Use of lactococcus lactis subsp. cremoris and products thereof in improving the effects of enteritis on body weight, gut and immunity

Abstract

This invention belongs to the field of microbial technology, specifically relating to the application of *Lactococcus lactis* subsp. *milkfat* and its products in improving the effects of enteritis on weight, intestinal health, and immunity. This invention provides a novel *Lactococcus lactis* subsp. *milkfat* strain, with the preservation number CGMCC No. 29363. Experiments have shown that supplementation with *Lactococcus lactis* subsp. *milkfat* LC-99 can increase the weight of patients with enteritis, improve intestinal inflammatory symptoms, and enhance immune function.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically relating to the application of Lactococcus lactis subsp. milk fat and its products in improving the effects of enteritis on weight, intestines and immunity. Background Technology

[0002] Enteritis is a relatively common intestinal disease. Globally, the incidence of inflammatory bowel disease (including ulcerative colitis and Crohn's disease, the two main types of enteritis) is on the rise. The incidence of infectious enteritis varies greatly depending on factors such as region, sanitation conditions, and season. It is significantly higher in areas with poor sanitation or during peak seasons for intestinal infectious diseases, such as summer. Enteritis severely impacts the quality of life of patients. Intestinal symptoms such as diarrhea and abdominal pain limit their daily lives, work, and social activities. For example, patients with inflammatory bowel disease may be unable to work normally due to frequent urination and may experience anxiety when traveling or participating in social activities. Furthermore, long-term illness can lead to complications such as malnutrition and anemia, further reducing their quality of life.

[0003] In the study of the pathogenesis of enteritis, an increasing number of studies have found that gut microbiota dysbiosis, genetic factors, and immune abnormalities play key roles in the development of enteritis. Despite these advances, the treatment of enteritis still faces many challenges. For example, inflammatory bowel disease is still not completely curable, drug treatment has certain side effects, and some patients do not respond well to drug treatment. For infectious enteritis, with the widespread use of antibiotics, the problem of drug-resistant bacterial infections is becoming increasingly serious, making treatment more difficult.

[0004] The gut microbiota, a complex ecosystem, is influenced by both host and environmental factors. It plays a crucial role in maintaining homeostasis and disrupting the balance between health and disease. The gut microbiota is vital for intestinal activities such as peristalsis, mucus production, intestinal epithelial cell differentiation, and intestinal immune responses, and is also essential for maintaining gut health and homeostasis. Studies have shown that intraepithelial neoplasia (IBD) can cause general changes in the gut microbiota structure, leading to decreased diversity and species abundance. These changes affect the normal functioning of the mucosal immune system, causing functional decline, and are closely linked to the development and progression of IBD. Research has found an increase in the types of harmful bacteria and a significant decrease in the types and abundance of beneficial bacteria in the gut microbiota of IBD patients, highlighting the important role of probiotics in IBD treatment in promoting gut microbiota balance.

[0005] Lactococcus lactis is generally considered to include three subspecies, of which the milk-derived subspecies is Lactococcus lactis (Lactococcus lactis). Lactococcus lactis subsp. lactis ) and Lactococcus lactis subsp. milk fat ( Lactococcus milk subsp .creamy ), and also *Lactococcus hominis* subsp. *hominis* isolated from leafhoppers ( Lactococcus lactis subsp.hordniae The main applications of *Lactococcus lactis* subsp. *milk fat* include: Food industry: As an important source of starter cultures in the production of probiotic fermented dairy products, it can be used to make yogurt, cheese, etc. It produces metabolites such as lactic acid, giving dairy products a good flavor and texture, while also imparting certain probiotic functions, helping to regulate intestinal flora, enhance immunity, etc. It can also be used to improve the flavor and quality of baked goods such as bread. Health product development: Due to its potential probiotic properties, it can be made into probiotic health products in the form of capsules, tablets, powders, etc., helping people maintain intestinal microecological balance and promote health. Pharmaceutical research: Related research is also deepening, and it is expected to play a role in disease prevention and treatment in the future, such as assisting in the treatment of some intestinal diseases by regulating intestinal immunity. However, currently, there is limited research on the use of *Lactococcus lactis* subsp. *milk fat* in the treatment of enteritis. Therefore, there is an urgent need to develop a *Lactococcus lactis* strain that can treat enteritis, increase the weight of enteritis patients, and enhance immune function. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides a strain of *Lactococcus lactis* subsp. *milk fat* (… Lactococcus milk subsp .creamy LC-99 can increase the weight of patients with enteritis, improve intestinal inflammation, and enhance immune function. This supplements existing research showing that *Lactococcus lactis* subsp. *milk fat* does not increase the weight of patients with enteritis.

[0007] On the one hand, the present invention provides a strain of Lactococcus lactis subsp. milk fat ( Lactococcus lactis subsp.cremoris The invention is characterized in that the preservation number of the lactococcus lactis subsp. milk fat is CGMCC No. 29363.

[0008] Specifically, this includes inoculating *Lactococcus lactis* subsp. milk fat onto a culture medium and culturing it.

[0009] More specifically, the inoculation methods include, but are not limited to, any one or more of the following: streak plate inoculation, slant inoculation, pour culture, puncture inoculation, and liquid inoculation.

[0010] More specifically, the vaccination dose can be 0.1%-20%, and in some cases, it can be higher or lower. Specifically, the vaccination dose can be 1%-20%, 2%-20%, 1%-15%, 1%-10%, 1%-5%, 1%-8%, 5%-15%, 5%-10%, 5%-8%, 8%-10%, 8%-15%, 5%-12%, or 2%-7%.

[0011] More specifically, the culture medium can be a solid culture medium, a semi-solid culture medium, or a liquid culture medium. More specifically, it can be any suitable culture medium type disclosed in the prior art, or a culture medium that is further improved on the culture medium type disclosed in the prior art to improve the performance of the strain, or a culture medium not disclosed in the prior art but that can be used for the culture of the aforementioned Lactococcus lactis subsp. milk fat.

[0012] Preferably, the culture medium includes, but is not limited to, MRS culture medium.

[0013] In another aspect, the present invention provides the use of the aforementioned *Lactococcus lactis* subsp. *lactofatica* in the preparation of products for the prevention, treatment and / or adjunctive treatment of enteritis.

[0014] Specifically, the product is a drug, pharmaceutical raw material, or health product.

[0015] More specifically, the drug, pharmaceutical raw material or health product includes one or more of the following: fermentation broth of Lactococcus lactis subsp. milk fat, fermentation broth supernatant, fermentation broth precipitate, live bacteria, dead bacteria, lyophilized powder and cell lysate.

[0016] More specifically, the fermentation broth refers to the liquid in which the microbial strain is inoculated into a culture medium and cultured for a period of time.

[0017] More specifically, the supernatant of the fermentation broth refers to the clear liquid at the top after centrifugation of the fermentation broth; it contains abundant metabolic products from the bacterial growth and reproduction process, as well as some bacterial cell fragments. The acidic substances and bacteriocins secreted by the bacteria have antagonistic and bactericidal effects on harmful bacteria. The amino acids and vitamins synthesized by the bacteria after decomposing food are also in the culture medium, as well as enzymes secreted by the bacteria that are useful to the human body. Some of the bacterial cell components also have an immune-boosting effect on the human body.

[0018] More specifically, the fermentation broth sediment refers to the liquid sediment obtained after centrifugation, including free proteins, residual bacterial cells, broken cells, and culture medium residues, mainly proteins and intracellular matrix.

[0019] More specifically, the live bacteria, also known as active bacteria, can colonize and multiply in the intestines, which helps increase the number of beneficial bacteria.

[0020] More specifically, the dead bacteria are microorganisms that have lost their vitality and are unable to grow and reproduce. The production process causes the probiotics to lose their vitality, such as high-temperature treatment or excessive drying.

[0021] More specifically, the lyophilized powder is obtained by lyophilizing the aforementioned culture medium. The lyophilized powder generally also includes a lyophilization protectant. The lyophilization protectant includes, but is not limited to, pH buffers, fillers, sugars, nonionic surfactants, ligands, etc. The pH buffer includes, but is not limited to, any one or more of Tris, amino acids or their salts, citric acid or its salts, acetic acid or its salts. The fillers include, but are not limited to, any one or more of mannitol, glycine, and bovine serum albumin. The sugars can be disaccharides, such as sucrose or trehalose, any one or more. The nonionic surfactants include, but are not limited to, Tween, such as Tween-20, Tween-60, Tween-80, etc. The lyophilization protectant may also include antioxidants, etc. Specifically, the lyophilization protectant may also include albumin, polyethylene glycol, etc.

[0022] More specifically, the cell lysis buffer can be obtained by lysing the bacterial cells cultured from the aforementioned culture. The lysis can be physical or chemical. Physical lysis includes, but is not limited to, grinding and ultrasonic disruption. Chemical lysis includes, but is not limited to, lysis with chemical reagents and enzymatic hydrolysis, where the enzymatic hydrolysis can be with hydrolases or oxidases. Lysis can also be achieved by increasing intracellular pressure to induce spontaneous cell rupture.

[0023] Specifically, the medicine or health product also includes pharmaceutically or food-grade excipients.

[0024] In another aspect, the present invention provides a drug comprising the aforementioned *Lactococcus lactis* subsp. *lactofattya*.

[0025] Specifically, the viable count of *Lactococcus lactis* subsp. *milk fat* in the drug can be no less than 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0026] Preferably, the viable count of *Lactococcus lactis* subsp. *lactofat* in the drug can be 1 × 10⁻⁶. 8 -1×10 12 CFU / g or 1×10 8 -1×10 12 CFU / mL. Specifically, it could be: 1×10⁻⁶ CFU / mL. 8 -1×10 9 CFU / g (CFU / mL), 1×10 8 -1×1010 CFU / g (CFU / mL), 1×10 8 -1×10 11 CFU / g (CFU / mL), 1×10 9 -1×10 12 CFU / g (CFU / mL), 1×10 9 -1×10 11 CFU / g (CFU / mL), 1×10 9 -1×10 10 CFU / g (CFU / mL), 1×10 10 -1×10 12 CFU / g (CFU / mL), 1×10 10 -1×10 11 CFU / g (CFU / mL), 1×10 11 -1×10 12 CFU / g (CFU / mL), 5×10 8 -1×10 12 CFU / g (CFU / mL), 8×10 8 -1×10 12 CFU / g (CFU / mL), 7×10 9 -2×10 11 CFU / g (CFU / mL) or 9×10 10 -6×10 11 CFU / g (CFU / mL).

[0027] Specifically, the drug also includes pharmaceutically acceptable excipients.

[0028] More specifically, the pharmaceutically acceptable excipients include, but are not limited to, any one or more of the following: excipients, stabilizers, diluents, binders, preservatives, lubricants, and antioxidants.

[0029] Specifically, the dosage form of the drug is tablets, liquids, capsules, powders, suppositories, or granules.

[0030] In another aspect, the present invention provides a probiotic preparation comprising the aforementioned Lactococcus lactis subsp. milk fat.

[0031] The technical effects achieved by this invention are as follows:

[0032] The *Lactococcus lactis* subsp. *milk fat* LC-99 provided by this invention has good effects in increasing the weight of patients with enteritis, improving intestinal inflammatory markers, and enhancing immune function. It can be used in products that prevent and avoid weight loss in patients with enteritis, improve intestinal inflammatory markers, and enhance immune function. Therefore, *Lactococcus lactis* subsp. *milk fat* LC-99 has great application potential in the preparation of products (such as food or pharmaceuticals) for preventing and / or reducing weight loss in patients with enteritis, and for preventing and / or improving obesity-related diseases.

[0033] In addition, the Lactococcus lactis subsp. milk fat LC-99 is a beneficial intestinal bacterium and is listed in the list of bacteria that can be used in food. Therefore, Lactococcus lactis subsp. milk fat LC-99 and products containing Lactococcus lactis subsp. milk fat LC-99 as the active ingredient have the advantage of food safety and are beneficial to human health with long-term use.

[0034] Preservation information:

[0035] Biomaterial: LC-99;

[0036] Classification and nomenclature: Lactococcus lactis subsp. milk fat ( Lactococcus lactis subsp.cremoris );

[0037] Accession number: CGMCC No. 29363;

[0038] Deposit date: December 19, 2023;

[0039] Preservation institution: China General Microbiological Culture Collection Center, China Committee on the Preservation and Management of Microbial Culture Collections;

[0040] Abbreviation of depositary institution: CGMCC;

[0041] Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Attached Figure Description

[0042] Figure 1 Microscopic image of Lactococcus lactis subsp. milk fat LC-99.

[0043] Figure 2 This represents the average nucleic acid consistency result.

[0044] Figure 3 This is a genome loop diagram.

[0045] Figure 4 The predicted results for probiotics.

[0046] Figure 5 This is the experimental design diagram.

[0047] Figure 6The percentage change in body weight for each group is represented by ***, where *** represents p < 0.001 and ** represents p < 0.01.

[0048] Figure 7 Disease Activity Index (DAI) scores were assigned to each group, with *** representing p<0.001 and ** representing p<0.01.

[0049] Figure 8 The values ​​represent immune markers, where A represents IL-10; B represents IL-6; C represents IL-9; and D represents TNF-α. *** represents p<0.001, ** represents p<0.01, and * represents p<0.05. Detailed Implementation

[0050] The present invention will be further described in detail below with reference to specific embodiments. The following embodiments are not intended to limit the present invention, but only to illustrate the present invention. Unless otherwise specified, the experimental methods used in the following embodiments are generally performed under conventional conditions. Unless otherwise specified, the materials and reagents used in the following embodiments are commercially available.

[0051] Example 1: Isolation and identification of *Lactococcus lactis* subsp. *milk fat*

[0052] The method for isolating and identifying *Lactococcus lactis* subsp. *milk fat* includes the following steps:

[0053] (1) Isolation, purification and preservation of Lactococcus lactis subsp. milk fat LC-99

[0054] One mL of whey sample was serially diluted with sterile physiological saline to the appropriate dilution and poured onto MRS solid agar plates. The plates were then inverted and incubated at 30°C for 24-48 h. Colonies of different morphologies were observed and selected. The selected colonies were inoculated onto MRS liquid agar and incubated at 30°C for 18-24 h. The purity of the picked colonies was checked by Gram staining and microscopic examination. Pure strains were obtained through repeated streak purification.

[0055] The bacterial strain was preserved by centrifuging the bacterial solution at 4000×g for 5 min, removing the supernatant, washing the obtained bacterial sludge three times with PBS, mixing it with skim milk in a certain ratio, and storing it at -80℃.

[0056] The results of microscopic examination of *Lactococcus lactis* subsp. *milk fat* LC-99 are as follows: Figure 1 Lactococcus lactis subsp. milk fat LC-99 is a Gram-positive bacterium that is non-motile and distributed in ellipsoidal chains.

[0057] (2) Genome analysis of Lactococcus lactis subsp. 99

[0058] ① Genome sequencing and assembly

[0059] The genome was sequenced using Nanopore Promethion 24 and Illumina Novaseq, the genome was assembled using NextDenove software, the genome was corrected using NextPolish software, and the genome was circularized using Circlator.

[0060] ② Comparison with GTDB genome database

[0061] The whole genome sequence of LC-99 was compared with GTDB, and the results showed that LC-99 is *Lactococcus lactis* subsp. *milk fat*. The results are shown in Table 1.

[0062] Table 1 GTDB alignment results

[0063]

[0064] ③ Calculation of average nucleotide identity (ANI)

[0065] At the genomic level, the ANI (Affinity Intent) value, obtained from homologous sequence alignment between strains, can be used to identify the phylogenetic relationship of strains. Generally, an ANI value greater than 95% is considered to indicate that they belong to the same species. (The text then abruptly shifts to a different topic: "Type strains...") Lactococcus milk subsp .creamy ATCC 19257 T With LC-99 and Lactococcus milk subsp .creamy The MG1363 was used to calculate the ANI value and construct a clustering heatmap. The results are shown (see...). Figure 2 ), LC-99 and Lactococcus lactis subsp .creamy ATCC 19257 T and Lactococcus milk subsp .creamy The ANI values ​​of MG1363 were all greater than 97%, indicating that LC-99 and Lactococcus milk subsp .creamy ATCC 19257 T They are the same species, that is, LC-99 is *Lactococcus lactis* subsp. *milk fat* ( Lactococcus lactis subsp .creamy ).

[0066] ④ Basic genome information

[0067] Analysis of the genome revealed that the genome of *Lactococcus lactis* subsp. *liquidambar* LC-99 is 2.49 Mb in size, with a GC content of 35.78%, containing 2828 CDS, 64 tRNAs, and 19 rRNAs. Its genome consists of one chromosome and two plasmids (see Table 2). A genome circumscopy map was constructed based on the *Lactococcus lactis* subsp. *liquidambar* LC-99 genome (see Table 2). Figure 3 ).

[0068] Table 2. Basic genomic information of Lactococcus lactis subsp. 99

[0069]

[0070] ⑤ Drug resistance gene annotation

[0071] Using the Perfect and Strict algorithms in RGI 5.2.1 software as screening criteria, *Lactococcus lactis* subsp. *limon* LC-99 was compared with the CARD (The Comprehensive Antibiotic Resistance Database) to predict antibiotic resistance-related genes in the strain's genome. The results showed that one vancomycin-resistant glycopeptide resistance gene, *vanY*, was detected in the *Lactococcus lactis* subsp. *limon* LC-99 genome, but its similarity was only 34.72% (similarity less than 40% is generally not considered to indicate identical function). Therefore, no antibiotic resistance genes were detected in *Lactococcus lactis* subsp. *limon* LC-99.

[0072] ⑥ Toxicity Factor Annotation

[0073] The genome of *Lactococcus lactis* subsp. *milk fat* LC-99 was compared with the VirulenceFinder database (https: / / cge.food.dtu.dk / services / VirulenceFinder / ). This currently includes comparisons with *Listeria* (…). Listeria) Staphylococcus aureus ( Streptococcus aureus ), Escherichia coli ( Escherichia coli ), Enterococcus ( Enterococcus The study investigated virulence-related genes in four types of microorganisms. The database's default values ​​(sequence similarity >90% and sequence coverage >60%) were used as screening criteria. Results showed that no potential virulence-related genes were detected in the genome of *Lactococcus lactis* subsp. *milkfat* LC-99.

[0074] ⑦ Probiotic Prediction

[0075] Probiotic prediction was performed on *Lactococcus lactis* subsp. *milk fat* LC-99 using the iProbiotics platform (which contains enriched genes for over 2282 probiotic strains). Analysis using the iProbiotics platform's Model 1: Probiotic Predictor revealed that *Lactococcus lactis* subsp. *milk fat* LC-99 had a 98.931% probability of being a probiotic. (See [link to iProbiotics platform]). Figure 4 (Green represents the probability of probiotics, and red represents the probability of non-probiotics. The prediction result includes one chromosome and two plasmids (plasmid 1 and plasmid 2)).

[0076] Results: Based on the ANI results, LC-99 was determined to belong to... Lactococcus lactis subsp .creamy The genome of *Lactococcus lactis* subsp. *limon* LC-99 consists of one chromosome and two plasmids, with a genome size of 2.49 Mb, a GC content of 35.78%, and contains 2828 CDS, 64 tRNAs, and 19 rRNAs. No drug resistance-related genes or potential virulence-related genes were detected in the *Lactococcus lactis* subsp. *limon* LC-99 genome. Its probability of being a probiotic is 98.931%.

[0077] Example 2: Effects of Lactococcus lactis subsp. milk fat LC-99 on intestinal and immune function in mice.

[0078] 1. Experimental Design

[0079] Forty male C57BL / 6J mice were administered 2.5% DSS (sodium dextran sulfate) in their drinking water from days 0-5, and sterile water from days 5-10. This process constituted one cycle, and a total of three cycles were performed to establish the model. The experimental group was fed *Lactococcus lactis* subsp. *milk fat* LC-99, control group 1 was fed *Lactococcus lactis* subsp. *milk fat* C09, and control group 2 was fed *Lactococcus lactis* subsp. *milk fat* C10. All control strains were isolated by our company. The bacterial concentration in each group was 3 × 10⁻⁶. 9 CFU / 0.2 mL / day; the model group was fed the same volume of sterile water. Eight animals were fed in each group for a total of 30 days. The experimental design was as follows: Figure 5 As shown.

[0080] 2. Measurement Indicators

[0081] The measurement indicators were: initial body weight and body weight during intervention, DAI score at different time points during intervention, and measurement of immune indicators.

[0082] 3. Measurement methods and results

[0083] (1) Weight changes

[0084] Experimental results are as follows Figure 6As shown, compared with the control group, the body weight of mice in all groups decreased significantly; only the LC-99 group had a significantly higher rate of body weight change than the model group ( P <0.01).

[0085] (2) DAI score

[0086] The Disease Activity Index (DAI) is used to assess the severity of IBD, and is the sum of three scores: Bodyweight Change (BWC) score, Stool Consistency score, and Fecal Occult Blood score. A higher DAI score indicates more severe IBD symptoms. The specific scoring criteria for Bodyweight Change (BWC) are: 0 = BWC ≥ 0%; 1 = -5% ≤ BWC < 0%; 2 = -10% ≤ BWC < -5%; 3 = -15% ≤ BWC < -10%; 4 = BWC < -15%. Fecal occult blood was detected using a fecal occult blood test kit (Shanghai Enzyme-Linked Biotechnology Co., Ltd., 48 / 96T). Scoring was based on the following criteria: 0 = Negative fecal occult blood test > 120s; 1 = 60s < Positive fecal occult blood test ≤ 120s; 2 = 30s < Positive fecal occult blood test ≤ 60s; 3 = 10s < Positive fecal occult blood test ≤ 30s; 4 = 0s < Positive fecal occult blood test ≤ 10s; 5 = Visible blood in the stool. Stool consistency was scored as follows: 0 = Normal; 1 = Moist / sticky stool; 2 = Soft stool; 3 = Diarrhea.

[0087] DAI score results are as follows Figure 7 As shown, the DAI score of the LC-99 group was significantly lower than that of other groups, but there was no significant difference compared with the model group.

[0088] (3) Immune indicators

[0089] Inflammatory response is a major symptom in patients with enteritis, and the levels of inflammatory factors are closely related to the severity of enteritis. This experiment used Luminex's xMAP® technology to determine the levels of immune factors in mouse serum, also known domestically as "liquid-phase suspension chip technology." 30 μL of serum sample was used for this experiment. The ProcartaPlex™ Multifactor Kit (catalog number: EPX170-26087-901) was used to quantitatively detect serum immune factors, including IL-6, IL-9, IL-10, and TNF-α. The experimental procedures were performed according to the kit instructions. The fluorescence intensity of each well in a 96-well plate was read using a Luminex 200 instrument. A five-parameter nonlinear regression method was used to fit a standard curve, and the concentrations of the four immune factors were calculated.

[0090] The results are as follows Figure 8 As shown, compared with the model group, the LC-99 group showed a significant increase in the anti-inflammatory marker IL-10 and a significant decrease in the inflammatory markers IL-6, IL-9, and TNF-α, with significant changes compared to Comparative Example 1 and Comparative Example 2. This indicates that LC-99 intake can enhance the immune function of mice.

[0091] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A strain of Lactococcus lactis subsp. milk fat ( Lactococcus lactis subsp.cremoris ), characterized in that, The preservation number of the Lactococcus lactis subsp. milk fat is CGMCC No. 29363.

2. The culture method of Lactococcus lactis subsp. cremoris according to claim 1, characterized in that, This includes inoculating *Lactococcus lactis* subsp. *milk fat* onto a culture medium for cultivation; the culture medium includes MRS medium.

3. The use of *Lactococcus lactis* subsp. *milk fat* as described in claim 1 in the preparation of products for the prevention, treatment and / or adjunctive treatment of enteritis; the specific method for modeling enteritis is to administer 2.5% sodium dextran sulfate to mice in drinking water for 0-5 days, and to administer sterile water to mice for 5-10 days. This process constitutes one cycle, and a total of three cycles are performed for modeling.

4. Use according to claim 3, characterized in that, The product is a drug or pharmaceutical raw material.

5. Use according to claim 4, characterized in that, The drug or pharmaceutical ingredient includes live Lactococcus lactis subsp. milk fat.

6. Use according to claim 4, characterized in that, The drug or pharmaceutical raw material includes fermentation broth or freeze-dried powder of Lactococcus lactis subsp. lactis.

7. A medicament, characterized by comprising a compound of the formula (I) or a pharmaceutically acceptable salt thereof. The drug comprises *Lactococcus lactis* subsp. *lactofattya* as described in claim 1.

8. The medicament according to claim 7, characterized in that, The viable cell number of Lactococcus lactis subsp. cremoris in the drug is not less than 1 x 10 6 CFU / mL or 1 x 10 6 CFU / g.

9. The drug according to claim 7 or 8, characterized in that, The drug also includes pharmaceutically acceptable excipients.

10. The medicament according to claim 9, characterized in that, The dosage form of the drug is tablets, liquids, capsules, powders, suppositories, or granules.

11. A probiotic preparation, characterized in that, The probiotic preparation includes the Lactococcus lactis subsp. milk fat as described in claim 1.

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