A wolfberry leaf extract, a preparation method thereof and application thereof in preparing a product for preventing and treating acne

Abstract

This invention discloses a wolfberry leaf extract and its application in the preparation of acne prevention and treatment products. The invention uses specific process steps to prepare different solvent extracts of wolfberry leaves. This wolfberry leaf extract can reduce the level of lipopolysaccharide-induced inflammatory factors in Hacat cells and significantly improve acne symptoms in a rat acne lesion model. The wolfberry leaf extract provided by this invention has the potential to become a therapeutic product for acne and exhibits good safety.

Description

Technical Field

[0001] This invention relates to a traditional Chinese medicine, specifically to a wolfberry leaf extract and its application in the prevention and treatment of acne, belonging to the field of pharmaceutical technology. Background Technology

[0002] Acne vulgaris (ACV) is a common skin disease characterized by chronic inflammation of the pilosebaceous unit. It typically occurs on the cheeks and forehead, followed by the chest, back, and shoulders. It often manifests as comedones, and in severe cases, as inflammatory papules, pustules, nodules, cysts, and even scarring. It is most prevalent during adolescence. Modern medicine has confirmed that the pathogenesis of acne is mainly related to four key factors: excessive sebum secretion, abnormal follicular keratinization, proliferation of the skin microbiota, and accompanying inflammatory responses. Current treatments primarily involve anti-androgens, isotretinoin, antibiotics, and corticosteroids, often combined with topical therapies such as chemical peels and photodynamic therapy. However, long-term oral anti-androgen medication carries the risk of inducing deep vein thrombosis. While isotretinoin can inhibit sebum secretion, it carries the risk of congenital malformations and may cause side effects such as dry skin and mucous membranes, elevated blood lipids, and liver damage. Long-term use of antibiotics easily leads to drug resistance. Topical retinoids may cause side effects such as skin redness, dryness, peeling, and photosensitivity.

[0003] Traditional Chinese medicine (TCM) views acne through various names such as "acne vulgaris," "lung wind acne," "facial acne," "facial sores," and "rosacea," which closely correspond to its typical location (mainly the face and nose), characteristics of skin lesions (bumps, pustules, scaling, redness, swelling, and pain), and prolonged course. Throughout ancient literature, the most prominent theme in the etiology and pathogenesis of acne is "heat": either external wind-heat, accumulated heat in the lungs and stomach, or stagnation of blood heat. This heat rises upwards and manifests on the face and head, while the closure of pores and stagnation of sebum (body fluids and oils) cause recurring lesions. Acne is recorded differently in ancient texts throughout history, with the understanding of "heat as the guiding principle" being further solidified in the Ming and Qing dynasty surgical system as "lung wind acne / lung meridian heat (blood heat)," forming one of the most representative TCM classification paradigms and influencing the "lung heat type acne" syndrome framework to this day.

[0004] Goji berry leaves are the tender stems and leaves of the Solanaceae plant *Lycium barbarum* or *Lycium barbarum*. From the development of herbal medicine knowledge, the medicinal use of goji berries did not initially strictly distinguish between different parts such as fruit, root, and leaves. Instead, it gradually evolved from "the whole plant can be used" to "each part is used for a specific purpose." In the earliest extant herbal texts, goji berries were first mentioned in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica), and later the *Mingyi Bielu* (Records of Famous Physicians) supplemented the descriptions of the properties and indications of different parts. From the Tang and Song dynasties onward, goji berry leaves began to be more clearly highlighted as an edible and medicinal product; by the Ming and Qing dynasties, the name "Tianjingcao" (Heavenly Essence Grass) gradually became fixed, and its characteristics in clearing heat, improving eyesight, detoxifying, and reducing swelling and sores became increasingly clear. Summary of the Invention

[0005] Purpose of the invention: The purpose of this invention is to study the effects of different solvent extracts of wolfberry leaves on improving acne or cell inflammation, and to obtain the wolfberry leaf extract with the best therapeutic effect, so as to provide a theoretical basis for the rational clinical use of wolfberry leaves to treat acne vulgaris, especially lung meridian wind-heat type acne and inflammatory skin diseases.

[0006] This invention screened the effects of different solvent extracts of Lycium barbarum leaves on Hacat cell inflammation and SD rat acne models. The results showed that the 70% ethanol extract of Lycium barbarum leaves was the most effective. Therefore, the 70% ethanol extract of Lycium barbarum leaves can be applied to the treatment of acne or cellular inflammatory diseases, achieving efficient and comprehensive utilization of Lycium barbarum leaf resources.

[0007] Technical solution: To achieve the above objectives, the technical solution adopted by this invention is as follows: The wolfberry leaf extract provided by this invention is prepared by the following method: weigh dried wolfberry leaves, add 5 to 10 times the mass of 70% ethanol, heat and reflux to extract 1 to 3 times, each extraction for 0.5 to 1 hour, to obtain wolfberry leaf ethanol extract, concentrate under reduced pressure, freeze dry to obtain wolfberry leaf ethanol extract.

[0008] As a preferred embodiment, the wolfberry leaf extract of the present invention is obtained by the following method: Wolfberry leaf ethanol extract: Weigh dried wolfberry leaves, add 10 times the mass of 70% ethanol, heat and reflux to extract twice, each time for 1 hour, to obtain wolfberry leaf ethanol extract, concentrate under reduced pressure, freeze dry to obtain wolfberry leaf ethanol extract.

[0009] The wolfberry leaf extract provided by this invention is prepared by the following method: weigh dried wolfberry leaves, add 5 to 10 times the mass of ultrapure water, heat and reflux to extract 1 to 3 times, each extraction for 0.5 to 1 hour, combine the filtrates to obtain wolfberry leaf aqueous extract, concentrate under reduced pressure, freeze dry to obtain wolfberry leaf aqueous extract.

[0010] As a preferred option, the water extract of wolfberry leaves is prepared by weighing 1 kg of dried wolfberry leaves, adding 10 times the mass of ultrapure water, heating and refluxing twice for 1 hour each time to obtain a water extract of wolfberry leaves, concentrating under reduced pressure, and freeze-drying to obtain an aqueous extract of wolfberry leaves.

[0011] This invention can formulate wolfberry leaf extract and pharmaceutically acceptable carriers into tablets, capsules, granules, oral liquids, pills, mixtures or ointments.

[0012] In this invention, when making tablets, a carrier lactose or corn starch is added to the wolfberry leaf extract, and a lubricant magnesium stearate is added if necessary. The mixture is then mixed evenly and compressed into tablets.

[0013] In this invention, when making capsules, wolfberry leaf extract and carrier lactose or corn starch are mixed evenly, granulated, and then encapsulated to make capsules.

[0014] In this invention, when preparing granules, wolfberry leaf extract and diluents such as lactose or corn starch are mixed evenly, granulated, and dried to form granules.

[0015] Beneficial effects: The wolfberry leaf extract provided by this invention has the following advantages compared with the prior art: This invention utilizes different extracts from Lycium barbarum leaves to treat Hacat cells and SD rats, and investigates the ameliorative effects of different solvent extracts of Lycium barbarum leaves on a lipopolysaccharide-induced Hacat cell inflammation model using CCK8 and qPCR methods. The protective effects of different solvent extracts of Lycium barbarum leaves on the SD rat model were examined by detecting auricular thickness and ear surface indices, pathological sections of lung and ear tissues, and levels of inflammatory factors in lung and ear tissues. This leads to the conclusion that Lycium barbarum leaves are effective extracts for improving acne. Hacat cell experiments showed that different solvent extracts of Lycium barbarum leaves could improve the lipopolysaccharide-induced Hacat cell inflammation model and downregulate the expression levels of ERK, p38, and JNK, thus improving the inflammatory condition, with the ethanol extract showing the best effect. In the acne rat model, different solvent extracts of Lycium barbarum leaves could improve ear acne in acne-prone rats and significantly reduce the levels of inflammatory factors in serum, ear tissue, and lung tissue. Therefore, the 70% ethanol extract of Lycium barbarum leaves screened in this invention can be used as a treatment and improvement product for acne. Attached Figure Description

[0016] Figure 1 The effects of different concentrations of lipopolysaccharide and different solvent extracts of wolfberry leaves on the survival rate of Hacat cells (CCK8).

[0017] Figure 2 The effects of different solvent extracts of Lycium barbarum leaves on the expression levels of inflammatory cytokine genes (IL-1β, IL-6) in Hacat cells induced by lipopolysaccharide damage.

[0018] Figure 3 The effect of ethanol extract of Lycium barbarum leaves on gene expression levels (p38, ERK, JNK) in Hacat cells with lipopolysaccharide-induced damage.

[0019] Figure 4 The effects of different solvent extracts of wolfberry leaves on the apparent parameters of Acne rat ears.

[0020] Figure 5 The effect of different solvent extracts of wolfberry leaves on the ear thickness of Acne rats.

[0021] Figure 6The effects of different solvent extracts of Lycium barbarum leaves on the levels of inflammatory factors in serum, ear tissue, and lung tissue of Acne rats.

[0022] Figure 7 The effects of different solvent extracts of wolfberry leaves on the histopathology of Acne rat ear tissue.

[0023] Figure 8 The effects of different solvent extracts of wolfberry leaves on the pathological characteristics of lung tissue in Acne rats. Detailed Implementation

[0024] The present invention can be better understood from the following embodiments. However, those skilled in the art will readily understand that the specific material ratios, process conditions, and results described in the embodiments are for illustrative purposes only and should not, and will not, limit the invention as described in detail in the claims.

[0025] The reagents used in the embodiments of this invention were commercially available.

[0026] The primers used in the embodiments of this invention include:

[0027] The animals and cells used in the embodiments of this invention include:

[0028] Hacat cells were obtained from the ATCC cell bank and cultured in DMEM complete medium containing 10% FBS (containing 1% antibiotics) in a cell culture incubator (37℃, 5% CO2), and passaged every 2-3 days.

[0029] SPF-grade male SD rats (8 weeks old), weighing (200±20) g, were purchased from Beijing Vital River Co., Ltd. They were housed in a sterile environment with a laboratory animal production license number of (21±1) ℃ and a relative humidity of (60±5)%, and all operations and research procedures were carried out in accordance with the "Regulations on the Management of Laboratory Animals".

[0030] Example 1: Preparation of different solvent-extracted fractions of wolfberry leaves

[0031] The wolfberry leaf extract of the present invention is obtained by the following method: Wolfberry leaf ethanol extract: Weigh 1 kg of dried wolfberry leaves, add 10 times the volume of 70% ethanol by weight of wolfberry leaves, heat and reflux to extract twice, each time for 1 hour, to obtain wolfberry leaf ethanol extract, concentrate under reduced pressure, freeze dry to obtain wolfberry leaf ethanol extract.

[0032] Lycium barbarum leaf aqueous extract: Weigh 1 kg of dried Lycium barbarum leaves, add 10 times the volume of ultrapure water, heat and reflux to extract twice, 1 hour each time, to obtain Lycium barbarum leaf aqueous extract, concentrate under reduced pressure, freeze dry to obtain Lycium barbarum leaf aqueous extract.

[0033] Example 2: Effect evaluation based on cell models

[0034] 1. Experimental Methods

[0035] 1.1 LPS-induced Hacat cell inflammation model

[0036] Preparation method of Lycium barbarum leaf active component solution: Weigh 1 mg each of the lyophilized powder of Lycium barbarum leaf ethanol extract and aqueous extract prepared in Example 1, dissolve them in 1 mL of DMEM medium to prepare a 1 mg / mL solution, dilute with DMEM medium to the required concentration before administration, and filter sterilize using a 22 μm filter membrane.

[0037] Hacat cells were cultured in DMEM medium containing 10% FBS and 1% penicillin / streptomycin at 37°C and 5% CO2. The modeling groups were treated with different concentrations of lipopolysaccharide for 24 h, and the optimal modeling concentration was observed. The drug-treated groups were simultaneously given corresponding concentrations of lyophilized Lycium barbarum leaf active components, and the concentration that had no significant effect on cell viability was selected as the modeling concentration.

[0038] 1.2 qPCR detection of the effects of different extraction solvents of Lycium barbarum leaves on gene expression levels in a lipopolysaccharide-induced Hacat cell inflammation model.

[0039] (1) Total RNA extraction

[0040] Cells were seeded in 6-well cell culture plates and divided into four groups: a blank control group (Con group), a model group (Mod group), a high-dose group of Lycium barbarum leaf aqueous extract (SH group), a low-dose group of Lycium barbarum leaf aqueous extract (SL group), a high-dose group of Lycium barbarum leaf ethanol extract (CH group), and a low-dose group of Lycium barbarum leaf ethanol extract (CL group). Cells were collected into EP tubes and centrifuged at 1000 rpm for 5 min, discarding the supernatant. 1 mL of Trizol reagent was added to each tube and vigorously pipetted for 15 s to mix thoroughly. The mixture was incubated at room temperature for 2-3 min, then rotated at 4°C for 1-2 h. 200 μL of chloroform was added, and the mixture was vortexed for approximately 15 s until no obvious layering was observed. The mixture was then centrifuged at 12000 g for 15 min. 500 μL of the supernatant was transferred to a nuclease-free EP tube, along with 500 μL of isopropanol. The mixture was vortexed to mix thoroughly and then placed on ice for 5-10 min. The tubes were then centrifuged at 12000 g for 10 min. Discard the supernatant, add 1 mL of 75% ethanol (pre-cooled to 4°C) prepared with DEPC water, gently swirl to mix, and centrifuge at 7500g for 5 min. Discard the supernatant, evaporate the remaining ethanol in a fume hood, add 25 μL of DEPC water, gently swirl to mix, and place on ice. After determining the concentration using an ultra-micro UV spectrophotometer, store at -80°C for later use.

[0041] (2) cDNA synthesis

[0042] cDNA was synthesized according to the kit instructions; the synthesis system is shown in Table 1. The reverse transcription procedure was as follows: the system was incubated at 42°C for 15 min, then inactivated at 95°C for 3 s. The product was stored at -20°C for later use.

[0043] Table 1 cDNA synthesis system

[0044] (3) PCR amplification

[0045] The PCR amplification system is shown in Table 2. The reaction mixture was thoroughly mixed, added to a 96-well PCR plate, centrifuged, and measured using a PCR machine. PCR reaction conditions are shown in Table 6. Gapdh was used as an internal control. After obtaining the sample CT value, 2... -ΔΔ The relative expression levels of p38, JNK, and ERK in the sample were calculated using the CT method.

[0046] Table 2 PCR amplification system

[0047] Table 3 PCR reaction conditions

[0048] 1.3 Statistical Analysis

[0049] Data analysis and plotting were performed using GraphPad Prism 9.4.0 software. All results are expressed as mean ± standard deviation (SD). One-way ANOVA was used to analyze data between groups. A p-value < 0.05 was considered statistically significant. (Compared with the model group, *P < 0.05; **P < 0.01; ***P < 0.001; Compared with the control group, #P < 0.05; ##P < 0.01; ###P < 0.001).

[0050] 2. Experimental Results

[0051] 2.1 Determination of LPS modeling and drug concentrations using the CCK8 method

[0052] This invention investigates the drug administration concentrations of LPS and different solvent extracts of wolfberry leaves to Hacat cells. The drug administration concentrations of different solvent extracts of wolfberry leaves and lipopolysaccharide were detected using a CCK8 assay kit.

[0053] Test results are shown Figure 1 The vertical axis represents cell viability. After drug administration, cell viability decreased with increasing concentration. Therefore, a concentration that had no significant effect on Hacat cell viability was chosen as the modeling and drug administration concentration.

[0054] 2.2 Effects of different solvent extracts of Lycium barbarum leaves on gene expression levels in an LPS-induced Hacat cell inflammation model

[0055] This invention investigates the effects of different extraction solvents from Lycium barbarum leaves on LPS-induced gene expression levels in Hacat cells. Changes in the inflammatory factors IL-1β and IL-6, as well as the expression levels of p38, ERK, and JNK genes in the MAPK pathway, were analyzed using qPCR.

[0056] qPCR results are shown below Figure 2 and Figure 3 The x-axis represents the group, and the y-axis represents the relative expression level of mRNA. After LPS-induced inflammation model, the relative expression levels of IL-1β, IL-6, p38, ERK, and JNK mRNA were significantly increased compared to the control group. After drug administration, the relative expression levels of IL-1β and IL-6 in different solvent extracts of Lycium barbarum leaves all increased, with the effect of Lycium barbarum leaf ethanol extract (CT) being more significant than that of Lycium barbarum leaf water extract (ST). Lycium barbarum leaf ethanol extract can significantly reduce the increase in relative expression levels of p38, ERK, and JNK mRNA in Hacat cells induced by modeling.

[0057] Example 3: Effect evaluation based on a rat model of pulmonary heat-type acne.

[0058] Acne due to wind-heat in the lung meridian falls under the category of "lung-wind acne" in Traditional Chinese Medicine (TCM). The core pathogenesis is that the lung governs the skin and hair; invasion by wind-heat pathogens leads to lung heat rising upwards, obstructing the skin, and causing stagnation of qi and blood. This case study uses LPS nasal drops combined with the traditional Kligman acne model to establish a acne model in SD rats. The nasal instillation of LPS simulates the TCM pathogenesis of external pathogens entering the lungs, causing impaired lung qi circulation, and resulting in stagnation and heat. The traditional Kligman acne model further simulates the upward movement of heat pathogens along the meridians to the face and hair follicles, aligning with the core pathogenesis of lung-heat acne: "lung heat rising upwards and stagnation in the skin." In terms of evaluation indicators, the model rats were systematically evaluated by observing their body weight, hair, mental state, redness and swelling of the auricle, measuring the levels of inflammatory factors in the rat's ear and lung tissue and serum, and observing the thickness of the ear epidermis, degree of keratinization, hair follicle dilation, dermal inflammatory infiltration, and edema and inflammatory cell infiltration of the lung tissue through HE pathological staining. This was done to better reflect the lung meridian wind-heat syndrome of acne in traditional Chinese medicine theory. The specific implementation method is as follows.

[0059] 1. Experimental Methods

[0060] 1.1 After 7 days of acclimatization, rats were divided into a blank control group (CON), a model group (MOD), a tanshinone capsule group, high and low dose groups of Lycium barbarum leaf ethanol extract prepared according to Example 1, and high and low dose groups of Lycium barbarum leaf aqueous extract. Six rats were randomly selected as the control group. The remaining rats were given 50 μL of LPS solution (4 mg / mL) via nasal drops on days 1, 4, and 7 of modeling. From days 7 to 21, a modified Kligman acne modeling method was used, in which 1.0 mL was evenly applied to the skin of the lateral auricle of the right ear of rats in the model group and each treatment group daily. Every other day after application, 50 μL of Propionibacterium acnes (1×10⁻⁶) was injected intradermally into the auricle of the rats. 9 (CFU / mL). Administration should begin on the day of model establishment and continue for 28 days.

[0061] 1.2 Effects of different solvent-extracted parts of wolfberry leaves on the apparent parameters of the ears of model rats.

[0062] During the modeling process, the auricular thickness of rats in each group was measured at the same location on the modeled and non-modeled sides using vernier calipers. The severity of auricular skin lesions in each group of rats was scored. The main evaluation indicators were formulated according to the "Chinese Guidelines for Acne Treatment (2019 Revised Edition)" and the "Specifications for the Preparation of Animal Models of Acne (Draft)". Details are shown in Table 4 below.

[0063] Table 4. Score Sheet for Phenotypic Indicators of Acne in Rats' Ears

[0064] 1.3 The effects of different solvent-extracted parts of Lycium barbarum leaves on ear and lung tissues and serum inflammatory factors in model rats were detected using the Elisa kit.

[0065] After the SD rat model was established, blood was collected from the abdominal aorta and centrifuged for 15 min at 3000 rpm / min, 4℃. The supernatant was then collected. At the same time, a certain mass of ear and lung tissue from each group of rats was collected into EP tubes, homogenized with PBS, centrifuged at 4000 rpm for 10 min, and the supernatant was collected. The levels of inflammatory factors (TNF-α, IL-1β, IL-6, IL-17) were detected according to the kit instructions.

[0066] 1.4 Pathological effects of different solvent extracts of Lycium barbarum leaves on ear and lung tissues of model rats

[0067] After the last administration, rats were euthanized by excessive blood collection from the abdominal aorta. Half of the right auricle and lung tissue were fixed in 4% paraformaldehyde, while the other half was flash-frozen in liquid nitrogen and then stored at -80°C for later use. The ear and lung tissues were embedded in paraformaldehyde, sectioned, stained, and then observed under a microscope for pathological changes such as the degree of epidermal thickening, hair follicle dilation and keratinization, and inflammatory cell infiltration in the lung tissue.

[0068] 1.5 Effects of Lycium barbarum leaf ethanol extract on barrier proteins and inflammatory cell infiltration in rat ear tissue

[0069] After the last administration, rats were sacrificed by excessive blood collection from the abdominal aorta. Auricular tissue from rats in the control group, the increased-dose group, and the low-dose group was embedded and fixed, and sections were prepared. The sections were sent to Wuhan Saiweier Biotechnology Co., Ltd. for immunofluorescence staining, and the results were observed under a fluorescence microscope.

[0070] 2. Statistical Analysis

[0071] Data analysis and plotting were performed using GraphPad Prism 9.4.0 software. All results are expressed as mean ± standard deviation (SD). One-way ANOVA was used to analyze data between groups. A p-value < 0.05 was considered statistically significant. (Compared with the model group, *P < 0.05; **P < 0.01; ***P < 0.001; Compared with the control group, #P < 0.05; ##P < 0.01; ###P < 0.001).

[0072] 3. Experimental Results

[0073] 3.1 Effects of different solvent-extracted fractions of Lycium barbarum leaves on the apparent parameters of the ears of model rats

[0074] This invention investigates the effects of different solvent-extracted fractions of Lycium barbarum leaves on the external parameters of rat ears. The appearance of the rat ears was observed during the feeding process, and the ear thickness of each group was measured using calipers after modeling and drug administration.

[0075] The external appearance of the ears of rats in each group is shown in the figure. Figure 4 Compared to the control group, the model group showed redness, swelling, and thickening of the auricular skin, with sebaceous scaling, significantly enlarged hair follicle pores, and some hair follicle openings becoming blocked into small cysts. The skin felt rough and hard to the touch. Measurements of the auricular region of rats in each group revealed significantly increased auricular thickness in both the model and control groups. The auricular thickness in each treatment group was also significantly greater than that in the model group. The changes in ear thickness in each group are shown in the table below. Figure 5 .

[0076] 3.2 Effects of different solvent-extracted parts of Lycium barbarum leaves on ear and lung tissues and serum inflammatory factors in model rats using ELISA kit

[0077] This invention investigates the effects of different solvent-extracted fractions of Lycium barbarum leaves on ear and lung tissues and serum inflammatory factors in model rats. ELISA kits were used to detect inflammatory factors (TNF-α, IL-1β, IL-6, IL-17) in rat ear and lung tissues and serum. Results are shown below. Figure 6Compared with the control group, the levels of TNF-α, IL-1β, IL-6, and IL-17 in the serum and ear tissues of rats in the model group, as well as the levels of TNF-α and IL-6 in the lung tissue, were significantly increased. Compared with the model group, the levels of TNF-α, IL-1β, IL-6, and IL-17 in the serum and ear tissues of all treatment groups, as well as the levels of TNF-α and IL-6 in the lung tissue, were significantly decreased, with the ethanol extract of Lycium barbarum leaves showing the most significant effect.

[0078] 3.3 Pathological effects of different solvent extracts of Lycium barbarum leaves on ear and lung tissues of model rats

[0079] This invention observes the effects of different solvent extracts of Lycium barbarum leaves on the pathological changes of ear and lung tissues in model rats to assess their efficacy in preventing and treating acne. HE pathological sections of the ear tissues from each group of rats are shown below. Figure 7 The results showed that the epidermis of the control group was intact, with sebaceous glands and hair follicles evenly distributed, and the collagen connective tissue density of the dermis was uniform, with no obvious inflammatory infiltration or hyperplasia. In the model group, moderate thickening of the granular and spinous layers of the epidermis, thickened keratin, incomplete keratinization, and a few microabscesses were observed. The dermis was thickened, with collagen hyperplasia and significant inflammatory cell infiltration. In all treatment groups, epidermal thickening was reduced, keratinization was alleviated, and inflammatory cell infiltration was improved. All treatment groups significantly improved hair follicle hyperplasia in rat ear acne tissue, reduced sebaceous gland enlargement, alleviated keratinization, and significantly reduced the number of infiltrating inflammatory cells in the dermis compared to the model group.

[0080] HE staining of lung tissue Figure 8 The results showed that the lung tissue of the normal group rats was structurally intact, with regular alveolar morphology, clear alveolar septa, and no obvious inflammatory cell infiltration or other abnormal pathological changes. The lung tissue of the model group rats showed more obvious pathological damage, mainly manifested as widened alveolar septa, mild interstitial edema, local vasodilation and congestion, and numerous inflammatory cell infiltrations around the bronchi and blood vessels, with unclear alveolar structure in some areas. Compared with the model group, the pathological damage to the lung tissue in each treatment group was reduced to varying degrees, manifested as a reduction in widened alveolar septa and inflammatory cell infiltration.

[0081] The above embodiments are only for illustrating the technical concept and features of the present invention, and are intended to enable those skilled in the art to understand the content of the present invention and implement it. They should not be used to limit the scope of protection of the present invention. All equivalent changes or modifications made in accordance with the spirit and essence of the present invention should be covered within the scope of protection of the present invention.

Claims

1. Application of wolfberry leaf extract in the preparation of products for the prevention and treatment of acne.

2. Application of Lycium barbarum leaf extract in the preparation of products for the prevention and treatment of lung-heat type acne.

3. Application of wolfberry leaf extract in the preparation of anti-skin inflammation products.

4. The application according to any one of claims 1 to 3, characterized in that, The wolfberry leaf extract was prepared by the following method: dried wolfberry leaves were weighed, water was added and heated under reflux to extract the wolfberry leaf aqueous extract, which was then concentrated under reduced pressure and freeze-dried to obtain the wolfberry leaf aqueous extract.

5. The application according to claim 4, characterized in that, The wolfberry leaf extract is prepared by the following method: weigh dried wolfberry leaves, add 5 to 10 times the mass of ultrapure water, heat and reflux to extract 1 to 3 times, each extraction for 0.5 to 1 hour, combine the filtrates to obtain wolfberry leaf aqueous extract, concentrate under reduced pressure, freeze dry to obtain wolfberry leaf aqueous extract.

6. The application according to any one of claims 1 to 3, characterized in that, The wolfberry leaf extract was prepared by the following method: dried wolfberry leaves were weighed, 70% ethanol was added, and the mixture was heated under reflux for extraction. The extracts were combined to obtain the wolfberry leaf ethanol extract, which was then concentrated under reduced pressure and freeze-dried to obtain the wolfberry leaf ethanol extract.

7. The application according to claim 6, characterized in that, The wolfberry leaf extract is prepared by the following method: weigh dried wolfberry leaves, add 5 to 10 times the mass of 70% ethanol, heat and reflux to extract 1 to 3 times, each extraction for 0.5 to 1 hour, to obtain wolfberry leaf ethanol extract, concentrate under reduced pressure, freeze dry to obtain wolfberry leaf ethanol extract.

8. The application according to any one of claims 1 to 3, wherein the product includes pharmaceutical products or daily chemical products.

9. The application according to claim 8, wherein the active extract of wolfberry leaves and a pharmaceutically acceptable carrier are formulated into tablets, capsules, granules, oral liquids, pills, mixtures or ointments.

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