Lactobacillus crispatus and application thereof in anti-inflammation, bacteriostasis and psoriasis improvement

Abstract

The invention discloses application of lactobacillus crispatus in resisting inflammation, inhibiting staphylococcus aureus and improving psoriasis, and belongs to the technical field of microorganisms and the technical field of medicines. The strain provided by the invention is a strain of Lactobacillus crispatus dipro-018, and the strain is a strain of Lactobacillus crispatus dipro-018. The strain lactobacillus crispatus provided by the invention has good anti-inflammatory performance and / or capability of inhibiting excessive proliferation of HaCaT cells, can effectively inhibit expression of proinflammatory factors of RAW246.7 cells under inflammatory stress, can also inhibit staphylococcus aureus, and can effectively improve skin and epidermal layer thickening symptoms in a psoriasis animal model.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and the technical field of medicine, and specifically relates to the application of a strain of Lactobacillus crispatus in anti-inflammation, inhibiting Staphylococcus aureus and improving psoriasis. Background Art

[0002] Psoriasis is a chronic inflammatory disease. The main visual symptoms are local erythema and papules covered with silvery white scales. The microscopic symptoms are mainly excessive proliferation of skin keratinocytes, parakeratosis and hyperkeratosis, accompanied by inflammatory cell infiltration. For many years, the cause of psoriasis has not been clarified. The most recognized cause is believed to be immune-mediated. A 2017 study on the epidemiology of psoriasis mentioned that in 20 countries, 0%-1.37% of children and 0.51%-11.43% of adults suffer from psoriasis.

[0003] As the largest organ in the human body, the skin is in direct contact with the external environment and is a platform for human skin-microorganism interaction. Staphylococcus aureus is a conditional pathogen that lives on the skin surface all year round. Its abundance changes when the skin microenvironment changes. A study has shown that the relative abundance of Staphylococcus aureus on the skin of patients with psoriasis increases both in lesions and healthy areas, and Staphylococcus aureus may affect the composition of the skin microbiome through interactions with other skin microorganisms, thereby affecting the skin immune response (see Chang HW, Yan D, et al. Alteration of the cutaneous microbiome in psoriasis and potential role in Th17 polarization. Microbiome. 2018 Sep 5; 6 (1): 154.).

[0004] Research on Lactobacillus crispatus is mostly focused on female reproductive tract health and is used as a microbial marker for vaginal health. It can produce antibacterial compounds such as bacteriocins and lactic acid to maintain vaginal homeostasis. Other studies have shown that Lactobacillus crispatus can inhibit rotavirus infection of human intestinal epithelial cells. However, there are few studies on the role of Lactobacillus crispatus in psoriasis. For example, in the Chinese invention patent with publication number CN112159778B, it is disclosed that an animal Bifidobacterium CCFM1148 can inhibit the excessive proliferation of human immortalized cortical cells, reduce the level of proinflammatory factors, and inhibit the activation of the NF-kB signaling pathway, but its role in psoriasis mice has not been verified, so further verification is needed; for example, in the invention patent with publication number CN116407565A, the mucoid Roseomonas CGMCC25967 can inhibit the proliferation of HaCaT cells and inflammatory factors, relieve psoriasis-like lesions on the back, spleen index and epidermal thickening, but it has not studied whether the inactivated bacteria of the mucoid Roseomonas CGMCC25967 have an effect on the proliferation of HaCaT cells and whether it has a good inhibitory effect on Staphylococcus aureus. Summary of the invention

[0005] Purpose of the invention: The technical problem to be solved by the present invention is to provide a strain of Lactobacillus crispatus dipro-018 which has anti-inflammatory and antibacterial functions, especially capable of inhibiting Staphylococcus aureus; and / or inhibiting excessive proliferation of keratinocytes, and can relieve psoriasis.

[0006] The technical problem that the present invention also solves is to provide a product containing the Lactobacillus crispatus dipro-018, which includes but is not limited to fermented products, foods, health products, medicines, feeds, protective agents, functional additives, auxiliary additives, drug carriers or pharmaceutical excipients.

[0007] The final technical problem to be solved by the present invention is to provide the application of the Lactobacillus crispatus dipro-018 and the product in the preparation of products for anti-inflammatory, antibacterial or prevention or treatment of excessive proliferation of keratinocytes.

[0008] Technical solution: In order to solve the above technical problems, the present invention provides a strain of Lactobacillus crispatus dipro-018, which was deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration on June 17, 2024, with a deposit number of CGMCC No.30968 and a deposit address of No. 1, Beichen West Road, Chaoyang District, Beijing, China.

[0009] The present invention also includes products containing the Lactobacillus crispatus dipro-018, and the products include but are not limited to fermented products, foods, health products, medicines, feeds, protective agents, functional additives, auxiliary additives, drug carriers or pharmaceutical excipients.

[0010] Among them, the products including but not limited to fermented products, foods, health products, medicines, feeds, protective agents, functional additives, auxiliary additives, drug carriers or pharmaceutical excipients include live bacteria or inactivated bacteria of Lactobacillus crispatus dipro-018.

[0011] The fermented product comprises the fermentation supernatant, fermentation precipitate or bacterial suspension of Lactobacillus crispatus dipro-018.

[0012] The dosage forms of the product include powder, capsule, tablet, pill, film-coated agent, aerosol, granule, liquid, liposome, transdermal agent, suppository or lyophilized powder injection.

[0013] Among them, the protective agent, functional additive, auxiliary additive, drug carrier or pharmaceutical excipient includes but is not limited to albumin, gelatin, soluble starch, dextrin, gravy, pectin, gum arabic, hydroxymethyl cellulose, algae, vitamin D, vitamin E, protein hydrolyzate, sodium thiosulfate, dextran and natural mixtures such as skim milk, serum, etc.

[0014] The present invention also includes the use of the Lactobacillus crispatus dipro-018 and the product in preparing products for anti-inflammatory, antibacterial or prevention or treatment of excessive proliferation of keratinocytes.

[0015] The present invention also includes the application of the Lactobacillus crispatus dipro-018 in the product having at least one of the following functions:

[0016] 1) Products that have the ability to inhibit Staphylococcus aureus;

[0017] 2) Products with the ability to reduce pro-inflammatory cytokines;

[0018] 3) Products with the ability of live bacteria and / or inactivated bacteria to inhibit excessive proliferation of keratinocytes;

[0019] 4) Products that have the ability to reduce visual damage to the skin of psoriasis mice;

[0020] 5) Products that have the ability to reduce skin thickness in psoriasis mice;

[0021] 6) Products that have the ability to reduce epidermal thickness in psoriasis mice;

[0022] 7) A product having the ability to reduce spleen index in psoriasis mice.

[0023] The Lactobacillus crispatus dipro-018 or its preparation has good antibacterial ability against Staphylococcus aureus ATCC6538.

[0024] Wherein, the Lactobacillus crispatus dipro-018 or its preparation can inhibit the production of nitric oxide (NO) by RAW246.7 cells induced by LPS.

[0025] The live bacteria or inactivated bacteria of Lactobacillus crispatus dipro-018 or the preparation thereof can inhibit the excessive proliferation of keratinocytes (HaCaT cells).

[0026] Wherein, the Lactobacillus crispatus dipro-018 or its preparation can alleviate and improve the visual damage of the skin of psoriasis mice induced by imiquimod (IMQ).

[0027] Wherein, the Lactobacillus crispatus dipro-018 or its preparation can reduce the increase in skin thickness of psoriasis mice induced by IMQ.

[0028] Wherein, the Lactobacillus crispatus dipro-018 or its preparation can reduce the increase in epidermal thickness of psoriasis mice induced by IMQ.

[0029] Wherein, the Lactobacillus crispatus dipro-018 or its preparation can reduce the increase of spleen index of psoriasis mice induced by IMQ.

[0030] Beneficial effects: Compared with the prior art, the present invention has the following advantages: Lactobacillus crispatus dipro-018 used in the present invention can significantly reduce the pro-inflammatory factor NO produced by RAW246.7 cells under LPS induction, and its live bacteria or inactivated bacteria can also significantly inhibit the excessive proliferation of HaCaT cells induced by TNF-α; Lactobacillus crispatus dipro-018 also has a good inhibitory ability against Staphylococcus aureus (Staphylococcusaureus ATCC6538); in the IMQ-induced psoriasis mouse model, dipro-018 can significantly improve the skin lesions, skin and epidermal thickness of psoriasis mice and reduce the spleen index. Therefore, the use of Lactobacillus crispatus dipro-018 strain for the preparation of anti-inflammatory, Staphylococcus aureus inhibition and psoriasis improvement products has a good prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Lactobacillus crispatus dipro-018 inhibits Staphylococcus aureus ATCC6538.

[0032] Figure 2 Lactobacillus crispatus dipro-018 inhibits LPS-induced RAW264.7 cells from producing the proinflammatory factor NO.

[0033] Figure 3 Live bacteria of Lactobacillus crispatus dipro-018 inhibit TNF-α-induced HaCaT cell overproliferation.

[0034] Figure 4 The inactivated bacteria of Lactobacillus crispatus dipro-018 inhibit TNF-α-induced excessive proliferation of HaCaT cells.

[0035] Figure 5 Lactobacillus crispatus dipro-018 can alleviate IMQ-induced visual damage to mouse skin. Ctrl: negative control, PBS gavage; IMQ group: positive control, IMQ treatment, PBS gavage;

[0036] IMQ+dipro-018 group: IMQ treatment, Lactobacillus crispatus dipro-018 (5×10 8 CFU) by intragastric administration.

[0037] Figure 6 Lactobacillus crispatus dipro-018 can reduce the increase in skin thickness induced by IMQ in mice. Ctrl: negative control, PBS gavage; IMQ group: positive control, IMQ treatment, PBS gavage;

[0038] IMQ+dipro-018 group: IMQ treatment, Lactobacillus crispatus dipro-018 (5×10 8 CFU) by intragastric administration.

[0039] Figure 7 Lactobacillus crispatus dipro-018 can reduce the increase in epidermal thickness induced by IMQ in mice. Ctrl: negative control, PBS gavage; IMQ group: positive control, IMQ treatment, PBS gavage;

[0040] IMQ+dipro-018 group: IMQ treatment, Lactobacillus crispatus dipro-018 (5×10 8 CFU) by intragastric administration. A: Mouse back skin tissue section, 20×; B: Epidermal thickness.

[0041] Figure 8 Lactobacillus crispatus dipro-018 can reduce the increase of spleen index in mice induced by IMQ. Ctrl: negative control, PBS gavage; IMQ group: positive control, IMQ treatment, PBS gavage; IMQ+dipro-018 group: IMQ treatment, Lactobacillus crispatus dipro-018 (5×10 8 CFU) by intragastric administration. DETAILED DESCRIPTION

[0042] The present invention is described in detail below in conjunction with the accompanying drawings and embodiments.

[0043] MRS medium was purchased from Guangdong Huankai Microbiology Technology Co., Ltd.; RAW264.7 cell line was purchased from Shanghai Cell Bank of the Chinese Academy of Sciences; HaCaT cell line was purchased from Wuhan Pronocell Life Science Technology Co., Ltd.;

[0044] DMEM culture medium, MEM culture medium, fetal bovine serum, penicillin-streptomycin and 0.25% trypsin were purchased from Gibco; LPS was purchased from Beijing Solebow Technology Co., Ltd.; cell proliferation-toxicity detection kit (CCK-8) was purchased from Tongren Chemical; TNF-α was purchased from PeproTech; NO detection kit was purchased from Beyotime Biotechnology Co., Ltd.; 7-week-old female BALB / c SPF mice were purchased from Jiangsu Jicui Pharmaceutical Kang Biotechnology Co., Ltd.; imiquimod cream (IMQ) was purchased from Sichuan Mingxin Pharmaceutical Co., Ltd.

[0045] Example 1 Isolation, Identification and Biological Characteristics of Lactobacillus crispatus dipro-018

[0046] The separation and identification method of Lactobacillus crispatus dipro-018 of the present invention is as follows:

[0047] 1. Isolation and identification of Lactobacillus crispatus dipro-018

[0048] Resuspend and dilute the fecal samples of children with sterile phosphate buffered saline (PBS, 10 mM, pH 7.2-7.4, the same below) or normal saline, and take 10 -4 , 10 -5 , 10 -6 The dilution gradient was spread on MRS+ agar medium (MRS agar medium supplemented with 0.05% cysteine), cultured anaerobically at 37°C for 24 hours, colonies were picked and streaked and purified for three generations, and then preliminarily identified using a MALDI mass spectrometer. Single colonies were picked and inoculated into MRS+ liquid medium (MRS liquid medium supplemented with 0.05% cysteine), cultured anaerobically at 37°C for 24 hours, and part of the bacterial solution was sent for whole genome sequencing. The strain was analyzed by whole genome sequencing and compared with the BLAST nucleic acid sequence of Lactobacillus crispatus through the NCBI gene database. The similarity between it and Lactobacillus crispatus was 97.652%, so it was named Lactobacillus crispatus dipro-018, and the safety of the strain in the genome (including whether it contained virulence genes and transferable resistance genes) was evaluated, and the rest of the bacterial solution was retained.

[0049] 2. Safety analysis of Lactobacillus crispatus

[0050] The genomic safety of the dipro-018 strain was evaluated based on whole genome sequencing. The results showed that the dipro-018 genome contained no genes related to transferable antibiotic resistance and no virulence genes.

[0051] 3. Hemolytic activity of Lactobacillus crispatus

[0052] Use an inoculation loop to dip the Dipro-018 strain after 3 generations of activation and streak it on Columbia blood agar plate. Incubate it anaerobically at 37°C for 24 hours. If there is no hemolytic ring around the colony, Dipro-018 has no hemolytic activity.

[0053] Example 2 Lactobacillus crispatus dipro-018 inhibits Staphylococcus aureus

[0054] Staphylococcus aureus ATCC6538 was activated for three generations in LB liquid medium, and the OD625 nm value was adjusted to 0.16-0.20 with sterile saline (the concentration of bacteria in the bacterial suspension was about 3×10 8 CFU / mL) for later use; dipro-018 in the exponential growth phase was inoculated in MRS+ liquid culture medium at a 2% (volume percentage, the same below) inoculation amount and anaerobically cultured for 24h for later use. Agar plates containing Staphylococcus aureus ATCC6538 were prepared by the pouring method, specifically, the corresponding agar culture medium was first prepared, cooled to 40°C after sterilization, 200μL of Staphylococcus aureus ATCC6538 bacterial solution was added and mixed, and then 30mL of each plate was poured into the plate, and after the agar solidified, holes were punched with an 8mm puncher, and only 200μL of MRS+ liquid culture medium was added to the wells of the control group, and 200μL of MRS+ liquid culture medium containing dipro-018 was added to the wells of the dipro-018 group, and cultured at 37°C for 16h, and the radius of the inhibition zone was measured.

[0055] The results are as follows Figure 1 As shown in the figure, the inhibitory ability of dipro-018 against Staphylococcus aureus ATCC6538 was significantly higher than that of the control group (P<0.01).

[0056] Example 3 Lactobacillus crispatus dipro-018 inhibits LPS-induced production of pro-inflammatory factor NO in RAW264.7 cells

[0057] 1. Activation of Lactobacillus crispatus dipro-018: Take out the cryopreserved tube, streak on MRS+ agar medium for anaerobically culture for 24 hours, then pick a single colony and culture it anaerobically in MRS+ liquid medium for 24 hours, and store it as seed liquid for later use. When using, take the bacterial liquid and inoculate it into MRS+ liquid medium at 2%. After the culture is completed, take the bacterial suspension and centrifuge it at 6000rpm / min for 5 minutes, discard the supernatant, resuspend it with PBS, centrifuge and wash it twice for later use.

[0058] 2. RAW246.7 cells were frozen in our laboratory. Recovery: Take out RAW246.7 cells from the liquid nitrogen tank, put them in a 37℃ constant temperature water bath to thaw quickly, transfer the thawed cells to 6mL DMEM culture medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin, mix well, and place in a 37℃, 5% CO2 incubator for culture. After the cells are completely attached to the wall, change the medium and continue to culture. When the cell density reaches about 80% to 90%, pass the cells. Passage: Discard the old DMEM culture medium, wash with 2mL PBS solution 3 times, gently blow the attached cells with a pipette to suspend them, add 6mL of fresh DMEM culture medium to a new culture bottle, add the cell suspension and mix well, place in a 37℃, 5% CO2 incubator for continued culture, and pass the cells for 3 generations in this way before conducting the experiment. Plating and testing (the culture medium used was DMEM culture medium without 10% fetal bovine serum and 1% penicillin-streptomycin): When RAW246.7 cells were growing well and reached the logarithmic growth phase, 1×10 5 / well were inoculated into 96-well plates and cultured until adhered to the wall. The DMEM culture medium was discarded, 100 μL DMEM culture medium was added to the negative wells as the Ctrl group, 100 μL DMEM culture medium containing 0.5 μg / mL LPS was added to the positive wells as the LPS group, and 100 μL DMEM culture medium containing 1×10 7 CFU Lactobacillus crispatus dipro-018 and 0.5 μg / mL LPS in DMEM culture medium were used as the dipro-018 group and incubated in a 37°C, 5% CO2 incubator for 18 hours. The supernatant after the above culture was collected and the NO concentration was detected using a kit.

[0059] The results are as follows Figure 2 After treatment, dipro-018 could inhibit the release of NO in RAW264.7 macrophages induced by LPS, and the amount of NO produced was significantly lower than that in the LPS group (P<0.001).

[0060] Example 4 Live bacteria of Lactobacillus crispatus dipro-018 inhibit TNF-α-induced excessive proliferation of HaCaT cells

[0061] 1. The culture method of Lactobacillus crispatus dipro-018 is basically the same as that in Example 3.

[0062] 2. HaCaT cells were frozen in our laboratory. Recovery: Take out the HaCaT cells from the liquid nitrogen tank, put them in a 37℃ constant temperature water bath to thaw quickly, transfer the thawed cells to 6mL of MEM culture medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin, mix well, and place them in a 37℃, 5% CO2 incubator for culture. After the cells are completely attached to the wall, change the medium and continue to culture. When the cell density reaches about 80% to 90%, pass the cells. Passage: Discard the old MEM culture medium, wash with 2mL PBS solution 3 times, gently blow the attached cells with a pipette to suspend them, add 6mL of fresh MEM culture medium to a new culture bottle, add the cell suspension and mix well, place it in a 37℃, 5% CO2 incubator for continued culture, and pass the cells for 3 generations in this way before conducting the experiment. Plating and testing (the culture medium used was MEM culture medium without 10% fetal bovine serum and 1% penicillin-streptomycin): When HaCaT cells were growing well and reached the logarithmic growth phase, 1.5×10 4 The cells were inoculated into 96-well plates at a density of 1 × 10 / well and cultured until adhered to the wall. The MEM culture medium was discarded. 100 μL MEM culture medium was added to the negative wells as the Ctrl group, 100 μL MEM culture medium containing 40 ng / mL TNF-α was added to the positive wells as the TNF-α group, and 100 μL viable 1×10 7 CFU Lactobacillus crispatus dipro-018 and MEM culture medium with 40ng / mL TNF-α were used as the dipro-018 live bacteria group and incubated in a 37°C, 5% CO2 incubator for 18 hours; then the liquid in each well was discarded, and 100μL MEM culture medium and 10μL cell proliferation-toxicity detection reagent were added to the negative well, positive well, and sample well, and only 100μL MEM culture medium and 10μL cell proliferation-toxicity detection reagent were added to the blank well without cells, and incubated in a 37°C, 5% CO2 incubator for 2 hours; the absorbance value of each well at 450nm was detected by an ELISA instrument, and the cell survival rate = (OD450nm value of the sample well or positive well - OD450nm value of the blank well) / (OD450nm value of the negative well - OD450nm value of the blank well)

[0063] The results are as follows Figure 3 As shown in Figure 3, the live bacteria of dipro-018 significantly inhibited the overproliferation of HaCaT cells induced by TNF-α, and the cell survival rate was only 1.49% (P<0.001).

[0064] Example 5 Inactivated Lactobacillus crispatus dipro-018 inhibits TNF-α-induced excessive proliferation of HaCaT cells

[0065] 1. The culture method of Lactobacillus crispatus dipro-018 was basically the same as that in Example 3, except that after washing with PBS twice, the mixture was resuspended in an appropriate amount of PBS and pasteurized (dipro-018 was placed in a water bath and heated to 70°C for 30 min) to a concentration of 1×10 8 CFU / mL of Lactobacillus crispatus dipro-018 bacteria were inactivated.

[0066] 2. HaCaT cells were frozen in our laboratory. Recovery: Take out the HaCaT cells from the liquid nitrogen tank, put them in a 37℃ constant temperature water bath to thaw quickly, transfer the thawed cells to 6mL of MEM culture medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin, mix well, and place them in a 37℃, 5% CO2 incubator for culture. After the cells are completely attached to the wall, change the medium and continue to culture. When the cell density reaches about 80% to 90%, pass the cells. Passage: Discard the old MEM culture medium, wash with 2mL PBS solution 3 times, gently blow the attached cells with a pipette to suspend them, add 6mL of fresh MEM culture medium to a new culture bottle, add the cell suspension and mix well, place it in a 37℃, 5% CO2 incubator for continued culture, and pass the cells for 3 generations in this way before conducting the experiment. Plating and testing (the culture medium used was MEM culture medium without 10% fetal bovine serum and 1% penicillin-streptomycin): When HaCaT cells were growing well and reached the logarithmic growth phase, 1.5×10 4 / well were inoculated into 96-well plates and cultured until adhered to the wall. The MEM culture medium was discarded, 100 μL MEM culture medium was added to the negative wells as the Ctrl group, 100 μL MEM solution containing 40 ng / mL TNF-α was added to the positive wells as the TNF-α group, and 100 μL inactivated 1×10 7 CFU Lactobacillus crispatus dipro-018 and 40ng / mL TNF-α MEM solution were used as the dipro-018 inactivated bacterial group and incubated in a 37°C, 5% CO2 incubator for 18 hours; then the liquid in each well was aspirated and discarded, and 100μL MEM culture medium and 10μL cell proliferation-toxicity detection reagent were added to the negative well, positive well, and sample well, and only 100μL MEM culture medium and 10μL cell proliferation-toxicity detection reagent were added to the cell-free blank well, and incubated in a 37°C, 5% CO2 incubator for 2 hours; the absorbance value of each well at 450nm was detected by an ELISA instrument, and the cell survival rate = (OD450nm value of the sample well or positive well - OD450nm value of the blank well) / (OD450nm value of the negative well - OD450nm value of the blank well)

[0067] The results are as follows Figure 4The inactivated dipro-018 bacteria after treatment significantly inhibited the excessive proliferation of HaCaT cells induced by TNF-α, and its cell survival rate was 107.53%, which was significantly lower than that of the TNF-α group (141.82%) (P<0.05).

[0068] Example 6 Lactobacillus crispatus dipro-018 improves skin ocular damage in psoriasis mice

[0069] Take the freeze-dried powder of dipro-018 and resuspend it in PBS to make the concentration of bacteria 2.5×10 9 CFU / mL. 7-week-old SPF female BALB / c mice were selected and randomly divided into 3 groups: control group (Ctrl), psoriasis model group (IMQ) and dipro-018 treatment group (IMQ+dipro-018), with 6 mice in each group. From day 1 to day 14, mice in the Ctrl and IMQ groups were gavaged with 200 μL PBS per day, and mice in the dipro-018 treatment group were gavaged with 200 μL dipro-018 freeze-dried bacterial suspension (5×10 8 CFU); on the 5th day, an electric shaver and depilatory cream were used to remove the hair of about 3cm×2cm (length×width) in the central area of ​​the back of the mice. From the 8th to the 14th day, 62.5mg IMQ was evenly applied to the hairless skin of the back in the IMQ and IMQ+dipro-018 groups. At the same time, the Ctrl group was applied with an equal amount of vaseline. The mice were gavaged before each application of vaseline or IMQ. On the 15th day, the skin symptoms of the mice were observed. The results are shown in Figure 5 The back skin of IMQ-induced psoriasis mice showed psoriasis-like inflammatory reactions, such as erythema and even bleeding, scaling and thickening; after dipro-018 treatment, the back skin of mice improved, with less bleeding and thickening. Lactobacillus crispatus dipro-018 can improve the visual damage of IMQ-induced psoriasis mice skin.

[0070] Example 7 Lactobacillus crispatus dipro-018 reduces skin thickness in psoriasis mice

[0071] The animal experiment treatment was basically the same as in Example 6, except that on the 15th day, the skin thickness was measured with a vernier caliper. The results are shown in Figure 6 Compared with the control group, the skin of IMQ-induced psoriasis mice was significantly thickened, while the skin thickness of dipro-018-treated mice was thinner than that of the IMQ group, and the difference was significant (P<0.05).

[0072] Example 8 Lactobacillus crispatus dipro-018 reduces epidermal thickness in psoriasis mice

[0073] The animal experiment treatment was basically the same as in Example 6, except that on the 15th day, the mice were killed by cervical dislocation, the skin tissue was collected and fixed in 4% paraformaldehyde solution, and then the tissue was embedded in paraffin and made into slices (5 μm) for hematoxylin-eosin (HE) staining and skin tissue pathology examination. The thickness of the epidermis was measured and statistically analyzed using ImageJ. The results are shown in Figure 7 IMQ-induced psoriasis mice showed psoriasis-like lesions, with significant thickening of the stratum corneum accompanied by hemorrhage and inflammatory cell infiltration, while the epidermal thickness of dipro-X-treated mice was thinner than that of untreated mice, and the difference was significant (P<0.05). In addition, dipro-X treatment also reduced the infiltration of inflammatory cells. Lactobacillus crispatus dipro-X can reduce the increase in epidermal thickness in IMQ-induced psoriasis mice.

[0074] Example 9 Lactobacillus crispatus dipro-018 reduces spleen index in psoriasis mice

[0075] The animal experiment treatment was basically the same as in Example 6, except that on the 15th day, the mice were killed by cervical dislocation, and the spleen tissue was collected and weighed. The spleen index = spleen weight mg / mouse body weight g. Figure 8 Compared with the control group, the spleen index of IMQ-induced psoriasis mice was significantly increased, while the spleen index of dipro-X-treated mice was lower than that of IMQ-treated mice and the difference was significant (P<0.01).

Claims

1. A strain of Lactobacillus crispatus ( Lactobacillus crispatus ) dipro-018, characterized in that The Lactobacillus crispatus ( Lactobacillus crispatus ) dipro-018 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on June 17, 2024, with the deposit number: CGMCC No. 30968.

2. Containing the Lactobacillus crispatus according to claim 1 ( Lactobacillus crispatus ) dipro-018 product, characterized in that, The products include fermented products, foods, health products, medicines or feeds.

3. The product according to claim 2, characterized in that The fermented product includes Lactobacillus crispatus ( Lactobacillus crispatus ) fermentation supernatant, fermentation precipitate or bacterial suspension of dipro-018.

4. The product according to claim 2, characterized in that The dosage forms of the product include powder, capsule, tablet, pill, film-coated agent, aerosol, granule, liquid, liposome, transdermal agent, suppository or lyophilized powder injection.

5. The product according to claim 2, characterized in that The product also includes a protective agent, a functional auxiliary agent / or an auxiliary additive, a drug carrier and / or a pharmaceutical excipient.

6. The product according to claim 2 or claim 5, characterized in that The fermented product, food, health product, medicine or feed, protective agent, functional additive and / or auxiliary additive, drug carrier and / or pharmaceutical excipient includes Lactobacillus crispatus ( Lactobacillus crispatus ) Live or inactivated bacteria of dipro-018.

7. The Lactobacillus crispatus according to claim 1 ( Lactobacillus crispatus ) dipro-018, use of the product according to any one of claims 2 to 6 in the preparation of products for anti-inflammatory, antibacterial or prevention or treatment of excessive proliferation of keratinocytes.

8. The use according to claim 7, characterized in that: Such diseases of excessive keratinocyte proliferation include psoriasis.

9. The use according to claim 7, characterized in that: The application of the product is a product having at least one of the following functions: 1) Products that have the ability to inhibit Staphylococcus aureus; 2) Products with the ability to reduce pro-inflammatory cytokines; 3) Products that have the ability to inhibit excessive proliferation of keratinocytes; 4) Products that have the ability to reduce visual damage to the skin of psoriasis mice; 5) Products with the ability to reduce skin thickness in psoriasis mice; 6) Products that have the ability to reduce epidermal thickness in psoriasis mice; 7) Products with the ability to reduce spleen index in psoriasis mice.

10. The use according to claim 9, characterized in that: The reducing of pro-inflammatory cytokines includes inhibiting the production of nitric oxide by RAW246.7 cells induced by LPS; the inhibiting of excessive proliferation of keratinocytes includes inhibiting excessive proliferation of keratinocyte HaCaT cells; the reducing of visual damage to the skin of psoriasis mice includes alleviating and improving the visual damage to the skin of psoriasis mice induced by imiquimod; the reducing of the skin thickness of psoriasis mice includes reducing the increase in skin thickness of psoriasis mice induced by IMQ; the reducing of the spleen index of psoriasis mice includes reducing the increase in spleen index of psoriasis mice induced by IMQ.

Images