compared with the model group, ** P <0.01.

[0026] Figure 5 This is the effect of three-herb antipruritic formula essential oil on the skin pathological state of AD mice.

[0027] Figure 6 The effect of three grass antipruritic formula essential oil on the levels of TNF-α and IL-6 in the serum of AD mice (n=6). Compared with the blank group, ##P <0.01; compared with the model group, ** P <0.01.

[0028] Figure 7 The effect of three-herb antipruritic formula essential oil on the serum IgE level of AD mice (n=6). Compared with the blank group, ## P <0.01; compared with the model group, ** P <0.01. DETAILED DESCRIPTION

[0029] The specific implementation modes of the present invention are further described in detail below in conjunction with the embodiments.

[0030] Example 1: Preparation of essential oils

[0031] Lithospermum essential oil: Grind the dried lithospermum into 4-6 mm fragments, weigh a certain amount of lithospermum fragments into a round-bottom flask, add 10 times the weight of ultrapure water, soak for 2 h, add zeolite and place the round-bottom flask in a heating mantle, connect the essential oil extraction device, and reflux extraction at 80-90°C (the condenser temperature is maintained at 15-25°C) until no more essential oil droplets appear, and collect the lithospermum essential oil.

[0032] Eclipta prostrata essential oil: Crush the dried Eclipta prostrata into 4-6 mm segments, weigh a certain amount of the broken Eclipta prostrata segments into a round-bottom flask, add 10 times the weight of ultrapure water, soak for 2 h, add zeolite and place the round-bottom flask in a heating mantle, connect the essential oil extraction device, and reflux extraction at 80-90°C (the condenser temperature is maintained at 15-25°C) until no more essential oil droplets appear, and collect the Eclipta prostrata essential oil.

[0033] Rubia essential oil: Grind the dried madder into 4-6 mm fragments, weigh a certain amount of the madder fragments into a round-bottom flask, add 10 times the weight of ultrapure water, soak for 2 h, add zeolite and place the round-bottom flask in a heating jacket, connect the essential oil extraction device, and reflux extraction at 80-90°C (the condenser temperature is maintained at 15-25°C) to obtain a yellow flaky solid, collect the yellow flaky solid and dry it with anhydrous sodium sulfate; weigh 30 mg of the dry flaky solid and dissolve it in 6 mL of sweet almond base oil to prepare madder essential oil.

[0034] Schisandra essential oil: Weigh a certain amount of Schisandra in a round-bottom flask, add 8 times the weight of ultrapure water, soak for 3 hours, add zeolite and place the round-bottom flask in a heating jacket, connect the essential oil extraction device, and reflux extraction at 80-90°C (the condenser temperature is maintained at 15-25°C) until no more essential oil droplets appear, and collect the Schisandra essential oil.

[0035] Black plum essential oil: Weigh a certain amount of black plum into a round-bottom flask, add 12 times the weight of ultrapure water, soak for 2 hours, add zeolite and place the round-bottom flask in a heating jacket, connect the essential oil extraction device, and reflux extraction at 80-90°C (the condenser temperature is maintained at 15-25°C) until no more essential oil droplets appear, and collect the black plum essential oil.

[0036] Embodiment 2: Three grass antipruritic formula essential oil

[0037] A three-herb antipruritic essential oil for external use against atopic dermatitis comprises the following raw materials: 3 mL of lithospermum erythrorhizon essential oil, 3 mL of eclipta prostrata essential oil, 3 mL of madder essential oil, 3 mL of schisandra chinensis essential oil, 2 mL of ebony essential oil and 3 g of borneol.

[0038] The preparation method is as follows: 3 mL of lithospermum erythrorhizon essential oil, 3 mL of Eclipta prostrata essential oil, 3 mL of madder essential oil, 3 mL of schisandra chinensis essential oil, and 2 mL of ebony essential oil are measured and mixed, and after diluting with an equal volume of sweet almond base oil (14 mL) of the mixed essential oil, 3 g of borneol is added to dissolve and mix well.

[0039] Example 3: Three Herbs Antipruritic Formula Essential Oil

[0040] The invention discloses an external anti-atopic dermatitis three-herb antipruritic essential oil, comprising the following raw materials: 2 mL of lithospermum erythrorhizon essential oil, 5 mL of eclipta root essential oil, 5 mL of madder essential oil, 2 mL of schisandra chinensis essential oil, 3 mL of ebony essential oil and 4 g of borneol.

[0041] The preparation method is the same as that in Example 2.

[0042] Embodiment 4: Three grass antipruritic formula essential oil

[0043] A three-herb antipruritic essential oil for external use against atopic dermatitis comprises the following raw materials: 5 mL of lithospermum erythrorhizon essential oil, 4 mL of eclipta prostrata essential oil, 2 mL of madder essential oil, 5 mL of schisandra chinensis essential oil, 4 mL of ebony essential oil and 2 g of borneol.

[0044] The preparation method is the same as that in Example 2.

[0045] Experimental Example 1: Cell Experiment

[0046] 1 Experimental Materials and Reagents

[0047] Mouse mononuclear RAW264.7 macrophages and fetal bovine serum (FBS) were purchased from Saiye Biotechnology Co., Ltd.; MTT and DMEM culture medium were purchased from Beijing Solebow Technology Co., Ltd.; mouse tumor necrosis factor-α (TNF-α) ELISA kit was purchased from MABTECH Swedish Biotechnology Co., Ltd.; lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.; Griess kit was purchased from Shanghai Biotech Co., Ltd.

[0048] 2 Experimental methods

[0049] 2.1 Cell culture

[0050] RAW264.7 macrophages were cultured in DMEM medium (high glucose) containing 10% (v / v) FBS in a constant temperature incubator at 37°C and 5% CO2. The medium was replaced every 24 hours. The cells were passaged when they grew to 80-90%. After the cells were stable, the formula essential oils were analyzed using the pro-inflammatory mediator NO and the pro-inflammatory factor TNF-α as indicators.

[0051] 2.2 Essential oil preparation

[0052] The essential oil prepared in Example 1 of the present invention (the dried flaky solid of the Rubia cordifolia essential oil was used as a sample) was prepared into single stock solutions of Lithospermum officinale (A), Rubia cordifolia (B), Eclipta prostrata (C), Schisandra chinensis (D) and Prunus mume (E) using DMEM culture medium, and the stock solutions were mixed in a mass ratio of A:B:C:D:E of 3:3:3:3:2 to prepare formula essential oil stock solutions of different concentrations.

[0053] 2.3 MTT method to evaluate the effect of essential oil formula on cell viability

[0054] RAW264.7 macrophages were divided into a cell-free blank group, a normal control group, and a formula essential oil administration group. Each group was replicated for 5 wells. Cells in the logarithmic growth phase were used to inoculate 5×10 4 100 μL of DMEM medium was added to the cell-free blank group and the normal control group, respectively; 100 μL of drug-containing DMEM medium with a total concentration of 23.5, 47.0, 70.5, 94.0, and 117.5 μg / mL of essential oil was added to the drug-treated group, respectively (taking 47.0 μg / mL as an example to illustrate the composition: containing 10.0 μg / mL of A, B, C, and D, and 7.0 μg / mL of E, respectively), and after continued cultivation for 24 h, 10 μL of 5 mg / mL MTT solution was added, and the culture was continued for 4 h. The solution in the 96-well plate was discarded, 150 μL of dimethyl sulfoxide (DMSO) was added, and the absorbance (OD) was measured at 490 nm.

[0055] Cell survival rate = [OD(drug group)- OD(blank group)] / [OD(control group)- OD(blank group)]×100%.

[0056] 2.4 Griess method for detecting NO content

[0057] RAW264.7 macrophages were divided into essential oil formula administration groups (including A, B, C, D, E, A+B, A+B+C, A+B+C+D, A+B+C+D+E), blank control group (CON), model group (LPS) and positive drug dexamethasone group (DEX), with 3 wells in each group. Cells in logarithmic growth phase were taken, and 2×10 5 Each well was inoculated in a 24-well plate and cultured overnight in a constant temperature incubator at 37°C and 5% CO2. The supernatant was discarded and 500 μL DMEM medium was added to the CON group, 500 μL DMEM medium containing 1.0 μg / mL LPS was added to the LPS group, and 500 μL essential oil treatment groups were added with 42.0 μg / mL A, 42.0 μg / mL B, 42.0 μg / mL C, 42.0 μg / mL D, 42.0 μg / mL E, 42.0 μg / mL (21.0 μg / mL A + 21.0 μg / mL B), 42.0 μg / mL (14.0 μg / mL A + 14.0 μg / mL B + 14.0 μg / mL C), 42.0 μg / mL (10.5 μg / mL A + 10.5 μg / mL B + 10.5 500 μL of DMEM medium containing 5 µg / mL DEX was added to the DEX group, and the culture was continued for 24 h. The supernatant was taken for later use. The NO concentration was determined using the Griess kit. 50 μL of cell culture supernatant was taken, and 50 μL of Griess I and 50 μL of Griess II were added respectively. The OD value was measured at a wavelength of 540 nm. A series of concentration samples were prepared with nitrite standards to make a standard curve, and the NO concentration was calculated by substituting the standard curve into the standard curve.

[0058] 2.5 ELISA for detection of TNF-α levels

[0059] Take the cell supernatant from part 2.4 and determine the TNF-α content in the cell supernatant according to the instructions of the TNF-α detection kit.

[0060] 3 Experimental results

[0061] 3.1 Effects of formulated essential oils on cell viability

[0062] The results are as follows Figure 1 As shown in Figure 2, compared with the control group, when the administration concentration increased to 117.5 μg / mL, the normal growth of cells was affected ( P <0.01), when the administration concentration was ≤94.0 μg / mL, there was almost no effect on cell survival rate. In view of the low solubility of high concentration with culture medium, 42.0 μg / mL was finally selected as the total concentration of essential oil for subsequent experiments.

[0063] 3.2 Effects of essential oil formulas on NO production in LPS-induced RAW264.7 cells

[0064] NO is an important regulatory factor in inflammatory and immune responses, and is produced by L-arginine catalyzed by nitric oxide synthase (NOS). During the course of immune inflammation, abnormal increases in NO levels in the body can activate the NF-κB signaling pathway and induce the production of pro-inflammatory factors such as TNF-α and IL-1β. Increased TNF-α can activate iNOS and produce more NO, thus forming a positive feedback loop and aggravating the inflammatory response. Therefore, in the LPS-induced macrophage model, the degree of NO inhibition is closely related to good anti-inflammatory efficacy. Based on this, the NO inhibitory effects of various essential oil formulas were analyzed. The results are shown in Table 1. The NO content in the LPS model group was much higher than that in the blank control group, and there was a significant difference ( P <0.01), indicating that the model was successfully constructed. Compared with the model group, when DEX and each single essential oil were added, the NO secretion was reduced ( P <0.01), but with the addition of single essential oils, the secretion of NO decreased significantly, and the compound three-herb antipruritic essential oil composed of A+ B+ C+D+E had the best effect.

[0065] 3.3 Effects of essential oil formulas on LPS-induced TNF-α in RAW264.7 cells

[0066] Inflammation is the body's natural defense mechanism. Generally, the severity of the disease is closely related to the amount of cytokines released by the cytokine storm. TNF-α is the earliest multifunctional cytokine produced in acute inflammation. Based on this, the anti-inflammatory effect of the essential oil formula was investigated using TNF-α as an indicator. The results are shown in Table 1. In the LPS model group, TNF-α increased sharply, with a significant difference ( P <0.01). Compared with the model group, the secretion of TNF-α decreased after administration of DEX and each single essential oil ( P<0.01), and as the single essential oils were added in sequence, the inhibitory effect of TNF-α was significantly enhanced, and the compound three-herb antipruritic essential oil composed of A+ B+ C+D+E had the best effect. It is also worth mentioning that the TNF-α effect of both single and combined essential oils is better than that of the positive drug group dexamethasone, highlighting the excellent pharmacodynamic effect of essential oils in anti-inflammatory and immunomodulation. Based on this, in subsequent in vivo experiments, the present invention selected the three-herb antipruritic formula essential oil composed of A+ B+ C+D+E for further research.

[0067] Table 1 Effects of essential oils in the formula on NO levels and TNF-α in cells (n=5)

[0068] Group NO (µmol / L) TNF-α (ng / mL) Control group 0.50±0.12 0.63 ±0.20 Model Group <![CDATA[7.00±2.11 ## ]]> <![CDATA[20.67±1.73 ## ]]> Positive group <![CDATA[1.82±0.98 ** ]]> <![CDATA[14.41 ±0.63 ** ]]> A <![CDATA[2.85±0.10 ** ]]> <![CDATA[6.18±0.30 ** ]]> B <![CDATA[3.10±0.21 ** ]]> <![CDATA[7.95±0.37 ** ]]> C <![CDATA[4.67±0.99 ** ]]> <![CDATA[10.30 ±1.03 ** ]]> D <![CDATA[4.30±0.75 ** ]]> <![CDATA[14.37±2.25 ** ]]> E <![CDATA[4.79±1.52 ** ]]> <![CDATA[13.30±1.09 ** ]]> A+B <![CDATA[2.26±0.42 ** ]]> <![CDATA[4.86 ±0.37 ** ]]> A+B+C <![CDATA[1.61±0.42 ** ]]> <![CDATA[3.69±0.43 ** ]]> A+B+C+D <![CDATA[1.20±0.45 ** ]]> <![CDATA[2.85 ±0.76 ** ]]> A+B+C+D+E <![CDATA[0.86±0.05 ** ]]> <![CDATA[2.40±0.16 ** ]]>

[0069] Note: Compared with the control group, ## P <0.01; compared with the model group, ** P <0.01.

[0070] Experimental Example 2: Animal Experiment

[0071] 1 Experimental Materials

[0072] 1.1 Experimental animals

[0073] BALB / c mice, SPF grade, 30 males, weighing 18-22 g, were provided by Shandong Jinan Pengyue Experimental Animal Breeding Co., Ltd., license number: SCXK (Lu) 20190003. The experimental unit use license number: SCXK (Yu) 2020-0004. They were raised in a constant temperature environment of 25°C, given sterile pure water and feed, and alternating day and night lighting.

[0074] 1.2 Experimental reagents and drugs

[0075] The three-herb antipruritic formula essential oil prepared in Example 2 of the present invention. Dinitrochlorobenzene (DNCB) was purchased from Sigma-Aldrich Company, batch number: BCCD8094; hydrocortisone butyrate cream was purchased from Tianjin Jinyao Pharmaceutical Co., Ltd., with the national medicine standard H10940095. Acetone was purchased from Yantai Shuangshuang Chemical Co., Ltd.; olive oil was purchased from Betis Olive Oil Company of Spain.

[0076] 2 Modeling and drug administration

[0077] The mice were randomly divided into 5 groups, namely, blank group (control group), model group, positive drug group, low-dose essential oil group, and high-dose essential oil group, with 6 mice in each group. After one week of adaptive feeding, the mice were shaved on the back, and 100 μL of 1% DNCB acetone-olive oil solution (4:1) was applied to the shaved area continuously on the first and second days (first prepare an acetone-olive oil solution with a volume ratio of 4:1, then weigh 200 mg of DNCB powder and dissolve it in 20 mL of acetone-olive oil (4:1, v / v) solution to obtain 1% DNCB acetone-olive oil solution). The blank group was not applied. The appearance of scabs, tissue fluid exudation, redness and swelling at the applied area indicated that the model was successfully established. On the 5th day, 20 μL of 0.5% DNCB acetone-olive oil solution (4:1) was applied to the inside and outside of the right ear of each group of mice for stimulation once. Drug treatment was given on the 5th to 8th day. The low-dose group was applied with 0.01 mL of compound essential oil, the high-dose group was applied with 0.02 mL, and the positive group was applied with 0.01 g of hydrocortisone butyrate cream. The model group and the blank group were not treated. Subsequently (starting from the 9th day), stimulation was continued once, and four medications were given (on the day of stimulation and the following three days). These 4 days were one enhanced stimulation cycle, and a total of 4 cycles were performed. The samples were collected on the day when the 4th cycle ended. After the 3rd and 4th enhanced stimulations, the number of times the mice in each group scratched the skin of the right ear, neck, and back within 10 minutes was recorded, and continuous scratching was counted as 1 time. 5 hours after the fourth enhanced stimulation, the thickness of the left and right ears of each group of mice was measured with a vernier caliper at the same position. The measurement was repeated 3 times in parallel to calculate the swelling degree of the mouse ears (ear swelling thickness = right ear thickness - left ear thickness). The mice were weighed before sampling, and dissected after blood was taken, and their spleen tissue was weighed. The organ index was obtained by dividing spleen weight by body weight. The blood was centrifuged and serum was obtained to determine the content of TNF-a, IL-6 and IgE; and pathological sections were made on the skin of the mouse lesions to observe the pathological changes.

[0078] 3 Experimental Results

[0079] 3.1 Effects of three-herb antipruritic formula essential oil on organ indexes of atopic dermatitis mice

[0080] In the DNCB-induced atopic dermatitis mouse model, hydrocortisone butyrate cream was used as the positive drug to compare the effects of high and low doses of three-herb antipruritic formula essential oil on animal organ indexes. Figure 2 As shown in Figure 2, after modeling, the lung tissue organ index of the model group increased significantly ( P <0.01); after administration, the organ indexes of the positive drug group, low essential oil dose group and high essential oil dose group were significantly reduced ( P <0.01), and there was no significant difference between the high and low essential oil dosage groups and the positive drug group, indicating that the three-herb antipruritic formula essential oil can effectively reduce the swelling of atopic dermatitis mice.

[0081] 3.2 Effect of three-herb antipruritic formula essential oil on the frequency of scratching in mice with atopic dermatitis

[0082] Skin itching is a typical feature of atopic dermatitis, so after a few days of treatment, the number of times each group of mice scratched the right ear, neck, and back skin within 10 minutes after the third and fourth booster stimulation was analyzed to examine the antipruritic effect of essential oils. Figure 3 As shown in Figure 2, after enhanced stimulation, the model group mice showed significant scratching compared with the blank group mice ( P <0.01, while the mice in the positive drug and high- and low-dose essential oil groups showed significant improvement ( P <0.01, and the high-dose essential oil group was superior to the positive drug group. At the same time, it is worth noting that after the fourth stimulation, the number of scratching times of mice in the high- and low-dose essential oil groups was significantly reduced compared to the third stimulation ( P <0.05), indicating that the three-herb antipruritic formula essential oil has excellent antipruritic effect.

[0083] 3.3 Effect of three-herb antipruritic formula essential oil on ear swelling in mice with atopic dermatitis

[0084] Ear swelling is an important indicator to measure the severity of animal allergies, so 5 hours after the fourth stimulation, the thickness of the same part of the left and right ears of each group of mice was measured with a vernier caliper to calculate the ear swelling of the mice. Figure 4 As shown, the degree of right ear swelling of mice in the model group was significantly higher than that in the blank group ( P <0.01); compared with the model group mice, the degree of right ear swelling in the positive drug group and the high and low dose groups of the three-herb antipruritic formula essential oil was significantly reduced ( P <0.01), and both the high and low dose essential oil groups were superior to the positive drug group.

[0085] 3.4 Effect of three-herb antipruritic formula essential oil on pathological changes of skin tissue in mice with atopic dermatitis

[0086] The epidermal cells of mice in the normal blank group were arranged in an orderly manner, with normal thickness and clear layers, and no obvious edema and inflammatory cell infiltration. Compared with the blank group, the epidermis of mice in the model group was significantly thickened, hyperkeratinized, acanthosis, edema of the dermis, and accompanied by a large number of inflammatory cell infiltrations. After treatment with the positive drug and the high and low dose essential oil groups, compared with the model group, tissue edema, inflammatory cell infiltration, keratinization and other phenomena were significantly improved, indicating that the formula essential oil can significantly improve the pathological state of atopic dermatitis ( Figure 5 ).

[0087] 3.5 Effect of three-herb antipruritic formula essential oil on TNF-α and IL-6 in the serum of atopic dermatitis mice

[0088] TNF-α in serum can promote lymphocytes to enter the body, release a large number of cytokines, and aggravate the body's inflammatory response; IL-6 is an important cytokine that regulates the body's immunity, which can induce the formation of C-reactive protein and the aggregation of inflammatory cells, thereby releasing more inflammatory mediators and aggravating the disease. Therefore, the levels of TNF-α and IL-6 in serum are important indicators for measuring the degree of atopic dermatitis. Based on this, TNF-α and IL-6 in mouse serum were analyzed, and the results are as follows: Figure 6 As shown in the figure, compared with the blank group, the levels of TNF-α and IL-6 in the model group mice were significantly increased ( P <0.01); after administration, compared with the model group, the positive drug group and the high and low dose groups of Sancao antipruritic essential oil reduced the contents of both ( P <0.01), and a significant callback phenomenon was observed, indicating that the three-herb antipruritic formula essential oil effectively controlled the inflammatory response in mice by inhibiting the expression of cytokines.

[0089] 3.6 Effect of three-herb antipruritic formula essential oil on serum IgE in atopic dermatitis mice

[0090] The causes of AD are complex and are related to genetics, immunity, and impaired skin barrier function. Multiple causes work together with allergens to activate immune pathways, leading to excessive release of IgE by B cells, which plays an important role in the pathogenesis of AD. Therefore, the IgE content in mouse serum is an important indicator for measuring the severity of AD. Figure 7 As shown, compared with the blank group, the IgE content of mice in the model group was significantly increased ( P <0.01); compared with the model group, the high and low dose groups of Sancao antipruritic essential oil significantly reduced the content of IgE ( P <0.01), and there was no significant difference between the positive drug group and the blank group, showing the anti-allergic efficacy of the three-herb antipruritic formula essential oil.

[0091] Experimental Example 3: Clinical Application

[0092] On the basis of animal experiments to ensure safety, clinical trials have shown very good results. The relevant information is as follows:

[0093] Thirty patients with atopic dermatitis diagnosed in the outpatient clinic, including 14 males and 16 females, were treated with a dose of 50 µL / cm 2 Apply the amount of the drug to the affected area twice a day for two weeks to evaluate the efficacy. Efficacy evaluation criteria: Significantly effective: the skin lesion area recovers by 80% or more, and symptoms such as itching disappear or basically disappear; Effective: The skin lesion area recovers by less than 80%, 30% or more, and symptoms such as itching are alleviated; Ineffective: The skin lesion recovers by less than 30%, and symptoms such as itching are not improved.

[0094] The results showed that after two weeks of treatment, 14 out of 30 patients showed significant therapeutic effects, accounting for 46.7% (significantly effective); 15 patients had reduced itching and obvious recovery of skin lesions, accounting for 50% (effective); there was 1 ineffective case, accounting for 3.3% (ineffective); the total effective rate was as high as 96.7%, indicating that the three-herb antipruritic formula essential oil used in the present invention has a good anti-atopic dermatitis effect.

[0095] in conclusion

[0096] From the above experimental results, it can be seen that the invention has abundant raw materials, simple and convenient production and preparation, good therapeutic effect, and has obvious effects of cooling blood and activating blood circulation, clearing rashes and removing spots, and anti-allergic. It is an innovation in the treatment of atopic dermatitis drugs and has a good market prospect.

Claims

1. A formula essential oil for external use against atopic dermatitis, characterized in that: The invention is composed of the following raw materials: 2-5 mL of lithospermum erythrorhizon essential oil, 3-5 mL of Eclipta prostrata essential oil, 2-5 mL of madder essential oil, 2-5 mL of schisandra chinensis essential oil, 2-4 mL of ebony essential oil, and 2-4 g of borneol; The preparation method of the madder essential oil comprises: crushing dried madder, weighing a certain amount of madder, adding ultrapure water to soak, refluxing extraction to obtain a yellow flaky solid, collecting the yellow flaky solid and drying it; dissolving the dried flaky solid in a base oil to prepare the madder essential oil; The ratio of dry flaky solid to base oil is 20-40 mg: 4-8 mL.

2. The essential oil according to claim 1, characterized in that It is composed of the following raw materials: 3 mL of Lithospermum officinale essential oil, 3 mL of Eclipta prostrata essential oil, 3 mL of Rubia cordifolia essential oil, 3 mL of Schisandra chinensis essential oil, 2 mL of Prunus mume essential oil, and 3 g of borneol.

3. A method for preparing the essential oil according to claim 1 or 2, characterized in that: The following steps are involved: (1) Preparation of essential oils a. Lithospermum essential oil, Eclipta essential oil, Schisandra essential oil, Plum essential oil: weigh a certain amount of Lithospermum, Eclipta, Schisandra or Plum, soak in ultrapure water, reflux extraction, collect Lithospermum essential oil, Eclipta essential oil, Schisandra essential oil or Plum essential oil; b. Madder essential oil: crushed dried madder, weighed a certain amount of madder, soaked in ultrapure water, refluxed to obtain a yellow flaky solid, collected the yellow flaky solid and dried; the dried flaky solid was dissolved in a base oil to prepare madder essential oil; (2) Take lithospermum erythrorhizon essential oil, Eclipta prostrata essential oil, Rubia cordifolia essential oil, Schisandra chinensis essential oil, and Prunus mume essential oil, mix them, add an equal volume of carrier oil to dilute the mixed essential oil, add borneol to dissolve and mix well.

4. The preparation method according to claim 3, characterized in that: The lithospermum officinale powder or Eclipta prostrata powder is first crushed into 4-6 mm, and then soaked in ultrapure water.

5. The preparation method according to claim 3, characterized in that: The amount of ultrapure water added is 8-12 times by weight.

6. The preparation method according to claim 3, characterized in that: The soaking time is 2-3 hours.

7. The preparation method according to claim 3, characterized in that: The reflux extraction temperature is 80-90°C.

8. The preparation method according to claim 3, characterized in that: The ratio of the dry flaky solid to the base oil in step b is 20-40 mg: 4-8 mL.

9. The preparation method according to claim 8, characterized in that: The ratio of the dried flaky solid to the base oil in step b is 30 mg:6 mL.

10. Use of the essential oil according to claim 1 or 2 in the preparation of an anti-atopic dermatitis drug.

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