CN113046274A

CN113046274A - Lactobacillus reuteri for relieving gastrointestinal injury caused by capsaicin and application thereof

Info

Publication number: CN113046274A
Application number: CN202110453834.8A
Authority: CN
Inventors: 崔树茂; 毛丙永; 项群然; 唐鑫; 翟齐啸; 刘小鸣; 赵建新; 陈卫
Original assignee: Jiangnan University
Current assignee: Jiangnan University
Priority date: 2021-04-26
Filing date: 2021-04-26
Publication date: 2021-06-29
Anticipated expiration: 2041-04-26
Also published as: CN113046274B

Abstract

The invention discloses lactobacillus reuteri for relieving gastrointestinal injury caused by capsaicin and application thereof, and belongs to the technical field of microorganisms. The invention provides lactobacillus reuteri CCFM1175, a composition, fermented food, application and a preparation method of a microbial inoculum thereof. The lactobacillus reuteri CCFM1175 can obviously improve the mental state of a mouse after the gavage of capsaicin, improve the activity of glutathione peroxidase and catalase in the liver of the mouse after the gavage of capsaicin and the content of glutathione, reduce the content of malondialdehyde, reduce the levels of P substances and calcitonin gene-related peptide in the serum of the mouse after the gavage of capsaicin, and improve the tissue damage such as the shedding of intestinal villi, the infiltration of colon inflammatory cells and the like caused by the capsaicin. The lactobacillus reuteri CCFM1175 is used for preparing a medicinal composition and a fermented food for relieving the piquancy and protecting the intestines and stomach, and has very wide application prospect.

Description

Lactobacillus reuteri for relieving gastrointestinal injury caused by capsaicin and application thereof

Technical Field

The invention relates to lactobacillus reuteri for relieving gastrointestinal injury caused by capsaicin and application thereof, and belongs to the technical field of microorganisms.

Background

Capsaicin is the main component of pepper, has the functional activities of analgesia, antioxidation, anti-obesity, bacteriostasis and the like, and is widely applied to diet and pharmacology. However, capsaicin is highly irritant, so that bad perceptions such as burning sensation, heartburn, abdominal pain and the like are often generated after the capsicum is eaten in daily life, and the main mechanism is that the capsaicin is combined with TRPV1 receptor to induce the transmission of pain mediators such as neuropeptide and the release of downstream proinflammatory factors, thereby causing gastrointestinal damage.

Common substances for relieving the piquancy in life comprise vinegar, milk and the like, wherein acetic acid in the vinegar can generate acid-base neutralization reaction with capsaicin, and fat-soluble substances in the milk wrap the capsaicin to enter the digestive tract from the oral cavity, so that the sense organ stimulation of the piquancy is relieved. However, the above mechanism of relieving the piquancy is based on the fat-soluble property of capsaicin, and the gastrointestinal tract injury caused by the capsaicin cannot be relieved.

In view of various problems of the existing method for relieving the piquancy, a new scheme for relieving the piquancy and protecting the intestines and stomach instead of the traditional food is particularly important. The research shows that part of probiotics can interact with capsaicin receptor TRPV1, but the relation of the probiotics and capsaicin induced gastrointestinal damage is not clear. Therefore, probiotic preparations or fermentation products for relieving gastrointestinal injury caused by capsaicin are also lacking in the market, and probiotics with the function of relieving the peppery taste are found to promote the matching application of the probiotics in daily peppery people.

Disclosure of Invention

The invention aims to provide a Lactobacillus reuteri (Lactobacillus reuteri) CCFM1175 which is preserved in Guangdong province microorganism strain preservation center at 19 months 3 in 2021, wherein the preservation number is GDMCC No: 61572.

in one embodiment, the lactobacillus reuteri CCFM1175 has the following properties:

(1) the microbial morphological characteristics are as follows: the cells are slightly irregular, single or short-chained, without flagella and spores.

(2) Culture properties: the colony cultured on MRS culture medium for 48 hr is milky white, smooth and raised, milky white and 1-2mm in diameter.

The invention also provides a composition containing the lactobacillus reuteri.

In one embodiment, the composition contains Lactobacillus reuteri in an amount of 1 × 10 or more⁵CFU/g or 1X 10⁵CFU/mL。

In one embodiment, the composition is a fermentation agent, the fermentation agent is a powder prepared by using a bacterial liquid containing the lactobacillus reuteri, and the powder contains 10¹¹More than CFU/g of viable Lactobacillus reuteri.

In one embodiment, the powder is prepared by conventional freeze-drying process or other processes of the bacterial liquid containing the active lactobacillus reuteri.

In one embodiment, the components of the lyoprotectant are 80-120 g/L skimmed milk powder, 80-140 g/L trehalose, 140-180g/L sucrose and water.

In one embodiment, the starter is prepared by the following steps:

(1) preparation of a culture medium: 10g/L of peptone, 10g/L of beef extract, 20g/L of glucose, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate₂PO₄·3H₂O 2.6g/L、MgSO₄·7H₂O 0.1g/L、MnSO₄0.05g/L, Tween 801 mL/L, cysteine hydrochloride 0.5g/L, and pH 6.8.

(2) Preparation of the protective agent: mixing water and protective agent raw materials to prepare a protective agent containing 130g/L of skimmed milk powder, 20g/L of sucrose and 20g/L of trehalose;

(3) inoculating lactobacillus reuteri CCFM1175 into the culture medium in the step (1), culturing at 37 ℃ for 18h, centrifugally collecting thalli, washing the thalli with phosphate buffer solution with pH 7.2 for 2-4 times, and re-suspending with the protective agent in the step (2) until the concentration reaches 10¹⁰And (5) performing freeze drying on the CFU/mL to obtain the leavening agent.

In one embodiment, the components of the lyoprotectant include skim milk powder, trehalose, sucrose, and water.

In one embodiment, the addition amount of the lyoprotectant in the resuspension solution is 2-4 times of the total weight of the bacterial sludge.

The second purpose of the invention is to provide the application of the lactobacillus reuteri in preparing medicines for relieving capsaicin-induced gastrointestinal injury.

In one embodiment, the application includes at least one of the following functions:

(1) improving mental state in a mammal;

(2) improving glutathione peroxidase activity, catalase activity and glutathione content of mammal liver, and reducing malondialdehyde content;

(3) reducing the level of substance P and calcitonin gene-related peptide in serum of mammals;

(4) can be used for treating mammal injuries such as villus desquamation, crypt hyperplasia, and colonic inflammatory cell infiltration.

In one embodiment, the product contains live lactobacillus reuteri CCFM1175 bacteria.

In one embodiment, the viable count of lactobacillus reuteri CCFM1175 in the product is not less than 1 × 10⁸CFU/mL。

In one embodiment, the product is a pharmaceutical product comprising lactobacillus reuteri, a pharmaceutical carrier and/or a pharmaceutical excipient.

In one embodiment, the drug carrier comprises microcapsules, microspheres, nanoparticles, and liposomes.

In one embodiment, the pharmaceutical excipient comprises an excipient and an additive.

In one embodiment, the pharmaceutical excipient comprises an anti-adhesive agent, a penetration enhancer, a buffering agent, a plasticizer, a surfactant, an antifoaming agent, a thickener, an encapsulating agent, an absorbent, a humectant, a solvent, a propellant, a solubilizer, a cosolvent, an emulsifier, a colorant, a pH adjuster, a binder, a disintegrant, a filler, a lubricant, a wetting agent, an integrating agent, an osmotic pressure adjuster, a stabilizer, a glidant, a flavoring agent, a preservative, a foaming agent, a suspending agent, a coating material, a fragrance, a diluent, a flocculating agent and a deflocculating agent, a filter aid, and a release retardant.

In one embodiment, the additive comprises microcrystalline cellulose, hydroxypropyl methylcellulose, and refined lecithin.

In one embodiment, the pharmaceutical product is in a dosage form comprising granules, capsules, tablets, pills or oral liquid.

In one embodiment, the product comprises a food, pharmaceutical or nutraceutical.

In one embodiment, the food product comprises a dairy product, a soy product, or a fruit and vegetable product produced using a lactobacillus reuteri-containing starter.

Has the advantages that: the lactobacillus reuteri CCFM1175 provided by the invention can obviously improve the mental state of a mouse after capsaicin gavage; the activities of glutathione peroxidase and catalase of the mouse liver and the content of glutathione after the gastric capsaicin administration are improved, and the content of malondialdehyde is reduced; can reduce the level of P substance and calcitonin gene related peptide in the blood serum of mice after the gavage of capsaicin; can improve tissue injury caused by capsaicin, such as small intestine villus desquamation, colon inflammatory cell infiltration, etc. Can be used for preparing a medicinal composition, fermented food or fermented beverage for relieving the piquancy and protecting the intestines and stomach, and has very wide application prospect.

Biological material preservation

Lactobacillus reuteri (Lactobacillus reuteri) CCFM1175, classified and named as Lactobacillus reuteri, which is deposited in the Guangdong province collection of microorganisms and strains in 3 and 19 months 2021, with the deposit number being GDMCC No: 61572.

drawings

FIG. 1 is a diagram of the mental state of different groups of mice after intragastric administration.

FIG. 2 shows the glutathione peroxidase activities of the livers of different groups of mice.

FIG. 3 shows the catalase activity of the livers of different groups of mice.

FIG. 4 is the glutathione content of the liver of different groups of mice.

FIG. 5 is the malondialdehyde content of the liver of different groups of mice.

FIG. 6 shows the serum content of substance P in mice of different groups.

FIG. 7 shows the amounts of calcitonin gene-related peptide in serum of different groups of mice.

FIG. 8 is jejunal tissue morphology (HE staining) of different groups of mice.

Figure 9 is ileal histomorphology (HE staining) of different groups of mice.

FIG. 10 is colon histomorphology (HE staining) of different groups of mice.

Detailed Description

The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and biomaterials, if not specifically indicated, are commercially available.

The media involved in the following examples are as follows:

MRS solid medium (g/L): peptone 10g/L, beef extract 10g/L, grape20g/L of sugar, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate₂PO₄·3H₂O 2.6g/L、MgSO₄·7H₂O 0.1g/L、MnSO₄0.05g/L, Tween 801 mL/L, agar 20g/L, cysteine hydrochloride 0.5g/L, and pH 6.8.

MRS liquid medium (g/L): 10g/L of peptone, 10g/L of beef extract, 20g/L of glucose, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate₂PO₄·3H₂O 2.6g/L、MgSO₄·7H₂O 0.1g/L、MnSO₄0.05g/L, tween 801 mL/L, cysteine hydrochloride 0.5g/L, pH 6.8.

Example 1: screening and identification of lactobacillus reuteri

1. Screening

Taking human excrement from Henan Xiuyi as a sample, pretreating the sample, storing the sample in a refrigerator at minus 80 ℃ in 30% glycerol, taking out the sample for thawing, uniformly mixing the sample, sucking 0.5mL of the sample, adding 4.5mL of 0.9% physiological saline for gradient dilution, selecting a proper gradient diluent to coat the gradient diluent on an MRS solid culture medium, culturing for 48 hours at 37 ℃, selecting a typical bacterial colony to an MRS plate, streaking and purifying, selecting a single bacterial colony, transferring to a liquid MRS liquid culture medium for enrichment, and preserving by 30% glycerol to obtain a strain CCFM 1175.

2. Identification

Extracting the whole genome DNA of the strain CCFM1175 to carry out 16S rDNA amplification, collecting the amplified DNA fragment for sequencing, carrying out nucleotide sequence comparison and evolutionary tree analysis on the sequence in NCBI, and displaying that the strain is Lactobacillus reuteri and is named as Lactobacillus reuteri CCFM 1175.

Example 2: culture of Lactobacillus reuteri

Lactobacillus reuteri (Lactobacillus reuteri) is inoculated into an MRS solid culture medium and cultured for 48 hours at 37 ℃, the colony is observed and the thallus is observed under a microscope, and the colony is milky white, irregular, round and convex and smooth, the shape of the thallus is slightly irregular and round, and the campylobacter with round ends exists usually in single, paired and small clusters.

Inoculating Lactobacillus reuteri (Lactobacillus reuteri) into MRS liquid culture medium, culturing at 37 deg.C for 18 hr, transferring into fresh MRS liquid culture medium, culturing under the same conditions for 18 hr, centrifuging at 6000rpm for 8min, washing with 0.9% physiological saline, centrifuging at 6000rpm for 5min again to obtain thallus, and resuspending with 30% sucrose solution to obtain thallus with bacterial concentration of 10¹¹CFU/mL, and freezing at 80 ℃ for later use.

Example 3: effect of Lactobacillus reuteri on capsaicin induced gastrointestinal injury mouse mental status

30 SPF-grade 6-week-old male C57BL/6 mice were housed in a constant temperature and humidity animal house, strictly following the 12h day and 12h night cycle standard, fed on a standard commercial formula, and fed freely. After 7 days of adaptive feeding, the animals were randomly divided into 3 groups of 10 animals, 3 groups were: control, capsaicin, CCFM1175, FHNXY12L7, DSM 17938.

The expected intragastric administration dryness is 14 days on 8-21 days, the intragastric administration dosage is 0.2 mL/patient, and the intragastric administration time is kept consistent every day. Wherein the control group and capsaicin group are perfused with normal saline, and the CCFM1175 group is perfused with Lactobacillus reuteri CCFM1175 (10)⁹CFU/CFU), FHNXY12L7 group lactobacillus gasseri FHNXY12L7 (10)⁹CFU/only), DSM17938 group lactobacillus reuteri DSM17938 (10)⁹CFU/r), wherein FHNXY12L7 is another strain of lactobacillus reuteri that was screened in the same batch as CCFM 1175. DSM17938 is a widely used commercial strain of Lactobacillus reuteri, which has been disclosed in the patent application with publication number "CN 104244736A", entitled "Lactobacillus reuteri DSM17938 for the development of the enteric nervous system".

Capsaicin molding is performed on the last 7 days, i.e., 15-21 days, of the intervention period. After two hours of gavage with physiological saline/bacterial suspension, the capsaicin group, CCFM1175 group, FHNXY12L7 group, DSM17938 group were gavaged with 60mg/kg bw capsaicin solution (solvent: 3% ethanol + 10% Tween 80+ 87% physiological saline, v/v), and the control group was gavaged with the solvent of capsaicin solution.

During the gavage period, the mental state of the mice after gavage was recorded as shown in fig. 1. Through observation of the mental state of the mice, compared with the control group of mice, the capsaicin group of mice shows the phenomena of digging pits and burying heads after gastric administration of capsaicin, shows the lying state after 2-3 minutes, has tachypnea and even shows the phenomenon of death of part of mice, and is presumably related to the sensory irritation of capsaicin to the oral cavity. The FHNXY12L7 group and DSM17938 group mice are prone after gavage and have rapid respiration, but the CCFM1175 group mice do not have the above state in the gavage process, and the strain is supposed to possibly relieve the strong irritation of capsaicin to the digestive tract, and the relieving effect is better than that of the Lactobacillus reuteri FHNXY12L7 and DSM 17938.

Example 4: influence of lactobacillus reuteri on liver oxidative stress of capsaicin-induced gastrointestinal injury mice

In vivo, there is a defense mechanism against oxidative stress injury, and when the organism is stimulated, the expression of downstream antioxidant enzymes such as glutathione peroxidase (GSH-Px), Catalase (CAT) and the like can be regulated, so as to inhibit inflammation and apoptosis reaction and play a role in protecting cells. Glutathione (GSH) is the most important non-enzymatic antioxidant in the body, and the decrease of this substance leads to impairment of mitochondrial function, further leading to excessive increase of active oxygen radicals, thereby causing oxidative damage. While MDA is considered to be an important product and indicator of lipid peroxidation in the body, changes in Malondialdehyde (MDA) content reflect the degree of oxidative damage to the body.

The animal model was established as in example 3, after fasting for 12 hours, the gavage-terminated mice were anesthetized and the eyeballs were quickly removed to collect blood, followed by cervical dislocation for sacrifice. Dissecting the sacrificed mouse, cutting a small opening on the left side of the abdominal midline by scissors, advancing to the sternal process, and making transverse incisions along the back edge of the rib to two sides to expose the abdominal cavity. Observing the normal position of organs in the abdominal cavity, picking the liver, adding precooled physiological saline with the weight 9 times that of the tissue block, fully grinding to homogenize the tissue, centrifuging at 6000rpm at 4 ℃ for 15 minutes, collecting supernatant to obtain tissue homogenate, and measuring the content of GSH-Px, CAT, GSH and MDA in the liver homogenate of each group of mice by adopting a Nanjing constructed kit.

The results of measuring the content of GSH-Px, CAT, GSH and MDA in liver homogenates of each group of mice are shown in figures 2-5.

As shown in fig. 2, the liver GSH-Px activity of the mice in the capsaicin group was significantly reduced by 22.5% compared with the control group, while the CCFM1175 group, FHNXY12L7 group and DSM17938 group of the mice in the lactobacillus reuteri intervention group were reduced by 5.2%, 19.8% and 17.3%, respectively. The capsaicin lavage shows that the activity of the antioxidant enzyme GSH-Px of the mouse liver is reduced, the lactobacillus reuteri CCFM1175 can effectively improve the injury, and the lactobacillus reuteri FHNXY12L7 and DSM17938 have no obvious improvement effect. As shown in fig. 3, the liver CAT activity of each group of mice also showed similar results, the CAT activity of the capsaicin group mice was reduced by 32.5% compared with the control group, the CAT activity of the CCFM1175 group mice was not significantly different from that of the control group, and the CAT activity of the FHXY12L7 group and DSM17938 group mice was reduced by 21.4% and 24.9% compared with the control group, respectively.

As a result of comparison of glutathione contents in mouse livers (FIG. 4), the glutathione contents in the mouse livers of the capsaicin group, the CCFM1175 group, the FHNXY12L7 group and the DSM17938 group were reduced by 64.0%, 14.8%, 50.2% and 33.2%, respectively, as compared with those in the control group (8.45. + -. 1.52. mu. mol/mg protein). The content of the oxidative damage marker MDA in each treatment group is respectively as follows: control group 0.44 ± 0.07nmol/mg protein, capsaicin group 0.67 ± 0.10nmol/mg protein, CCFM1175 group 0.47 ± 0.10nmol/mg protein, FHNXY12L7 group 0.79 ± 0.07nmol/mg protein, DSM17938 group 0.51 ± 0.06nmol/mg protein. After the lactobacillus reuteri CCFM1175 and the lactobacillus reuteri DSM17938 are perfused, the MDA content of the liver of the mouse has no significant difference with that of a control group, and the MDA content of the mouse in a capsaicin group and a lactobacillus reuteri FHNXY12L7 group is significantly increased (figure 5).

The results show that the lactobacillus reuteri CCFM1175 can effectively relieve the liver oxidative damage caused by capsaicin, specifically recover the activity of GSH-PX and CAT and the level of GSH, effectively reduce the MDA content in the tissues, and the relieving effect is obviously higher than that of the lactobacillus reuteri FHNXY12L7 and the lactobacillus reuteri DSM 17938.

Example 5: effect of Lactobacillus reuteri on capsaicin induced gastrointestinal injury mouse serum neuropeptide

Capsaicin receptor TRPV1 in the gastrointestinal tract mediates crosstalk between the nervous system and the immune system by modulating the release of neuropeptides, Substance P (SP), calcitonin gene-related peptide (CGRP), two neuropeptides involved in nociceptive transmission and play an important role in pain and inflammatory responses.

The animal model was constructed as in example 3, except that after fasting for 12 hours, the gavage-terminated mice were anesthetized and the eyes were quickly removed to bleed blood, followed by cervical dislocation to be sacrificed. Collecting blood samples of the mice after sacrifice, standing the blood samples at room temperature for 3h, centrifuging at 4000rpm for 15min, carefully collecting upper serum, and measuring the contents of SP and CGRP in the mouse serum according to the instructions of the Shanghai enzyme-linked reagent kit.

As shown in FIGS. 6 and 7, the contents of neuropeptide SP and CGRP in the serum of the mice in the capsaicin group are respectively increased by 35.8% and 47.4% compared with the control group, which shows that the mice can promote the release of neuropeptide substances after being stimulated by capsaicin, thereby increasing the pain transmission. After the lactobacillus reuteri CCFM1175 is perfused, compared with the mice in the capsaicin group, the levels of SP and CGRP in the serum are reduced, and are only increased by 9.7 percent and 5.2 percent compared with the control group, which shows that the lactobacillus reuteri CCFM1175 can relieve the visceral hyperalgesia caused by capsaicin. The serum SP levels of mice in FHNXY12L7 group and DSM17938 group were increased by 37.4% and 22.2% respectively compared with the control group, and the CGRP levels were increased by 38.4% and 23.9% respectively compared with the control group, which indicates that the two strains of Lactobacillus reuteri could not relieve the visceral hyperalgesia caused by capsaicin.

Example 6: effect of Lactobacillus reuteri on gastrointestinal tissue morphology of mice with capsaicin-induced gastrointestinal injury

The animal model was constructed as in example 3, except that after fasting for 12 hours, the gavage-terminated mice were anesthetized and the eyes were quickly removed to bleed blood, followed by cervical dislocation to be sacrificed. Dissecting the sacrificed mouse, cutting a small opening on the left side of the abdominal midline by scissors, advancing to the sternal process, and making transverse incisions along the back edge of the rib to two sides to expose the abdominal cavity. Observing the normal position of organs in the abdominal cavity, picking the jejunum, the ileum and the colon, slightly rinsing with normal saline, and fixing in 4% paraformaldehyde fixing solution. After fixation for 48h, paraffin embedding and slicing are carried out, and the morphology of jejunum, ileum and colon tissues is observed by hematoxylin-eosin (HE) staining.

As shown in FIG. 8, the jejunum villi structure of the control group was intact and well-arranged. The villi of the capsaicin group, FHNXY12L7 group and DSM17938 group were irregularly arranged, and were accompanied by local villi breakage and shedding, and crypt proliferation appeared, but there was no massive inflammatory cell infiltration. The structure and the shape of jejunum villi of the lactobacillus reuteri CCFM1175 group are recovered to a certain degree.

As shown in FIG. 9, the ileal villi of the capsaicin group had irregular arrangement, a large amount of villi were broken and shed, and crypt hyperplasia appeared, while those of FHNXY12L7 and DSM17938 group had shed, but had no crypt hyperplasia. The villus structure of ileum of the CCFM1175 group is complete and similar to that of the control group.

As shown in FIG. 10, the control mice had intact colon structure, aligned mucosa and villi, healthy crypt structure, abundant goblet cells, and no inflammatory cell infiltration or mucosal damage. The mice of the capsaicin group and the FHNXY12L7 group have a small amount of inflammatory cell infiltration, the caliform cells of the intestinal tissues of the mice of the DSM17938 group are greatly reduced, and the colon tissue cells of the mice of the CCFM1175 group have similar forms to those of the control group, are rich in the caliform cells and have no inflammatory cell infiltration and mucosal injury.

By combining the results of the tissue morphology of the jejunum, ileum and colon of mice in different treatment groups, the inventor finds that the ingestion of 60mg/kg/day of capsaicin can cause the villi of the jejunum and ileum of the mice to fall off, the crypt proliferation phenomenon occurs, meanwhile, the colon has inflammatory cell infiltration and goblet cell reduction, and the gastrointestinal injury of the mice is caused to a certain extent. Lactobacillus reuteri CCFM1175 significantly ameliorates this damage to a greater extent than Lactobacillus reuteri FHNXY12L7 and Lactobacillus reuteri DSM 17938.

Example 7: use of Lactobacillus reuteri CCFM1175 in preparation of instant lactobacillus powder

Preparation of a culture medium: 10g/L of peptone, 10g/L of beef extract, 20g/L of glucose, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate₂PO₄·3H₂O 2.6g/L、MgSO₄·7H₂O 0.1g/L、MnSO₄0.05g/L, tween 801 mL/L, cysteine hydrochloride 0.5g/L, pH 6.8.

Preparation of the protective agent: preparing a protective agent solution containing 130g/L of skimmed milk powder, 20g/L of sucrose and 20g/L of trehalose.

The specific preparation process of the fungus powder is as follows:

inoculating lactobacillus reuteri CCFM1175 into the culture medium according to the inoculation amount (by volume) of 2%, then culturing for 18h at the temperature of 37 ℃, washing for 2-4 times by phosphate buffer solution with pH 7.2, and re-suspending by protective agent to reach the concentration of 10¹⁰And (5) performing freeze drying on the CFU/mL to obtain the instant lactic acid bacteria powder. The powder can also be used as leaven.

Optionally, in the preparation process of the bacterial powder, the bacterial powder can be mixed with other species and genera of lactic acid bacteria to prepare a mixed bacterial liquid, and then the mixed bacterial liquid is subjected to freeze drying.

Example 8: lactobacillus reuteri CCFM1175 for preparing fermented milk

The specific preparation process of the fermented milk comprises the following steps:

heat sterilizing cow milk or reconstituted milk at 95 deg.C for 20min, cooling to 37 deg.C, adding Lactobacillus reuteri CCFM1175 or the starter containing Lactobacillus reuteri CCFM1175 prepared in example 7 to make the thallus concentration reach 10⁷More than CFU/mL, and adding appropriate amount of commercial leaven (containing Streptococcus thermophilus and Lactobacillus bulgaricus) to make total viable count after inoculation reach 10⁴～10⁵CFU/mL, wherein the number ratio of streptococcus thermophilus to lactobacillus bulgaricus in the commercial starter is 1: (1-2). Fermenting the inoculated cow milk or reconstituted milk at 37 ℃ for 24h, and refrigerating at 4 ℃ for 12h to obtain the fermented milk containing the live lactobacillus reuteri.

Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. Lactobacillus reuteri (Lactobacillus reuteri) CCFM1175, which has been deposited with the Guangdong province culture Collection in 19 months 3 in 2021 with the deposit number GDMCC No: 61572.

2. a composition comprising lactobacillus reuteri CCFM1175 of claim 1.

3. The composition according to claim 2, wherein the content of Lactobacillus reuteri in the composition is not less than 1 x 10⁵CFU/g or 1X 10⁵CFU/mL。

4. A starter culture comprising the Lactobacillus reuteri CCFM1175 of claim 1, wherein the starter culture comprises 10 or more¹¹CFU/g live Lactobacillus reuteri CCFM 1175.

5. The fermentation agent according to claim 4, wherein the fermentation agent is prepared by freeze-drying a bacterial liquid containing Lactobacillus reuteri CCFM 1175.

6. A fermented product characterized in that the fermented food is a fermented food or a fermented drink produced by using the Lactobacillus reuteri according to claim 1 or the fermentation agent according to any one of claims 4 to 5.

7. Use of lactobacillus reuteri CCFM1175 according to claim 1 for the preparation of a medicament for the prevention and/or alleviation of capsaicin-induced gastrointestinal damage.

8. Use according to claim 7, characterized in that it comprises at least one of the following functions:

(1) improving mental state in a mammal;

(2) improving GSH-PX, CAT activity and GSH content of mammal liver, and reducing MDA content;

(3) reducing the levels of SP, CGRP in the serum of a mammal;

(4) improve mammal empty, ileum villus shed, crypt hyperplasia, colon inflammatory cell infiltration injury.

9. Use according to claim 7 or 8, wherein the product is a pharmaceutical product; the medicine contains lactobacillus reuteri CCFM1175, a medicine carrier and/or a pharmaceutic adjuvant.

10. The use according to claim 7 or 8, wherein the product is a dairy product, a soy product or a fruit and vegetable product containing Lactobacillus reuteri CCFM 1175.