Application of Lactobacillus casei Zhang in preparing preparations for treating skin diseases

Abstract

The present invention discloses the use of Lactobacillus casei Zhang in preparing a preparation for treating skin diseases, relating to the technical field of microbial probiotic preparations. The present invention provides the use of Lactobacillus casei Zhang in preparing a preparation for treating or alleviating psoriasis. The Lactobacillus casei Zhang has a deposit number of CGMCC No. 5469. The Lactobacillus casei Zhang of the present invention has an alleviating effect on the symptoms of psoriasis, can inhibit the weight loss trend of psoriasis patients, and reduce the spleen index. It can be used to prepare a preparation for alleviating psoriasis. It can also be used as a microecological preparation in medicines, fermented foods, or health products, providing a potential option for developing new psoriasis treatment methods.

Description

Technical Field

[0001] The present invention relates to the technical field of microbial probiotic preparations, and in particular to application of Lactobacillus casei Zhang in preparing preparations for treating skin diseases. Background Art

[0002] Psoriasis is a chronic, immune-mediated skin disease with a long course and high recurrence rates. Symptoms include erythema and scaling, with itching and pain being the primary symptoms. Psoriasis, commonly referred to as psoriasis, is clinically categorized into psoriasis vulgaris, pustular psoriasis, erythrodermic psoriasis, and arthritic psoriasis.

[0003] At present, the main clinical methods for treating psoriasis are topical medication, phototherapy and systemic treatment. Topical medication is a common treatment method for patients with mild psoriasis, but the effect is not very obvious for patients with moderate and severe psoriasis. Its advantages are that it can directly reach the affected area, has a fast onset of action, is easy to use, and has few side effects on other parts of the body. However, long-term use may cause local side effects. Ultraviolet light therapy is a common treatment for psoriasis. It can promote the apoptosis of T cells and delay the proliferation of lymphocytes. However, ultraviolet light therapy may cause side effects such as itching, desquamation, and redness and swelling.

[0004] Psoriasis is often considered a disease that manifests solely on the skin. However, studies have shown that gut microbial disturbances are associated with the development and / or progression of chronic low-grade inflammation. Furthermore, serum levels of psoriasis patients are found to harbor a high number of dysregulated cytokines and inflammatory markers. Therefore, the systemic inflammation associated with psoriasis may be closely linked to imbalances in the gut and skin microbiota. Currently, exploring the relationship between health and disease from a microbial perspective is becoming a growing focus and a hot topic. Probiotics can colonize the intestine, forming a natural biological barrier with the intestinal mucosa by regulating the gut microbiota and increasing the expression of adhesion proteins. Probiotics have been shown to suppress the inflammatory response in atopic dermatitis and have a clear therapeutic effect, providing insights into the potential use of probiotics in the treatment of psoriasis.

[0005] Chinese patent CN115708835A discloses a strain of Bifidobacterium breve that can alleviate psoriasis and its application. The deposit number of Bifidobacterium breve CCFM683 described in the invention is CGMCC No. 11828. The Bifidobacterium breve CCFM683 of the invention has the effect of alleviating psoriasis caused by imiquimod, which is specifically reflected in: (1) significantly reducing the weight loss of psoriasis mice; (2) significantly improving the pathological characteristics of psoriasis-like mice such as scaling, thickening and redness; (3) regulating the level of immune factors in the skin of psoriasis-like mice; and (4) significantly inhibiting the proliferation of keratinocytes in the skin of psoriasis mice.

[0006] Chinese patent CN112812989A discloses a bifidobacterium adolescentis capable of relieving psoriasis and its application. The deposit number of the bifidobacterium adolescentis in the invention is GDMCC No. 61349. The bifidobacterium adolescentis provided by the invention ( Bifidobacterium adolescentis )CCFM667 has the effect of alleviating psoriasis, alleviating the skin lesions of psoriasis-like mice and inhibiting the thickening of the epidermis of the skin of psoriasis-like mice, reducing the spleen weight of psoriasis-like mice, and reducing the levels of IL-22 and IL-23 in the skin of psoriasis-like mice by 30.2%.

[0007] Currently, there are few studies on probiotics that can alleviate psoriasis, and there is an urgent need to provide a probiotic that can effectively alleviate psoriasis. Summary of the Invention

[0008] The present invention aims to provide an application of Lactobacillus casei Zhang in preparing a preparation for treating psoriasis.

[0009] In order to achieve the above-mentioned object of the invention, the technical solution of the present invention is as follows:

[0010] In one aspect, the present invention provides a use of Lactobacillus casei Zhang in preparing a preparation for treating or alleviating psoriasis. The Lactobacillus casei Zhang has a deposit number of CGMCC No. 5469.

[0011] Specifically, the taxonomic name of the Lactobacillus casei Zhang is Lacticaseibacillus casei It has been deposited in the General Microbiology Center of China Culture Collection Administration of Microorganisms, with the deposit number CGMCC No.5469 and the deposit date being November 18, 2011.

[0012] Specifically, the psoriasis includes psoriasis vulgaris, pustular psoriasis, erythrodermic psoriasis and arthritic psoriasis.

[0013] Specifically, the viable count of Lactobacillus casei Zhang in the preparation is 1×10 6 -1×10 11 CFU / mL or 1×10 6 -1×10 11 CFU / g.

[0014] For example, 1×10 6 CFU / mL (CFU / g), 1×10 7 CFU / mL (CFU / g), 1×10 8 CFU / mL (CFU / g), 1×10 9 CFU / mL (CFU / g), 1×10 10CFU / mL (CFU / g), 1×10 11 CFU / mL (CFU / g), etc. Other point values ​​within this numerical range can be selected.

[0015] Specifically, the preparation includes Lactobacillus casei Zhang fermentation liquid supernatant, Lactobacillus casei Zhang live bacteria or Lactobacillus casei Zhang freeze-dried powder.

[0016] Furthermore, the live bacteria, also known as active flora, can colonize and multiply in the intestine, which is beneficial to increasing the number of beneficial bacteria.

[0017] Furthermore, the freeze-dried powder refers to the powder obtained by freeze-drying the strain.

[0018] Furthermore, the preparation includes the supernatant of Lactobacillus casei Zhang fermentation broth.

[0019] Furthermore, the dosage of the fermentation broth supernatant is 2.625 mg / kg, 5.25 mg / kg or 10.5 mg / kg.

[0020] Specifically, the preparation method of the Lactobacillus casei Zhang fermentation broth supernatant comprises the following steps:

[0021] (1) Lactobacillus casei Zhang was divided into (1-10) × 10 6 The inoculum size of CFU / g was inoculated into the fermentation medium;

[0022] (2) Cultivate at 35-38°C until the pH reaches 4-6;

[0023] (3) Centrifuge to obtain the fermentation liquid supernatant.

[0024] Furthermore, the inoculum size in step (1) is 6×10 6 CFU / g.

[0025] Furthermore, the fermentation medium in step (1) is selected from at least one of MRS medium, M17 medium, PDA medium, LB medium, TB medium, SOB medium, MEM medium, DMEM medium, and RPMI1640 medium.

[0026] Furthermore, the fermentation medium in step (1) is MRS medium.

[0027] Furthermore, the culture conditions of step (2) are: culturing at 37°C until the pH reaches 4.5.

[0028] Furthermore, the fermentation broth supernatant in step (3) also includes a filtration process using a 0.22 μm filter membrane.

[0029] Furthermore, the preparation also includes a pharmaceutically acceptable carrier.

[0030] Furthermore, the pharmaceutically acceptable carrier is selected from one or more of an excipient, a stabilizer, a diluent, a binder, a preservative, and a lubricant.

[0031] Specifically, the dosage form of the drug includes, but is not limited to, a gastrointestinal dosage form and a parenteral dosage form according to the administration method.

[0032] Furthermore, the dosage forms for administration via the gastrointestinal tract include but are not limited to tablets, powders, granules, solutions, capsules, emulsions, suspensions, and oils.

[0033] Furthermore, the non-gastrointestinal dosage forms include but are not limited to: injection dosage forms, respiratory tract dosage forms, skin dosage forms, mucosal dosage forms and cavity dosage forms.

[0034] Furthermore, the injectable dosage forms include but are not limited to intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, and intracavitary injection.

[0035] Furthermore, the respiratory tract administration dosage forms include but are not limited to sprays, aerosols and powder sprays.

[0036] Furthermore, the skin administration dosage forms include but are not limited to lotions, ointments, external solutions, plasters, pastes, and patches.

[0037] Furthermore, the mucosal administration dosage form includes but is not limited to eye drops, nasal drops, eye ointments, sublingual tablets, and patches.

[0038] Furthermore, the intracavitary administration dosage forms include but are not limited to suppositories, aerosols, effervescent tablets, drops and pills.

[0039] Furthermore, the preparation is in the form of skin administration.

[0040] Specifically, the preparation has the following effects:

[0041] (1) It can inhibit the weight loss trend of psoriasis patients;

[0042] (2) Reduce spleen index;

[0043] (3) Alleviate erythema, epidermal desquamation and infiltration symptoms on the back skin.

[0044] Specifically, the preparation includes a drug.

[0045] In another aspect, the present invention provides a preparation for treating or alleviating psoriasis, wherein the preparation comprises Lactobacillus casei Zhang, and the deposit number of the Lactobacillus casei Zhang is CGMCC No. 5469.

[0046] Specifically, the preparation includes Lactobacillus casei Zhang fermentation broth supernatant, Lactobacillus casei Zhang live bacteria, and Lactobacillus casei Zhang freeze-dried powder.

[0047] Furthermore, the preparation includes the supernatant of Lactobacillus casei Zhang fermentation broth.

[0048] Furthermore, the dosage of the fermentation broth supernatant is 2.625 mg / kg, 5.25 mg / kg or 10.5 mg / kg.

[0049] Specifically, the preparation method of the Lactobacillus casei Zhang fermentation broth supernatant comprises the following steps:

[0050] (1) Lactobacillus casei Zhang was divided into (1-10) × 10 6 The inoculum size of CFU / g was inoculated into the fermentation medium;

[0051] (2) Cultivate at 35-38°C until the pH reaches 4-6;

[0052] (3) Centrifuge to obtain the fermentation liquid supernatant.

[0053] Furthermore, the inoculum size in step (1) is 6×10 6 CFU / g.

[0054] Furthermore, the fermentation medium in step (1) is selected from at least one of MRS medium, M17 medium, PDA medium, LB medium, TB medium, SOB medium, MEM medium, DMEM medium, and RPMI1640 medium.

[0055] Furthermore, the fermentation medium in step (1) is MRS medium.

[0056] Furthermore, the culture conditions of step (2) are: culturing at 37°C until the pH reaches 4.5.

[0057] Furthermore, the fermentation broth supernatant in step (3) also includes a filtration process using a 0.22 μm filter membrane.

[0058] Furthermore, when the preparation is a medicine, the medicine also includes a pharmaceutically acceptable carrier.

[0059] Furthermore, the pharmaceutically acceptable carrier is selected from one or more of an excipient, a stabilizer, a diluent, a binder, a preservative, and a lubricant.

[0060] Specifically, the dosage form of the drug includes, but is not limited to, a gastrointestinal dosage form and a parenteral dosage form according to the administration method.

[0061] Furthermore, the dosage forms for administration via the gastrointestinal tract include but are not limited to tablets, powders, granules, solutions, capsules, emulsions, suspensions, and oils.

[0062] Furthermore, the non-gastrointestinal dosage forms include but are not limited to: injection dosage forms, respiratory tract dosage forms, skin dosage forms, mucosal dosage forms and cavity dosage forms.

[0063] Furthermore, the injectable dosage forms include but are not limited to intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, and intracavitary injection.

[0064] Furthermore, the respiratory tract administration dosage forms include but are not limited to sprays, aerosols and powder sprays.

[0065] Furthermore, the skin administration dosage forms include but are not limited to lotions, ointments, external solutions, plasters, pastes, and patches.

[0066] Furthermore, the mucosal administration dosage form includes but is not limited to eye drops, nasal drops, eye ointments, sublingual tablets, and patches.

[0067] Furthermore, the intracavitary administration dosage forms include but are not limited to suppositories, aerosols, effervescent tablets, drops and pills.

[0068] Furthermore, the dosage form of the drug is for skin administration.

[0069] The beneficial effects of the present invention are:

[0070] The present invention proves through experiments that Lactobacillus casei Zhang intervention can inhibit the weight loss trend of psoriasis mice and reduce the spleen index. At the same time, it can effectively reduce the severity of psoriasis in mice and alleviate psoriasis symptoms. BRIEF DESCRIPTION OF THE DRAWINGS

[0071] Figure 1 This study was designed to investigate the efficacy of Lactobacillus casei Zhang active ingredients in alleviating psoriasis.

[0072] Figure 2 The figure shows the effect of Lactobacillus casei Zhang active substance on the spleen volume of mice. From left to right in the figure are the normal group, model group, low-dose group, medium-dose group, and high-dose group.

[0073] Figure 3 The effect of Lactobacillus casei active substances on the body weight of mice.

[0074] Figure 4 This is the effect of Lactobacillus casei Zhang active substance on the spleen index of mice.

[0075] Figure 5 This figure shows the effects of Lactobacillus casei Zhang active substances on skin damage in psoriasis mice. From top to bottom in the figure are the normal group, model group, low-dose group, medium-dose group, and high-dose group.

[0076] Figure 6 This is the effect of Lactobacillus casei Zhang active substances on the PASI scores of mice.

[0077] Figure 7 The effects of Lactobacillus casei Zhang, Lactobacillus casei LC09 active substances and body weight of mice in the model group are shown in the figure: a, b, c represent the differences between the groups ( P <0.05). DETAILED DESCRIPTION

[0078] In order to make the technical means, creative features, purpose and efficacy of the present invention easy to understand, the present invention is further illustrated below in conjunction with specific examples, but the following examples are only preferred embodiments of the present invention, not all. Based on the examples in the implementation manner, other embodiments obtained by those skilled in the art without making creative work are all within the scope of protection of the present invention. In the following examples, unless otherwise specified, the operating methods used are all conventional operating methods, the equipment used are all conventional equipment, and the equipment and materials used in each embodiment are all the same.

[0079] The Lactobacillus casei Zhang in the present invention has been disclosed in patent CN105802876A.

[0080] Example 1 Preparation of Lactobacillus casei Zhang active substance

[0081] Lactobacillus casei Zhang was added at a rate of 6×10 6 After inoculating an inoculum of 100 CFU / g into MRS liquid culture medium, the culture was cultured at 37° C. until the pH reached 4.5 to obtain a fermentation broth. The fermentation broth was centrifuged and the supernatant was collected, which was then filtered through a 0.22 μm filter membrane to obtain the Lactobacillus casei Zhang active substance.

[0082] Example 2: Alleviating Effect of Lactobacillus casei Zhang Active Ingredient on Psoriasis

[0083] 1 Experimental Materials

[0084] Forty SPF-grade male C57BL / 6 mice, 6-8 weeks old, weighing 20-25 g, were used as experimental animals. Animal feed, maintenance feed, and rat bedding were purchased from Sbefore Biotechnology (Beijing) Co., Ltd. [SCXK (Beijing) 2019-0014]. Mice were housed in individual cages at a temperature of 22 ± 2°C, a humidity of 55%-75%, and a 12-h light-dark cycle. Water and food were freely available. All mice were acclimated to the environment for one week before the experiment.

[0085] Imiquimod cream was purchased from Mingxinlidi, and vaseline cream was purchased from Zhaonuo.

[0086] 2 Experimental Design

[0087] Forty 6-week-old BALB / c male mice were selected. After one week of adaptation, the hair in the central area of ​​the back of the mice was shaved to form a 3×3 cm 2 The mice were randomly divided into 5 groups (n=8) based on the area of ​​​​the skin, including normal group, model group, low-dose group (Lactobacillus casei Zhang active substance solution 2.625 mg / kg), medium-dose group (Lactobacillus casei Zhang active substance solution 5.25 mg / kg), and high-dose group (Lactobacillus casei Zhang active substance solution 10.5 mg / kg). The normal group was applied with vaseline (90 mg) on ​​the back of the mice once a day, and the other 4 groups were applied with imiquimod cream (5% IMQ, 90 mg) in the same way to establish a psoriasis model. The experimental design is as follows: Figure 1 shown.

[0088] 2.1 Calculation of mouse spleen index

[0089] Weigh the mouse, isolate the spleen, remove excess surrounding tissue, rinse with saline to remove blood, and blot dry with filter paper. Weigh the spleen using an electronic balance and calculate the spleen index. Spleen index = spleen mass (mg) / mouse mass (g).

[0090] 2.2 Patient severity index (PASI) score

[0091] Daily observation and recording of skin lesion changes were performed. The PASI scoring system was used to assess the erythema, epidermal desquamation, and degree of infiltration of the lesions. The scoring scale was 0 (no symptoms); 1 (mild); 2 (significant); 3 (severe); and 4 (very severe). The sum of the scores for these three components constituted the total PASI score. The PASI scoring system is shown in Table 1.

[0092] Table 1 PASI score

[0093]

[0094] 3 Experimental Results

[0095] Mouse spleen volume Figure 2 As shown in the figure, compared with the normal group, the spleen of the mice in the model group became darker and swollen significantly; compared with the model group, the spleen volume of the mice in the low, medium and high dose groups of the active substance was significantly reduced. Figure 3 As shown in the results, on the 9th day, the body weight of mice in the high-dose active substance group was significantly higher than that in the model group (P < 0.01). Both the low- and medium-dose groups had a certain improvement effect, but there was no statistical difference. The results showed that low, medium and high doses of active substances could inhibit the weight loss trend of psoriasis mice.

[0096] The spleen can produce a variety of immune-active cytokines. The increase in spleen index indicates that the body's inflammatory immune response has activated spleen cell proliferation, which reflects the body's immune function abnormality to a certain extent. The spleen index is obtained by combining the spleen mass and body mass of mice. The results are as follows: Figure 4 As shown, compared with the model group, the spleen index of mice in the high-dose active substance group was significantly reduced, indicating that the active substance intervention effectively alleviated psoriasis-related skin lesions and systemic inflammatory responses.

[0097] like Figure 5 As shown in the results, compared with the model group, the thickness of skin lesions, erythema and scaling on the back of mice were significantly reduced after the active substance intervention. The PASI (Patient Area and Severity Index) score was used to assess the severity of psoriasis. Figure 6 As shown in the data, starting from the 5th day, the daily PASI scores of mice in the low, medium and high dose groups were significantly lower than those in the model group (P < 0.0001). The results showed that low, medium and high doses of the active substance can effectively reduce the severity of psoriasis in mice and relieve psoriasis symptoms.

[0098] In summary, intervention with Lactobacillus casei Zhang active substances can inhibit the weight loss trend of psoriasis mice and reduce the spleen index. At the same time, it can effectively reduce the severity of psoriasis in mice and alleviate psoriasis symptoms.

[0099] Comparative Example 1

[0100] The back application method for mice in this comparative example was the same as in Example 2, except that the Lactobacillus casei Zhang active substance was replaced with the Lactobacillus casei LC09 active substance, which was prepared using the same method as in Example 1. Weight changes in each group of mice were recorded according to the time points of the original experiment. The average weight of each group of mice at different time points was calculated and compared with that of the model group and the high-dose Lactobacillus casei Zhang group.

[0101] The experimental results are as follows Figure 7As shown, compared with the model group, the weight loss of mice was significantly alleviated after intervention with Lactobacillus casei LC09 active substance, but the effect was worse than that of Lactobacillus casei Zhang active substance. By the 9th day, the weights of the three groups of mice were Lactobacillus casei Zhang active substance group (Zhang group), Lactobacillus casei LC09 active substance (LC09 group), and model group, and there were significant differences ( P <0.05), indicating that the effect of Lactobacillus casei Zhang active substance in improving psoriasis is better than that of Lactobacillus casei LC09 active substance.

[0102] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. Application of Lactobacillus casei Zhang in preparing a preparation for treating or alleviating psoriasis, characterized in that, The deposit number of the Lactobacillus casei Zhang is CGMCC No. 5469; The psoriasis includes psoriasis vulgaris, pustular psoriasis, erythrodermic psoriasis and arthritis psoriasis; The preparation comprises Lactobacillus casei Zhang fermentation liquid supernatant, Lactobacillus casei Zhang live bacteria or Lactobacillus casei Zhang freeze-dried powder; The preparation method of the described Lactobacillus casei Zhang fermentation broth supernatant comprises the following steps: (1) Lactobacillus casei Zhang was added at a rate of 1×10 6 -1×10 7 The inoculum size of CFU / g was inoculated into the fermentation medium; (2) Cultivate at 35-38°C until the pH reaches 4-6; (3) Centrifuge to obtain the fermentation supernatant, and filter with a 0.22 μm filter membrane.

2. The use according to claim 1, characterized in that The preparation also includes a pharmaceutically acceptable carrier.

3. The use according to claim 2, characterized in that The pharmaceutically acceptable carrier is selected from excipients.

4. The use according to claim 2, characterized in that The pharmaceutically acceptable carrier is selected from one or more of a stabilizer, a diluent, a binder, a preservative, and a lubricant.

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