Phytobacterium plantarum XJSN202504 and application thereof in acute alcoholism

Abstract

The invention relates to the technical field of microorganisms, and discloses a plant lactobacillus XJSN202504 and application thereof in acute alcoholism, the plant lactobacillus XJSN202504 is preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC NO.34746, and the preservation time is June 3, 2025. The invention further discloses a preparation method of the plant lactobacillus XJSN202504. The lactobacillus plantarum XJSN202504 disclosed by the invention can be used for improving liver injury related indexes in serum and expression of inflammation related genes, and also can be used for adjusting the activity of antioxidant enzymes in the liver and relieving alcohol-induced oxidative stress injury; moreover, gastric mucosa, small intestine and colon injuries caused by a large amount of alcohol can be effectively prevented, the anti-inflammatory activity and the antioxidant activity are better, the anti-alcohol function can be achieved through gastrointestinal tracts, and the effects of protecting liver and digestive tract mucosa are achieved.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and more specifically, to a plant lactobacillus XJSN202504 and its application in acute alcohol poisoning. Background Technology

[0002] Acute alcohol poisoning (AAI), also known as acute ethanol poisoning, is a central nervous system abnormality caused by the ingestion of a large amount of alcohol in a short period of time. It can lead to varying degrees of motor incoordination, altered consciousness, and respiratory depression; in severe cases, it can cause multiple organ damage and even endanger life. Studies have shown that acute alcohol poisoning is a leading cause of cirrhosis and liver failure worldwide, and alcohol dependence and abuse have become a global issue causing economic stress and health problems.

[0003] The main treatment approaches for AAI are: slowing down the absorption rate of alcohol in the body, enhancing the activity of alcohol-related enzymes in the body and improving antioxidant capacity to remove free radicals in the body; clinically, measures such as diuresis to promote ethanol excretion, maintaining the balance of water and electrolytes in the body, and infusion of glucose and vitamin C are usually given; for patients with acute severe alcohol poisoning, naloxone or combined with metadoxine is commonly used, but it is easy to cause adverse reactions such as hypertension, drowsiness, and irritability, and most of the treatments are post-treatment relief and cannot prevent it in advance [4]. Other treatment methods are supportive, such as disulfiram, acampic acid and other drugs, which can be used as adjunctive therapy to reduce the dependence of alcohol poisoning patients in the short term, but the treatment scope is limited and the side effects are large. Therefore, the development of traditional natural active substances or other health care therapies to prevent and alleviate AAI in advance has become the focus of global attention.

[0004] The pathogenesis of acute alcoholic liver disease (AAI) is complex. Normally, some ethanol is metabolized by alcohol dehydrogenase into acetaldehyde, which is then converted to acetic acid by acetaldehyde dehydrogenase, and finally broken down into H2O and CO2. Excessive alcohol consumption at one time can lead to the accumulation of acetaldehyde, an intermediate product that can interfere with the activity of various enzymes, DNA replication, and lipid metabolism, resulting in tissue damage and even death. Gut microorganisms contain these two enzymes. Probiotics can effectively inhibit the colonization of pathogenic bacteria by maintaining gut microbiota balance, thereby regulating the effects of alcohol dehydrogenase and acetaldehyde dehydrogenase on ethanol metabolism. They can also exert an oxidizing effect, reducing endotoxin production. Increasing research demonstrates that the intake of various microorganisms and probiotics can significantly alleviate alcohol-induced liver damage. Some studies, focusing on the correlation between alcohol metabolism capacity and the alcohol metabolism activity of gut microbiota, have used strains with high alcohol metabolism activity as subjects. Representative strains were screened, and a mouse model of acute alcohol poisoning was established, confirming that *Bifidobacterium pseudosporidis*, *Saccharomyces cerevisiae*, and *Lactobacillus plantarum* with high enzyme levels significantly alleviated acute alcohol poisoning. Therefore, probiotics have great potential for development in the prevention and relief of alcohol poisoning due to their advantages in anti-inflammatory and antioxidant properties, as well as their ability to regulate liver function through the gut-liver axis.

[0005] Therefore, providing a *Lactobacillus plantarum* XJSN202504 and its application in acute alcohol poisoning has important practical significance. Summary of the Invention

[0006] In view of this, the present invention proposes a *Lactobacillus plantarum* XJSN202504 and its application in acute alcohol poisoning, aiming to solve at least one of the problems in the background art.

[0007] This invention proposes a plant lactobacillus XJSN202504, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.34746 and deposit date of June 3, 2025.

[0008] The present invention also provides a *Lactobacillus plantarum* inoculant, which includes *Lactobacillus plantarum* XJSN202504 as described in the above technical solution.

[0009] The present invention also provides a composition comprising *Lactobacillus plantarum* XJSN202504 or *Lactobacillus plantarum* inoculum as described in the above-described technical solutions.

[0010] The present invention also provides an application of the above-mentioned Lactobacillus plantarum XJSN202504, specifically the application of Lactobacillus plantarum XJSN202504 in the preparation of drugs for treating acute alcohol poisoning.

[0011] The present invention also provides an application of the *Lactobacillus plantarum* agent described in the above technical solution, specifically the application of the *Lactobacillus plantarum* agent in the preparation of a drug for treating acute alcohol poisoning.

[0012] The present invention also provides an application of the composition described in the above technical solution, specifically the application of the composition in the preparation of a drug for treating acute alcohol poisoning.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: The *Lactobacillus plantarum* XJSN202504 described in this invention can effectively prevent and improve AAI-induced liver tissue cell damage and the resulting changes in body weight and liver index, and improve the expression of liver damage-related indicators and inflammation-related genes in serum. Furthermore, the *Lactobacillus plantarum* XJSN202504 of the present invention can also regulate the activity of antioxidant enzymes in the liver, reduce alcohol-induced oxidative stress damage, and effectively prevent damage to the gastric mucosa, small intestine, and colon caused by large amounts of alcohol. It has good anti-inflammatory and antioxidant activities, can exert its alcohol-detoxifying function through the gastrointestinal tract, and has the effect of protecting the liver and digestive tract mucosa. Attached Figure Description

[0014] Various other advantages and benefits will become apparent to those skilled in the art upon reading the following detailed description of preferred embodiments. The accompanying drawings are for illustrative purposes only and are not intended to limit the invention. Furthermore, the same reference numerals denote the same parts throughout the drawings. In the drawings: Figure 1 A schematic diagram showing the effects of Lactobacillus plantarum XJSN202504 on body weight and liver index in AAI mice; Figure 2 A schematic diagram showing the effects of Lactobacillus plantarum XJSN202504 on the intoxication rate, tolerance time, and intoxication time of AAI mice; Figure 3 A schematic diagram showing the effects of Lactobacillus plantarum XJSN202504 on liver index, liver alcohol metabolism enzyme activity, and blood ethanol concentration in AAI mice; Figure 4 A schematic diagram showing the effect of Lactobacillus plantarum XJSN202504 on liver damage indicators in the serum of AAI mice; Figure 5A schematic diagram showing the effects of Lactobacillus plantarum XJSN202504 on the morphological characteristics of liver tissue in AAI mice; Figure 6 Schematic diagram showing the effect of Lactobacillus plantarum XJSN202504 on the activity of antioxidant factors in serum and liver of AAI mice; Figure 7 A schematic diagram showing the effect of Lactobacillus plantarum XJSN202504 on the expression of inflammatory genes in the liver of AAI mice; Figure 8 A schematic diagram showing the effects of Lactobacillus plantarum XJSN202504 on the morphological characteristics of gastric tissue in AAI mice. Figure 9 This is a schematic diagram showing the effects of Lactobacillus plantarum XJSN202504 on the morphological characteristics of intestinal tissue in AAI mice. Detailed Implementation

[0015] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention. It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the present invention.

[0016] Furthermore, regarding the numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included within this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0017] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0018] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0019] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0020] This invention proposes a plant lactobacillus XJSN202504, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.34746 and deposit date of June 3, 2025.

[0021] The present invention also provides a *Lactobacillus plantarum* inoculant, which includes *Lactobacillus plantarum* XJSN202504 as described in the above technical solution.

[0022] The present invention also provides a composition comprising *Lactobacillus plantarum* XJSN202504 or *Lactobacillus plantarum* inoculum as described in the above-described technical solutions.

[0023] The present invention also provides an application of the above-mentioned Lactobacillus plantarum XJSN202504, specifically the application of Lactobacillus plantarum XJSN202504 in the preparation of drugs for treating acute alcohol poisoning.

[0024] The present invention also provides an application of the *Lactobacillus plantarum* agent described in the above technical solution, specifically the application of the *Lactobacillus plantarum* agent in the preparation of a drug for treating acute alcohol poisoning.

[0025] The present invention also provides an application of the composition described in the above technical solution, specifically the application of the composition in the preparation of a drug for treating acute alcohol poisoning.

[0026] Example 1: Screening yielded Lactobacillus plantarum XJSN202504 I. Experimental Materials Traditional naturally fermented yogurt was collected from the homes of herders in Kashgar, Xinjiang. After the yogurt was thoroughly stirred with a sterile spoon, 50 mL was drawn into a sterilized centrifuge tube using a sterile syringe, placed in a low-temperature food sampling box, and brought back to the laboratory for freezing and storage in an ultra-low temperature freezer at -80°C for later use.

[0027] II. Experimental Methods 1. Isolation and Identification of Bacteria 1.1 Isolation and purification of bacteria Take 1 mL of yogurt sample and serially dilute it 10-fold with sterile physiological saline to 10⁻⁶. Then, take 100 μL of bacterial suspensions at three gradients (10⁻⁴, 10⁻⁵, and 10⁻⁶) and spread them on plates. Incubate at 37℃ for 24-48 h, and observe and record the colony morphology. Pick colonies of different morphologies from the plates and streak them for isolation. After incubating at 37℃ for 48 h, pick single colonies of different morphologies from the plates again and streak them for isolation. Repeat this process multiple times until pure single colonies with consistent morphology are obtained.

[0028] 1.2 Bacterial DNA Extraction The purified suspected target strain was inoculated into MRS broth and incubated at 37°C for 18-24 hours. DNA was then extracted using a bacterial genomic DNA extraction kit. The extracted DNA was numbered and stored at -20°C for later use.

[0029] 1.3 Genomic DNA PCR amplification and agarose gel electrophoresis detection The extracted DNA was subjected to PCR amplification. The mixture consisted of 1 μL of upstream primer 27F (5'-AGA GTT TGA TCC TGGCTCAG-3'), 1 μL of downstream primer 1495R (5'-CTA CGG CTA CCTTGT TAC GA-3'), 12.5 μL of 2×Taq plus Buffer, and 1 μL of template DNA. The volume was brought to 25 μL with sterile dd H2O. Sterile ultrapure water was used as a negative control instead of template DNA. The amplification conditions were: 94℃ for 5 min; 94℃ for 30 s, 55℃ for 30 s, 72℃ for 1 min, for a total of 29 cycles, with a final extension at 72℃ for 5 min.

[0030]

[0031] Example 2: Verification of the efficacy of Lactobacillus plantarum XJSN202504 1. Preparation of bacterial culture The bacteria were activated using MRS medium and incubated at 37°C for 48 hours until the logarithmic growth phase. The number of colony-forming units (CFU) was standardized by measuring optical density (wavelength 600 nm). After centrifugation, bacterial suspensions were prepared with PBS at high concentrations of 10⁹ CFU / (kg mb·d) and low concentrations of 10⁸ CFU / (kg mb·d) for later use.

[0032] 2. Animal models 2.1 Animal grouping Fifty male SPF-grade C57 mice, aged 8-10 weeks and weighing 30-35g, were used. All experimental procedures were conducted in accordance with the requirements of the Laboratory Animal Ethics Committee. The experiment was divided into 5 groups, with 10 mice in each group: normal control group (NC group), acute alcohol poisoning model group (AAI group), metadoxine-positive control group (MTDX group), high-concentration *Lactobacillus plantarum* XJSN202504 experimental group (XJSN202504-H group), and low-concentration *Lactobacillus plantarum* XJSN202504 experimental group (XJSN202504-L group).

[0033] 2.2 AAI Model Establishment and Intervention After the start of the experiment, at 9:00 AM every day, mice in the XJSN202504-H group and the XJSN202504-L group were administered bacterial culture via gavage at doses of 10⁹ CFU / (kg mb·d) and 10⁸ CFU / (kg mb·d), respectively. Mice in the remaining NC group, AAI group, and positive control MTDX group were administered the same volume of PBS via gavage (0.2 ml / mouse / day for all mice). The gavage intervention continued for one week, and mice were weighed daily. For the last gavage, mice in the MTDX group were administered Metadoxine at a concentration of 20 g / kg. The other groups maintained the previous gavage procedure. Thirty minutes after the completion of the gavage procedure, except for the blank group which was administered physiological saline, mice in the other groups were administered baijiu (56%, v / v) via gavage (5.6 g / kg body weight / day). The disappearance time, recovery time, and sleep time of the righting reflex were observed and recorded.

[0034] 2.3 Righting Reflection Experiment Mice were administered 56% alcohol by gavage and observed for approximately 20 minutes. When a mouse began to wobble or become sluggish, it was turned over so that its back was facing down. If the mouse could not roll back to its back within 30 seconds and the righting reflex disappeared, it was considered intoxicated. When the mouse could roll back to its back and the righting reflex reappeared, it was considered sober. The number of mice that lost the righting reflex was observed and recorded. Mice that were not intoxicated were recorded as 0 min, and mice that were still not sober after 7 hours were recorded as 420 min.

[0035] 2.4 Serum collection and tissue homogenization At the end of the experiment, peripheral blood from mice was collected using the enucleation method into 1.5 ml sterile centrifuge tubes. After standing at room temperature for 2 hours and then incubating overnight at 4°C, the tubes were centrifuged at 3500 r / min for 10 min. The supernatant serum was collected and stored at -80°C for later analysis. An appropriate amount of liver tissue was minced and added to Tris-HCl solution at pH 7.4 at a ratio of 1:9 (w:v). The tissue was homogenized and centrifuged at 3500 r / min for 10 min. The precipitate was discarded, and the supernatant was collected to obtain the liver tissue homogenate.

[0036] 2.5 Detection of alcohol concentration in mouse serum and metabolic enzymes in liver Biochemical reagent kits were used to detect serum alcohol content, the activities of alcohol-metabolizing enzymes ADH and ALDH in liver tissue homogenate, and the activities of antioxidants SOD, MDA, GSH, and GSH-Px in serum and liver. Additionally, the activities of ALT, ALP, AST, and GGT in liver tissue homogenate were measured to verify the level of liver damage. Following the instructions, blank wells, zeroing wells, and parallel controls were set up. Absorbance (A) values ​​were measured at 450 nm, and the levels of the measured indicators were calculated by plotting a standard curve.

[0037] 2.6 Detection of antioxidant factors in mouse serum and liver tissue The levels of antioxidant factors such as GSH, GSH-Px, SOD, and MDA in serum and liver tissue homogenates were detected according to the instructions using a biochemical reagent kit. To prepare a 10% tissue homogenate: mouse liver tissue was taken, and 0.2-1g of tissue was rinsed with ice-cold physiological saline to remove blood. After drying with filter paper, pre-cooled 0.86% physiological saline was added. The amount of physiological saline should be 9 times the tissue weight. The tissue was quickly minced with ophthalmic scissors and homogenized. The homogenate was then centrifuged at 3000 rpm for 10-15 minutes, and the supernatant was collected for analysis.

[0038] 2.7 Histological observation Mice were euthanized by cervical dislocation after blood collection. Fresh liver, stomach, small intestine, and colon tissues were collected and fixed in 4% paraformaldehyde for 12-24 hours or more until further processing. After routine tissue dehydration, the tissues were dried, cleared, embedded in paraffin, and sectioned to a thickness of 4-5 μm. Hematoxylin and eosin (HE) staining was performed, and histopathological changes were observed under an optical microscope.

[0039] 2.8 RT-qPCR detection of gene expression Immediately after collection, fresh mouse liver tissue was frozen at -80℃ for later use. When extracting RNA from the tissue, a certain amount of tissue was taken and excised. Following the instructions of the RNA extraction and reverse transcription kit, whole-genome RNA was extracted. After determining the concentration, the RNA was reverse transcribed into cDNA. The purity of the cDNA was then checked and used as a sample. The RT-qPCR process and parameters were performed according to the reagent instructions. Reverse transcriptase and other reactants were added, along with primers and SYBR Green reagent for qPCR. β-actin was used as an internal reference gene. The primer sequences for all detected genes are shown in Table 1. Table 1 Primer sequence listing

[0040] Note: IL-1β, interleukin-1β; NF-κB, nuclear factor-kappa B; TNF-α, tumor necrosis factorα; Nrf-2, nuclear factor erythroid 2-related factor 2; β-actin.

[0041] 2.9 Results and Analysis (1) Effects of Lactobacillus plantarum XJSN202504 on body weight and liver index in AAI mice as follows Figure 1 As shown, where Figure 1 Figure A shows the effect of *Lactobacillus plantarum* XJSN202504 on the body weight of AAI mice. Figure 1 Figure B shows the effect of Lactobacillus plantarum XJSN202504 on liver index in AAI mice. based on Figure 1It was found that there was no significant difference in body weight among the groups of mice within one week of gavage intervention, proving that high and low concentrations of XJSN202504 administered by gavage, like PBS, had no significant effect on mouse body weight. However, after three days of alcohol administration, differences in body weight began to appear among the groups. Compared with the normal group, the AAI group mice showed a significant decrease in body weight after three days of alcohol administration, while the MTDX group mice showed a significantly smaller decrease in body weight. The body weight of the XJSN202504-H group mice was similar to that of the MTDX group mice, while the body weight of the XJSN202504-L group mice showed the same trend and was similar to that of the AAI group. This proves that XJSN202504 intake helps alleviate AAI-induced body weight loss, with high concentrations showing a significant effect and low concentrations showing a less significant effect. Simultaneously, the liver index (considered one of the indicators of alcoholic liver disease) of the AAI mice in each group increased to varying degrees due to continuous alcohol intake, but the MTDX group and XJSN202504-H group showed the least increase in liver index, and the XJSN202504-L group was less effective than the XJSN202504-H group.

[0042] (2) Effects of *Lactobacillus plantarum* XJSN202504 on the intoxication rate, tolerance time, and intoxication duration in AAI mice. Figure 2 As shown, where, Figure 2 In diagram A, the intoxication rate of mice after oral administration of alcohol is shown. Figure 2 Figure B shows the tolerance time and intoxication results of mice after oral administration of alcohol in each group; based on Figure 2 As shown in Figure A, the alcohol intoxication rate in the MTDX model group was 63.33±4.00, which was significantly lower than that in the AAI model group (96.67±6.33) (P<0.05). The alcohol intoxication rates in the XJSN202504-H and XJSN202504-L groups were also lower than those in the model group, but there was no statistically significant difference. Among them, the alcohol intoxication rate in the high-concentration XJSN202504-H group was significantly lower than that in the low-concentration XJSN202504-L group. based on Figure 2B shows that the tolerance time and intoxication time of the AAI model group mice were 12.00±5.54 min and 320.83±55.90 min, respectively. Compared with the AAI model group, the MTDX group mice significantly prolonged the tolerance time and effectively shortened the intoxication time (P<0.05). The XJSN202504-H group and the XJSN202504-L group could prolong the tolerance time of mice (P>0.05) and significantly shorten the sleep time (P<0.05), and it can be seen that the XJSN202504-H group was slightly better than the XJSN202504-L group. The results indicate that XJSN202504 can reduce the intoxication rate of AAI mice to a certain extent, prolong the tolerance time, and especially shorten the sobriety time. The high concentration group of XJSN202504 (109 CFU / (kg mb·d)) was more effective than the low concentration group (108 CFU / (kg mb·d)).

[0043] (3) The effects of *Lactobacillus plantarum* XJSN202504 on liver index, liver alcohol metabolism enzyme activity, and blood ethanol concentration in AAI mice are as follows: Figure 3 As shown, where, Figure 3 The diagram in Figure A shows the results of ethanol concentration in the blood of AAI mice. Figure 3 Image B is a schematic diagram of the liver index results in AAI mice. Figure 3 The diagram in middle C shows the results of alcohol metabolism enzyme activity in the liver of AAI mice; based on Figure 3 As shown in Figure A, compared with the NC control group, the blood alcohol concentration in the AAI alcohol treatment model group was significantly increased (P<0.05). Compared with the AAI model group, the blood alcohol concentration in the MTDX group, XJSN202504-H group, and XJSN202504-L group was significantly decreased (P<0.05), with the XJSN202504-H group showing better results than the XJSN202504-L group, while the effect was not different from that of the MTDX group. These results indicate that high concentrations of XJSN202504 can significantly reduce the blood alcohol content in mice with acute alcohol poisoning.

[0044] based on Figure 3 According to the BC assay, compared with the blank control group, mice in the AAI model group showed a significant increase in both ADH and ADH activity in the liver due to alcohol intake (P<0.05), and the ADH activity in the MTDX group and the XJSN202504-H treatment group was slightly higher than that in the model group (P>0.05). This demonstrates that XJSN20250401 pretreatment can enhance ADH activity, with higher concentrations showing a more significant effect.

[0045] (4) The effects of *Lactobacillus plantarum* XJSN202504 on liver damage markers in the serum of AAI mice are as follows: Figure 4As shown, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are important transaminases in hepatocytes that produce amino acid degradation. Hepatocyte death leads to elevated serum ALT and AST activity. Along with alkaline phosphatase (ALP) and glucagon (GGT), these are also important indicators of hepatocyte damage. Figure 4 The results showed that, compared with the NC control group, alcohol treatment significantly increased the activities of AST, ALT, GGT, and ALP in the serum of mice in the AAI group (P<0.05). In contrast, AST in the serum of mice in the MTDX group and those pretreated with high concentration XJSN202504-H significantly decreased to normal levels. While the activities of ALT, ALP, and GGT decreased in all three treatment groups, there was no statistically significant difference compared to the AAI group. Furthermore, XJSN202504-H was more effective than XJSN202504-L. These results indicate that alcohol gavage damages hepatocytes, and pretreatment with high concentration XJSN202504 can, to some extent, prevent and alleviate liver damage caused by alcohol poisoning.

[0046] (5) Results of the effects of Lactobacillus plantarum XJSN202504 on liver tissue morphology in AAI mice are as follows: Figure 5 As shown, based on Figure 5 It was found that in the normal NC group mice, hepatocytes were neatly arranged, forming radial streaks to form liver plates. The hepatocyte structure was intact, the nucleus structure was clear, and the boundaries were well-defined. In contrast, in the AAI model group, hepatocytes were irregularly arranged, swollen, with enlarged intercellular spaces (black arrows), enlarged hepatic sinusoids filled with blood cells (yellow arrows), some hepatocyte nuclei were atrophied, deformed, or even disappeared, cell boundaries were blurred, and large areas of necrosis were visible (within the black box). Numerous inflammatory cells infiltrated the blood vessel edges and surrounding tissues. Mice pretreated with Metadoxine and XJSN202504 showed reduced inflammatory infiltration in the liver, without the extensive cell necrosis seen in the model group. The morphology and arrangement of hepatocytes tended to be normal, with the XJSN202504-H group showing better results than the XJSN202504-L group. These results indicate that alcohol causes severe liver damage, and high concentrations of XJSN202504 can play a good role in prevention and relief.

[0047] (6) The effects of *Lactobacillus plantarum* XJSN202504 on the activity of antioxidant factors in the serum and liver of AAI mice are as follows: Figure 6 As shown, based on Figure 6It was found that, compared with the NC group, the serum and liver tissue levels of AAI mice were significantly reduced (P<0.05). All three intervention groups increased the serum and liver levels of GSH, GSH-px, and SOD. Although Metadoxine was generally more effective than XJSN202504-H, which was superior to XJSN202504-L, XJSN202504-H and XJSN202504-L showed better effects on liver GSH-px. Furthermore, compared with the NC normal group, the serum and liver tissue levels of the AAI model group were significantly increased (P<0.05), while all three intervention groups inhibited serum MDA levels with no significant difference between groups (P>0.05). The XJSN202504-L group had no significant effect on liver MDA.

[0048] (7) The effects of *Lactobacillus plantarum* XJSN202504 on the expression of inflammatory genes in the liver of AAI mice are as follows: Figure 7 As shown, by detecting the mRNA expression levels of TNF-α, IL-1β, Nrf-2, and NF-κB, the degree of inflammatory response in the liver induced by alcohol (AAI) can be determined. Figure 7 It was found that the expression levels of TNF-α, IL-1β, and NF-κB mRNA in the AAI acute alcohol model group were significantly increased (P<0.05). The gene expression trends of the three treatment groups were opposite to those of the AAI model group, demonstrating that XJSN202504 has the same effect as Metadoxine in reducing the expression of inflammatory genes in the liver and is positively correlated with bacterial concentration. That is, the effect of the XJSN202504-H group is closer to that of the MTDX group, and its effect is better than that of the XJSN202504-L group. In contrast to the above gene expression trends, the expression level of Nrf-2 mRNA in the AAI model group was significantly decreased compared with the NC normal group (P<0.05). However, XJSN202504-H can significantly inhibit the decrease of Nrf-2 mRNA expression level in liver tissue of AAI mice, and its effect is better than that of the MTDX group and the XJSN202504-L group.

[0049] (8) Results of the effects of Lactobacillus plantarum XJSN202504 on the morphology of gastric tissue in AAI mice are as follows: Figure 8 As shown, Figure 8 A is a gross image of the isolated stomach of each group of mice; B is a schematic diagram of the gastric mucosal layer thickness of each group of mice; and C is a schematic diagram of the gastric wall muscle layer thickness of each group of mice. based on Figure 8As shown in Figure A, the stomach in the normal NC group was pink with no visible damage. In the AAI model group, obvious bleeding and swelling and thinning of the lower gastric antrum were observed. Gastric bleeding was significantly reduced in all three intervention groups. Pathological section HE staining results showed that AAI alcohol modeling caused significant damage to the gastric wall, increased the separation distance between the mucosa and submucosa, inflammatory cell infiltration, thickening of the muscular layer (marked by black lines), and thinning of the gastric intrinsic mucosa with damage and loss of upper cells (marked by yellow lines). based on Figure 8 BC analysis revealed that the Metadoxine and XJSN202504 treatment groups showed less damage to the gastric wall morphology. The MTDX and XJSN202504-H groups significantly prevented and alleviated alcohol-induced thinning of the gastric mucosa and inflammatory fibrosis thickening of the gastric wall muscle layer. Therefore, XJSN202504 has a certain protective effect against alcohol-induced gastric damage in AAI model mice, with XJSN202504-H showing better efficacy than XJSN202504-L.

[0050] (9) Results of the effects of Lactobacillus plantarum XJSN202504 on the morphology of intestinal tissue in AAI mice are as follows: Figure 9 As shown, based on Figure 9 It was found that, compared with the normal group, the mice in the AAI group, which were administered alcohol by gavage, suffered severe intestinal damage. The small intestine and colon were significantly thinner and shorter in diameter, and a large number of deeply stained inflammatory cells infiltrated and clumped between the submucosa and muscularis propria. The small intestinal villi were shrunken and broken in large numbers, and the intervillous spaces were enlarged. Erythrocyte infiltration was obvious in the colonic lumen, and the mucosa and muscularis propria separated. Compared with the AAI group, no obvious inflammatory cells were found in the small intestine of the MTDX group, and the villi were arranged regularly without obvious breakage. The inflammatory infiltration of the small intestine in the XJSN202504-H group was significantly reduced compared with the AAI group, and the small intestinal villi breakage was reduced, showing better results than the XJSN202504-L group. Compared with the AAI group, the colons of the MTDX group and XJSN202504-H group were significantly thicker, with fewer inflammatory cells and congestion in the colonic submucosa, and less mucosal tissue damage, showing significantly better results than the XJSN202504-L group.

[0051] In summary, compared to the AAI group, pretreatment with *Lactobacillus plantarum* XJSN202504 significantly reduced blood alcohol concentration in AAI mice by regulating the activity of alcohol-metabolizing enzymes in the liver, thus reducing the rate of intoxication, prolonging tolerance time, and especially shortening sobriety time. It also effectively improved liver, stomach, and intestinal tissue damage in AAI mice, as well as the resulting changes in body weight and liver index; improved the expression of serum liver injury-related indicators and inflammation-related genes; and alleviated alcohol-induced oxidative stress damage by regulating the activity of antioxidant enzymes in the liver.

[0052] 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 it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the specific implementation of the present invention. Any modifications or equivalent substitutions that do not depart from the spirit and scope of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. A type of *Lactobacillus plantarum* XJSN202504, characterized in that, The *Lactobacillus plantarum* XJSN202504 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.34746 on June 3, 2025.

2. A plant lactobacillus inoculant, characterized in that, The *Lactobacillus plantarum* inoculant includes *Lactobacillus plantarum* XJSN202504 as described in claim 1.

3. A composition, characterized in that, The composition comprises *Lactobacillus plantarum* XJSN202504 as described in claim 1 or *Lactobacillus plantarum* inoculum as described in claim 2.

4. An application of *Lactobacillus plantarum* XJSN202504 as described in claim 1, characterized in that, The specific application is the use of *Lactobacillus plantarum* XJSN202504 in the preparation of drugs for treating acute alcohol poisoning.

5. The application of the *Lactobacillus plantarum* inoculant according to claim 2, characterized in that, The specific application is the use of the *Lactobacillus plantarum* agent in the preparation of drugs for treating acute alcohol poisoning.

6. An application of the composition according to claim 3, characterized in that, The specific application is the use of the composition in the preparation of a medicine for treating acute alcohol poisoning.

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