Traditional Chinese medicine composition for treating rash, medicinal preparation and preparation method thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating rash, a medicinal preparation and a preparation method thereof, and belongs to the technical field of medicines. The traditional Chinese medicine composition comprises the following components: radix rehmanniae, radix ophiopogonis, radix scutellariae, fried tribulus terrestris, fructus kochiae, cortex lycii radicis, radix scrophulariae, radix puerariae, rhizoma cimicifugae, radix paeoniae alba, radix saposhnikoviae and liquorice. The traditional Chinese medicine composition and the medicinal preparation provided by the invention have the effects of removing heat to cool blood and nourishing yin to dispel wind, and can be used for treating rash, especially yin deficiency and internal heat type chronic rash with symptoms of dry and erythema of skin, severe itching at night and red tongue with little tongue fur.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a traditional Chinese medicine composition for treating skin rashes, a pharmaceutical preparation and its preparation method. Background Technology

[0002] A rash refers to a localized change in skin color and shape, possibly accompanied by symptoms such as itching and pain; it is a manifestation of skin damage. Rashes are very common clinically and come in many types, including macules and papules. Different rashes have different patterns and morphological characteristics, which can help in the differential diagnosis of different diseases. Rashes are often caused by systemic diseases, such as allergic reactions, infectious skin diseases, and connective tissue diseases, but can also be caused by factors such as drugs, environment, and lifestyle.

[0003] Treatment methods vary depending on the cause of the rash. For allergic reactions, antihistamines and corticosteroid creams are commonly used; rashes caused by infectious diseases usually require antibiotics; and rashes caused by autoimmune diseases require immunosuppressants. In addition to medication, physical therapies such as semiconductor laser irradiation and infrared irradiation can also be used. At home, it is important to keep the skin clean, avoid irritation, and pay attention to diet and rest.

[0004] In Traditional Chinese Medicine (TCM), the rash is diagnosed as a blood deficiency and wind-dryness syndrome. TCM treatments for rashes vary. For example, Xiaofeng San (from *Wai Ke Zheng Zong*) contains Angelica sinensis, Rehmannia glutinosa, Saposhnikovia divaricata, Cicadae Periostracum, Anemarrhena asphodeloides, Sophora flavescens, Schizonepeta tenuifolia, Atractylodes lancea, Arctium lappa, Gypsum fibrosum, Glycyrrhiza uralensis, Akebia trifoliata, and Sesamum indicum. Its effects focus on dispelling wind and dampness, clearing heat and nourishing blood (preferably for cases of wind-damp-heat syndrome). It is suitable for acute eczema / urticaria of the wind-damp-heat type, characterized by red rashes, significant exudation, and intense itching. This medicine contains Atractylodes lancea and Sophora flavescens, which are drying herbs and can easily damage yin fluids. Akebia trifoliata and Gypsum fibrosum have strong heat-clearing properties, but long-term use may damage the spleen and stomach. Shengma Gegen Tang (from *Yan's Treatise on Pediatric Prescriptions*) contains Shengma (Cimicifuga rhizome), Gegen (Pueraria root), Shaoyao (Peony root), and Gancao (Licorice root). Its effects focus on relieving muscle tension and promoting rash eruption, dispersing external pathogens (especially for cases of unresolved external pathogens). It is suitable for early-stage measles or when the rash is not fully erupting, accompanied by fever, cough, and a floating, rapid pulse. This medicine is simple and potent, characterized by its ability to promote rash eruption and dispel pathogens, and contains no yin-nourishing ingredients. Zengye Tang (from *Treatise on Warm Diseases*) contains Xuanshen (Scrophularia root), Maidong (Ophiopogon root), and Shengdi (Rehmannia root). Its effects focus on replenishing fluids and moistening dryness, nourishing yin and generating fluids (for pure yin deficiency and fluid depletion syndrome). It is suitable for yin deficiency constipation, accompanied by dry mouth and tongue, and dry, scaly skin. This medicine purely nourishes yin and moistens dryness, without the effects of dispelling wind, relieving itching, or promoting rash eruption. Dang Gui Yin Zi contains Dang Gui (Angelica sinensis), Sheng Di (Rehmannia glutinosa), Bai Shao (Paeonia lactiflora), Jing Jie (Schizonepeta tenuifolia), Fang Feng (Saposhnikovia divaricata), Huang Qi (Astragalus membranaceus), He Shou Wu (Polygonum multiflorum), and Chuan Xiong (Ligusticum chuanxiong). Its effects focus on nourishing blood and moisturizing dryness, dispelling wind and relieving itching, and tonifying qi and consolidating the exterior. It is suitable for symptoms of blood deficiency and wind dryness combined with qi and blood deficiency, characterized by "dryness, itching, and deficiency". The medicine is mild in nature and focuses on "nourishing blood, tonifying qi, and dispelling wind", making it suitable for those with weak constitution, wind dryness, and prolonged illness.

[0005] Since the causes and symptoms of rashes vary, and different Chinese medicines have different focuses of efficacy, more Chinese medicines for treating rashes still need to be developed in order to better address different types of diseases. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the present invention aims to provide a traditional Chinese medicine composition, pharmaceutical preparation, and preparation method for treating skin rashes. This traditional Chinese medicine composition and pharmaceutical preparation possess the effects of clearing heat and cooling blood, nourishing yin and dispelling wind, and can be used to treat skin rashes.

[0007] To achieve this objective, the present invention adopts the following technical solution:

[0008] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating skin rashes, the traditional Chinese medicine composition comprising the following components:

[0009] Rehmannia glutinosa, Ophiopogon japonicus, Scutellaria baicalensis, Tribulus terrestris (fried), Kochia scoparia, Lycium chinense root bark, Scrophularia ningpoensis, Pueraria lobata, Cimicifuga foetida, Paeonia lactiflora, Saposhnikovia divaricata, and Glycyrrhiza uralensis.

[0010] In some preferred embodiments of the present invention, the traditional Chinese medicine composition comprises the following components in parts by weight:

[0011] 20-30 parts of Rehmannia glutinosa, 28-32 parts of Ophiopogon japonicus, 18-22 parts of Scutellaria baicalensis, 13-17 parts of stir-fried Tribulus terrestris, 13-17 parts of Kochia scoparia, 13-17 parts of Lycium chinense root bark, 22-26 parts of Scrophularia ningpoensis, 12-18 parts of Pueraria lobata, 8-12 parts of Cimicifuga foetida, 13-17 parts of Paeonia lactiflora, 10-14 parts of Saposhnikovia divaricata, and 4-6 parts of Glycyrrhiza uralensis.

[0012] The weight of raw rehmannia root can be any value within the range of 20-30 parts, such as 20 parts, 20.5 parts, 21 parts, 21.5 parts, 22 parts, 22.5 parts, 23 parts, 23.5 parts, 24 parts, 24.5 parts, 25 parts, 25.5 parts, 26 parts, 26.5 parts, 27 parts, 27.5 parts, 28 parts, 28.5 parts, 29 parts, 29.5 parts, or 30 parts, etc.

[0013] The weight of Ophiopogon japonicus can be any value within the range of 28-32 parts, such as 28 parts, 28.2 parts, 28.5 parts, 28.8 parts, 29 parts, 29.2 parts, 29.5 parts, 29.8 parts, 30 parts, 30.2 parts, 30.5 parts, 30.8 parts, 31 parts, 31.2 parts, 31.5 parts, 31.8 parts, or 32 parts, etc.

[0014] The weight of Scutellaria baicalensis can be any value within the range of 18-22 parts, such as 18 parts, 18.2 parts, 18.5 parts, 18.8 parts, 19 parts, 19.2 parts, 19.5 parts, 19.8 parts, 20 parts, 20.2 parts, 20.5 parts, 20.8 parts, 21 parts, 21.2 parts, 21.5 parts, 21.8 parts, or 22 parts, etc.

[0015] The weight of roasted tribulus terrestris can be any value within the range of 13-17 parts, such as 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts, or 17 parts, etc.

[0016] The weight of Kochia scoparia seeds can be any value within the range of 13-17 parts, such as 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts, or 17 parts, etc.

[0017] The weight of Scrophularia can be any value within the range of 22-26 parts, such as 22 parts, 22.2 parts, 22.5 parts, 22.8 parts, 23 parts, 23.2 parts, 23.5 parts, 23.8 parts, 24 parts, 24.2 parts, 24.5 parts, 24.8 parts, 25 parts, 25.2 parts, 25.5 parts, 25.8 parts, or 26 parts, etc.

[0018] The weight of kudzu can be any value in the range of 12-18 parts, such as 12 parts, 12.5 parts, 13 parts, 13.5 parts, 14 parts, 14.5 parts, 15 parts, 15.5 parts, 16 parts, 16.5 parts, 17 parts, 17.5 parts, or 18 parts, etc.

[0019] The weight of Cimicifuga can be any value within the range of 8-12 parts, such as 8 parts, 8.2 parts, 8.5 parts, 8.8 parts, 9 parts, 9.2 parts, 9.5 parts, 9.8 parts, 10 parts, 10.2 parts, 10.5 parts, 10.8 parts, 11 parts, 11.2 parts, 11.5 parts, 11.8 parts, or 12 parts, etc.

[0020] In some preferred embodiments of the present invention, the traditional Chinese medicine composition comprises the following components in parts by weight:

[0021] 24 parts of Rehmannia glutinosa, 30 parts of Ophiopogon japonicus, 20 parts of Scutellaria baicalensis, 15 parts of stir-fried Tribulus terrestris, 15 parts of Kochia scoparia, 15 parts of Lycium chinense root bark, 24 parts of Scrophularia ningpoensis, 15 parts of Pueraria lobata, 10 parts of Cimicifuga foetida, 15 parts of Paeonia lactiflora, 12 parts of Saposhnikovia divaricata, and 5 parts of Glycyrrhiza uralensis.

[0022] The efficacy of each component in the traditional Chinese medicine composition provided by this invention is as follows:

[0023] Rehmannia glutinosa: Nourishes Yin and cools the blood, clears and resolves latent heat in the blood;

[0024] Ophiopogon japonicus: Nourishes Yin and moistens dryness, clears the heart and relieves irritability;

[0025] Scutellaria baicalensis: Clears heat and dries dampness, drains excess heat from the lungs and stomach;

[0026] Stir-fried Tribulus terrestris: dispels wind and relieves itching, soothes the liver and unblocks the meridians;

[0027] Kochia scoparia: Clears heat and promotes diuresis, dispels wind and relieves itching;

[0028] Lycium bark: Clears deficiency heat, cools the blood and eliminates steaming heat;

[0029] Scrophularia: Nourishes Yin and reduces fire, detoxifies and dissipates nodules;

[0030] Kudzu root: relieves muscle tension and reduces fever, promotes saliva production and relieves rashes;

[0031] Cimicifuga rhizome: Clears heat and detoxifies, promotes yang and relieves rashes;

[0032] White peony root: nourishes blood and astringes yin, softens the liver and relieves spasms;

[0033] Fangfeng: dispels wind and releases the exterior, eliminates dampness and relieves pain;

[0034] Licorice: harmonizes the effects of other herbs, protects the stomach and regulates the middle jiao (digestive system).

[0035] The compatibility mechanism of each component is as follows:

[0036] The principal herbs are Rehmannia glutinosa, Scrophularia ningpoensis, and Ophiopogon japonicus. Rehmannia glutinosa is sweet and cold, entering the heart, liver, and kidney meridians; it cools the blood, nourishes yin, clears heat, and generates fluids. Scrophularia ningpoensis is bitter, salty, and cold, nourishing yin, reducing fire, detoxifying, and dissipating nodules. The combination of these two herbs can clear latent heat in the blood, relieving symptoms such as rashes, erythema, desquamation, and burning sensations. Ophiopogon japonicus is sweet, slightly bitter, and cold, nourishing yin, moistening the lungs, benefiting the stomach, and generating fluids. Together with Rehmannia glutinosa and Scrophularia ningpoensis, it enhances the yin-nourishing and dryness-moistening effects, making it suitable for those with rashes

[0037] Assistant herbs: Scutellaria baicalensis, Lycium chinense root bark, Pueraria lobata root, and Cimicifuga foetida rhizome. Scutellaria baicalensis is bitter and cold, clearing heat from the upper lungs, purging fire, and detoxifying; Lycium chinense root bark is sweet and cold, cooling the blood, eliminating steaming heat, clearing the lungs, and reducing fire. The combination of these two herbs can clear lung heat and toxins, relieving redness, itching, and cough due to lung heat. Pueraria lobata root and Cimicifuga foetida rhizome assist the principal herbs in releasing stagnant heat from the skin surface, making them suitable for rashes caused by wind-heat or blood-heat.

[0038] Adjuvant herbs: Saposhnikovia divaricata, stir-fried Tribulus terrestris, Kochia scoparia, and Paeonia lactiflora. Saposhnikovia divaricata is pungent and warm in nature, dispelling wind and relieving exterior symptoms, as well as itching and pain, making it an essential herb for rubella and itching. Stir-fried Tribulus terrestris calms the liver, dispels wind, relieves itching, and disperses nodules; the two herbs work together to dispel wind pathogens and alleviate itching and tingling sensations. Kochia scoparia is bitter and cold, clearing heat and dampness, dispelling wind and relieving itching, and is effective for damp-heat type rashes. Paeonia lactiflora is sour and cold, nourishing blood and astringing yin, softening the liver and relieving spasms, harmonizing the body's defenses, and reducing the recurrence of rashes.

[0039] Used herb: Licorice. It is sweet and neutral in nature, clearing heat and detoxifying, harmonizing other herbs, and simultaneously alleviating the irritation of bitter and cold herbs on the spleen and stomach.

[0040] The traditional Chinese medicine composition provided by this invention takes "nourishing yin and cooling blood" as its root and "clearing and dispelling wind" as its branch, integrating the four methods of nourishing yin, clearing heat, promoting skin function, and relieving itching, forming a formulation style that is "moistening without being greasy, clearing without being cold, and dispersing without being drying." This traditional Chinese medicine composition works synergistically to clear heat and cool blood, nourish yin and dispel wind (primarily nourishing yin and clearing heat, while also dispelling wind, relieving itching, cooling blood, and promoting rash eruption). It can be used to treat rashes, especially chronic rashes of the yin deficiency and internal heat type (such as senile pruritus and chronic urticaria), with symptoms such as dry, erythematous skin, severe itching at night, and a red tongue with little coating.

[0041] In a second aspect, the present invention provides a pharmaceutical preparation for treating rashes, the pharmaceutical preparation comprising the active ingredients of the traditional Chinese medicine composition as described in the first aspect.

[0042] In some embodiments of the present invention, the pharmaceutical preparation further includes excipients.

[0043] In some embodiments of the present invention, the dosage form of the pharmaceutical preparation is granules, tablets, capsules, or decoction.

[0044] Thirdly, the present invention provides a method for preparing a pharmaceutical preparation as described in the second aspect, the method comprising the following steps:

[0045] (1) The traditional Chinese medicine composition described in the first aspect is subjected to water extraction to obtain an aqueous extract;

[0046] (2) The aqueous extract is prepared into the dosage form of the drug preparation.

[0047] In some embodiments of the present invention, the water extraction step includes:

[0048] The herbal composition is decocted with water at least twice (e.g., twice, three times, four times, or five times), each time for 1-4 hours (e.g., 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, or 4 hours). After each decoction, the mixture is filtered, and the filtrates are combined to obtain the aqueous extract.

[0049] In some embodiments of the present invention, in the water extraction step, the total mass of water is 18-35 times the mass of the traditional Chinese medicine composition; for example, it can be 18 times, 19 times, 20 times, 22 times, 23 times, 25 times, 26 times, 28 times, 30 times, 32 times, 33 times or 35 times, etc.

[0050] In some embodiments of the present invention, the water extraction step involves two decoctions. For the first decoction, the mass of water is 10-20 times the mass of the herbal composition (e.g., 10, 12, 13, 15, 16, 18, or 20 times, etc.), and the decoction time is 2-3 hours (e.g., 2 hours, 2.2 hours, 2.3 hours, 2.5 hours, 2.6 hours, 2.8 hours, or 3 hours, etc.). For the second decoction, the mass of water is 8-15 times the mass of the herbal composition (e.g., 8, 9, 10, 11, 12, 13, 14, or 15 times, etc.), and the decoction time is 1-2 hours (e.g., 1 hour, 1.2 hours, 1.3 hours, 1.5 hours, 1.6 hours, 1.8 hours, or 2 hours, etc.).

[0051] Compared with the prior art, the present invention has the following beneficial effects:

[0052] This invention utilizes a scientific combination of Rehmannia glutinosa, Ophiopogon japonicus, Scutellaria baicalensis, Tribulus terrestris (fried), Kochia scoparia, Lycium chinense root bark, Scrophularia ningpoensis, Pueraria lobata, Cimicifuga foetida, Paeonia lactiflora, Saposhnikovia divaricata, and Glycyrrhiza uralensis to obtain a traditional Chinese medicine composition and pharmaceutical preparation with the effects of clearing heat and cooling blood, nourishing yin and dispelling wind (mainly nourishing yin and clearing heat, while also dispelling wind, relieving itching, cooling blood and promoting rash eruption). It can be used to treat rashes, especially chronic rashes of the yin deficiency and internal heat type (such as senile pruritus and chronic urticaria), with symptoms such as dry skin with erythema, severe itching at night, and red tongue with little coating. Attached Figure Description

[0053] Figure 1A Photographs of skin lesions on the backs of mice in different groups at different drug administration times;

[0054] Figure 1B Bar chart showing the back injury scores of mice in each group at different drug administration times;

[0055] Figure 2 A bar chart showing the number of scratches by mice in each group on day 15 after drug administration;

[0056] Figure 3A Bar graph showing the thickness of the left ear of mice in each group on day 15 after drug administration;

[0057] Figure 3B Bar graph showing the thickness of the right ear of mice in each group on day 15 after drug administration;

[0058] Figure 4A Bar chart showing serum IL-1β levels in mice of each group on day 15 after drug administration;

[0059] Figure 4B A bar chart showing the serum IL-4 levels in mice of each group on day 15 after drug administration;

[0060] Figure 4C Bar chart showing serum TNF-α levels in mice of each group on day 15 after drug administration;

[0061] Figure 5 HE staining images of the back skin tissue of mice in each group;

[0062] Figure 6 HE staining images of ear tissue from each group of mice. Detailed Implementation

[0063] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. Those skilled in the art should understand that the specific embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention.

[0064] Example 1

[0065] This embodiment provides a traditional Chinese medicine composition for treating skin rashes, comprising the following components in parts by weight:

[0066] 20 parts of Rehmannia glutinosa, 32 parts of Ophiopogon japonicus, 18 parts of Scutellaria baicalensis, 17 parts of stir-fried Tribulus terrestris, 13 parts of Kochia scoparia, 13 parts of Lycium chinense root bark, 22 parts of Scrophularia ningpoensis, 18 parts of Pueraria lobata, 8 parts of Cimicifuga foetida, 13 parts of Paeonia lactiflora, 10 parts of Saposhnikovia divaricata, and 4 parts of Glycyrrhiza uralensis.

[0067] This embodiment also provides a decoction for treating skin rashes, the preparation method of which is as follows:

[0068] Weigh the medicinal materials according to the proportions of the Chinese herbal composition, add water and decoct twice. For the first decoction, add 20 times the amount of water as the medicinal materials and decoct for 3 hours. For the second decoction, add 15 times the amount of water as the medicinal materials and decoct for 2 hours. After each decoction, filter, combine the filtrates, and concentrate under reduced pressure to obtain a decoction for treating rashes.

[0069] Example 2

[0070] This embodiment provides a traditional Chinese medicine composition for treating skin rashes, comprising the following components in parts by weight:

[0071] 30 parts of Rehmannia glutinosa, 28 parts of Ophiopogon japonicus, 22 parts of Scutellaria baicalensis, 13 parts of stir-fried Tribulus terrestris, 17 parts of Kochia scoparia, 17 parts of Lycium chinense root bark, 26 parts of Scrophularia ningpoensis, 12 parts of Pueraria lobata, 12 parts of Cimicifuga foetida, 17 parts of Paeonia lactiflora, 14 parts of Saposhnikovia divaricata, and 6 parts of Glycyrrhiza uralensis.

[0072] This embodiment also provides a decoction for treating skin rashes, the preparation method of which is as follows:

[0073] Weigh the medicinal materials according to the proportions of the Chinese herbal composition, add water and decoct twice. For the first decoction, add 10 times the amount of water as the medicinal materials and decoct for 2 hours. For the second decoction, add 8 times the amount of water as the medicinal materials and decoct for 1 hour. After each decoction, filter, combine the filtrates, and concentrate under reduced pressure to obtain a decoction for treating rashes.

[0074] Example 3

[0075] This embodiment provides a traditional Chinese medicine composition for treating skin rash, comprising the following components in parts by weight:

[0076] 24 parts of Rehmannia glutinosa, 30 parts of Ophiopogon japonicus, 20 parts of Scutellaria baicalensis, 15 parts of stir-fried Tribulus terrestris, 15 parts of Kochia scoparia, 15 parts of Lycium chinense Mill., 24 parts of Scrophularia ningpoensis, 15 parts of Pueraria lobata, 10 parts of Cimicifuga foetida, 15 parts of Paeonia lactiflora, 12 parts of Saposhnikovia divaricata, and 5 parts of Glycyrrhiza uralensis.

[0077] This embodiment also provides a decoction for treating skin rash, and its preparation method is as follows:

[0078] Weigh the medicinal materials according to the ratio of the traditional Chinese medicine composition, decoct them with water twice. For the first decoction, add 15 times the mass of water of the medicinal materials and decoct for 2.5 hours; for the second decoction, add 10 times the mass of water of the medicinal materials and decoct for 1.5 hours; filter after each decoction, combine the filtrates, and concentrate under reduced pressure to 2.0 g of the traditional Chinese medicine composition / mL to obtain the decoction for treating skin rash, and store it at 4°C for later use.

[0079] Therapeutic efficacy test

[0080] The decoction for treating skin rash prepared in Example 3 was used for animal experiments to confirm its therapeutic effect on the skin rash model of mice.

[0081] 1. Test materials

[0082] 1.1. Test animals

[0083] SPF-grade male C57BL / 6 mice, 4 - 6 weeks old, were purchased from Spf (Suzhou) Biotechnology Co., Ltd., with the production license number SCXK (Su) 2022 - 0006 and the certificate number A202410180252. Feeding environmental conditions: The feeding environmental conditions are standard, referring to the national standard GB14925 - 2010 of the People's Republic of China; the temperature is 20 - 26°C (daily temperature difference ≤ 4°C); the relative humidity is 40 - 70%; the pressure in the feeding room ≥ 10 Pa; 12 / 12 hours of day and night light and dark alternation.

[0084] 1.2. Test reagents

[0085] The decoction for treating skin rash prepared in Example 3 was diluted to 0.375 g / mL, 0.75 g / mL, and 1.5 g / mL respectively with sterile injection water. This concentration refers to the mass of the traditional Chinese medicine composition corresponding to the decoction contained in each 1 mL of the solution.

[0086] Xiaoyin capsules, with a specification of 0.3 g / capsule, batch number 230501, stored in a sealed manner in a cool place, produced by Shaanxi Modemeide Qixuehe Pharmaceutical Co., Ltd.; the product approval number is国药准字 Z2 = 0000110. Preparation method: Dilute the contents of the Xiaoyin capsules with sterile injection water to 94.6 mg / mL.

[0087] 2,4-Dinitrofluorobenzene (DNFB), 5g / bottle, batch number JW297036, should be stored in a tightly sealed container at low temperature. Manufacturer: Shanghai Yuanye Biotechnology Co., Ltd. Preparation method: Dilute 2,4-dinitrofluorobenzene to 0.5% (0.5g / 100mL) with a mixed solvent of acetone and olive oil. The volume ratio of acetone to olive oil in the mixed solvent is 3:1.

[0088] Acetone, 500mL / bottle, batch number 20231219, store in a cool, dark place, manufactured by Sinopharm Chemical Reagent Co., Ltd.

[0089] Olive oil, 500mL / bottle, batch number C13659019, store away from light at room temperature, manufacturer is Shanghai Maclean Biochemical Technology Co., Ltd.

[0090] Sterile water for injection, 500 mL / bottle, batch number 240621, to be stored in a sealed container, manufactured by Shaanxi Sheng'ao Animal Pharmaceutical Co., Ltd.; product approval number is Veterinary Drug No. 270071791.

[0091] This is a general-purpose tissue fixative, available in 500mL vials, batch number GP24103081634. Store at room temperature. The manufacturer is Wuhan Saiweier Biotechnology Co., Ltd.

[0092] 1.3. Instruments

[0093] The pipette is manufactured by Eppendorf and has a model number of 200μL.

[0094] The electronic balance is manufactured by Shanghai Yaoxin Electronic Technology Co., Ltd., and its model number is LQ-C12001.

[0095] The benchtop high-speed refrigerated microcentrifuge is manufactured by Thermo and its model number is Sorvall Legand Micro 17R.

[0096] Mini mixing centrifuge, manufacturer: LABGIC, model: L-CM-MINI;

[0097] The microplate reader is manufactured by Thermo and is model VARIOSKAN LUX.

[0098] The dehydrator is manufactured by DIAPATH and its model is Donatello.

[0099] The embedding machine is manufactured by Wuhan Junjie Electronics Co., Ltd., and its model number is JB-P5.

[0100] The freezing station is manufactured by Wuhan Junjie Electronics Co., Ltd., and its model number is JB-L5.

[0101] The pathology slider is manufactured by Leica Instruments Shanghai Co., Ltd., and its model number is RM2016.

[0102] The sheet spreader is manufactured by Zhejiang Jinhua Kedi Instrument Equipment Co., Ltd., and its model is KD-P.

[0103] The oven is manufactured by Tianjin Laiborui Instrument Equipment Co., Ltd., and its model is GFL-230.

[0104] An upright optical microscope, manufactured by Nikon Japan, model Nikon Eclipse E100;

[0105] The imaging system is manufactured by Nikon of Japan, and its model is NIKON DS-U3.

[0106] An upright white light photographic microscope, manufactured by Nikon Japan, model Eclipse Ci-L.

[0107] 2. Test Methods

[0108] 2.1. Modeling, grouping, and drug administration

[0109] After being fed a normal diet for one week, the mice were randomly divided into 6 groups: blank group, model group, low-dose prescription group, medium-dose prescription group, high-dose prescription group, and Xiaoyin capsule group. The model group consisted of 10 mice, and the other groups consisted of 8 mice each.

[0110] The control group was fed normally, while mice in all other groups were used to establish a mouse rash model. The day before the experiment, the hair on the backs of the mice was removed using depilatory cream, covering an area of ​​2.0cm × 1.5cm. On days 1, 2, 6, and 7 of the experiment, 0.5wt% DNFB solution was applied dropwise to the bare skin on the backs of the mice (100μL / time) and also applied to the inner skin of the ears. On days 11, 14, 18, 21, and 25 of the experiment, 0.5% DNFB solution (50μL / time) was applied dropwise to the bare skin on the backs of the mice and also applied to the inner skin of the ears to induce dermatitis. Mice in the control group were only shaved. Simultaneously, starting on day 14 of the experiment, except for mice in the control group which were treated with sterile water for injection, mice in all other groups were administered the drug twice daily via application to the ears, back, and ear area, and by gavage, for 15 consecutive days.

[0111] The types and dosages of drugs administered orally to mice in each group are as follows:

[0112] Blank group and model group: physiological saline, administration volume 100μL / 10g;

[0113] Low-dose prescription group: a diluted solution of the decoction prepared in Example 3 with a concentration of 0.375 g / mL, administered at a dose of 3.75 g / kg, with an administration volume of 100 μL / 10 g;

[0114] The dosage group in the prescription was a diluted solution of the decoction prepared in Example 3 with a concentration of 0.75 g / mL, administered at a dose of 7.5 g / kg and a volume of 100 μL / 10 g.

[0115] High-dose prescription group: a diluted solution of the decoction prepared in Example 3 with a concentration of 1.5 g / mL, administered at a dose of 15 g / kg, with an administration volume of 100 μL / 10 g;

[0116] Xiaoyin capsule group: Xiaoyin capsules diluted at a concentration of 94.6 mg / mL, administered at a dose of 0.946 g / kg, with an administration volume of 100 μL / 10 g.

[0117] 2.2. Detection Indicators and Methods

[0118] 2.2.1. Number of scratches

[0119] Scratching of the ears and head with the forepaws, scratching of the trunk and back with the hind paws, and biting of various parts of the body with the mouth were used as indicators of itching. The number of scratches by the mice within 10 minutes on the last day of the experiment was observed. Continuous scratching was counted as 1 time. Each mouse was observed separately.

[0120] 2.2.2. Back injury score

[0121] The Alzheimer's disease (AD) skin lesion scoring system in mice is a sum of scores based on the severity of skin lesions on the back. Back skin lesions include four symptoms: erythema, edema / papules, epidermal desquamation, and dryness. Each lesion is scored from 0 to 3 points on a four-point scale, with 0 points for no symptoms, 1 point for mild, 2 points for moderate, and 3 points for severe. The severity of skin lesions was scored on days 4, 7, 14, 21, and 28 of the experiment. On day 28, the back skin lesions were compared between groups and photographed for record-keeping.

[0122] 2.2.3. Thickness of the left and right ears

[0123] The thickness of the left and right ears of mice was measured on day 15 after drug administration.

[0124] 2.2.4. HE staining: back skin and local tissues of the auricle

[0125] (1) Paraffin slice preparation

[0126] 1) Sample collection: After the experiment, the mice were euthanized, and full-thickness skin from the back lesion and ear tissue were collected and fixed with fixative for more than 24 hours. The tissue was removed from the fixative and trimmed with a scalpel in a fume hood. The trimmed tissue and corresponding labels were placed in a dehydration box.

[0127] 2) Dehydration and wax impregnation: The dehydration box is placed in a dehydrator for gradient alcohol dehydration. The steps are as follows: 75% alcohol for 4 hours, 85% alcohol for 2 hours, 90% alcohol for 2 hours, 95% alcohol for 1 hour, anhydrous ethanol I for 30 minutes, anhydrous ethanol II for 30 minutes, benzyl alcohol for 10 minutes, xylene I for 10 minutes, xylene II for 10 minutes, 65℃ melted paraffin I for 1 hour, 65℃ melted paraffin II for 1 hour, and 65℃ melted paraffin III for 1 hour.

[0128] 3) Embedding: The paraffin-impregnated tissue is embedded in an embedding machine. First, the molten paraffin is placed into the embedding frame. Before the paraffin solidifies, the tissue is removed from the dehydration box, placed into the embedding frame according to the embedding surface requirements, and labeled accordingly. The tissue is then cooled on a -20°C freezing stage. After the paraffin solidifies, the paraffin block is removed from the embedding frame and trimmed.

[0129] 4) Sectioning: Place the trimmed wax block on a -20℃ freezing stage to cool, then place the cooled wax block on a paraffin microtome to section to a thickness of 4μm. Float the sections on 40℃ warm water in a slide to flatten the tissue, then lift the tissue onto a glass slide and bake in a 60℃ oven. After the wax has melted in the water, remove the slide and store it at room temperature for later use.

[0130] (2) Staining

[0131] 1) Dewaxing paraffin sections to water: Place the sections in xylene I for 20 min, xylene II for 20 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, 75% ethanol for 5 min, and then wash with tap water.

[0132] 2) Hematoxylin staining: After dewaxing, the sections were stained with hematoxylin solution for 5 minutes, washed with tap water, differentiated with differentiation solution, washed with tap water, blued with blue solution, and rinsed with running water.

[0133] 3) Eosin staining: After hematoxylin staining, the sections were dehydrated in 85% and 95% alcohol for 5 minutes each, and then stained in eosin staining solution for 5 minutes.

[0134] 4) Dehydration and mounting: After eosin staining, the sections were sequentially immersed in anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, anhydrous ethanol III for 5 min, xylene I for 5 min, xylene II for 5 min, and then mounted with clear neutral resin.

[0135] 5) Microscopic examination, image acquisition and analysis.

[0136] 2.2.5. Serum TNF-α, IL-4, and IL-1β levels

[0137] The detection was performed using an ELISA kit, and the steps are as follows:

[0138] (1) Sample collection: Use test tubes free of pyrogens and endotoxins, and avoid any cell stimulation during the operation. After the experiment, take mouse blood and centrifuge at 3000 rpm for 10 minutes to quickly and carefully separate serum and red blood cells.

[0139] (2) Remove the required strips from the aluminum foil bag after equilibration at room temperature for 20 minutes, and seal the remaining strips in a self-sealing bag and return them to 4℃.

[0140] (3) Set up standard wells and sample wells, and add 50 μL of standard of different concentrations to each standard well.

[0141] (4) Add 10 μL of the sample to be tested to the sample well first, and then add 40 μL of sample diluent; do not add to the blank well.

[0142] (5) Except for the blank wells, add 100 μL of horseradish peroxidase (HRP) labeled detection antibody to each of the standard wells and sample wells, seal the reaction wells with sealing film, and incubate at 37°C in a water bath or incubator for 60 min.

[0143] (6) Discard the liquid, pat dry on absorbent paper, fill each well with washing liquid, let stand for 1 minute, shake off the washing liquid, pat dry on absorbent paper, and repeat the washing process 5 times (or use a plate washer).

[0144] (7) Add 50 μL of substrate A and B to each well and incubate at 37°C in the dark for 15 min.

[0145] (8) Add 50 μL of stop solution to each well and measure the OD value of each well at a wavelength of 450 nm within 15 min.

[0146] (9) Plot the standard curve: plot the linear regression curve of the standard with the concentration of the standard on the x-axis and the corresponding OD value on the y-axis, and calculate the concentration value of each sample according to the curve equation.

[0147] 2.3. Data Analysis and Processing

[0148] GraphPad 9.5.0 and SPSS 26 software were used for plotting and statistical analysis. Data are expressed as mean ± SEM. One-way ANOVA was used for statistical analysis to test for significant differences between the two groups. P < 0.05 was considered statistically significant.

[0149] 3. Test Results

[0150] 3.1. Effects of the formulation of this invention on skin lesions on the back of mice with rash, the number of scratches, and the thickness of the left and right ears.

[0151] Figure 1A Photos of skin lesions on the backs of mice in different groups at different drug administration times (day 0 represents before drug administration). Figure 1B Bar chart showing the back injury scores of mice in each group at different drug administration times; Figure 2 A bar chart showing the number of scratches by mice in each group on day 15 after drug administration; Figure 3A This is a bar chart showing the thickness of the left ear of mice in each group on day 15 after drug administration. Figure 3B This is a bar chart showing the right ear thickness of mice in each group on day 15 of drug administration. Compared to the control group, *** p < 0.001; compared with the model group, # p < 0.05.

[0152] from Figure 1A and Figure 1B It can be seen that, compared with the blank group, the model group mice had obvious skin damage on their backs; after administration, the skin damage on the backs of the mice was significantly reduced and the recovery was faster.

[0153] from Figure 2 It can be seen that, compared with the blank group, the number of scratches by mice in the model group increased significantly (P<0.001); compared with the model group, the number of scratches by mice in each dosage group of the prescription of the present invention decreased significantly (P<0.05), and there was no significant difference in the number of scratches by mice in the Xiaoyin capsule group.

[0154] As shown in Figure 3, compared with the blank group, the thickness of both ears of mice in the model group was significantly increased (P < 0.001); compared with the model group, the thickness of the left ear of mice in the medium and high dose groups of the prescription of the present invention was significantly decreased (P < 0.05), and the thickness of the right ear of mice in the high dose group of the prescription of the present invention was significantly decreased (P < 0.05).

[0155] 3.2. Effects of the formulation of this invention on serum inflammatory markers in mice with skin rash

[0156] TNF-α (tumor necrosis factor-α) and IL-1β (interleukin-1β) are pro-inflammatory cytokines, while IL-4 (interleukin-4) is an anti-inflammatory factor. Elevated levels of TNF-α and IL-1β and decreased levels of IL-4 in serum suggest an imbalance between pro-inflammatory and anti-inflammatory activity, indicating an inflammatory response in the body.

[0157] The serum TNF-α, IL-4, and IL-1β levels of mice in each group on day 15 of drug administration are shown in Table 1.

[0158] Table 1

[0159] Group IL-1β (pg / mL) IL-4 (pg / mL) TNF-α (pg / mL) Blank group 37.72±13.25 56.40±18.44 217.57±84.46 Model group <![CDATA[75.89±22.82 *** ]]> <![CDATA[30.63±10.54 *** ]]> <![CDATA[465.45±149.27 *** ]]> Low-dose prescription group <![CDATA[63.43±21.48 ###@@@ ]]> <![CDATA[42.62±14.97 ###@@@ ]]> <![CDATA[374.63±132.06 ###@@@ ]]> Medium-dose group of prescription <![CDATA[58.35±20.08 ###@@ ]]> <![CDATA[39.27±13.02 ##@@@ ]]> <![CDATA[358.09±34.21 ###@@@ ]]> High-dose prescription group <![CDATA[50.31±17.79 ### ]]> <![CDATA[47.64±16.21 ###@ ]]> <![CDATA[274.71±94.89 ### ]]> Xiaoyin capsule group <![CDATA[47.14±16.53 ### ]]> <![CDATA[54.39±18.45 ### ]]> <![CDATA[242.87±84.92 ### ]]>

[0160] Figure 4A This is a bar chart showing the serum IL-1β levels in mice of each group on day 15 after drug administration. Figure 4B This is a bar chart showing the serum IL-4 levels of mice in each group on day 15 after drug administration. Figure 4CThe bar chart shows the serum TNF-α levels in mice of each group on day 15 after drug administration. Compared to the control group, *** p < 0.001; compared with the model group, ## p < 0.01, ### p < 0.001; compared with the Xiaoyin capsule group, @ p < 0.05 @@ p < 0.01, @@@ p < 0.001.

[0161] From Table 1 and Figures 4A-4C It can be seen that, compared with the blank group, the serum IL-1β level in the model group mice was significantly increased (P < 0.001). Compared with the model group, the serum IL-1β level in all prescription dose groups and the positive drug Xiaoyin capsule group was significantly decreased (P < 0.001). Compared with the positive drug Xiaoyin capsule group, the serum IL-1β level in the low-dose prescription group was significantly increased (P < 0.001); the serum IL-1β level in the medium-dose prescription group was significantly increased (P < 0.01); and there was no significant difference in the serum IL-1β level in the high-dose prescription group (P > 0.05).

[0162] Compared with the control group, the serum IL-4 level in the model group mice was significantly decreased (P < 0.001). Compared with the model group, the serum IL-4 level in the low-dose prescription group, high-dose prescription group, and positive control drug Xiaoyin capsule group was significantly increased (P < 0.001), and the serum IL-4 level in the medium-dose prescription group was significantly increased (P < 0.01). Compared with the positive control drug Xiaoyin capsule group, the serum IL-4 level in the low-dose prescription group and medium-dose prescription group was significantly decreased (P < 0.001); the serum IL-4 level in the low- and medium-dose prescription groups was slightly decreased (P < 0.05).

[0163] Compared with the control group, the serum TNF-α level in the model group mice was significantly increased (P < 0.001). Compared with the model group, the serum TNF-α level in all prescription dose groups and the positive control drug Xiaoyin capsule group was significantly decreased (P < 0.001). Compared with the positive control drug group, the serum TNF-α level in the low- and medium-dose prescription groups was significantly increased (P < 0.001); there was no significant difference in the high-dose group (P > 0.05).

[0164] The above results show that low, medium, and high doses of the prescription of this invention and the positive control drug Xiaoyin capsules can inhibit the levels of IL-1β and TNF-α in mouse serum and increase the level of IL-4 to varying degrees, indicating that low, medium, and high doses of the prescription of this invention and the positive control drug Xiaoyin capsules can reduce the inflammatory response in mice with rashes and have a good therapeutic effect on rashes.

[0165] 3.3. Effects of the formulation of this invention on the dorsal skin and auricular tissue of mice with skin rash model

[0166] Figure 5 HE staining images (200×) of the dorsal skin tissue of mice in each group. Figure 6 HE staining images (200×) of mouse auricle tissue from each group. Among them, A is the blank group, B is the model group, C is the low-dose prescription group, D is the medium-dose prescription group, E is the high-dose prescription group, and F is the Xiaoyin capsule group.

[0167] Pathological examination, such as Figure 5 As shown, no obvious pathological changes were observed in the dorsal skin tissue of mice in the control group. Compared with the control group, inflammatory cell infiltration was observed in the dorsal skin tissue of mice in the model group. A large number of samples showed epidermal thickening, necrosis, and vascular congestion, while a significant number of samples showed connective tissue hyperplasia, and a small number of samples showed elongation of the skin ridges and hemorrhage. Compared with the model group, the proportion of samples with hemorrhage and congestion in the dorsal skin tissue of mice in the positive control group (Xiaoyin capsules) decreased, with no other significant differences. In the high-dose prescription group, the epidermal thickness of mice was thinner, the degree of inflammatory cell infiltration was reduced, the proportion of samples with necrosis, hemorrhage, and congestion decreased, and the necrotic area was smaller. The proportion of samples with connective tissue hyperplasia increased, with no other significant differences. In the medium-dose prescription group, the area of ​​necrosis in the dorsal skin of mice was smaller, the degree of inflammatory cell infiltration was reduced, and the proportion of samples with congestion decreased, with no other significant differences. In the low-dose prescription group, the proportion of samples with necrosis, elongation of the skin ridges, and hemorrhage decreased, while the proportion of samples with connective tissue hyperplasia increased. Compared with the positive drug Xiaoyin capsule group, the proportion of mice with bleeding and ecchymosis on their backs was slightly lower in the high-dose prescription group, with no other significant differences; there were no significant differences in the back skin tissue of mice in the low-dose prescription group and the positive drug Xiaoyin capsule group.

[0168] Pathological examination, such as Figure 6As shown, in the control group, a large number of ear skin samples showed minor pigmentation, with some samples exhibiting thickened epidermis, focal necrosis, minimal connective tissue hyperplasia in the dermis, significant inflammatory cell infiltration, and extensive vascular congestion. Compared to the control group, the model group showed an increased proportion of samples with thickened epidermis, necrosis, connective tissue hyperplasia, inflammatory cell infiltration, and congestion, a decreased proportion of samples with pigmentation, and a few samples showing slight hemorrhage. Compared with the model group, the proportion of mice in the positive control group (Xiaoyin capsules) showing epidermal thickening, necrosis, connective tissue hyperplasia, inflammatory cell infiltration, and congestion in their ear skin decreased, and the degree of necrosis and inflammatory cell infiltration was reduced, while the proportion of samples with pigmentation increased; other differences were not significant. In the medium and high dose groups, the proportion of mice in the ear skin showing epidermal thickening, connective tissue hyperplasia, inflammatory cell infiltration, and congestion decreased, and the degree of inflammatory cell infiltration was reduced, while the proportion of samples with pigmentation increased; no significant necrosis was observed, and other differences were not significant. In the prescription dose group, the proportion of mice in the ear skin showing necrosis and pigmentation decreased, while other differences were not significant. Compared with the positive control group (Xiaoyin capsules), the proportion of mice in the low dose group showing inflammatory cell infiltration and congestion in their ear skin increased, and the degree of necrosis and inflammatory cell infiltration was aggravated; there was no significant difference in the degree of epidermal and inflammatory cell infiltration in the medium dose group; and the proportion of mice in the high dose group showing inflammatory cell infiltration and congestion decreased slightly.

[0169] In summary, mice with rashes treated with the prescription of this invention and the positive control drug Xiaoyin capsules showed reduced scratching frequency, decreased inflammatory response, and significantly improved pathological results of the skin on the back and auricle. The efficacy of high-dose prescription of this invention is comparable to that of the positive control drug Xiaoyin capsules.

[0170] The above description is merely a specific embodiment of this disclosure, enabling those skilled in the art to understand or implement it. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of this disclosure. Therefore, this disclosure is not to be limited to the embodiments described herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A traditional Chinese medicine composition for treating skin rashes, characterized in that, The traditional Chinese medicine composition includes the following components: Rehmannia glutinosa, Ophiopogon japonicus, Scutellaria baicalensis, Tribulus terrestris (fried), Kochia scoparia, Lycium chinense root bark, Scrophularia ningpoensis, Pueraria lobata, Cimicifuga foetida, Paeonia lactiflora, Saposhnikovia divaricata, and Glycyrrhiza uralensis.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The traditional Chinese medicine composition comprises the following components in parts by weight: 20-30 parts of Rehmannia glutinosa, 28-32 parts of Ophiopogon japonicus, 18-22 parts of Scutellaria baicalensis, 13-17 parts of stir-fried Tribulus terrestris, 13-17 parts of Kochia scoparia, 13-17 parts of Lycium chinense root bark, 22-26 parts of Scrophularia ningpoensis, 12-18 parts of Pueraria lobata, 8-12 parts of Cimicifuga foetida, 13-17 parts of Paeonia lactiflora, 10-14 parts of Saposhnikovia divaricata, and 4-6 parts of Glycyrrhiza uralensis.

3. The traditional Chinese medicine composition according to claim 1 or 2, characterized in that, The traditional Chinese medicine composition comprises the following components in parts by weight: 24 parts of Rehmannia glutinosa, 30 parts of Ophiopogon japonicus, 20 parts of Scutellaria baicalensis, 15 parts of stir-fried Tribulus terrestris, 15 parts of Kochia scoparia, 15 parts of Lycium chinense root bark, 24 parts of Scrophularia ningpoensis, 15 parts of Pueraria lobata, 10 parts of Cimicifuga foetida, 15 parts of Paeonia lactiflora, 12 parts of Saposhnikovia divaricata, and 5 parts of Glycyrrhiza uralensis.

4. A pharmaceutical preparation for treating skin rashes, characterized in that, The pharmaceutical preparation contains the active ingredient of the traditional Chinese medicine composition as described in any one of claims 1-3.

5. The pharmaceutical preparation according to claim 4, characterized in that, The pharmaceutical preparation also includes excipients.

6. The pharmaceutical preparation according to claim 4 or 5, characterized in that, The dosage form of the pharmaceutical preparation is granules, tablets, capsules, or decoction.

7. A method for preparing a pharmaceutical formulation as described in any one of claims 4-6, characterized in that, The preparation method includes the following steps: (1) The traditional Chinese medicine composition according to any one of claims 1-3 is subjected to water extraction to obtain an aqueous extract; (2) The aqueous extract is prepared into the dosage form of the drug preparation.

8. The preparation method according to claim 7, characterized in that, The water extraction step includes: The herbal composition is decocted with water at least twice, each time for 1-4 hours. After each decoction, the mixture is filtered, and the filtrates are combined to obtain the aqueous extract.

9. The preparation method according to claim 7 or 8, characterized in that, In the water extraction step, the total mass of water is 18-35 times the mass of the traditional Chinese medicine composition.

10. The preparation method according to claim 8, characterized in that, In the water extraction step, the decoction is performed twice. In the first decoction, the mass of water is 10-20 times the mass of the Chinese herbal composition, and the decoction time is 2-3 hours. In the second decoction, the mass of water is 8-15 times the mass of the Chinese herbal composition, and the decoction time is 1-2 hours.

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