Application of cibocephalum in preparation of product for relieving or treating alcoholic liver disease

Abstract

The invention relates to the field of application of cilexellatus, provides application of cilexellatus in preparation of a product for relieving or treating alcoholic liver diseases, and proves that cilexellatus can be used as a raw material for preparing the product for relieving or treating alcoholic liver diseases by constructing an alcoholic liver model mouse and feeding with a suspension of cilexellatus.

Description

Technical Field

[0001] The present invention relates to the application field of Dubosie bacteria, and in particular to the application of Dubosie bacteria in preparing a product for alleviating or treating alcoholic liver disease. Background Art

[0002] Alcoholic liver disease (ALD) is a condition in which liver tissue is damaged by chronic alcohol abuse. Alcohol-induced liver damage encompasses alcoholic fatty liver disease, alcoholic hepatitis, alcoholic liver fibrosis, and alcoholic cirrhosis, posing a serious threat to human health. Recent studies have shown that, based on data from the Global Burden of Disease (GBD) study, the global prevalence of alcoholic liver disease (ALD) increased by 38.68% between 2000 and 2021, and alcohol-related primary liver cancer increased by 94.12%. This indicates a significant upward trend in the global prevalence of ALD. The pathogenesis of ALD is a complex process involving the interplay of multiple factors, including oxidative stress, altered gut microbiota, inflammatory responses, and autophagy dysfunction. The gut-liver axis plays a key role in the pathogenesis of ALD. Alcohol consumption alters the composition of the gut microbiome, reducing the abundance of beneficial bacteria and increasing the proportion of harmful bacteria. This change increases intestinal permeability, allowing bacterial endotoxins (such as LPS) to more easily enter the bloodstream and reach the liver, causing liver damage. Currently, treatments for alcoholic liver injury primarily include alcohol abstinence, nutritional support, antioxidants, and anti-inflammatory drugs. However, existing treatment options have numerous limitations, including poor efficacy and severe side effects. In recent years, beneficial bacteria have gained increasing attention as therapeutic approaches. Beneficial bacteria can regulate the balance of the intestinal microbiota, reduce the production and absorption of intestinal toxins, and alleviate liver inflammation, thereby potentially improving alcoholic liver injury. For example, Lactobacillus helveticus can effectively alleviate alcoholic liver injury, primarily by regulating the intestinal microbiome, enhancing intestinal barrier integrity, and suppressing systemic inflammatory responses. Heat-killed Lactobacillus johnsonii (HKLJ) effectively alleviates liver injury caused by alcohol-related liver disease by activating the innate intestinal immune system. Lactobacillus rhamnosus NKU FL1-8 significantly alleviates acute alcoholic liver injury in mice by improving the liver's antioxidant system, reducing liver and colon inflammation, maintaining intestinal epithelial integrity, and regulating gut microbiota homeostasis. These beneficial bacteria have a positive role in the treatment of alcoholic liver disease.

[0003] Dubosiella newyorkensis (DN) is a Gram-positive, obligate anaerobic bacterium first isolated from the intestines of mice at a New York hospital in the United States. On solid culture media, the colonies are round, raised, and off-white to pale yellow. This obligate anaerobe has an optimal growth temperature of 37°C and an optimal pH of approximately 7.0. It can ferment a variety of carbohydrates, such as glucose, lactose, and maltose, producing acid without producing gas. Dubosiella newyorkensis has shown significant advantages in the treatment of metabolic diseases. Summary of the Invention

[0004] The purpose of the present invention is to provide a use of Duboisia in preparing a product for alleviating or treating alcoholic liver disease.

[0005] The embodiments of the present invention are achieved through the following technical solutions: A use of Duboisia in preparing a product for alleviating or treating alcoholic liver disease.

[0006] Preferably, the products include: products with anti-alcoholic liver activity and products with liver protective function.

[0007] Preferably, the products having anti-alcoholic liver activity include: products for alleviating or treating liver lipid abnormalities and products for alleviating or treating liver cell oxidative damage.

[0008] Preferably, the product for alleviating or treating liver lipid abnormalities includes: a product for improving abnormal lipid metabolism.

[0009] Preferably, the product for improving abnormal lipid metabolism includes: a product for improving cholesterol metabolism disorder.

[0010] Preferably, the product for alleviating or treating oxidative damage to liver cells includes: a product that enhances the ability to clear acetaldehyde.

[0011] Preferably, the products that exert a protective function on the liver include: products that improve and / or protect the intestinal structure, products that affect the abundance of intestinal flora, products that increase the diversity of intestinal flora, and products that maintain the balance of intestinal flora.

[0012] Preferably, the product that affects the abundance of intestinal flora includes: a product that reduces the abundance of Proteobacteria.

[0013] Preferably, the products for improving and / or protecting the intestinal structure include: products that affect the expression of tight junction proteins in the intestine, and products that restore the proliferation and regeneration capacity of intestinal epithelial cells.

[0014] Preferably, the products include: medicines and foods.

[0015] The present invention has at least the following beneficial effects: The present invention provides a new application of Dubosie bacteria, and by constructing alcoholic liver disease model mice and feeding them with a suspension of Dubosie bacteria, it is proved that Dubosie bacteria can be used as a raw material for preparing products for alleviating or treating alcoholic liver disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0017] Figure 1 This is a graph showing the test results of liver index and liver pathology analysis; Figure 2 The graph shows the test results of ALT and AST, ALDH and ADH, TC and TG, HDL and LDL, TNF-α and IL-6, MDA, and LPS; Figure 3 This is the detection result diagram of the changes in intestinal flora abundance; Figure 4 The figure shows the results of intestinal damage and Western Blotting. Figure 5 This is the result of immunofluorescence immunohistochemistry of ileum. DETAILED DESCRIPTION

[0018] In order to make the purpose, method scheme and advantages of the embodiments of the present invention clearer, the method scheme in the embodiments of the present invention is clearly and completely described. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0019] Example 1: Use of Duboisia in preparing a product for alleviating or treating alcoholic liver disease.

[0020] Example 2: The products include: products with anti-alcoholic liver activity and products with liver protective function.

[0021] Example 3: The products with anti-alcoholic liver activity include: products for alleviating or treating liver lipid abnormalities and products for alleviating or treating liver cell oxidative damage.

[0022] Example 4: The product for alleviating or treating abnormal liver lipids includes: a product for improving abnormal lipid metabolism.

[0023] Example 5: The product for improving abnormal lipid metabolism includes: a product for improving cholesterol metabolism disorder.

[0024] Example 6: The product for alleviating or treating oxidative damage to liver cells includes: a product that enhances the ability to clear acetaldehyde.

[0025] Example 7: The products that exert protective functions on the liver include: products that improve and / or protect the intestinal structure, products that affect the abundance of intestinal flora, products that increase the diversity of intestinal flora, and products that maintain the balance of intestinal flora.

[0026] Example 8: The product that affects the abundance of intestinal flora includes: a product that reduces the abundance of Proteobacteria.

[0027] Example 9: The products for improving and / or protecting the intestinal structure include: products that affect the expression of tight junction proteins in the intestine, and products that restore the proliferation and regeneration capacity of intestinal epithelial cells.

[0028] Example 10: The products include: medicines and foods.

[0029] In practice, Duboisia can be used in various pharmaceutical formulations. Certain formulations affect the rate at which the drug enters the patient's bloodstream. Therefore, some formulations are immediate-release, while others are delayed-release, sustained-release, or extended-release. Drugs can be granules, powders, tablets, capsules, sprays, solutions, emulsions, and injections, among others.

[0030] A validation trial of the effectiveness of Duboisia against alcoholic liver disease Dubosiella was purchased from Beijing Biobowei Biotechnology Co., Ltd., and the product name is bio-115795 Dubosiella newyorkensis.

[0031] 1. Culture of Duboisia nycticola GAM medium: Contains 5.0g peptone, 5.0g quinone, 5.0g soytone, 2.5g yeast extract powder, 2.5g beef powder, 10g digested serum powder, 1.2g beef liver extract powder, 0.5g glucose, 2.5g potassium dihydrogen phosphate, 3.0g sodium chloride, 5.0g soluble starch, 0.3g L-cysteine, 1.0g L-arginine, 0.2g L-tryptophan, 0.3g sodium thioglycolate, pH 7.3±0.1. Autoclave at 121°C for 15 minutes, cool to room temperature, add 1ml of sterile 0.1% vitamin K1 solution and 2mg of sterile hemin per 200ml of medium, mix well, and set aside. Liquid GAM differs from solid GAM medium in the presence or absence of agar.

[0032] Culture of Dubosiella nycticola: Add 500 μl of Dubosiella culture medium to every 10 ml of liquid culture medium, loosen the cap of the bacterial tube, place it in an anaerobic bag, and incubate at 37°C for 48 hours.

[0033] Preparation of bacterial suspension: Suspend Duboisia nycnicotinella in PBS buffer to a bacterial concentration of 5*108 CFU, which is the daily gavage dosage (mix thoroughly before gavage to avoid bacterial sedimentation that affects concentration deviation between gavage individuals).

[0034] 2. Establishment and Grouping of Animal Models Thirty-six SPF C57BL / 6 healthy male mice were randomly divided into three groups of 12 mice each after one week of adaptive feeding. The body weights were measured and recorded every three days from the beginning of the experiment. The mice were given oral administration: the blank control group received daily oral administration of normal saline; the alcoholic liver injury model group received oral administration of normal saline; and the drug intervention group received oral administration of 5×10 Duboisia nycopene. 8 CFU. The alcoholic liver disease model was established using a liquid feed method. After the experimental mice were grouped, the blank control group was first given a liquid control feed. The model and intervention groups were each given a mixture of control feed and ethanol feed (prepared at a volume ratio of 2:1, 1:1, and 1:2, respectively). Each concentration was fed for two days, for a total of six days. For the following 10 days, the blank control group was fed only the control feed, while the model and intervention groups were fed a 5% (v / v) ethanol feed. At 8:00 AM on the second day after modeling, the model group was gavage-administered with 31.5% (v / v) ethanol, while the blank control group was gavage-administered with 45% dextrin. Nine hours later, the mice were anesthetized with an intraperitoneal injection of 2% sodium pentobarbital. Body weights were measured, blood was collected from the orbitals, and liver and small intestinal tissue samples were collected. Liver weights were measured, and liver-to-body weight ratios were calculated.

[0035] 3. Pathological observation of liver tissue: The left lobe of mouse liver was fixed with 4% paraformaldehyde solution, embedded in paraffin, sliced, and then stained with hematoxylin and eosin dyes. The tissues were observed and photographed under a microscope.

[0036] 4. Serum-related index detection The effect of Duboisia nycronii in treating chronic alcoholic liver injury was evaluated in vivo by measuring the activity of serum transaminases, alcohol metabolism-related enzymes, liver fat density indicators, inflammatory cytokine content, etc. in mice, and performing pathological observation of liver tissue.

[0037] Serum sample preparation: Blood was collected and allowed to stand overnight at 4°C. Centrifuged at 2500 rpm for 15 minutes, and the supernatant was collected as serum. After collection, the serum was aliquoted and stored at -80°C to avoid repeated freezing and thawing.

[0038] Biochemical indexes determination: Mouse serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), acetaldehyde dehydrogenase (ALDH), alcohol dehydrogenase (ADH), low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglyceride (TG), total cholesterol (TC), malondialdehyde (MDA), lipopolysaccharide (LPS) were performed strictly according to the specific operating procedures of the corresponding kit instructions.

[0039] Determination of inflammatory factor expression: The expression levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the liver were detected by ELISA.

[0040] 5. Western Blotting After serum was collected at the end of the experiment on the 16th day, the mice were killed and dissected, and the colon was collected. Total protein was extracted according to the instructions of the BCA protein quantification kit, and the expression of occludin protein in the mouse colon was detected by Western Blot.

[0041] 6. Detection of changes in intestinal flora abundance On the 14th day of the experiment, feces of mice in each group were collected and quick-frozen in liquid nitrogen. DNA extraction, PCR amplification and high-throughput sequencing were performed by Shanghai Meiji Biotechnology Co., Ltd.

[0042] 7. Ileum immunohistochemistry: Mouse ileum tissues stored at -80°C were collected to determine the changes in ZO-1 and Occludin protein levels among different groups. The detection was performed by Sichuan Saiinster Biotechnology Co., Ltd.

[0043] 8. Ileal immunofluorescence detection: Mouse ileum tissues stored at -80°C were collected to determine the changes in LGR5+, LYZ1, and MUC2 protein levels among different groups. The detection was performed by Sichuan Saiinster Biotechnology Co., Ltd.

[0044] Experimental results Liver index: In the development of alcoholic liver disease, an increase in liver index usually means the occurrence of liver enlargement and fatty degeneration. By dynamically monitoring the liver index, the extent of liver damage and its development trend can be understood. Changes in liver index can be used as an important indicator for evaluating the effectiveness of treatment. In experiments using drugs or other interventions to treat alcoholic liver disease, a decrease in liver index usually means that the treatment is effective. Figure 1 As shown in Figures A and B, the liver index in the model group was significantly higher than that in the blank control group, indicating that the alcoholic liver disease model was successfully established. In the treatment groups, each drug showed a statistically significant decrease in liver index compared to the model group, indicating that Duboisia nycronii can effectively improve liver index.

[0045] Liver pathology analysis: By HE staining and Oil Red O fat staining of liver tissue sections, the pathological changes and fat accumulation of mouse liver tissue can be observed intuitively. Figure 1 As shown in Figures C, D, and E, in the blank control group, the hepatic lobule structure was clear, hepatocytes were arranged radially and neatly, the hepatic sinusoids were normal, and the cell nuclei were clearly structured. In contrast, the model group mice showed severe liver damage, and the alcohol model group mice showed disorganized hepatocytes, narrowed and dispersed hepatic sinusoids, hepatocyte necrosis, and inflammatory cell infiltration, among other pathological changes. Oil-red results revealed that the model group had a large amount of lipid accumulation in the liver. Combining the liver HE pathology score and the Oil-red-positive area rate, it was found that the use of Dubosiebacteriaceae significantly improved these pathological changes, further demonstrating that Dubosiebacteriaceae can effectively alleviate alcohol-induced hepatocyte damage and abnormal lipid accumulation in the liver.

[0046] ALT and AST analysis results: ALT and AST are mainly present in liver cells. When liver cells are damaged, these enzymes are released into the blood. Therefore, the increase of ALT and AST usually reflects the degree of damage to liver cells. The effect of Duboisia nycopene on the ALT and AST activity content in mice with alcoholic liver disease is shown in Figure 2. Figure 2 As shown in A and B, in the model group, serum ALT and AST levels were significantly higher than those in the blank control group, with statistically significant differences. Compared with the model group, serum ALT and AST levels in the treatment group were significantly lower, demonstrating that Duboisia nycator infection effectively reduces serum ALT and AST levels and alleviates liver damage.

[0047] Analysis of ALDH and ADH results: ADH and ALDH are key enzymes in alcohol metabolism. First, ADH catalyzes the conversion of ethanol into acetaldehyde, and then ALDH catalyzes the conversion of acetaldehyde into acetic acid, which finally enters the tricarboxylic acid cycle to produce carbon dioxide and water. Since the generated acetaldehyde has a direct toxic effect on liver cells, excessive intake of ethanol will inhibit the activity of liver ADH and ALDH to a certain extent. The results of the effect of the treatment group on the activity of ADH and ALDH in the liver of mice are shown in Figure 2,C,D. Compared with the blank control group, the ALDH activity in the liver of mice in the model group decreased significantly, with statistical significance. This indicates that Duboisia nycronii alleviates hepatocyte oxidative damage by enhancing acetaldehyde scavenging ability. The liver oxidative stress marker MDA decreased and glutathione (GSH) levels recovered, suggesting that the drug alleviates hepatocyte oxidative damage by enhancing acetaldehyde scavenging ability. In addition, the ADH level decreased, but not significantly, indicating that the lack of a significant increase in ADH activity may avoid the mismatch between acetaldehyde production and ALDH detoxification capacity caused by the rapid ethanol metabolism rate, further proving the rationality of the Duboisia nycronii intervention strategy.

[0048] Analysis of TC and TG results: In alcoholic liver disease, TC (total cholesterol) and TG (triglycerides) are common biochemical indicators, which have certain significance in liver health and disease development. TC is one of the important lipids in the body. It is mainly synthesized by the liver and circulates in the body. In alcoholic liver disease, liver cell damage may affect the synthesis and metabolism of cholesterol, leading to changes in TC levels in the blood. TG is the main form of lipid storage and also plays an important role in the liver. In alcoholic liver disease, abnormal liver lipid metabolism may lead to the accumulation of TG in the liver, which in turn causes fatty liver and other lesions. The results of the effects of the treatment group on TC and TG activity in the mouse liver are shown in Figure 2. Figure 2 E, F, Compared with the blank control group, the TC and TG levels in the livers of mice in the model group increased significantly, reaching statistical significance. Compared with the model group, the TC level of D. nycopene in the treatment group decreased significantly, reaching statistical significance. TG level decreased, but was not statistically significant. This suggests that D. nycopene may preferentially improve cholesterol metabolism disorders, while having limited effects on alcohol-induced TG accumulation, requiring further experimental verification.

[0049] Analysis of HDL and LDL results: In alcoholic liver disease, changes in HDL and LDL reflect abnormal liver function and lipid metabolism. A decrease in HDL may increase the risk of atherosclerosis and cardiovascular disease, while an increase in LDL may exacerbate the development of atherosclerosis. The results of the effects of the treatment group on HDL and LDL activity in mouse livers are shown in Figure 2 G, H. Compared with the blank control group, the model group showed a statistically significant decrease in HDL and a significant increase in LDL levels, demonstrating the successful establishment of this model. Compared with the model group, the treatment groups showed a statistically significant increase in HDL and a decrease in LDL levels. This suggests that Duboisia nycronii can improve alcohol-induced lipid metabolism abnormalities, effectively increasing HDL and reducing LDL levels.

[0050] Analysis of TNF-α and IL-6 results: TNF-α and IL-6 are generally considered to be inflammatory cytokines secreted by body tissues. Figure 2As shown in Figures I and J, TNF-α and IL-6 levels in the livers of mice in the model group increased significantly, while in the drug-treated group, treatment significantly reduced these levels. This result suggests that Duboisia nycronii can inhibit the alcohol-induced increase in inflammatory cytokines in the liver.

[0051] MDA analysis: MDA is the end product of lipid peroxidation and reflects the degree of oxidative damage to polyunsaturated fatty acids in cell membranes. Increased levels directly indicate cell membrane damage caused by free radicals (such as ROS). Alcohol exposure significantly increases liver MDA levels ( Figure 2 K), which was statistically different from the blank control group, indicating that ethanol-induced oxidative stress led to lipid peroxidation damage. The MDA level in the treatment group was significantly decreased, indicating that Duboisia nycronii inhibited the progression of this process.

[0052] LPS result analysis: LPS is a component of the outer membrane of Gram-negative bacteria. Alcohol can destroy intestinal tight junction proteins, leading to increased intestinal permeability, thereby affecting liver damage through the liver-gut axis. Figure 2 As shown in Figure L, Duboisia nycsec protects the intestine and indirectly protects the liver by affecting the expression of tight junction proteins in the intestine.

[0053] Analysis of intestinal flora abundance changes: 16S microbial diversity detection results are shown in Figure 3 From the Chao Index analysis ( Figure 3 A) The model group showed a higher bacterial richness than the blank control group, suggesting that alcohol may induce an increase in certain bacterial communities. With the intervention of Duboisia nycopene, the bacterial richness returned to normal. Figure 3 The Simpson index of the blank control group and the drug-treated group in B was higher, indicating that the intestinal flora diversity of these groups was higher, that is, the species richness and uniformity were higher. The Simpson index of the model group was lower, indicating that the intestinal flora diversity of the alcoholic liver model group was lower, which may be due to the influence of alcohol causing the number of certain flora to decrease or disappear. Combined with PCOA analysis ( Figure 3 C) Samples from the blank control group, the drug-treated group, and the model group were clearly clustered and separated, indicating significant differences in the bacterial community structure between the blank control group and the drug-treated group and the model group. The treatment group and the alcohol group had significant overlap, and the two data sets had some similarity in multidimensional space. Figure 3 D, E analyze the changes of microorganisms in each group at the phylum and genus levels respectively. Figure 3Figures F, G, H, I, and J show that the abundance of Firmicutes was slightly increased in the model group compared to the blank control group, while the abundance of Bacteroidetes decreased, but the differences between the groups did not reach statistical significance. Although no statistically significant differences were observed in Bacteroidetes and Firmicutes, the abundance of Proteobacteria was significantly increased. Overgrowth of Proteobacteria is a hallmark of alcoholic liver disease, activating hepatic inflammatory pathways by releasing lipopolysaccharide. Treatment with Duboisia nycronii significantly altered this phenomenon, potentially through preferential inhibition of Proteobacteria or modulation of specific genera rather than direct intervention in the balance between Firmicutes and Bacteroidetes. At the genus level, Lactobacillus levels in the treatment group returned to normal levels, achieving statistical significance. Compared to the model group, the abundance of Akkermansia decreased in the treatment group, but the difference was not significant. This indicates that although therapeutic intervention alleviates liver damage, intestinal mucosal repair may lag behind the improvement of liver function, resulting in delayed recovery of Akkermansia spp.

[0054] Analysis of intestinal damage and Western Blotting results: HE staining of the ileum and colon can more intuitively reveal the effects of alcohol on the intestine. Figure 4 As shown in A, in the blank control group, the structure of the ileum was intact, the intestinal villi were thin and long, arranged neatly, and the goblet cells were abundant. The colonic crypts were complete and clear, there was no inflammatory infiltration inside the structure, and the glands were neatly arranged. In contrast, the mice in the model group showed obvious intestinal damage. The ileal mucosal structure of the mice in the model group was damaged, the crypt structure partially disappeared, the glands were arranged in disorder, the villi were shortened, and large areas of villi were missing. Crypts appeared in the colon accompanied by inflammatory cell infiltration and a decrease in goblet cells. Combined with the pathological scores of the ileum and colon ( Figure 4 B, C, D, E), as well as changes in the length of intestinal villi and crypt depth in the ileum. These results indicate that the model group mice not only have structural destruction of the intestinal mucosal mechanical barrier (villous atrophy, crypt damage), but also impaired immune barrier function (inflammatory cell infiltration, goblet cell depletion), verifying the success of the model construction. The use of New York Duboisia can significantly improve intestinal damage and has a good protective effect on the intestinal barrier. Occludin is an important component of tight junction proteins and plays a key role in maintaining tight junctions and barrier function between cells. In alcoholic liver disease, alcohol and its metabolites may cause liver cell damage and inflammatory response, thereby destroying the tight junction structure and affecting the expression and function of Occludin. Figure 4As shown in Figure F, decreased occludin was observed in the model group, potentially leading to increased intercellular permeability, allowing harmful substances to more easily enter cells and exacerbate liver damage. Increased ocludin levels in the treatment group suggest that during treatment of alcoholic liver disease, ocludin expression and tight junction restoration improve the liver's barrier function.

[0055] Analysis of ileal immunofluorescence immunohistochemistry results: ZO-1 and Occludin are key proteins of tight junctions between intestinal epithelial cells. Their expression levels and distribution characteristics directly reflect the integrity of the intestinal barrier function. In alcoholic liver disease, alcohol damages the intestinal barrier, leading to increased intestinal permeability ("leaky gut"), promoting the translocation of bacterial endotoxins (such as LPS) to the liver, activating Kupffer cells and triggering liver inflammation and fibrosis. The degree of intestinal barrier damage was assessed by immunohistochemical detection of ileal ZO-1 and Occludin expression ( Figure 5 A, B, C). Compared with the blank control group, the model group showed decreased ZO-1 / Occludin expression, suggesting structural disruption of tight junctions. Combined with the reduction in serum LPS levels and improved liver pathology, this suggests that Duboisia nycronii may mitigate the progression of alcoholic liver injury by protecting the integrity of the intestinal barrier and inhibiting endotoxin translocation.

[0056] exist Figure 5 Duboisia nycronii was found in D, E, F, and G to exert multiple protective effects against alcohol-induced intestinal barrier damage. LGR5+ (an intestinal stem cell marker) reflects the proliferation and regenerative capacity of intestinal epithelial stem cells. In alcoholic liver disease, intestinal barrier damage is often accompanied by suppressed stem cell function, leading to impaired epithelial repair. Compared with the blank control group, the number of LGR5+ cells decreased in the model group, while the number of LGR5+ cells in the drug-treated group increased significantly, suggesting that Duboisia nycronii can restore stem cell proliferation to a certain extent. LYZ1 (lysozyme, a Paneth cell marker) Paneth cells secrete antimicrobial peptides (such as lysozyme) to maintain intestinal microbial balance. Alcohol may disrupt Paneth cell function and weaken innate immune defenses. LYZ1 expression was decreased in the model group, while the drug-treated group may restore Paneth cell activity through anti-inflammatory or antioxidant effects. MUC2 (mucin secreted by goblet cells) is a major component of the intestinal mucus layer, preventing direct contact between pathogens and the epithelium. Alcohol can reduce the number of goblet cells, leading to a thinning of the mucus layer. Compared with the model group, the expression of MUC2 in the drug-treated group increased significantly, promoting mucus secretion and repairing the mucus barrier.

[0057] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Various modifications and variations are readily apparent to those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. Use of Duboisia in preparing a product for alleviating or treating alcoholic liver disease.

2. The use according to claim 1, characterized in that The products include: products with anti-alcoholic liver activity and products that exert protective functions on the liver.

3. The use according to claim 2, characterized in that The products with anti-alcoholic liver activity include: products that alleviate or treat liver lipid abnormalities, products that alleviate or treat liver cell oxidative damage, and products that inhibit the increase in the content of inflammatory factors in the liver.

4. The use according to claim 3, characterized in that The products for alleviating or treating abnormal liver lipids include products for improving abnormal lipid metabolism.

5. The use according to claim 4, characterized in that The products for improving abnormal lipid metabolism include: products for improving cholesterol metabolism disorders.

6. The use according to claim 3, characterized in that The products for alleviating or treating oxidative damage to liver cells include products that enhance the ability to clear acetaldehyde.

7. The use according to any one of claims 1 to 6, characterized in that The products that exert protective functions on the liver include: products that improve and / or protect intestinal structure, products that affect the abundance of intestinal flora, products that increase the diversity of intestinal flora, and products that maintain the balance of intestinal flora.

8. The use according to claim 7, characterized in that The products that affect the abundance of intestinal flora include: products that reduce the abundance of Proteobacteria.

9. The use according to claim 7, characterized in that The products for improving and / or protecting the intestinal structure include: products that affect the expression of tight junction proteins in the intestine, and products that restore the proliferation and regeneration capacity of intestinal epithelial cells.

10. The use according to claim 7, characterized in that The products include: medicines and food.

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