Lactobacillus brevis for dispelling effects of alcohol and protecting liver and application of lactobacillus brevis

Abstract

The invention provides lactobacillus breviscapus for dispelling effects of alcohol and protecting liver and application thereof, and relates to the technical field of biological fermentation, the lactobacillus breviscapus is lactobacillus breviscapus, the strain number of the lactobacillus breviscapus is KMHD10-222, the registration number of the lactobacillus breviscapus in the China General Microbiological Culture Collection Center is CGMCC No.32358, and the number of the lactobacillus breviscapus in the China General Microbiological Culture Collection Center is KMHD10-222. The degradation rate of the lactobacillus brevicornu KMHD10-222 to ethanol with the concentration of 3% reaches 36.05%, the degradation rate of the lactobacillus brevicornu KMHD10-222 to ethanol with the concentration of 7% reaches 19.74%, and the lactobacillus brevicornu KMHD10-222 can degrade ethanol; the lactobacillus brevis KMHD10-222 microbial inoculum has the effect of relieving the acute alcoholic liver injury. The technical problem that in the prior art, probiotics capable of degrading ethyl alcohol and having the effects of dispelling the effects of alcohol and protecting the liver are lacked is solved.

Description

Technical Field

[0001] This invention relates to the field of bio-fermentation technology, and in particular to a type of Lactobacillus curvilinearis that can relieve hangovers and protect the liver, and its applications. Background Technology

[0002] Moderate consumption of alcoholic beverages can promote blood circulation, clear the meridians, and relieve fatigue. Drinking some fruit wines and medicinal wines can also have certain auxiliary therapeutic effects. However, excessive consumption can seriously harm health. When alcohol is consumed in excess, the body's metabolic burden increases, and the liver is unable to metabolize most of the alcohol, leading to damage to the liver from ethanol and its harmful metabolites. Therefore, long-term heavy drinking or consuming excessive amounts of alcohol at once can cause alcoholic liver disease.

[0003] Currently, many commercially available hangover remedies and liver-protecting medications suffer from issues such as unclear ingredients, slow effectiveness, and significant side effects. Probiotics, on the other hand, are characterized by high safety, strong functionality, and no obvious side effects. Studies have shown that probiotics can improve alcoholic liver damage by regulating gut microbiota and increasing liver enzyme activity.

[0004] In view of this, the present invention is hereby proposed. Summary of the Invention

[0005] One of the objectives of this invention is to provide a curved widely distributed Lactobacillus to solve the technical problem of the lack of a probiotic in the prior art that can degrade ethanol and has the effect of relieving hangovers and protecting the liver.

[0006] The second objective of this invention is to provide a fermentation broth.

[0007] The third objective of this invention is to provide a microbial inoculant.

[0008] The fourth objective of this invention is to provide the application of the above-mentioned *Lactobacillus curvularis*, the above-mentioned fermentation broth, and the above-mentioned microbial agent in the degradation of ethanol or the preparation of products for the degradation of ethanol.

[0009] The fifth objective of this invention is to provide the application of the above-mentioned *Lactobacillus curvularis*, the above-mentioned fermentation broth, and the above-mentioned microbial agent in the preparation of a drug for relieving hangovers and protecting the liver.

[0010] The sixth objective of this invention is to provide a drug for relieving hangovers and protecting the liver.

[0011] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted: In a first aspect, the present invention provides *Lactobacillus curvilinearus*, wherein the *Lactobacillus curvilinearus* is *Lactobacillus curvilinearus* (… Latilactobacillus curvatusThe strain number is KMHD10-222, and its registration number at the China General Microbiological Culture Collection Center is CGMCC No. 32358.

[0012] In a second aspect, the present invention provides a fermentation broth comprising the aforementioned *Lactobacillus curvilinearis* KMHD10-222.

[0013] Thirdly, the present invention provides a microbial inoculant, comprising the aforementioned *Lactobacillus curvularis* KMHD10-222 and / or the aforementioned fermentation broth.

[0014] Furthermore, the *Lactobacillus curvilinearis* KMHD10-222 exists in the form of living cells.

[0015] Fourthly, the present invention provides the application of the above-mentioned *Lactobacillus curvularis*, the above-mentioned fermentation broth, and the above-mentioned microbial agent in the degradation of ethanol or the preparation of products for the degradation of ethanol.

[0016] Fifthly, the present invention provides the use of the above-mentioned *Lactobacillus curvularis*, the above-mentioned fermentation broth, and the above-mentioned microbial agent in the preparation of a drug for relieving hangovers and protecting the liver.

[0017] Furthermore, the sobering and liver-protecting effects include reducing liver damage.

[0018] Furthermore, the reduction of liver damage includes at least one of reducing hepatocyte damage or necrosis, inhibiting inflammatory responses, improving the liver's antioxidant capacity, inhibiting inflammatory factors, or improving spleen indices.

[0019] In a sixth aspect, the present invention provides a drug for relieving hangovers and protecting the liver, comprising at least one of the above-mentioned *Lactobacillus curvularis*, the above-mentioned fermentation broth, and the above-mentioned microbial inoculant.

[0020] Furthermore, the viable count of *Lactobacillus curvularis* KMHD10-222 in the drug is 1 × 10⁻⁶. 8 ~ 5×10 9 CFU / ml.

[0021] The *Lactobacillus curvularis* strain provided by this invention achieves a degradation rate of 36.05% for 3% ethanol and 19.74% for 7% ethanol, demonstrating its ability to degrade ethanol. The *Lactobacillus curvularis* KMHD10-222 bacterial agent can alleviate acute alcoholic liver injury. This invention provides a strain capable of degrading ethanol and alleviating acute alcoholic liver injury, solving the technical problem of the lack of a probiotic strain in the prior art that can degrade ethanol and has hangover-relieving and liver-protecting effects. Attached Figure Description To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0022] Figure 1 The effect of Lactobacillus curvaturei KMHD10-222 provided by the present invention on the AST and AST expression levels in rat serum; Figure 2 The effect of Lactobacillus curvilinearis KMHD10-222 provided by the present invention on changes in MDA and its content in rat liver; Figure 3 The effects of Lactobacillus curvaturei KMHD10-222 provided by this invention on liver and spleen indices in rats. Detailed Implementation

[0023] Unless otherwise defined herein, the scientific and technical terms used in conjunction with this invention shall have the meanings commonly understood by one of ordinary skill in the art. The meaning and scope of terms shall be clear; however, in any case of potential ambiguity, the definitions provided herein shall prevail over any dictionary or foreign definitions. In this application, unless otherwise stated, the use of "or" means "and / or". Furthermore, the use of the term "comprising" and other forms is non-limiting.

[0024] Unless otherwise stated, the methods and techniques of the present invention are generally carried out according to conventional methods well known in the art and as described in various general and more specific references, which are cited and discussed throughout this specification.

[0025] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0026] This invention provides, in one aspect, *Lactobacillus curvilinearus*, wherein *Lactobacillus curvilinearus* is *Lactobacillus curvilinearus* (… Latilactobacillus curvatus The strain number is KMHD10-222, and its registration number at the China General Microbiological Culture Collection Center is CGMCC No. 32358.

[0027] Preservation instructions for Lactobacillus curvaturei KMHD10-222 Strain name: Lactobacillus curvilinearis KMHD10-222; Latin name: Latilactobacillus curvatus ; Preservation institution: China General Microbiological Culture Collection Center, China Committee on the Preservation and Management of Microbial Cultures; The abbreviation for the depository institution is CGMCC. Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing; Deposit date: October 25, 2024; Collection Center Accession Number: CGMCC No. 32358.

[0028] Lactobacillus curvatureis KMHD10-222 exhibits a degradation rate of 36.05% for 3% ethanol and 19.74% for 7% ethanol, demonstrating its ability to degrade ethanol. Furthermore, the Lactobacillus curvatureis KMHD10-222 bacterial agent can alleviate acute alcoholic liver injury. This invention provides a bacterial strain capable of degrading ethanol and alleviating acute alcoholic liver injury, solving the technical problem of the lack of a probiotic strain in the prior art that can degrade ethanol and possess hepatoprotective and alcohol-degrading effects.

[0029] According to another aspect of the invention, a fermentation broth is also provided, comprising the aforementioned *Lactobacillus curvilinearis* KMHD10-222.

[0030] In some specific embodiments, the fermentation conditions of the fermentation broth include static culture under anaerobic conditions at 37°C to obtain the fermentation broth, wherein the culture time can be controlled according to the required viable bacteria concentration of the fermentation broth and the culture time is 18~24h.

[0031] According to another aspect of the present invention, a microbial inoculant is also provided, comprising the aforementioned *Lactobacillus curvularis* KMHD10-222 and / or the aforementioned fermentation broth.

[0032] In some specific embodiments, the microbial agent is in the form of a liquid agent or a solid agent; in some specific embodiments, the preparation method of the solid agent includes centrifuging the fermentation broth to collect the precipitate, mixing the freeze-drying protectant and the wet weight of the precipitate at a 1:1 volume ratio, and freeze-drying to obtain the solid agent, wherein the freeze-drying conditions are -50℃ to -70℃ for 36 hours. In some specific embodiments, after centrifuging to collect the precipitate, the precipitate is further washed with physiological saline at least 1 to 3 times, preferably 2 times.

[0033] In some specific embodiments, the *Lactobacillus curvilinearis* KMHD10-222 exists in the form of live cells.

[0034] According to another aspect of the present invention, the use of the above-described Lactobacillus curvularis, the above-described fermentation broth, and the above-described microbial agent in the degradation of ethanol or the preparation of products for the degradation of ethanol is also provided.

[0035] According to another aspect of the present invention, the use of the above-mentioned *Lactobacillus curvularis*, the above-mentioned fermentation broth, and the above-mentioned microbial agent in the preparation of a drug for relieving hangovers and protecting the liver is also provided.

[0036] In some specific embodiments, the alcohol-relieving and liver-protecting effects include reducing liver damage; in some specific embodiments, reducing liver damage includes at least one of reducing hepatocyte damage or necrosis, inhibiting inflammatory responses, improving the liver's antioxidant capacity, inhibiting inflammatory factors, or improving spleen indices. Improving spleen indices includes alleviating the decrease in spleen coefficient caused by alcohol.

[0037] According to another aspect of the present invention, a drug for relieving hangovers and protecting the liver is also provided, comprising at least one of the above-mentioned *Lactobacillus curvularis*, the above-mentioned fermentation broth, and the above-mentioned microbial inoculant.

[0038] In some specific embodiments, the viable count of *Lactobacillus curvularis* KMHD10-222 in the drug is 1 × 10⁻⁶. 8 ~ 5×10 9 CFU / ml.

[0039] The present invention will be further illustrated by the following examples. Unless otherwise specified, the materials in the examples are prepared according to existing methods or purchased directly from the market.

[0040] MRS medium: Casein digest 10 g / L, beef extract 10 g / L, yeast extract 4 g / L, triammonium citrate 2 g / L, sodium acetate 5 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, dipotassium hydrogen phosphate 2 g / L, glucose 20 g / L, Tween-80 1.08 g / L, 1‰ L-cysteine ​​hydrochloride anhydrous, 2% agar powder.

[0041] Bromocresol green medium: Casein digest 10 g / L, beef extract 10 g / L, yeast extract 4 g / L, triammonium citrate 2 g / L, sodium acetate 5 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, dipotassium hydrogen phosphate 2 g / L, glucose 20 g / L, Tween-80 1.08 g / L, bromocresol green 6.5 mL / L, 1‰ L-cysteine ​​hydrochloride anhydrous, 2% agar powder.

[0042] Example 1: Isolation, Identification, and Preservation of Strains 1. Isolation and purification of strains Weigh 1g of fermented bean curd sample and place it in a sterile bag. Add 9mL of sterile saline and homogenize twice in a homogenizer for 2 minutes each time to prepare 10 samples. -1 Diluent. Add 10 -1 The diluent was serially diluted 10-fold to 10. -7 A series of dilutions were plated and inverted in an anaerobic incubator at 37°C for 48 hours. Single colonies that turned bromocresol green agar yellow were picked and streaked again on MRS agar, and incubated anaerobically at 37°C. The obtained colonies were purified by streaking multiple times to obtain single bacteria.

[0043] 2. Identification of strains (1) Morphological identification The morphology of single bacterial colonies after multiple purifications was observed. The *Lactobacillus curvilinearis* KMHD10-222 strain was round and milky white, with a slightly transparent outer ring, a moist, smooth, and raised surface, and regular edges. Gram staining was purple, enzyme contact reaction was negative, and microscopic examination revealed rod-shaped colonies.

[0044] (2) 16S identification Using colony DNA as a template, the 16S rRNA gene fragment was amplified using universal primers 27F and 1492R for bacterial sequencing. The PCR amplification reaction system and amplification procedure are shown in Tables 1 and 2.

[0045] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' (SEQ ID NO. 1).

[0046] 1492R: 5'-TACGGCTACCTTGTTACGACTT-3' (SEQ ID NO. 2).

[0047] Table 1 PCR reaction system

[0048] Table 2 PCR reaction procedure

[0049] Amplicon sequence alignment was performed, and the PCR products were sent to the company for 16S rRNA gene amplicon sequencing. The 16S sequence is as follows: The base sequence from the assay results was entered into the NCBI database (http: / / blast.ncbi.nlm.nih.gov / Blast.cgi) for comparison, and strain KMHD10-222 was identified as *Lactobacillus curvilinearis*. Latilactobacillus curvatus The strain KMHD10-222 was inoculated into glycerol tubes and cryopreserved. It was deposited at the China General Microbiological Culture Collection Center (CGMCC) with the registration number CGMCC No. 32358.

[0050] Example 2: Preparation of Fermentation Broth and Microbial Agent 1. Preparation of fermentation broth (1) A single colony of Lactobacillus curvaturei KMHD10-222 was inoculated into MRS liquid medium and cultured statically at 37℃ for 20 h under anaerobic conditions to obtain seed liquid 1; (2) Seed liquid 1 was inoculated into MRS liquid medium at an inoculation amount of 1% and cultured statically at 37℃ for 20h under anaerobic conditions to obtain seed liquid 2. (3) Inoculate seed liquid 2 into MRS liquid medium with an inoculation amount of 1, and incubate under anaerobic conditions at 37℃ for 20h to obtain fermentation liquid.

[0051] 2. Microbial agents (1) Centrifuge the fermentation broth prepared in step 1 (4℃, 5000rpm, 5min), discard the supernatant, wash the cells twice with physiological saline, and weigh the wet weight of the cells after centrifugation.

[0052] (2) The freeze-drying protectant and the wet weight of the bacterial cells were mixed and resuspended at a volume ratio of 1:1, and then frozen at -80℃ overnight. The mixture was then frozen at -50℃ to -70℃ for 36 hours in a freeze dryer to obtain freeze-dried bacterial powder.

[0053] Example 3: In vitro ethanol degradation capacity test of Lactobacillus curvaturei KMHD10-222 1. Materials Potassium dichromate-sulfuric acid solution: Take 12.5 mL of 80 g / L potassium dichromate solution, add 12.5 mL of pure water, mix well, and then slowly add 4 mL of concentrated sulfuric acid while stirring.

[0054] Preparation of standard solutions: Prepare 1 / 2 MRS standard solutions with ethanol content of 0.5%, 1%, 2%, 3%, 4%, and 5% by volume, as shown in Table 3.

[0055] Table 3

[0056] 2. Preparation of Standard Curve: Add 0.05 mL of 1 / 2 MRS standard solution with 0%, 0.5%, 1%, 2%, 3%, 4%, and 5% ethanol contents to glass test tubes, respectively. Add 0.5 mL of potassium dichromate-sulfuric acid solution to each test tube, shake thoroughly, and cap. Place the test tubes in a 100℃ gold bath for 10 min to allow the ethanol to fully evaporate and be fully oxidized by the potassium dichromate-sulfuric acid solution. Remove the test tubes and measure the absorbance of the potassium dichromate-sulfuric acid solution at a wavelength of 610 nm. Prepare a standard curve with ethanol content as the ordinate and absorbance as the abscissa. Calculate the ethanol volume fraction based on the standard curve.

[0057] MRS containing ethanol was prepared by filtering anhydrous ethanol through a 0.22 nm filter membrane. MRS media containing 3% and 7% anhydrous ethanol were then prepared and placed in anaerobic tubes. Two generations of activated *Lactobacillus curvularia* KMHD10-222 bacterial culture (prepared according to the method in Example 2) were inoculated at a rate of 2% (v / v). Samples were taken at 0 h and 19 h, centrifuged, and the supernatant was used to test the ethanol content. The residual ethanol was determined using the potassium dichromate-concentrated sulfuric acid method and a standard curve. The calculated degradation rate of 3% ethanol was 36.05%, and the degradation rate of 7% ethanol was 19.74%.

[0058] Example 4: Tolerance test of the strain 1. Materials Simulated gastric juice (SGF): Weigh 2.0g NaCl + 3.2g pepsin, add 7.0ml concentrated HCl, and dilute to 1000ml with ddH2O. Mix well. Take 50ml of this solution, adjust the pH to 2.5, filter and sterilize for later use.

[0059] Simulated intestinal fluid (SIF): Weigh 6.8g of potassium dihydrogen phosphate (dH2O) and bring the volume to 250ml; measure 77ml of 0.2M NaOH and bring the volume to 500ml with dH2O. Mix 250ml of potassium dihydrogen phosphate with 500ml of NaOH solution, add 10.0g of trypsin and 3g of bile salts, adjust the pH to 6.80, bring the volume to 1000ml, filter and sterilize for later use.

[0060] Bacterial powder: Weigh the bacterial powder (lyophilized bacterial powder prepared according to the method in Example 2), dissolve it in sterile physiological saline, and prepare 3×10⁻⁶ saturated saline solution. 9 CFU / ml bacterial solution.

[0061] 2. Experimental Procedure Strain simulated gastric juice (SGF) experiment: Centrifuge 1 ml of bacterial culture (5000 rpm for 5 min) and discard the supernatant. Resuspend the bacterial cells in an equal volume of simulated gastric juice (SGF). Take bacterial cultures incubated at 37℃ for 0 h and 2 h respectively and dilute them 10-fold serially, i.e., add 0.1 mL of bacterial culture to 0.9 mL of sterile physiological saline and mix thoroughly to obtain 10... -1 Diluent; then replace with a new sterile pipette tip, starting from 10... -1 Take 0.1 mL from the first tube and add it to 0.9 mL of sterile saline in the second tube. Mix well to obtain 10 mL of the first tube. -2 Diluent; and so on, prepare 10 -3 Dilution, 10 -4 Dilution, 10 -5 Dilution and 10 -6 Dilution and plate colony counting.

[0062] Strain simulated intestinal fluid (SIF) experiment: Centrifuge 1 ml of bacterial culture (5000 rpm for 5 min) and discard the supernatant. Resuspend the bacterial cells in an equal volume of simulated intestinal fluid (SIF). Take bacterial cultures incubated at 37℃ for 0 h and 3 h respectively and dilute them 10-fold serially, i.e., add 0.1 mL of bacterial culture to 0.9 mL of sterile physiological saline and mix thoroughly to obtain 10... -1 Diluent; then replace with a new sterile pipette tip, starting from 10... -1 Take 0.1 mL from the first tube and add it to 0.9 mL of sterile saline in the second tube. Mix well to obtain 10 mL of the first tube. -2 Diluent; and so on, prepare 10 -3 Dilution, 10 -4 Dilution, 10 -5 Dilution and 10 -6 Dilution and plate colony counting.

[0063] 3. Results After being treated with simulated gastric and intestinal fluids for a certain period of time, the viable count of *Lactobacillus curvaturei* KMHD10-222 reached 1×10⁻⁶. 6 The CFU / mL indicates that the strain can survive in simulated gastric and intestinal fluids, resisting the influence of the gastrointestinal fluid environment and maintaining the strain's activity.

[0064] Example 5 Safety Evaluation 1. Antibiotic sensitivity Lactobacillus curvaturei KMHD10-222 bacterial powder (lyophilized bacterial powder prepared according to the method in Example 2) was prepared to a concentration of 1×10⁻⁶. 6CFU / mL bacterial suspension was evenly spread on the surface of MRS solid medium plates. After standing at room temperature for 10 minutes, drug sensitivity test discs were placed on top. The plates were then incubated at 37°C for 24 hours. After incubation, the diameter of the inhibition zone around each drug sensitivity test disc was measured using calipers. Each antibiotic was tested in triplicate, including tetracycline, ampicillin, clindamycin, and clarithromycin. The results were used to determine the susceptibility of the strains to the drugs according to the standards of the National Committee for Clinical Laboratory Standards (NCCLS), and the results were classified into three levels: sensitive (S), intermediate (I), and resistant (R). The results showed that *Lactobacillus curvularis* KMHD10-222 was sensitive (S) to each antibiotic, indicating that *Lactobacillus curvularis* KMHD10-222 is safe.

[0065] 2. Hemolytic After reviving and activating *Lactobacillus curvatureensis* strain KMHD10-222 for three generations, it was streaked onto Columbia blood agar plates and incubated at 37°C for 24 hours. The presence of hemolytic zones around the colonies was then observed. α-hemolysis was characterized by a narrow, grass-green hemolytic zone; β-hemolysis by a wide, well-defined, completely transparent hemolytic zone; and γ-hemolysis by no hemolysis, i.e., no hemolytic zone was observed.

[0066] The plate test results showed γ-hemolysis, i.e., no hemolysis, indicating that Lactobacillus curvaturei KMHD10-222 is safe.

[0067] Example 6: Verification of the alleviating effect of Lactobacillus curvaturei KMHD10-222 on acute alcoholic liver injury in rats. 1. Experimental Operation Sixty Wistar rats, 8 weeks old and weighing 250-270g, were used and acclimatized for one week. The rats were randomly divided into 6 groups of 10 rats each, labeled as Control group, Model group, Low-dose group (LK, 5×10⁻⁶), and other groups. 8 CFU / kg), medium-dose group (MK, 5×10) 9 CFU / kg), high-dose group (HK, 5×10) 10 The alcohol concentration was CFU / kg. Fasting was allowed for 12 hours prior to the experiment, but water was permitted. The control and model groups were administered 3 ml of distilled water by gavage daily. The low-dose, medium-dose, and high-dose groups were administered 3 ml of the drug and 3 ml of the bacterial agent by gavage, respectively, for 14 consecutive days. Starting on day 15, 0.5 hours after each group was administered the corresponding solution by gavage, the model, low-dose, medium-dose, and high-dose groups were administered 56° Red Star Erguotou (10 mL / kg) by gavage. The control group received an equal volume of distilled water instead of the alcohol. Alcohol was administered by gavage once every 12 hours, for a total of 6 times.

[0068] Blood parameters: The levels of ALT and AST in rat serum were measured using a kit.

[0069] The ALT detection kit was purchased from Nanjing Jiancheng Biotechnology Research Institute Co., Ltd., catalog number C009-2-1; the AST detection kit was purchased from Nanjing Jiancheng Biotechnology Research Institute Co., Ltd., catalog number C010-2-1.

[0070] Liver index testing: The liver is removed and weighed, and the liver index is calculated. Liver index (%) = (liver wet weight ÷ animal body weight) × 100%.

[0071] The levels of MDA and GSH in liver tissue were measured according to the kit instructions.

[0072] The MDA detection kit was purchased from Nanjing Jiancheng Biotechnology Research Institute Co., Ltd., catalog number A003-1-1; the GSH detection kit was purchased from Nanjing Jiancheng Biotechnology Research Institute Co., Ltd., catalog number A006-2-1.

[0073] Spleen index testing: The spleen is removed and weighed, and the spleen index is calculated. Spleen index (%) = (spleen wet weight ÷ animal body weight) × 100%.

[0074] 2. Test Results (1) Changes in ALT and AST enzyme activities under the action of Lactobacillus curvaturei KMHD10-222 are as follows: Figure 1 As shown, compared with the blank group, the serum ALT and AST levels of rats in the model group were significantly increased. After intervention with Lactobacillus curvularis KMHD10-222, the serum ALT and AST levels of rats were reduced, with significant differences (p<0.05). This indicates that Lactobacillus curvularis KMHD10-222 can reduce hepatocellular inflammation, damage or necrosis and alleviate alcohol-induced liver injury.

[0075] (2) Changes in MDA and GSH content under the action of Lactobacillus curvaturei KMHD10-222 are as follows: Figure 2 As shown, compared with the control group, the MDA level in the liver of rats in the model group was significantly increased (p < 0.05), and *Lactobacillus curvilinearis* KMHD10-222 significantly reduced (p < 0.05) the MDA concentration in the liver of rats. This indicates that *Lactobacillus curvilinearis* KMHD10-222 can improve the antioxidant capacity of the liver and significantly reduce its lipid peroxidation. Compared with the control group, the GSH content in the liver of rats in the model group was increased, indicating that alcohol gavage increased the degree of oxidative stress in rats, and *Lactobacillus curvilinearis* KMHD10-222 can significantly reduce the level of oxidative stress.

[0076] (3) Changes in liver and spleen indices under the action of Lactobacillus curvaturei KMHD10-222 are as follows: Figure 3As shown, there was no significant difference in liver index changes among the groups; compared with the blank group, the spleen index of the liver of rats in the model group was significantly reduced (p<0.05), and Lactobacillus curvilinearis KMHD10-222 could significantly alleviate the decrease in spleen index, indicating that Lactobacillus curvilinearis KMHD10-222 can alleviate the decrease in spleen index caused by alcohol when improving alcoholic liver injury, and at the same time improve liver function indicators.

[0077] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. *Lactobacillus curvilinearus*, characterized in that, The *Lactobacillus curvilinearus* is *Lactobacillus curvilinearus* (… Latilactobacillus curvatus The strain number is KMHD10-222, and its registration number at the China General Microbiological Culture Collection Center is CGMCC No. 32358.

2. A fermentation broth, characterized in that, Includes *Lactobacillus curvilinearis* KMHD10-222 as described in claim 1.

3. A microbial inoculant, characterized in that, Includes Lactobacillus curvaturei KMHD10-222 as described in claim 1 or / and the fermentation broth as described in claim 2.

4. The microbial agent according to claim 3, characterized in that, The *Lactobacillus curvilinearis* KMHD10-222 exists in the form of live cells.

5. The use of the *Lactobacillus curvularis* of claim 1, the fermentation broth of claim 2, and the microbial agent of claim 3 or 4 in the degradation of ethanol or the preparation of products for the degradation of ethanol.

6. The use of the *Lactobacillus curvularis* of claim 1, the fermentation broth of claim 2, and the microbial agent of claim 3 or 4 in the preparation of a drug for relieving hangovers and protecting the liver.

7. The application according to claim 6, characterized in that, The so-called hangover relief and liver protection includes reducing liver damage.

8. The application according to claim 7, characterized in that, The reduction of liver damage includes at least one of reducing hepatocyte damage or necrosis, inhibiting inflammatory responses, improving the liver's antioxidant capacity, inhibiting inflammatory factors, or improving spleen indices.

9. A drug for relieving hangovers and protecting the liver, characterized in that, It includes at least one of the following: Lactobacillus curvularis as described in claim 1, the fermentation broth as described in claim 2, and the microbial agent as described in claim 3 or 4.

10. The medicament according to claim 9, characterized in that, The viable count of *Lactobacillus curvularis* KMHD10-222 in the drug is 1×10⁻⁶. 8 ~5×10 9 CFU / ml.

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