Phytobacterium plantarum and application thereof in immunoregulation and intestinal environment improvement

Abstract

The invention discloses plant lactobacillus and application thereof in immunoregulation and intestinal environment improvement, and relates to the technical field of microorganisms. The invention provides a strain of plant lactobacillus LIHUO 05, and the preservation number of the plant lactobacillus LIHUO 05 is CGMCC (China General Microbiological Culture Collection Center) No.36240. The invention also provides a preparation method of the plant lactobacillus LIHUO 05. The plant lactobacillus LISUO 05 can be used for promoting macrophages to secrete TNF (Tumor Necrosis Factor)-alpha and IL-1beta; the thermally inactivated strain LIHUO 05 has a certain immunoregulation effect on the body weight, thymus index and TNF-alpha content in serum of immunocompromised mice. Meanwhile, the phytobacterium plantarum LIHUO 05 can change the colon flora structure in the intestinal tract, improve the relative abundance of beneficial bacteria and adjust the variety and number of colon metabolites in the intestinal tract, and has a certain effect of improving the intestinal tract environment.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbial technology, in particular to Lactobacillus plantarum and its application in immune regulation and improvement of intestinal environment. BACKGROUND

[0002] The immune system, which is composed of immune organs, immune cells and immune factors, can significantly weaken the body's ability to defend against pathogens, remove metabolic waste and maintain homeostasis when its function is imbalanced. The cyclophosphamide-induced immunocompromised mouse model, as an important tool for studying immune regulation mechanisms, has been widely verified for its application value. For the functional reconstruction of immunocompromised organisms, existing clinical intervention methods often face the dual dilemma of over-activated immune response or insufficient targeting. It is worth noting that recent studies have shown that animal bifidobacterium, Bifidobacterium breve and other probiotic strains can enhance the immune function of immunocompromised individuals through specific mechanisms, and by regulating the expression level of key inflammatory factors such as TNF-α and IL-1β, they can simulate the immune warning signal in the natural infection process under the condition of limited dose and intervention time. This regulatory strategy can effectively avoid tissue damage caused by cytokine storm, and through promoting lymphocyte differentiation and other ways, it can achieve the mild reconstruction of immune surveillance function. This intervention paradigm of simulation-induction-balance breaks through the limitations of traditional immune enhancement therapy and provides a theoretical basis for the development of new immunomodulators.

[0003] As a kind of human-derived lactic acid bacteria, breast milk-derived lactic acid bacteria have great potential in functional mining and application due to their good safety and human adaptability, and have become a precious microbial resource. Studies have shown that these strains have unique advantages in immune regulation and intestinal health regulation. Lactobacillus plantarum is a widely used probiotic bacteria, which can affect the body's immunity through various ways such as changing the structure and composition of the host's intestinal flora, regulating the level of short-chain fatty acids in feces, etc.

[0004] In prior art 1 (Dhanani AS, Bagchi T. Lactobacillus plantarum CS24.2 prevents Escherichia coli adhesion to HT-29 cells and also down-regulates enteropathogen-induced tumor necrosis factor-alpha and interleukin-8 expression [J]. Microbiol Immunol. 2013 Apr;57(4):309-15. doi: 10.1111 / 1348-0421.12038.) it is disclosed that Lactobacillus plantarum CS24.2 can protect intestinal epithelial cells, and by studying HT-29 cells infected with Escherichia coli O26:H11, it is found that the strain of probiotics can reduce the expression of tumor necrosis factor and interleukin 8 in HT-29 cells. In prior art 2 (Wang Yuan, Guo Yilin, Xie Qiong, et al. Lactobacillus plantarum ZDY2013 and Bifidobacterium bifidum WBIN03 for the treatment of TNBS-induced colitis in mice [J]. Chinese Journal of Microecology, 2017, 29 (03):279-283. DOI:10.13381 / j.cnki.cjm.201703007.) it is disclosed that after mice with ulcerative colitis were continuously gavaged with Lactobacillus plantarum ZDY2013 and Bifidobacterium bifidum WBIN03 for one week, it was found that the two strains of probiotics promoted the expression of Superoxide dismutase 1 (SOD1), SOD2, SOD3 and other genes in mice, reduced the production of TNF-α, and activated immune cells such as T lymphocytes and B lymphocytes, and enhanced the host immunity.

[0005] In Chinese patent CN104480032A, a Lactobacillus plantarum with enhanced immune activity is disclosed, and the strain is Lactobacillus plantarum (D-92). Lactobacillus plantarum The strain has the ability to improve the body's immunity, and the composition is in the form of a food or a pharmaceutical composition. The invention also discloses a method for screening and determining the strain of lactic acid bacteria, and a method for improving the body's immunity by improving the organ index, non-specific immune response and specific immune response of the body. However, the strain is derived from pickles, and has poor safety.

[0006] At present, no researchers have systematically focused on the influence of breast milk-derived Lactobacillus plantarum on the immune system of immunocompromised hosts and the intestinal environment. Therefore, providing a Lactobacillus plantarum with immune regulation and intestinal environment improvement functions can provide excellent strain resources for the development of breast milk-derived probiotics. SUMMARY

[0007] The application aims to provide a breast milk source plant lactobacillus LIHUO 05 and application thereof in immune regulation and improvement of intestinal environment.

[0008] To achieve the above application purposes, the technical solutions of the application are as follows. In a first aspect, the application provides a plant lactobacillus LIHUO 05, which is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC No.36240. Lactiplantibacillus plantarum Specifically, the 16s rDNA sequence of the plant lactobacillus LIHUO 05 is as shown in SEQ ID NO:1-2.

[0009] >27F (SEQ ID NO:1):

[0010] >27F (SEQ ID NO:1): NNGCNNNNGGGCGGCGTGCCTATACATGCAAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGTTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGAGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAAGGCTGAAACTCAAAGGAATTGACAGGGGCCCGC.

[0011] >1492R (SEQ ID NO: 2): CNNANNNNNTGTCACCTTAGGCGGCTGGTTCCTAAAAGGTTACCCCACCGACTTTGGGTGTTACAAACTCTCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCATGTAGGCGAGTTGCAGCCTACAATCCGAACTGAGAATGGCTTTAAGAGATTAGCTTACTCTCGCGAGTTCGCAACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCACCAGAGTGCCCAACTTAATGCTGGCAACTGATAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTATCCATGTCCCCGAAGGGAACGTCTAATCTCTTAGATTTGCATAGTATGTCAAGACCTGGTAAGGTTCTTCGCGTAGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGCCTTGCGGCCGTACTCCCCAGGCGGAATGCTTAATGCGTTAGCTGCAGCACTGAAGGGCGGAAACCCTCCAACACTTAGCATTCATCGTTTACGGTATGGACTACCAGGGTATCTAATCCTGTTTGCTACCCATACTTTCGAGCCTCAGCGTCAGTTACAGACCAGACAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACTCAAGTTTCCCAGTTTCCGATGCACTTCTTCGGTTGAGCCGAAAGCTTTCACATCAGACTTAAAAAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCCGGACAACGCTTG.

[0012] Specifically, the Lactobacillus plantarum LIHUO 05 is derived from the breast milk of a healthy female.

[0013] Specifically, the Lactobacillus plantarum LIHUO 05 has a colony on MRS solid medium, which is milky white, semicircular, smooth and moist, and has a neat edge.

[0014] In a second aspect, the present application provides a preparation comprising the Lactobacillus plantarum LIHUO 05.

[0015] Specifically, the preparation comprises a fermentation broth of the Lactobacillus plantarum LIHUO 05, a supernatant of the fermentation broth of the Lactobacillus plantarum LIHUO 05, a precipitate of the fermentation broth of the Lactobacillus plantarum LIHUO 05, live bacteria of the Lactobacillus plantarum LIHUO 05, and dead bacteria of the Lactobacillus plantarum LIHUO 05.

[0016] Specifically, the fermentation broth refers to a liquid after inoculation of a bacterial strain into a culture medium.

[0017] Further specifically, the supernatant of the fermentation broth refers to a clear liquid in the upper layer after centrifugation of the fermentation broth, which contains rich metabolic products and a part of bacterial fragments in the process of bacterial growth and reproduction.

[0018] Further specifically, the precipitate of the fermentation broth refers to a liquid precipitate after centrifugation, which comprises free proteins, residual bacterial bodies, broken cells, and residues of culture medium.

[0019] According to some embodiments of the present application, the dead bacteria of the Lactobacillus plantarum LIHUO 05 are heat-inactivated bacterial suspension of the Lactobacillus plantarum LIHUO 05.

[0020] According to some embodiments of the present application, the preparation of the heat-inactivated bacterial suspension of the Lactobacillus plantarum LIHUO 05 comprises the following steps: (1) inoculating the activated liquid of the Lactobacillus plantarum LIHUO 05 into MRS liquid medium for culture to obtain a bacterial liquid; (2) centrifuging the bacterial liquid to obtain bacterial bodies of the Lactobacillus plantarum; (3) resuspending the bacterial bodies of the Lactobacillus plantarum in PBS to obtain a bacterial suspension; (4) inactivating the bacterial suspension at 80-100℃ for 15-30 min, and obtaining the heat-inactivated bacterial suspension.

[0021] Specifically, the preparation method of the activated liquid of the Lactobacillus plantarum LIHUO 05 comprises the following steps: streaking on MRS solid medium and culturing at 37℃ for 48 h to obtain single colonies; inoculating the single colonies into MRS liquid medium and culturing at 37℃ for 13-18 h for activation; and continuously activating for two generations to obtain the activated liquid.

[0022] According to some embodiments of the present application, the inoculation amount in step (1) is 1-5% (v / v); preferably, the inoculation amount in step (1) is 2% (v / v).

[0023] Specifically, the culture condition in step (1) is 37°C for 13-18h.

[0024] Specifically, the centrifugation condition in step (2) is 4°C, 6000g for 5min.

[0025] Further, the inactivation condition in step (4) is 100°C for 30min.

[0026] In a third aspect, the present application provides a microbial agent, which comprises the above-mentioned Lactobacillus plantarum LIHUO 05 or the above-mentioned preparation.

[0027] Specifically, the microbial agent is a powder or a liquid preparation.

[0028] Specifically, the microbial agent can be in a solid, liquid, semi-solid or any physical form that can be obtained by current or future possible technology and contains necessary active ingredients to achieve any application, function or effect of the present application.

[0029] For some embodiments, the microbial agent is in a solid form. Specifically, it can be a freeze-dried powder.

[0030] Specifically, the preparation method of the freeze-dried powder can use the current conventional method in the art, or other methods that can appear in the future to obtain a solid form of the product of the microorganism or the microbial culture. It should be clarified that the preparation method of the freeze-dried powder should not be a condition to limit the scope of protection of the present application.

[0031] In a fourth aspect, the present application provides the use of the above-mentioned Lactobacillus plantarum LIHUO 05 or the above-mentioned preparation or the above-mentioned microbial agent in the preparation of a product for enhancing immunity.

[0032] Specifically, the number of viable Lactobacillus plantarum LIHUO 05 in the product is 1×10 5 -1×10 12 CFU / mL or 1×10 5 -1×10 12 CFU / g, specifically 1×10 5 CFU / mL (CFU / g), 1×10 6 CFU / mL (CFU / g), 1×10 7 CFU / mL (CFU / g), 1×10 8 CFU / mL (CFU / g), 1×109 CFU / mL (CFU / g), 1x10 10 CFU / mL (CFU / g), 1x10 11 CFU / mL (CFU / g), 1x10 12 CFU / mL (CFU / g), 1x10

[0033] Further, the number of viable bacteria of Lactobacillus plantarum LIHUO 05 in the product is 1x10 7 1x10 11 CFU / mL or 1x10 7 1x10 11 CFU / g.

[0034] Further, the number of viable bacteria of Lactobacillus plantarum LIHUO 05 in the product is 1x10 8 1x10 10 CFU / mL or 1x10 8 1x10 10 CFU / g.

[0035] Specifically, the product includes a medicine and a health food.

[0036] Specifically, the product has at least one of the following effects: (1) promoting the secretion of TNF-α and IL-1β in macrophages; (2) promoting the proliferation of lymphocytes; (3) capable of increasing the body weight, thymus index and TNF-α content in serum of an immunocompromised subject; (4) changing the intestinal colon flora structure of an immunocompromised subject, and increasing the relative abundance of beneficial bacteria Alistipes and Alloprevotella; (5) capable of regulating the types and quantities of intestinal colon metabolites in an immunocompromised subject, increasing the abundance of immune-related metabolites Soyasaponin V and Soyasaponin IV, and reducing the abundance of Murideoxycholic acid and Nylidrin.

[0037] Specifically, when the product is a medicine, the medicine includes a pharmaceutically acceptable carrier.

[0038] Further, the pharmaceutically acceptable carrier is selected from one or more of excipients, stabilizers, diluents, binders, preservatives, and lubricants.

[0039] Specifically, the dosage form of the drug is drops, mixture, tincture, injection, tablet, powder, oral liquid, capsule, granule, ointment, suspension, powder, emulsion, solution, drop pill, pill, buccal tablet, lyophilized powder injection, gel, suppository or aerosol.

[0040] Further, the health food further comprises health food conventional adjuvants, and the adjuvants include but are not limited to fillers, flavoring agents, binders, disintegrants, lubricants, antacids and nutritional fortifiers.

[0041] Further, the health food comprises emulsion products, solution products, powder products and solid products.

[0042] In a fifth aspect, the present application provides the use of the above-mentioned Lactobacillus plantarum LIHUO 05 or the above-mentioned preparation or the above-mentioned bacterial agent in the preparation of a product for regulating the intestinal flora environment.

[0043] Specifically, the product contains 1×10 5 -1×10 12 CFU / mL or 1×10 5 -1×10 12 CFU / g, specifically 1×10 5 CFU / mL (CFU / g), 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 10 CFU / mL (CFU / g), 1×10 11 CFU / mL (CFU / g), 1×10 12 CFU / mL (CFU / g), etc., and other point values within the numerical range can be selected.

[0044] Further, the product contains 1×10 7 -1×10 11 CFU / mL or 1×10 7 -1×10 11 CFU / g.

[0045] Still further, the product contains 1×10 8 -1×10 10 CFU / mL or 1×10 8 -1×1010 CFU / g.

[0046] Specifically, the product includes a medicine and a health food.

[0047] Specifically, the product has at least one of the following effects: (1) At the door level, the relative abundance of Bacteroidota, Campylobacterota, Verrucomicrobiota is reduced, and the relative abundance of Bacillota_A, Bacillota is increased. (2) At the genus level, the relative abundance of CAG-485, Alistipes, Duncaniella is reduced, and the relative abundance of Alloprevotella, UBA3263 is increased.

[0048] Specifically, when the product is a medicine, the medicine includes a pharmaceutically acceptable carrier.

[0049] Further, the pharmaceutically acceptable carrier is selected from one or more of excipients, stabilizers, diluents, binders, preservatives, lubricants.

[0050] Specifically, the dosage form of the medicine is drops, mixture, tincture, injection, tablet, powder, oral liquid, capsule, granule, ointment, suspension, powder, emulsion, solution, drop pill, pill, buccal tablet, freeze-dried powder injection, gel, suppository or aerosol.

[0051] Further, the health food further includes a health food conventional auxiliary material, and the auxiliary material includes but is not limited to a filler, a flavoring agent, a binder, a disintegrant, a lubricant, an antacid and a nutritional fortifier.

[0052] Further, the health food includes emulsion products, solution products, powder products and solid products.

[0053] In a sixth aspect, the present application provides a product for enhancing immunity or regulating intestinal flora, wherein the product contains the above-mentioned plant lactobacillus LIHUO 05 or the above-mentioned preparation or the above-mentioned bacterial agent.

[0054] Specifically, the product contains 1x10 5 -1x10 12 CFU / mL or 1x10 5 -1x10 12 CFU / g.

[0055] Further, the number of viable bacteria of the plant lactobacillus LIHUO 05 in the product is 1×10 7 -1×10 11 CFU / mL or 1×10 7 -1×10 11 CFU / g.

[0056] Further, the number of viable bacteria of the plant lactobacillus LIHUO 05 in the product is 1×10 8 -1×10 10 CFU / mL or 1×10 8 -1×10 10 CFU / g.

[0057] Specifically, the product includes medicines and health foods.

[0058] Specifically, when the product is a medicine, the medicine includes a pharmaceutically acceptable carrier.

[0059] Further, the pharmaceutically acceptable carrier is selected from one or more of excipients, stabilizers, diluents, binders, preservatives, and lubricants.

[0060] Specifically, the dosage form of the medicine is a drop, a mixture, a tincture, an injection, a tablet, a powder, an oral liquid, a capsule, a granule, an ointment, a suspension, a powder, an emulsion, a solution, a dripping pill, a pill, a buccal tablet, a freeze-dried powder injection, a gel, a suppository, or an aerosol.

[0061] Further, the health food further includes conventional health food auxiliaries, and the auxiliaries include, but are not limited to, fillers, flavoring agents, binders, disintegrants, lubricants, antacids, and nutritional fortifiers.

[0062] Further, the health food includes emulsion products, solution products, powder products, and solid products.

[0063] The beneficial effects of the present application are: The present application screens a plant lactobacillus LIHUO 05 with the functions of regulating immunity and improving intestinal environment, and the specific functions are: (1) The viable bacteria of LIHUO 05 can increase the content of TNF-α secreted by macrophages under normal conditions by 46.96 times and the content of IL-1β by 118.30%. (2) In a mouse primary epithelial cell-spleen lymphocyte co-culture system, heat-inactivated LIHUO 05 can promote the proliferation of lymphocytes by 1.90%.

[0064] (3) LIHUO 05 viable bacteria can increase the body weight of immunocompromised mice by 6.76%, increase the thymus index by 41.25%, and increase the serum TNF-α content by 24.69%. (4) LIHUO 05 viable bacteria can change the intestinal colon flora structure of immunocompromised mice, and increase the relative abundance of beneficial bacteria Alistipes and Alloprevotella.

[0065] It can regulate the types and quantities of colon metabolites in the intestines of immunocompromised mice, significantly increase the abundance of immune-related metabolites Soyasaponin V and Soyasaponin IV, and reduce the abundance of Murideoxycholic acid and Nylidrin.

[0066] Deposit Description Deposit number: CGMCC No. 36240; Classification name: Plantamolibacter lathyri Lactiplantibacillus plantarum Deposit date: October 17, 2025; Preservation unit: China General Microbiological Culture Collection Center; Abbreviation of preservation unit: CGMCC; Preservation address: No. 3, Yikhina Road, Chaoyang District, Beijing, China Institute of Microbiology. Brief Description of the Drawings

[0067] Figure 1 The colony morphology of Plantamolibacter lathyri LIHUO 05.

[0068] Figure 2 The effect of Plantamolibacter lathyri JZ-LP02 on TNF-α of normal macrophages, compared with the normal group, p <0.0001.

[0069] Figure 3 The effect of Plantamolibacter lathyri LIHUO 05 on IL-1β of normal macrophages, compared with the normal group, p <0.001.

[0070] Figure 4 The effect of heat-inactivated Plantamolibacter lathyri LIHUO 05 on the proliferation of lymphocytes in the mouse primary epithelial cell-spleen lymphocyte co-culture system, compared with the normal group, p <0.01.

[0071] Figure 5 The final body weight of mice, compared with the model group, p <0.01, p ​<0.0001.

[0072] Figure 6 For the thymus index of mice, compared with the model group, p <0.01.

[0073] Figure 7 For serum TNF-α content, compared with the model group, p <0.05, p <0.01, p <0.0001.

[0074] Figure 8 For the relative abundance of the door level.

[0075] Figure 9 For the relative abundance of the genus level.

[0076] Figure 10 For the relative abundance of Allpprevotella and Alistipes at the genus level.

[0077] Figure 11 For the partial least squares-discriminant analysis (PLS-DA) diagram of metabolites.

[0078] Figure 12 For the KEGG enrichment histogram of differential metabolites (up).

[0079] Figure 13 For the KEGG enrichment histogram of differential metabolites (down).

[0080] Figure 14 For the box plot of immune-related differential metabolites (up).

[0081] Figure 15 For the box plot of immune-related differential metabolites (down).

[0082] Figure 16 For the LIHUO 05 genome circle diagram.

[0083] Figure 17 For the LIHUO 05 GO function annotation classification statistical chart.

[0084] Figure 18 For the LIHUO 05 KEGG annotation classification statistical chart.

[0085] Figure 19 For the LIHUO 05 eggNOG function gene function classification statistical chart. DETAILED DESCRIPTION

[0086] In order to make the technical means, creative features, purposes and effects of the present application easy to understand, the following specific embodiments are further described to clarify the present application. However, the following embodiments are only preferred embodiments of the present application, not all. Based on the embodiments in the embodiments, other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the present application. In the following embodiments, if not otherwise specified, the operation methods used are conventional operation methods, the equipment used is conventional equipment, and the equipment materials used in each embodiment are the same.

[0087] The purchase manufacturers of experimental materials are as follows: The mouse monocyte macrophage RAW 264.7 was purchased from the Shanghai Cell Library of the Chinese Academy of Sciences.

[0088] The mouse primary small intestinal mucosal epithelial cells and mouse primary spleen lymphocytes were purchased from Wuhan Punsun Life Science and Technology Co., Ltd.

[0089] The culture media involved in the following embodiments are as follows: MRS culture medium formula (1L): proteose peptone 10g, beef extract 10g, yeast powder 5g, glucose 20g, K2HPO42g, diammonium citrate 2g, sodium acetate 2g, Tween 80 1mL, MgSO4·7H2O 0.5g, MnSO4·4H2O 0.25g, pH 7.2-7.4; sterilized at 121℃ for 15 min.

[0090] MRS solid culture medium formula (1L): proteose peptone 10g, beef extract 10g, yeast powder 5g, glucose 20g, K2HPO42g, diammonium citrate 2g, sodium acetate 2g, Tween 80 1mL, MgSO4·7H2O 0.5g, MnSO4·4H2O 0.25g, agar 20g, pH 7.2-7.4; sterilized at 121℃ for 15 min.

[0091] The culture method of macrophage RAW 264.7 involved in the following embodiments is as follows: After the macrophage is recovered, the cell adhesion is observed, if the cell growth state is good and the density reaches about 80%, the cell is subcultured. The culture medium in the culture bottle is aspirated, 5 mL PBS is added to rinse 1-2 times, and the residual culture medium is removed; after the rinsed PBS is aspirated, 1 mL trypsin is added, the trypsin is shaken to uniformly cover the bottom of the bottle, and is placed in a culture box at 37°C for digestion; during the digestion, it is taken out to observe every 3-5 min, until the edge of part of the cells begins to fall off, the trypsin being digested is aspirated to blow the cells and collect them into a centrifugal tube containing 3-4 mL complete culture medium, the cell suspension in the centrifugal tube is gently blown to disperse the cells; the cells are collected by centrifugation at 1000 rpm for 3 min; the cells are gently resuspended with fresh DMEM medium, the cell suspension is uniformly inoculated into a culture dish, and is mixed by a cross method or an 8-shaped method, and then is placed in a culture box for continuous culture, and the cell state is observed. After 2-3 times of subculture, subsequent experiments are carried out.

[0092] The culture method of mouse primary small intestinal mucosal epithelial cells involved in the following examples is as follows: The culture medium in the culture bottle is aspirated, the cells are rinsed once with PBS. 1 mL of 0.25% trypsin digestion solution is added to the T25 culture bottle, the culture bottle is slightly rotated until the digestion solution covers the entire bottom of the culture bottle, then the excess trypsin is aspirated, and is incubated at 37°C for 1-3 min; inverted microscope observation is performed, and after the cells retract, 5 mL of complete culture medium is added to terminate the digestion. The pipette is gently blown to mix, inoculated into a T25 culture bottle according to the subculture ratio, and then fresh complete culture medium is supplemented to 5 mL, and is placed in a 37°C, 5% CO2, saturated and moderate cell culture box for static culture. After the cells completely adhere, the culture is observed, and is used for experiments. Then, the fresh complete culture medium is replaced according to the frequency. When the cells grow to 80%-90% of the bottom of the bottle, 0.25% trypsin is used to digest at 37°C for 2-3 min, and then the digestion is terminated, the cells are resuspended and counted after centrifugation, and are used. The mouse primary small intestinal mucosal epithelial cells are inoculated into a Transwell culture chamber, 100 μL of cell suspension (the number of cells is 1.0×10 5 6.0×10 4. The cells are cultured for 21 days.

[0093] The culture method of mouse spleen lymphocytes involved in the following examples is as follows: Collect the cells in the culture flask into a 15 mL centrifuge tube, centrifuge at 1200 rpm for 3 min, discard the supernatant, collect the cell precipitate. Add 5 mL of fresh lymphocyte special culture medium (Punoxi), gently blow and mix with a pipette to disperse the cells; adjust the dispersed cells to the appropriate density and inoculate into a culture vessel, and place in a 37℃, 5% CO2, saturated and moderate cell culture box for incubation. After the cell state is stable, culture observation is carried out for experiments. Then, the fresh complete culture medium is replaced according to the replacement frequency.

[0094] The preparation method of the bacterial suspension involved in the following examples is as follows: The streaks are cultured on MRS solid medium at 37℃ for 48h to obtain single colonies; the single colonies are inoculated in MRS liquid medium and cultured at 37℃ for 13-18h for activation, and two generations of continuous activation are carried out to obtain an activation liquid; the activation liquid is inoculated in MRS liquid medium at an inoculation amount of 2%(v / v) and cultured at 37℃ for 13-18h to obtain a bacterial liquid; the bacterial liquid is centrifuged at 4℃ and 6000g for 5min to obtain Lactobacillus plantarum bacteria; the Lactobacillus plantarum bacteria are washed with 0.01M PBS and resuspended in 0.01M PBS to a bacterial concentration of 1×10 8 CFU / mL to obtain a bacterial suspension.

[0095] The preparation method of the heat-inactivated bacterial suspension involved in the following examples is as follows: The streaks are cultured on MRS solid medium at 37℃ for 48h to obtain single colonies; the single colonies are inoculated in MRS liquid medium and cultured at 37℃ for 13-18h for activation, and two generations of continuous activation are carried out to obtain an activation liquid; the activation liquid is inoculated in MRS liquid medium at an inoculation amount of 2%(v / v) and cultured at 37℃ for 13-18h to obtain a bacterial liquid; the bacterial liquid is centrifuged at 4℃ and 6000g for 5min to obtain Lactobacillus plantarum bacteria; the Lactobacillus plantarum bacteria are washed with 0.01M PBS and resuspended in 0.01M PBS to a bacterial concentration of 1×10 8 CFU / mL to obtain a bacterial suspension. The bacterial suspension is inactivated at 100℃ for 30min, a small amount of bacterial liquid is taken and spread on an MRS solid plate, and the plate is cultured in a 37℃ constant temperature incubator for 24-48h to observe whether colonies are generated.

[0096] Example 1: Screening, isolation and identification of Lactobacillus plantarum strain The strain involved in the present application is a Lactobacillus screened from breast milk of healthy people, which is identified as Lactobacillus plantarum based on whole genome molecular biology, Figure 1 is a colony morphology diagram of the Lactobacillus plantarum strain in the examples of the present application.

[0097] The specific steps are as follows: (1) Dilution coating Aseptic operation to take 1 mL sample (human milk) and add to 9 mL sterilized water to mix into 1:10 uniform dilution, repeat the above dilution step, and get 10 -2 , 10 -3 , 10 -4 , 10 -5 and 10 -6 dilutions in turn. Use aseptic gun head to take 100 μL of the above 10 -4 , 10 -5 and 10 -6 dilutions, and use a coating rod to coat into MRS solid culture medium, and incubate at 37°C for 48 h to get dilution plates.

[0098] (2) Purified culture Take dilution coating plates with colony numbers in the range of 30-300, and randomly pick 10-20 single colonies of different sizes with white or light color from each sample, and perform three-area streaking on MRS solid culture medium, and incubate at 37°C for 48 h to get single colonies. Take the single colonies on the above streaking plates and inoculate into MRS liquid culture medium, and incubate at 37°C for 12 h to get secondary purified culture solution.

[0099] (3) Strain preservation and identification Mix the secondary purified culture solution, and take the bacterial bodies (incubated at 37°C for 13 h) into 2 mL clean strain preservation tubes, centrifuge to discard the supernatant, resuspend the bacterial bodies with 1 mL 30% glycerol, and place in a -80°C refrigerator. At the same time, send the strain to EUBIO Co., Ltd. for strain identification, which is identified as Lactiplantibacillus plantarum, named LIHUO 05, and the preservation work is completed. Lactiplantibacillus plantarum CICC 21004 is used for comparison, and the strain is purchased from China Industrial Microbial Culture Collection Center.

[0100] Example 2 Effect of Lactiplantibacillus plantarum on secretion of inflammatory factors TNF-α and IL-1β by macrophages I. Experimental steps Macrophages were cultured in DMEM medium added with 10% (v / v) fetal bovine serum, and the culture condition was 37°C, 5% CO2. Cells were inoculated on a 48-well cell culture plate at a number of 1×10 5 cells / well, and after overnight culture, fresh medium was replaced to start experimental treatment.

[0101] The experimental groups are as follows: LIHUO 05 group: 100 μL LIHUO 05 bacterial suspension was added to each well; CICC group: 100 μL CICC 21004 bacterial suspension was added to each well; Blank group: Add 100 μL of PBS to each well.

[0102] Each group had three replicates. After processing, the plate was incubated in a 37℃ CO2 incubator for 2 hours. After incubation, the supernatant from each well was aspirated into a 1.5 mL EP tube and centrifuged at 6000g for 5 minutes to obtain the incubation culture supernatant, which was then stored at -80℃. The levels of TNF-α (Mouse TNF-α Quantikine ELISA kit, R&D, catalog number MTA00B-1) and IL-1β (Mouse IL-1β DuoSet ELISA kit, R&D, catalog number DY401-05) in the incubation culture supernatant were detected according to their respective instructions.

[0103] II. Experimental Results like Figure 2 As shown, LIHUO 05 (1106.00±39.60 pg / mL) can increase the content of TNF-α secreted by macrophages in the blank group (23.06±10.11 pg / mL) by 46.96 times, and the promoting effect is better than that of CICC 21004 (1031.00±66.99 pg / mL).

[0104] like Figure 3 As shown, LIHUO 05 (10.50±1.35 pg / mL) can increase the IL-1β secretion content of macrophages in the blank group (4.81±0.44 pg / mL) by 118.30%, and the promoting effect is significantly better than that of CICC 21004 (4.56±0.23 pg / mL).

[0105] Example 3: Effect of heat-inactivated Lactobacillus plantarum LIHUO 05 on lymphocyte proliferation in a mouse primary small intestinal mucosal epithelial cell-splenic lymphocyte co-culture system. I. Experimental Procedure The methods for culturing mouse primary small intestinal mucosal epithelial cells and spleen lymphocytes are as described above. After the mucosal epithelial cells have formed a dense monolayer, they are seeded in the lower chamber of a Transwell cell line with 10 cells per 100 cells. 5 One spleen lymphocyte was added to 500 μL of spleen lymphocyte-specific culture medium. After the cells adhered and grew, they were transferred to the upper chamber of the mucosal epithelial cells that formed the barrier, and then replaced with fresh complete culture medium.

[0106] The experimental groups are as follows: LIHUO 05 group: 200 μL of special culture solution for mucosal epithelial cells and 100 μL of LIHUO 05 heat-inactivated bacterial suspension were added to each well; CICC group: 200 μL of special culture solution for mucosal epithelial cells and 100 μL of CICC 21004 heat-inactivated bacterial suspension were added to each well; Blank group: 200 μL of special culture solution for mucosal epithelial cells and 100 μL of PBS were added to each well.

[0107] Each group had 3 repeats, and was incubated at 37°C in a 5% CO2 incubator for 2 hours. After incubation, the upper chamber was removed, 50 μL of CCK-8 solution was added to the lower chamber, and it was incubated at 37°C in a 5% CO2 incubator for 4 hours. The OD450nm was detected, and the lymphocyte proliferation rate was calculated.

[0108] Proliferation rate = (OD 实验组 - OD 空白组 ) / OD 空白组 x 100%.

[0109] II. Experimental results Figure 4 As shown in the figure, LIHUO 05 (1.90 ± 0.85%) can promote the proliferation of spleen lymphocytes, with an increase of 1.90%, and the promotion effect is obviously better than that of CICC 21004 (0.63 ± 0.35%).

[0110] Example 4: Effect of Plantarum LIHUO 05 on the symptoms of immunocompromised mice induced by cyclophosphamide I. Experimental steps (1) Preparation of Plantarum LIHUO 05 bacterial suspension The bacterial strain of Plantarum LIHUO 05 was inoculated into MRS liquid medium, and incubated at 37°C anaerobically for 18 hours. After 3 activations, the bacterial strain was centrifuged to discard the supernatant, and the bacterial body was washed twice with PBS solution. The bacterial body was resuspended with an appropriate amount of PBS solution and adjusted to a concentration of 1.0 x 10 9 CFU / mL to prepare the bacterial suspension.

[0111] (2) Construction of immunocompromised mouse model and intervention of bacterial strain 5-6 week old male Balb / c mice (Changsha Tianqin Biotechnology Co., Ltd.) weighing 18-22 g were adaptively fed for one week, and then randomly divided into 4 groups, 10 in each group, which were blank group, model group, LIHUO 05 group, and CICC group, respectively. Except for the blank group, the other groups were subcutaneously injected with 50 mg / Kg BW / d of cyclophosphamide (McLin, C16482048) to establish an immunocompromised mouse model. After continuous gavage for 2 weeks, the blank group and the model group were given sterile PBS, the LIHUO 05 group was given LIHUO 05 bacterial suspension, and the CICC group was given CICC 21004 bacterial suspension. All samples were gavaged at a dose of 200 μL, and water and feed were supplemented regularly every day. After gavage was completed, the animals were observed, and the animal state was recorded in a timely manner. The bedding was changed twice a week.

[0112] (3) Evaluation of the final body weight and thymus index of mice After the last gavage, the mice were fasted for 12 h without water, their body weight was measured, and they were euthanized using isoflurane anesthesia. After the eyeball of all mice was removed to collect blood samples, the mouse abdomen was disinfected with 75% ethanol solution, the chest was incised along the midline incision using standard dissecting scissors, the thymus was removed and weighed, and the thymus index was calculated according to the following formula: Thymus index = thymus weight / body weight x 100%.

[0113] (4) Detection of serum inflammatory cytokine TNF-α After the mouse blood sample was allowed to stand and stratify, serum was obtained by centrifugation at 4°C and 3000 r / min for 15 min. The concentration of TNF-α in the serum of each group of mice was determined using a TNF-α (Elabscience Company, item number E-EL-M3063) enzyme-linked immunoassay kit.

[0114] The experimental results are as follows: Due to the intervention of cyclophosphamide, the body weight of the model group (22.94 ± 0.84 g) was significantly decreased by 9.54% compared to the normal group (25.36 ± 1.31 g), and the body weight of the LIHUO 05 group (24.49 ± 1.15 g) was increased by 6.76% compared to the model group, which was better than the CICC group (23.50 ± 1.11 g), indicating that the intervention of plant Lactobacillus plantarum LIHUO 05 alleviated the decrease in body weight of immunocompromised mice induced by cyclophosphamide to a certain extent Figure 5 ).

[0115] Due to the fact that cyclophosphamide can cause atrophy of the thymus in mice, the thymus index of the model group (0.087 ± 0.013 %) was decreased by 27.5% compared to the normal group (0.12 ± 0.024 %), and the thymus index of the LIHUO 05 group (0.12 ± 0.032 %) was increased by 41.25% compared to the model group, which was better than the CICC group (0.11 ± 0.018 %), indicating that the intervention of plant Lactobacillus plantarum LIHUO 05 alleviated the shrinkage of the thymus of immunocompromised mice induced by cyclophosphamide to a certain extent Figure 6 ).

[0116] The content of serum TNF-α in the mice decreased due to cyclophosphamide, the thymus index of the model group (26.20 ± 3.51 pg / mL) decreased by 25.52% compared with the normal group (35.18 ± 2.47 pg / mL), the LIHUO 05 group (32.67 ± 4.15 pg / mL) increased by 24.69% compared with the model group, and the effect was better than that of the CICC group (30.97 ± 3.67 pg / mL), indicating that the intervention of the plant Lactobacillus LIHUO 05 alleviated the decrease of the content of serum TNF-α in the cyclophosphamide-induced immunocompromised mice to a certain extent Figure 7 ).

[0117] Example 5 Effect of plant Lactobacillus LIHUO 05 on the colon intestinal flora of cyclophosphamide-induced immunocompromised mice The experimental steps are as follows: The grouping, modeling and treatment of the mice were the same as in Example 4, and the colon content samples of the mice were collected under sterile conditions.

[0118] Six mice in each group were randomly selected for colon content to be sent to Eurobiolab Technology Co., Ltd. for microbial metagenome detection.

[0119] The experimental steps are as follows: The DNA extraction kit (QIAamp® Fast DNA Stool Mini Kit, (Qiagen, Hilden, Germany)) was used for total microbial DNA extraction of the samples. NanoDrop2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and agarose gel electrophoresis were used for DNA concentration determination and integrity analysis. S220 focused ultrasonic instrument (Focused-ultrasonicators) (Covaris, USA) was used for DNA fragmentation treatment, and Agencourt AMPureXP beads (Beckman Coulter Co., USA) was used for purification. TruSeq Nano DNA LT Sample Prepararion Kit (Illumina, San Diego, CA, USA) kit was used for library construction. Library construction, sequencing and data analysis were completed by Shanghai Eurobiolab Technology Co., Ltd.

[0120] The experimental results are as follows: Intestinal flora can reflect the degree of body health to some extent. The present application analyzes the structure of mouse colon microbial population at the door level and genus level, finds that the microbial structure and relative abundance of different groups are different, such as Figure 8 and Figure 9 Compared with the model group, the LIHUO 05 group can reduce the relative abundance of Bacteroidota, Campylobacterota and Verrucomicrobiota at the door level, and increase the relative abundance of Bacillota_A and Bacillota. At the same time, it is found that LIHUO 05 can further down-regulate the relative abundance of CAG-485, Alistipes and Duncaniella at the genus level, and increase the relative abundance of Alloprevotella and UBA3263, among which the regulation of beneficial bacteria Alistipes and Alloprevotella is significant, and the effect is better than CICC 21004, as shown in Figure 10 .

[0121] Example 6 Effect of plant lactobacillus LIHUO 05 on colon metabolites of immunocompromised mice induced by cyclophosphamide The experimental steps are as follows: The grouping, modeling and treatment of mice are the same as in Example 4, and the colon content samples of mice are collected under sterile conditions.

[0122] 6 mice in each group were randomly selected for colon content to be detected by Eurobio Technology Co., Ltd.

[0123] The experimental steps are as follows: weigh 20 mg of colon content and put it into a 1.5 mL EP tube; add two small steel balls and 300 μL of methanol-water (V:V=4:1, containing mixed internal standard); pre-cool in a-40℃ refrigerator for 2 min, then put into a grinder (45 Hz, 2 min); ultrasonic extraction in ice water bath for 10 min, overnight at-40℃; centrifuge for 20 min (12000 rpm, 4℃), take 150 μL of supernatant and load into a LC-MS sample vial with a foot lining tube for analysis; the quality control sample is prepared by mixing all sample extracts in equal volume. The analysis instrument is liquid chromatography-mass spectrometry Waters ACQUITY UPLC I-Class plus / Thermo QE HF. The data analysis part is completed by Shanghai Eurobio Biomedical Technology Co., Ltd.

[0124] The experimental results are as follows: Compared with the blank group and the model group, the LIHUO 05 group can change the level of colon metabolites, form a difference with the metabolic structure of the two groups, as shown in Figure 11As shown. And by analyzing the KEGG enrichment bar chart of differential metabolites ( Figure 12 and Figure 13 Compared to the model group, LIHUO 05 decreased nearly 17 products in 8 metabolism-related pathways, while increasing nearly 39 products in 15 other pathways; decreased 5 metabolites in 5 biological system-related pathways, while increasing 4 metabolites in 3 other pathways; decreased 3 metabolites in 2 environmental information processing pathways, while increasing 4 metabolites in another pathway; decreased 2 metabolites in 1 human disease-related pathway, while increasing 2 metabolites in another pathway; and decreased 4 metabolites in 4 cellular process-related pathways. LIHUO 05 significantly increased the abundance of immune-related metabolites Soyasaponin V and Soyasaponin IV, and decreased the abundance of Murideoxycholic acid and Nylidrin, such as... Figure 14 and Figure 15 As shown.

[0125] Example 7: Genome sequencing and functional annotation analysis of *Lactobacillus plantarum* LIHUO 05 (1) Whole genome sequencing of Lactobacillus plantarum LIHUO 05 The complete genome of *Lactiplantibacillus plantarum* LIHUO 05 was obtained through sequencing and genome assembly using Nanopore third-generation sequencing and second-generation sequencing platforms. Figure 16 This is a visualized genome map of the bacterium, which has a genome size of 3,164,934 bp and a GC ratio of 45.16%.

[0126] (2) Annotation of the whole genome of Lactobacillus plantarum LIHUO 05 in the Gene Ontology database The highest-level functional nodes in the GO database include three: (1) Cellular Component: used to describe subcellular structures, locations, and macromolecular complexes; (2) Molecular Function: used to describe the function of genes and individual gene products; and (3) Biological Process: used to describe the biological processes in which gene-encoded products participate.

[0127] In this invention, during the specific annotation process, the software Blast2GO v2.5 is used to identify these corresponding nodes. The annotation information corresponding to these nodes is the annotation information of the genes in the sequenced genome, and the results are as follows: Figure 17 As shown in the figure. The horizontal axis represents the contents of each GO category, and the vertical axis represents the number of genes. Figure 18The display is the gene enrichment of each secondary function of GO in the whole genetic background, which reflects the position of each secondary function in the background.

[0128] (3) Lactobacillus plantarum LIHUO 05 whole genome KEGG (Kyoto Encyclopedia of Genes and Genomes) database annotation KEGG is a comprehensive database that collects genome, pathway and compound information of organisms.

[0129] In the specific annotation of the application, by BLAST comparison of the protein sequence of the gene and the protein sequence included in the KEGG database, the most similar sequence in the KEGG database is found, and the KEGG annotation of the gene is performed, and then classified according to the KEGG metabolic pathways they participate in, and the results are shown in Figure 18 The horizontal coordinate in the figure is the number of genes annotated under the Pathway classification; the vertical coordinate is the Pathway classification, and different colors represent different large classifications.

[0130] (4) Lactobacillus plantarum LIHUO 05 whole genome eggNOG database annotation EggNOG is a database that collects orthologous gene clusters of organisms, and is continuously updated on the basis of the COG (Cluster of Orthologous Groups of proteins) database.

[0131] In the specific annotation of the application, by BLAST comparison of the protein sequence of the gene and the protein sequence included in the KEGG database, the most similar sequence in the KEGG database is found, and the KEGG annotation of the gene is performed, and then classified according to the KEGG metabolic pathways they participate in, and the results are shown in Figure 19 .

[0132] (5) Lactobacillus plantarum LIHUO 05 whole genome CAZy database annotation After the whole genome of Lactobacillus plantarum LIHUO 05 described in the application is annotated by the special database, it is found that the annotation of the carbohydrate active enzyme in the CAZy database is shown in Table 1.

[0133] Table 1 CAZy annotation

[0134] The above only describes the preferred embodiments of the application and should not be used to limit the application, and any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the application should be included in the protection scope of the application.

Claims

1. A Lactobacillus plantarum (L. Lactiplantibacillus plantarum ) LIHUO 05, characterized in that, The plant lactiplantibacillus LIHUO 05 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 36240.

2. The Lactiplantibacillus halli LIHUO 05 according to claim 1, characterized in that, The 16s rDNA sequence of the plant lactiplantibacillus LIHUO 05 is as shown in SEQ ID NO: 1-2.

3. A preparation comprising the plant lactiplantibacillus LIHUO 05 of any one of claims 1-2.

4. The preparation according to claim 3, characterized in that, The preparation includes a fermentation broth of the plant lactiplantibacillus LIHUO 05, a supernatant of the fermentation broth of the plant lactiplantibacillus LIHUO 05, a precipitate of the fermentation broth of the plant lactiplantibacillus LIHUO 05, live bacteria of the plant lactiplantibacillus LIHUO 05, and dead bacteria of the plant lactiplantibacillus LIHUO 05.

5. The preparation according to claim 4, characterized in that, The dead bacteria of the plant lactiplantibacillus LIHUO 05 are heat-inactivated plant lactiplantibacillus LIHUO 05 bacterial suspension.

6. The preparation according to claim 5, characterized in that, The preparation of the heat-inactivated plant lactiplantibacillus LIHUO 05 bacterial suspension includes the following steps: (1) inoculating the activated plant lactiplantibacillus LIHUO 05 activation liquid into MRS liquid medium for culture to obtain a bacterial liquid; (2) centrifuging the bacterial liquid to obtain plant lactiplantibacillus bacteria; (3) resuspending the plant lactiplantibacillus bacteria in PBS to obtain a bacterial suspension; (4) inactivating the bacterial suspension at 80-100℃ for 15-30min to obtain the heat-inactivated plant lactiplantibacillus LIHUO 05 bacterial suspension.

7. An inoculant characterized in that, The bacterial agent includes the plant lactiplantibacillus LIHUO 05 of any one of claims 1-2 or the preparation of any one of claims 3-6.

8. Use of the plant lactiplantibacillus LIHUO 05 of any one of claims 1-2 or the preparation of any one of claims 3-6 or the bacterial agent of claim 7 in the preparation of a product for enhancing immunity.

9. Use according to claim 8, characterized in that, The product contains 1×10 5 -1×10 12 CFU / mL or 1×10 5 -1×10 12 CFU / g of live bacteria of Lactobacillus plantarum LIHUO 05.

10. Use according to claim 8, characterized in that, The product includes a medicine and a health food.

11. Use according to claim 8, characterized in that, The product has at least one of the following effects: (1) promoting the secretion of TNF-α and IL-1β in macrophages; (2) promoting the proliferation of lymphocytes; (3) increasing the body weight, thymus index, and TNF-α content in serum of an immunocompromised subject; (4) changing the intestinal colon flora structure of an immunocompromised subject and increasing the relative abundance of Alistipes and Alloprevotella; (5) regulating the types and amounts of intestinal colon metabolites in an immunocompromised subject, increasing the abundance of Soyasaponin V and Soyasaponin IV, and reducing the abundance of Murideoxycholic acid and Nylidrin.

12. The use according to claim 10, characterized in that, When the product is a medicine, the medicine includes a pharmaceutically acceptable carrier.

13. Use of the plant lactiplantibacillus LIHUO 05 of any one of claims 1-2 or the preparation of any one of claims 3-6 or the bacterial agent of claim 7 in the preparation of a product for regulating the intestinal flora environment.

14. Use according to claim 13, characterized in that, The product contains 1×10 5 -1×10 12 CFU / mL or 1×10 5 -1×10 12 CFU / g of live Lactobacillus plantarum LIHUO05.

15. The use according to claim 13, characterized in that, The product has at least one of the following effects: (1) At the door level, the relative abundance of Bacteroidota, Campylobacterota, Verrucomicrobiota is reduced, and the relative abundance of Bacillota_A, Bacillota is increased; (2) At the genus level, the relative abundance of CAG-485, Alistipes, Duncaniella is down-regulated, and the relative abundance of Alloprevotella, UBA3263 is increased.

16. A product for enhancing immunity or modulating gut flora, characterized in that, The product contains the Lactobacillus plantarum LIHUO 05 of any one of claims 1-2 or the preparation of any one of claims 3-6 or the microbial inoculant of claim 7.

17. The product of claim 16, wherein, The product contains 1×10 5 -1×10 12 CFU / mL or 1×10 5 -1×10 12 CFU / g.

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