Application of staphylococcus epidermidis H62-3 viable bacterium preparation in preparation of medicine for preventing or treating psoriasis

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to application of a staphylococcus epidermidis H62-3 viable bacterium preparation in preparation of a medicine for preventing or treating psoriasis. Experimental results show that the staphylococcus epidermidis H62-3 viable bacterium preparation improves skin erythema and scale symptoms of a psoriasis mouse model, remarkably reduces the skin lesion thickness, reduces the mRNA expression level of cytokines related to psoriasis at the skin lesion position of the psoriasis mouse model, and improves the psoriasis resistance of the psoriasis mouse model. And the cell factors comprise interleukin-17A, interleukin-17F and interleukin-23. The invention also discloses a preparation method of the cell factor. Therefore, the staphylococcus epidermidis H62-3 viable bacterium preparation has the prospect of being developed into the medicine for preventing or treating psoriasis.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and specifically relates to an application of a live bacterial preparation of Staphylococcus epidermidis H62-3 in preparing a medicine for preventing or treating psoriasis. Background Art

[0002] Psoriasis is a chronic inflammatory skin disease characterized by erythema, localized scaly papules and plaques. Psoriasis causes itching, irritation, burning and stinging. Its pathogenesis is complex and is believed to be the result of the combined action of genetic, epigenetic and environmental factors. The most common type of psoriasis is psoriasis vulgaris, which accounts for nearly 85-90% of all psoriasis cases. Pathologically, psoriasis manifests as excessive proliferation and abnormal differentiation of keratinocytes, dilation of new capillaries, and inflammatory infiltration of the dermis. Clinically, psoriasis manifests as recurrent multi-layered silvery-white dry scales, with extensive infiltrative erythema and hemorrhages under the scales, which may be accompanied by itching symptoms.

[0003] At present, the treatment of psoriasis is mainly divided into traditional therapy and biological targeted therapy according to the type of drugs. Traditional treatment can only achieve short-term clinical efficacy, is difficult to cure, and most are accompanied by adverse reactions of varying degrees; biological targeted therapy has higher efficacy and better safety than traditional therapy, but there are also some patients with poor treatment effects or adverse reactions. In addition, some biological agents are expensive. Psoriasis cannot be cured at present, but management should aim to minimize physical and psychological damage by treating patients early in the disease process, identifying and preventing related multi-morbidities, changing lifestyles, and adopting personalized treatment methods. For the treatment of psoriasis patients, the development of new safe, effective and economical psoriasis treatment drugs is of great significance.

[0004] Bacteria, fungi and viruses colonized on the surface of the skin constitute the skin microbiome, also known as the skin flora. Studies in recent years have shown that the skin flora of normal people interact with the host, and healthy skin flora is essential for maintaining the skin barrier function. A relative balance is maintained between pathogenic bacteria and beneficial bacteria on the surface of healthy skin. Once the balance is broken, it may cause skin inflammation, itching and even disease. For example, when the sebaceous glands secrete vigorously and sebum discharge is impaired, microorganisms in the hair follicles, especially Propionibacterium acnes, will multiply in large numbers. It secretes lipase, protease and hyaluronidase, destroys tissues and causes infection, forms pustules and acne, and thus affects personal appearance and life. At the same time, there are also a large number of beneficial bacteria on healthy skin. For example, beneficial bacteria represented by Staphylococcus epidermidis can secrete antimicrobial peptides, decompose sebum to produce metabolites such as short-chain fatty acids, and have antibacterial, immunomodulatory, and barrier repair promotion functions.

[0005] Studies have found that psoriasis patients have severe skin microbiome imbalance. Psoriasis-associated bacterial communities show higher diversity and heterogeneity than healthy skin bacterial communities, and specific microbial characteristics are associated with psoriasis lesional skin, psoriasis non-lesional skin, and healthy skin. Staphylococcus aureus is more abundant in both non-lesional and lesional skin of psoriasis. In contrast, the incidence of Staphylococcus epidermidis and Propionibacterium acnes in psoriasis lesions is lower than that in healthy skin, and Proteus is over-enriched in psoriasis skin. Based on the close association of skin flora with inflammatory skin diseases such as psoriasis, topical probiotics with therapeutic potential are screened, and pharmaceutical ointments or creams with the addition of specific probiotics are prepared to antagonize the growth of other harmful bacteria, or interact with the immune system to enhance its ability to repair lesions and maintain skin health. Summary of the invention

[0006] In view of the deficiencies in the prior art, the purpose of the present invention is to provide an application of a live bacterial preparation of Staphylococcus epidermidis H62-3 in the preparation of a drug for preventing or treating psoriasis, thereby providing a new idea for the effective treatment of psoriasis with topical probiotic strains or topical drugs for skin.

[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions: In a first aspect, the present invention provides a use of Staphylococcus epidermidis H62-3 in the preparation of a medicament for preventing or treating psoriasis.

[0008] The present invention first collects facial skin flora of healthy people of different ages, and isolates the effective Staphylococcus epidermidis strain H62-3 through culture omics. The classification of the Staphylococcus epidermidis is named Staphylococcus epidermidis ( Staphylococcus epidermidis ), which was deposited in the General Microbiology Center of China Microorganism Culture Collection Committee (CGMCC) on February 21, 2023. The deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 26640. For relevant information, please refer to CN116286543A.

[0009] This strain is Gram-positive, with small white colonies on TSA plates, forming round protrusions with a smooth and opaque surface. It is spherical under a microscope, has no flagella, cannot move, and does not produce spores. Its optimal growth temperature is 32-38°C. It grows rapidly in TSB liquid culture medium, enters the logarithmic phase in about 2 hours, and enters the stable phase in about 12 hours.

[0010] In some embodiments of the present invention, the Staphylococcus epidermidis H62-3 improves the skin erythema and scaling symptoms of psoriasis mouse models and significantly reduces the thickness of skin lesions.

[0011] In some embodiments of the present invention, the Staphylococcus epidermidis H62-3 is used to reduce the expression level of cytokine mRNA associated with psoriasis in damaged skin.

[0012] In some embodiments of the present invention, the cytokines include interleukin-17A, interleukin-17F, and interleukin-23.

[0013] In some embodiments of the present invention, the psoriasis is at least one of psoriasis vulgaris, erythrodermic psoriasis, articular psoriasis or pustular psoriasis.

[0014] In a second aspect, the present invention provides a use of a live bacterial preparation of Staphylococcus epidermidis H62-3 in the preparation of a medicament for preventing or treating psoriasis, wherein the live bacterial preparation of Staphylococcus epidermidis H62-3 is prepared from the Staphylococcus epidermidis H62-3 described in the first aspect.

[0015] In some embodiments of the present invention, the live Staphylococcus epidermidis H62-3 preparation improves the skin erythema and scaling symptoms of psoriasis mouse models and significantly reduces the thickness of skin lesions.

[0016] In some embodiments of the present invention, the live bacterial preparation of Staphylococcus epidermidis H62-3 is used to reduce the expression level of cytokine mRNA associated with psoriasis in damaged skin.

[0017] In some embodiments of the present invention, the cytokines include interleukin-17A, interleukin-17F, and interleukin-23.

[0018] In some embodiments of the present invention, the psoriasis is at least one of psoriasis vulgaris, erythrodermic psoriasis, articular psoriasis or pustular psoriasis.

[0019] In some embodiments of the present invention, the preparation method of the live bacterial preparation of Staphylococcus epidermidis H62-3 comprises the following steps: picking a single colony of the activated Staphylococcus epidermidis strain H62-3 and inoculating it in a TSB liquid culture medium, placing it in a 32-38°C shaker, and shaking and culturing it for 14-18 hours to serve as a seed liquid; inoculating the seed liquid into the TSB liquid culture medium at an inoculation rate of 3%-5% by volume, shaking and culturing it in a 32-38°C shaker for 14-18 hours to obtain a fermentation liquid; centrifuging the fermentation liquid, resuspending the bacterial precipitate with a TSB liquid culture medium, and preparing a 1х10 9 CFU / mL of live Staphylococcus epidermidis preparation.

[0020] The present invention first collects facial skin flora of healthy people of different age stages, isolates single strains through culture omics, and screens and verifies the efficacy of the strains. The live bacterial preparation of Staphylococcus epidermidis H62-3 can improve the symptoms of skin erythema, scaling and the like in a psoriasis mouse model, significantly reduce the thickness of skin lesions, and reduce the mRNA expression level of cytokines associated with psoriasis in the skin lesions of the psoriasis mouse model. The cytokines include interleukin-17A, interleukin-17F, and interleukin-23. The preparation has the prospect of being developed into a drug for preventing or treating psoriasis.

[0021] The beneficial effects of the present invention are: (1) The present invention provides a use of a live bacterial preparation of Staphylococcus epidermidis H62-3 in the preparation of a drug for preventing or treating psoriasis. The live bacterial preparation of Staphylococcus epidermidis H62-3 can improve symptoms such as skin erythema and scaling in a psoriasis mouse model, significantly reduce the thickness of skin lesions, and reduce the mRNA expression level of cytokines associated with psoriasis in the skin lesions of the psoriasis mouse model. The cytokines include interleukin-17A, interleukin-17F, and interleukin-23. The live bacterial preparation of Staphylococcus epidermidis H62-3 has the prospect of being developed into a drug for preventing or treating psoriasis.

[0022] (2) The present invention provides an application of Staphylococcus epidermidis H62-3 in the preparation of a drug for preventing or treating psoriasis, which provides new ideas for the safe and effective treatment of psoriasis with topical live bacteria and topical probiotic strains for the skin. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings: Figure 1 This is the colony morphology of Staphylococcus epidermidis H62-3; Figure 2 This is the Gram stain of Staphylococcus epidermidis H62-3; Figure 3 is the growth curve of Staphylococcus epidermidis H62-3; Figure 4 The ANI analysis results of Staphylococcus epidermidis H62-3; Figure 5 The effect of Staphylococcus epidermidis H62-3 on imiquimod-induced psoriasis lesions in C57BL / 6 mice; Figure 6 The effect of Staphylococcus epidermidis H62-3 on the PASI score of psoriasis lesions induced by imiquimod in C57BL / 6 mice; Figure 7The effect of Staphylococcus epidermidis H62-3 on the psoriasis-like skin pathology induced by imiquimod in C57BL / 6 mice; Figure 8 The effect of Staphylococcus epidermidis H62-3 on the thickness of stratum spinosum of psoriasis lesions induced by imiquimod in C57BL / 6 mice; Fig. 9 The effect of Staphylococcus epidermidis H62-3 on the spleen index of C57BL / 6 mice induced by imiquimod; Fig.10 To investigate the effects of Staphylococcus epidermidis H62-3 on imiquimod-induced psoriasis lesions in C57BL / 6 mice and the transcriptional levels of psoriasis-related cytokines; Fig.11 The effect of Staphylococcus epidermidis H62-3 on Ki67 protein level in imiquimod-induced psoriasis lesions of C57BL / 6 mice. DETAILED DESCRIPTION

[0024] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

[0025] The bacterial culture medium used in the following examples is as follows: Tryptone soy broth liquid culture medium (TSB): tryptone 17.0 g / L, soy peptone 3.0 g / L, sodium chloride 5.0 g / L, potassium dihydrogen phosphate 2.5 g / L, glucose 2.5 g / L, autoclave at 121°C for 15 min and set aside.

[0026] Tryptone soy broth solid medium (TSA): tryptone 17.0 g / L, soy peptone 3.0 g / L, sodium chloride 5.0 g / L, dipotassium hydrogen phosphate 2.5 g / L, glucose 2.5 g / L, agar powder 15 g / L, autoclave at 121°C for 15 min, pour into plates and set aside.

[0027] Example 1: This example is the separation, screening and purification of Staphylococcus epidermidis H62-3 described in the present invention (1) Sample collection ① Volunteers of different ages (18-90 years old) with healthy facial skin were recruited. They were required to have no acne, pustules, inflammation, erythema, etc., and had not taken oral antibiotics in the past 2 months or applied topical antibiotics in the past 2 weeks. The face was not cleaned or washed within 12 hours of sampling.

[0028] ② The sampler wears a mask and disposable sterile gloves, soaks a sterile sampling cotton swab with sterile saline, and uses a 2cm cotton swab to make a 4cm long cut at the sampling site. Wipe back and forth 50 times for about 30 seconds, covering an area of ​​about 8cm 2 After wiping, place the cotton swab in a glycerin storage tube, seal it tightly with the lid, and record the number.

[0029] (2) Cultivation, isolation, purification and preservation of skin flora ① Take 0.05 mL of glycerol preservative solution and add 0.15 mL of sterile water to dilute it. Spread the dilution on a TSA solid plate and place it in a 37°C constant temperature incubator and culture it upside down for 24 hours.

[0030] ② According to the colony characteristics of Staphylococcus epidermidis, single colonies were picked and separated by four-zone lines, and placed in an inverted incubator at 37°C for 24 hours.

[0031] ③ Perform 16S rRNA sequencing and determine that the single colony is Staphylococcus epidermidis and store it in glycerol at -80℃.

[0032] (3) Screening of functional bacteria The whole genome RNA of bacteria from different strains of Staphylococcus epidermidis was extracted and reverse transcribed into cDNA. qRT-PCR experiments were performed to screen out the Staphylococcus epidermidis strain H62-3 that highly expressed genes closely related to moisturizing efficacy.

[0033] Example 2: This example is the microbiological identification of Staphylococcus epidermidis H62-3 (1) Colony characteristics ① The colony morphology of the strain on TSA plate is as follows Figure 1 As shown, the colonies are white and small, forming round protrusions with a smooth and opaque surface and no hemolysis zone.

[0034] ② Gram stain positive, such as Figure 2 As shown in the figure, the bacteria are spherical, some are arranged in grape-like shapes, some are arranged singly, without flagella, and do not produce spores.

[0035] (2) Cultivation characteristics ①The optimum growth temperature of this strain is 32-38℃, and the optimum pH is 5.0-8.0.

[0036] ② Pick a single colony of the activated strain and inoculate it into TSB liquid culture medium, place it in a 32-38℃ shaker, and shake and culture it for 14-18h as seed liquid; inoculate the seed liquid into TSB liquid culture medium with an inoculum volume fraction of 3%-5%, shake and culture it in a 37℃ shaker for 24h, measure the OD value of the culture solution at 600nm every 1h, and draw the growth curve of the strain with OD600 as the vertical axis and time as the horizontal axis, as shown in the figure. Figure 3As shown, it can be found that Staphylococcus epidermidis H62-3 grows rapidly in TSB liquid culture medium, enters the logarithmic phase in about 2 hours, and enters the stable phase in about 12 hours.

[0037] (3) Genetic characteristics The genomic DNA of strain H62-3 was extracted, and the sample was sequenced using the Illumina Hiseq 2000 sequencing platform to obtain second-generation data, and PacBio sequencing was used to obtain third-generation data. After quality control, Denovo assembly was performed, and gene prediction was performed after splicing and correction into a complete genome. The predicted gene protein was aligned to various databases such as the NR library to obtain the species information of the strain. As shown in Table 1, it can be determined that strain H62-3 is Staphylococcus epidermidis.

[0038] Table 1: Sequence alignment results of Staphylococcus epidermidis H62-3

[0039] Based on the above-mentioned characteristics of colony, culture, genetics, etc., strain H62-3 was determined to be Staphylococcus epidermidis. Since the ANI of Staphylococcus epidermidis H62-3 and the model strain of Staphylococcus epidermidis CGMCC 1.4260 is 97.1%, Figure 4 As shown, it can be determined that the two are not the same strain. Single nucleotide diversity (SNP) and insertion and deletion (InDel) analysis found that many bases of Staphylococcus epidermidis H62-3 mutated, increased or deleted, resulting in changes in the protein and affecting its biological function. The strain was deposited in the China General Microbiological Culture Collection Center (CGMCC) on February 21, 2023, and its deposit number is CGMCC No. 26640.

[0040] Example 3: This example is an experiment on improving the psoriasis skin lesions of C57BL / 6 mice induced by imiquimod using a live bacterial preparation of Staphylococcus epidermidis H62-3 Feeding environment: SPF grade; mice: C57BL / 6 female mice, 7 weeks old; modeling method: After a week of stable feeding, the purchased 7-week-old C57BL / 6 female mice of uniform size were depilated from the back by 2 cm х 2 cm and randomly divided into 4 groups, 5 mice in each group. 62.5 mg of imiquimod was evenly applied to the back of each mouse every morning for 6 consecutive days. Four treatment methods were set up in the experiment, and 5 biological replicates were set for each treatment method. The drug administration was unified on the day of modeling, with local administration, modeling every morning, and drug administration 12 hours later. The treatment method is as follows: ① Treatment 1 (control), normal control mice, without any treatment; ② Treatment 2 (imiquimod): 62.5 mg of imiquimod was evenly applied to the back of each mouse every morning, and 12 hours later, a blank matrix of topical live bacterial preparation was given for 6 consecutive days, which was the psoriasis model group; ③ Treatment 3 (calcipotriol): 62.5 mg of imiquimod was evenly applied to the back of each mouse every morning, and 12 hours later, 12 mg / 4 cm of calcipotriol ointment was applied topically. 2 , for 6 consecutive days, which was the positive control group; ④ Treatment 4 (H62-3): 62.5 mg of imiquimod was evenly applied to the back of each mouse every morning, and 2×10 live Staphylococcus epidermidis H62-3 preparation was applied topically 12 hours later. 8 CFU / 4 cm 2 , for 6 consecutive days, which was the experimental group.

[0041] The mice in each group were photographed every other day and the PASI score of psoriasis lesions was calculated. The mice were killed on the 7th day, and the back lesions were taken for HE staining analysis of the lesions and pathology. Figure 5 As shown in the figure, compared with normal control mice, imiquimod induced typical psoriasis lesions, which were manifested as thickening of the skin on the back of mice, erythema and scaling. Compared with psoriasis model mice, the group with topical Staphylococcus epidermidis H62-3 live bacterial preparations improved erythema, scaling and skin thickening, and the efficacy was comparable to that of the calcipotriol positive control group. However, the mice in the calcipotriol positive control group generally lost weight and had obvious side effects. Therefore, topical Staphylococcus epidermidis H62-3 live bacterial preparations for the treatment of psoriasis is a new safe and low-toxic therapy.

[0042] Example 4: This example is an experiment on the improvement of PASI score of psoriasis lesions in C57BL / 6 mice induced by imiquimod by live bacterial preparation of Staphylococcus epidermidis H62-3 Mice were killed according to Example 3, and the erythema, scaling and thickening of the back skin lesions were scored. The severity of the back skin lesions, including skin erythema, scaling and infiltration thickness, was evaluated according to the Psoriasis Area and Severity Index (PASI) scoring system (see Table 2). Each parameter was independently scored on a scale of 0 to 4, where 0 = no symptoms, 1 = mild symptoms, 2 = moderate symptoms, 3 = obvious symptoms, and 4 = very obvious symptoms, as shown in Table 2. The cumulative score represents the statistical results of the severity of the disease. Compared with normal control mice, after continuous application of imiquimod for 6 days, obvious erythema, scaling and skin thickening appeared on the back of the mice. Both the calcipotriol positive control group and the Staphylococcus epidermidis H62-3 live bacterial preparation group significantly alleviated the back erythema, scaling and skin thickening induced by imiquimod, as shown in Table 2. Figure 6 shown.

[0043] Table 2: Scoring details of mouse psoriasis-like dermatitis severity (PASI)

[0044] Example 5: This example is an experiment on the improvement of imiquimod-induced psoriasis skin lesions in C57BL / 6 mice by live bacterial preparation of Staphylococcus epidermidis H62-3 The skin lesions of the mice killed in Example 3 were stained with HE. The typical pathological results are as follows: Figure 7 Compared with normal control mice, after continuous application of imiquimod for 6 days, the mice in the modeling group showed phenotypes such as epidermal thickening, inflammatory cell infiltration, hyperkeratosis, incomplete keratosis, reduced granular layer, and elongated epidermal protrusions. The calcipotriol positive control group and the Staphylococcus epidermidis H62-3 live bacteria preparation group can effectively alleviate the phenotypes of epidermal thickening, inflammatory cell infiltration, hyperkeratosis, incomplete keratosis, reduced granular layer, and elongated epidermal protrusions induced by imiquimod. Figure 7 shown.

[0045] Example 6: This example is an experiment on the improvement of the spinous layer thickness of psoriasis lesions in C57BL / 6 mice induced by imiquimod by the live bacterial preparation of Staphylococcus epidermidis H62-3 The skin lesions of the mice killed in Example 3 were taken for HE staining, observed under a microscope, scanned and photographed, and histopathological analysis was performed, and the acanthosis and dermal inflammatory cell infiltration indicators were quantified. The quantification method of acanthosis is to select the visual field of the scanned image under the microscope after HE staining, fix the magnification (×20), and use Adobe Photoshop2023 software to select all epidermal areas in the visual field in the visual field to obtain the pixel area of ​​the epidermal area, and the degree of acanthosis = epidermal pixels / total pixels in the visual field. The quantification method of dermal inflammatory cell infiltration is to select the visual field of the scanned image under the microscope after HE staining, fix the magnification (×20), and use Adobe Photoshop 2023 software in the visual field to randomly select 5 non-overlapping square areas (the side length is fixed to 1 unit) in the dermis area, and use the counting tool to calculate the number of inflammatory cells in each area, then the dermal inflammatory cell infiltration = average unit area inflammatory cell count. Compared with normal control mice, after continuous application of imiquimod for 6 days, the model group mice had acanthosis and dermal inflammatory cell infiltration. The calcipotriol positive control group and the Staphylococcus epidermidis H62-3 live bacteria preparation group could effectively alleviate the acanthosis and dermal inflammatory cell infiltration induced by imiquimod in mice. Figure 8 shown.

[0046] Example 7: This example is an experiment on the improvement of spleen index of C57BL / 6 mice with psoriasis induced by imiquimod by live bacterial preparation of Staphylococcus epidermidis H62-3 The spleens of the mice sacrificed in Example 3 were taken, and the weight and spleen size of the mice were counted. Compared with the normal control mice, the spleen index of the mice increased significantly after continuous application of imiquimod for 6 days. The calcipotriol positive control group and the Staphylococcus epidermidis H62-3 live bacteria preparation group could slightly alleviate the increase in the spleen index of mice induced by imiquimod, such as Fig. 9 shown.

[0047] Example 8: This example is an experiment on the expression of transcriptional levels of cytokines related to psoriasis in C57BL / 6 mice induced by imiquimod by live bacterial preparation of Staphylococcus epidermidis H62-3 The skin lesions of the mice killed in Example 3 were taken, and the total RNA was extracted by the Trizol method, which was reverse transcribed into cDNA, and then qPCR was performed to detect the expression of three inflammatory factor genes. Compared with normal control mice, continuous application of imiquimod for 6 days can induce psoriasis-like dermatitis in mice, involving the release of cytokines and cell infiltration related to psoriasis in the skin lesions. Compared with normal control mice, the levels of psoriasis-related cytokines IL-17A, IL-17F, and IL-23 in the skin lesions of mice in the imiquimod modeling group were significantly increased. The calcipotriol positive control group and the Staphylococcus epidermidis H62-3 live bacterial preparation group can reduce the transcription levels of the above cytokines to varying degrees, such as Fig.10 As shown. In the calcipotriol positive control group, the inhibition rates of IL-17A, IL-17F, IL-23, and IL-23 after local administration can reach 70.8%, 67.5%, and 67.2%, respectively. In the Staphylococcus epidermidis H62-3 live bacteria preparation group, the inhibition rates of IL-17A, IL-17F, and IL-23 after local administration can reach 55.9%, 55.0%, and 53.2%, respectively.

[0048] Example 9: This example is an experiment on the improvement of Ki67 protein expression level in imiquimod-induced psoriasis lesions of C57BL / 6 mice by live bacterial preparation of Staphylococcus epidermidis H62-3 The skin lesions of the mice killed in Example 3 were fixed, paraffin-embedded, sliced, and stained with Ki67 immunohistochemistry. They were observed under a microscope and scanned and photographed. Compared with the normal control mice, the expression level of Ki67 in the skin lesions of the mice in the imiquimod modeling group was significantly increased, and the calcipotriol positive control group and the Staphylococcus epidermidis H62-3 live bacterial preparation group could effectively reduce the expression level of Ki67, such as Fig.11 shown.

[0049] The present invention provides an application of a live bacterial preparation of Staphylococcus epidermidis H62-3 in the preparation of a drug for treating psoriasis. The live bacterial preparation of Staphylococcus epidermidis H62-3 can improve the symptoms of skin erythema and scaling in a psoriasis mouse model, significantly reduce the thickness of skin lesions, and reduce the expression level of cytokine mRNA related to psoriasis in the skin lesions of the psoriasis mouse model. The inflammatory factors include interleukin-17A, interleukin-17F, and interleukin-23. The live bacterial preparation of Staphylococcus epidermidis H62-3 has the prospect of being developed into a drug for treating psoriasis. The present invention provides an application of a live bacterial preparation of Staphylococcus epidermidis H62-3 in the preparation of a drug for treating psoriasis, providing new ideas for the treatment of psoriasis with topical live bacteria and the safe and effective treatment of psoriasis with topical probiotic strains for skin.

[0050] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. Use of live Staphylococcus epidermidis H62-3 bacteria in the preparation of a drug for preventing or treating psoriasis.

2. Use of Staphylococcus epidermidis H62-3 according to claim 1 in the preparation of a medicament for preventing or treating psoriasis, characterized in that: The Staphylococcus epidermidis H62-3 improves the skin erythema and scaling symptoms of the psoriasis mouse model and significantly reduces the thickness of skin lesions.

3. Use of Staphylococcus epidermidis H62-3 according to claim 1 in the preparation of a medicament for preventing or treating psoriasis, characterized in that: The Staphylococcus epidermidis H62-3 is used to reduce the expression level of cytokine mRNA associated with psoriasis in damaged skin.

4. Use of Staphylococcus epidermidis H62-3 as claimed in claim 3 in the preparation of a medicament for preventing or treating psoriasis, characterized in that: The cytokines include interleukin-17A, interleukin-17F, and interleukin-23.

5. Use of Staphylococcus epidermidis H62-3 according to claim 1 in the preparation of a medicament for preventing or treating psoriasis, characterized in that: The psoriasis is at least one of psoriasis vulgaris, erythrodermic psoriasis, articular psoriasis or pustular psoriasis.

6. Use of a live bacterial preparation of Staphylococcus epidermidis H62-3 in the preparation of a drug for preventing or treating psoriasis, characterized in that: The live bacteria preparation of Staphylococcus epidermidis H62-3 is prepared from the Staphylococcus epidermidis H62-3 described in claim 1.

7. Use of the live bacterial preparation of Staphylococcus epidermidis H62-3 according to claim 6 in the preparation of a medicament for preventing or treating psoriasis, characterized in that: The live bacterial preparation of Staphylococcus epidermidis H62-3 improves the skin erythema and scaling symptoms of a psoriasis mouse model and significantly reduces the thickness of skin lesions.

8. Use of the live bacterial preparation of Staphylococcus epidermidis H62-3 according to claim 6 in the preparation of a medicament for preventing or treating psoriasis, characterized in that: The live bacteria preparation of Staphylococcus epidermidis H62-3 is used to reduce the expression level of cytokine mRNA associated with psoriasis in damaged skin, and the cytokines include interleukin-17A, interleukin-17F, and interleukin-23.

9. Use of the live bacterial preparation of Staphylococcus epidermidis H62-3 according to claim 6 in the preparation of a medicament for preventing or treating psoriasis, characterized in that: The psoriasis is at least one of psoriasis vulgaris, erythrodermic psoriasis, articular psoriasis or pustular psoriasis.

10. Use of the live bacterial preparation of Staphylococcus epidermidis H62-3 according to any one of claims 6 to 9 in the preparation of a medicament for preventing or treating psoriasis, characterized in that: The preparation method of the Staphylococcus epidermidis H62-3 live bacteria preparation comprises the following steps: The activated Staphylococcus epidermidis strain H62-3 was selected and inoculated into TSB liquid culture medium, placed in a shaking incubator at 32-38°C, and cultured for 14-18 hours to obtain a seed solution; the seed solution was inoculated into TSB liquid culture medium at a volume fraction of 3%-5%, and cultured for 14-18 hours in a shaking incubator at 32-38°C to obtain a fermentation solution; the fermentation solution was centrifuged, and the bacterial precipitate was resuspended in TSB liquid culture medium to prepare 1х10 9 CFU / mL of live Staphylococcus epidermidis preparation.

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