Composition for preventing and / or treating alcoholic liver disease, product and application thereof

Abstract

The invention discloses a composition for preventing and / or treating alcoholic liver diseases, a product and application thereof. The present invention relates to a composition comprising: breast milk oligosaccharides and vitamins; the breast milk oligosaccharide comprises fucosyllactose and sialic acid lactose; the composition can provide nutritional support for patients with alcoholic liver diseases, repair intestinal mucosal barrier, reduce endotoxin entering blood, improve abundance of beneficial bacteria in intestinal tracts and reduce inflammatory response, so as to achieve the purpose of preventing and / or treating alcoholic liver diseases induced by alcohol.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to a composition, article, and application thereof for the prevention and / or treatment of alcoholic liver disease. Background Technology

[0002] Alcoholic liver disease (ALD) is a general term for a series of liver diseases caused by long-term heavy drinking. It usually manifests as fatty liver in the early stages, and can then develop into alcoholic hepatitis, liver fibrosis, cirrhosis, and even liver failure or hepatocellular carcinoma. ALD is one of the most common liver diseases worldwide.

[0003] The liver-gut axis is a crucial interactive mechanism for maintaining health. Excessive alcohol intake disrupts the intestinal barrier function, increases intestinal permeability, and allows microbial products such as lipopolysaccharides to enter the portal vein, thereby triggering liver inflammation and damaging the liver. Impaired intestinal barrier function also leads to decreased levels of short-chain fatty acids. Simultaneously, alcohol intake causes gut microbiota dysbiosis, and the reduction of probiotics further exacerbates the progression of ALD. Furthermore, alcohol alters the distribution and function of intestinal immune cells, increasing the risk of systemic inflammation. Therefore, repairing the intestinal barrier, restoring microbiota balance, and enhancing intestinal immune function are essential for controlling the progression of ALD. Human milk oligosaccharides (HMOs) are naturally occurring prebiotics in breast milk. Previous studies have found that HMOs have functions such as regulating gut microbiota, inhibiting pathogens, enhancing immune regulation, improving metabolic function, improving the intestinal barrier, and reducing endotoxin entry into the bloodstream; however, their application in the prevention and / or treatment of ALD is rarely reported.

[0004] Currently, personalized precision nutrition therapy is not yet widespread, and while routine nutritional interventions (such as vitamin supplementation) can improve nutritional status, they are not sufficiently targeted at liver-gut axis disorders, necessitating the exploration of more effective and universally applicable solutions. Therefore, research on HMOs in the prevention and / or treatment of ALD is of practical significance. Summary of the Invention

[0005] This invention provides a composition comprising: human milk oligosaccharides and vitamins; the human milk oligosaccharides include fucoidanol and sialylated lactose; the composition provides nutritional support to patients with alcoholic liver disease while repairing the intestinal mucosal barrier, reducing endotoxin entry into the blood, increasing the abundance of beneficial intestinal bacteria, and reducing inflammatory response, thereby achieving the purpose of preventing and / or treating alcohol-induced alcoholic liver disease.

[0006] The technical solution adopted in this invention is: The present invention provides a composition comprising: human milk oligosaccharides and vitamins; wherein the human milk oligosaccharides include fucoidanolose and sialylated lactose.

[0007] Preferably, the fucoidan is selected from 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), difucosyllactose (DFL), lactose-N-fucopentose I (LNFP I), lactose-N-fucopentose II (LNFP II), lactose-N-fucopentose III (LNFP III), lactose-N-fucopentose V (LNFP V), lactose-N-difucohexasose I (LNDFH I), lactose-N-difucohexasose II (LNDFH II), fucoidan-N-hexasose I (F-LNH I), fucoidan-N-hexasose II (F-LNH II), fucoidan-N-hexasose III (F-LNH III), trifucosyllactose-N-hexasose (TF-LNH), and difucosyllactose-N-hexasose a (DF-LNH-I). a) One or more of the following: 2'-fucosyllactose-N-hexasaccharide b (DF-LNH-I b), 2'-fucosyllactose-N-hexasaccharide II (DFLNH II), 2'-fucosyllactose-N-neohexose I (F-LNnH I), 2'-fucosyllactose-N-neohexose II (F-LNnH II), 2'-fucosyllactose-N-neohexose I (DFLNnH I), 2'-fucosyllactose-N-neohexose II (DFLNnH II), preferably 2'-fucosyllactose and / or 3'-fucosyllactose.

[0008] Preferably, the sialylated lactose is selected from 3'-sialylated lactose and / or 6'-sialylated lactose.

[0009] Preferably, the vitamin is selected from one or more of vitamin A, vitamin B, vitamin C, vitamin D, vitamin E, vitamin K, and vitamin P; more preferably, it is selected from one or more of vitamin B and vitamin D; more preferably, it is selected from vitamin B1, vitamin B2, vitamin B6, and vitamin B1. 12 One or more of vitamins D1 and D2; more preferably vitamins B1 and / or vitamin D3.

[0010] Preferably, the human milk oligosaccharides include 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose, and 6'-sialylated lactose.

[0011] Preferably, the composition comprises: 2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, vitamin B1, and vitamin D3.

[0012] Preferably, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is (25~35):(12~18):(12~18):(12~18), more preferably (27~32):(13~16):(13~16):(13~16), further preferably (29~30):(14~15):(14~15):(14~15), and even more preferably 29.1:14.55:14.55:14.55.

[0013] Preferably, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (25~35):(12~18):(12~18):(12~18):(0.1~3):(1*10) -4 ~1*10 -3 The preferred ratio is (27~32):(13~16):(13~16):(13~16):(1~2):(3*10). -4 ~7*10 -4 ), further preferably (29~30):(14~15):(14~15):(14~15):(1~1.5):(4*10 -4 ~6*10 -4 A further preferred ratio is 29.1:14.55:14.55:14.55:1:5*10. -4 .

[0014] Preferably, the human milk oligosaccharide further includes lactose-N-neotetrasaccharide.

[0015] Preferably, the human milk oligosaccharides include 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, and 6'-sialylated lactose.

[0016] Preferably, the composition comprises: 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3.

[0017] Preferably, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose and 6'-sialylated lactose is (25~35):(12~18):(20~30):(12~18):(12~18), more preferably (27~32):(13~16):(22~27):(13~16):(13~16), further preferably (29~30):(14~15):(24~25):(14~15):(14~15), and even more preferably 29.1:14.55:24.25:14.55:14.55.

[0018] Preferably, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (25~35):(12~18):(20~30):(12~18):(12~18):(0.1~3):(1*10) -4 ~1*10 -3 The preferred order is (27~32):(13~16):(22~27):(13~16):(13~16):(1~2):(3*10) -4 ~7*10 -4 ), further preferably (29~30):(14~15):(24~25):(14~15):(14~15):(1~1.5):(4*10 -4 ~6*10 -4 A further preferred ratio is 29.1:14.55:24.25:14.55:14.55:1:5*10. -4 .

[0019] Preferably, the composition further includes zinc.

[0020] Preferably, the zinc element is selected from one or more of zinc sulfate, zinc gluconate, zinc glycinate, zinc pyridinecarboxylate, zinc oxide, zinc lactate, zinc citrate, zinc chloride, zinc acetate, zinc carbonate, and zinc citrate trihydrate, with zinc gluconate being the most preferred.

[0021] Preferably, the composition comprises: 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, vitamin D3, and zinc gluconate.

[0022] Preferably, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, vitamin D3, and zinc gluconate is (25~35):(12~18):(20~30):(12~18):(12~18):(0.1~3):(1*10) -4 ~1*10 -3 ): (0.1~6), preferably (27~32): (13~16): (22~27): (13~16): (13~16): (1~2): (3*10 -4 ~7*10 -4 ): (2~4), further preferably (29~30): (14~15): (24~25): (14~15): (14~15): (1~1.5): (4*10 -4 ~6*10 -4 (2~3), and more preferably 29.1:14.55:24.25:14.55:14.55:1:5*10 -4 :2.

[0023] The present invention also provides an article comprising the composition described in any one of the preceding claims.

[0024] The present invention also provides the use of the compositions or articles described in any of the above claims in the preparation of products for the prevention and / or treatment of alcoholic liver disease.

[0025] Due to the adoption of the above technical solutions, the present invention has the following advantages: The compositions of this invention are effective in preventing and / or treating alcohol-induced alcoholic liver disease and have broad applicability. Specifically, HMOs can regulate gut microbiota, inhibit pathogens, reduce inflammatory responses, improve the intestinal barrier, and reduce endotoxin entry into the bloodstream; vitamin B1 can prevent nervous system damage and peripheral neuropathy; vitamin D3 can prevent osteoporosis and reduce the risk of infection; and zinc gluconate can stabilize intestinal barrier function, reduce endotoxemia, reduce the production of pro-inflammatory cytokines, reduce oxidative stress, and alleviate apoptotic hepatocyte death.

[0026] The composition of this invention contains a specific ratio and type of complex human milk oligosaccharides, which can effectively regulate intestinal flora, increase the abundance of beneficial intestinal flora, improve intestinal mucosal barrier damage, reduce endotoxin entry into the blood, reduce serum transaminase levels, reduce liver lipid levels, and reduce liver inflammatory response. In particular, the addition of LNnT significantly enhances the preventive and therapeutic effects. At the same time, it contains a specific ratio and type of micronutrients (such as vitamins and zinc), and its universality makes up for the limitation that individualized nutritional support is difficult to fully popularize. Among them, the addition of zinc gluconate further stabilizes intestinal barrier function, reduces the production of pro-inflammatory cytokines, and has a synergistic effect with human milk oligosaccharides and vitamins. Attached Figure Description

[0027] Figure 1 The effect of the composition in Example 2 of this invention on serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in ALD mice; Figure 2 The effect of the composition in Example 2 of this invention on the levels of total triglycerides (TG) and total cholesterol (TC) in the liver of ALD mice; Figure 3 The effect of the composition in Example 2 of this invention on the levels of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) in the liver of ALD mice; Figure 4 The effect of the composition in Example 2 of this invention on the levels of lipopolysaccharide (LPS) in the serum and feces of ALD mice; Figure 5 The effect of the composition in Example 2 of this invention on the Chao 1 index and Shannon index of the intestinal flora of ALD mice; Figure 6 This describes the effect of the composition in Example 2 of the present invention on the phylum level of the intestinal flora of ALD mice. Figure 7 This describes the effect of the composition in Example 2 of the present invention on the gut microbiota of ALD mice at the genus level. This application Figures 1 to 7 In the figure, * indicates that compared with the control group, *P<0.05, **P<0.01, ***P<0.001; # indicates that compared with the model group, #P<0.05, ##P<0.01, ###P<0.001. Detailed Implementation

[0028] To make the technical solution and beneficial effects of the present invention more apparent and understandable, a detailed description is provided below by listing specific embodiments. The accompanying drawings are not necessarily drawn to scale, and local features may be enlarged or reduced to more clearly show the details of the local features; unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical field to which this application pertains.

[0029] In this specification, the range of values ​​referred to as "value A to value B" refers to the range including the endpoint values ​​A and B.

[0030] In this specification, the numerical range indicated by "above" or "below" refers to the numerical range that includes the stated number.

[0031] In this specification, the word "may" has two meanings: to perform a certain process and not to perform a certain process.

[0032] The present invention provides a composition comprising: human milk oligosaccharides and vitamins; wherein the human milk oligosaccharides include fucoidanolose and sialylated lactose.

[0033] In some embodiments, the fucoidan is selected from 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), difucosyllactose (DFL), lactose-N-fucopentose I (LNFP I), lactose-N-fucopentose II (LNFP II), lactose-N-fucopentose III (LNFP III), lactose-N-fucopentose V (LNFP V), lactose-N-difucohexasose I (LNDFH I), lactose-N-difucohexasose II (LNDFH II), fucoidan-N-hexasose I (F-LNH I), fucoidan-N-hexasose II (F-LNH II), fucoidan-N-hexasose III (F-LNH III), trifucosyllactose-N-hexasose (TF-LNH), and difucosyllactose-N-hexasose a (DF-LNH-I). a) One or more of the following: fucosyllactose-N-hexasaccharide b (DF-LNH-I b), fucosyllactose-N-hexasaccharide II (DFLNH II), fucosyllactose-N-neohexose I (F-LNnHI), fucosyllactose-N-neohexose II (F-LNnH II), fucosyllactose-N-neohexose I (DFLNnH I), and fucosyllactose-N-neohexose II (DFLNnH II).

[0034] In some embodiments, the fucosyllactose is selected from 2'-fucosyllactose and / or 3-fucosyllactose.

[0035] In some embodiments, the fucosyllactose is selected from 2'-fucosyllactose and 3-fucosyllactose.

[0036] In some embodiments, the sialyl lactose is selected from 3'-sialyl lactose and / or 6'-sialyl lactose.

[0037] In some embodiments, the sialyl lactose is selected from 3'-sialyl lactose and 6'-sialyl lactose.

[0038] In some embodiments, the vitamin is selected from one or more of vitamin A, vitamin B, vitamin C, vitamin D, vitamin E, vitamin K, and vitamin P; preferably one or more of vitamin B and vitamin D.

[0039] In some embodiments, the vitamin is selected from vitamin B1, vitamin B2, vitamin B6, and vitamin B1. 12 One or more of vitamin D3.

[0040] In some embodiments, the vitamin is selected from vitamin B1 and / or vitamin D3.

[0041] In some embodiments, the human milk oligosaccharides include 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose, and 6'-sialylated lactose.

[0042] In some embodiments, the composition includes: 2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, vitamin B1, and vitamin D3.

[0043] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is (25~35):(12~18):(12~18):(12~18).

[0044] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is (27~32):(13~16):(13~16):(13~16).

[0045] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is (29~30):(14~15):(14~15):(14~15).

[0046] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is 29.1:14.55:14.55:14.55.

[0047] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (25~35):(12~18):(12~18):(12~18):(0.1~3):(1*10) -4 ~1*10 -3 ).

[0048] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (27~32):(13~16):(13~16):(13~16):(1~2):(3*10) -4 ~7*10 -4 ).

[0049] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (29~30):(14~15):(14~15):(14~15):(1~1.5):(4*10) -4 ~6*10 -4 ).

[0050] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is 29.1:14.55:14.55:14.55:1:5*10 -4 .

[0051] In some embodiments, the human milk oligosaccharide further includes lactose-N-neotetrasaccharide.

[0052] In some embodiments, the human milk oligosaccharides include 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, and 6'-sialylated lactose.

[0053] In some embodiments, the composition includes: 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3.

[0054] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose and 6'-sialylated lactose is (25~35):(12~18):(20~30):(12~18):(12~18).

[0055] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose and 6'-sialylated lactose is (27~32):(13~16):(22~27):(13~16):(13~16).

[0056] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose and 6'-sialylated lactose is (29~30):(14~15):(24~25):(14~15):(14~15).

[0057] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, and 6'-sialylated lactose is 29.1:14.55:24.25:14.55:14.55.

[0058] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (25~35):(12~18):(20~30):(12~18):(12~18):(0.1~3):(1*10) -4 ~1*10 -3 ).

[0059] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (27~32):(13~16):(22~27):(13~16):(13~16):(1~2):(3*10) -4 ~7*10 -4 ).

[0060] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (29~30):(14~15):(24~25):(14~15):(14~15):(1~1.5):(4*10)-4 ~6*10 -4 ).

[0061] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is 29.1:14.55:24.25:14.55:14.55:1:5*10 -4 .

[0062] In some embodiments, the composition further includes zinc.

[0063] In some embodiments, the zinc element is selected from one or more of zinc sulfate, zinc gluconate, zinc glycinate, zinc pyridinecarboxylate, zinc oxide, zinc lactate, zinc citrate, zinc chloride, zinc acetate, zinc carbonate, and zinc citrate trihydrate.

[0064] In some embodiments, the zinc element is selected from zinc gluconate.

[0065] In some embodiments, the composition includes: 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, vitamin D3, and zinc gluconate.

[0066] In some embodiments, the composition comprises 2'-fucosyllactose, 3-fucosyllactose, lactose-N-neotetrasaccharide, 3'-sialyllactose, 6'-sialyllactose, vitamin B1, vitamin D3, and zinc gluconate.

[0067] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, vitamin D3, and zinc gluconate is (25~35):(12~18):(20~30):(12~18):(12~18):(0.1~3):(1*10) -4 ~1*10 -3 ): (0.1~6).

[0068] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, vitamin D3, and zinc gluconate is (27~32):(13~16):(22~27):(13~16):(13~16):(1~2):(3*10) -4 ~7*10 -4 ): (2~4).

[0069] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, vitamin D3, and zinc gluconate is (29~30):(14~15):(24~25):(14~15):(14~15):(1~1.5):(4*10) -4 ~6*10 -4 ): (2~3).

[0070] In some embodiments, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, vitamin D3, and zinc gluconate is 29.1:14.55:24.25:14.55:14.55:1:5*10 -4 :2.

[0071] The present invention also provides an article comprising the composition described in any one of the preceding claims.

[0072] The present invention also provides the use of the compositions or articles described in any of the above claims in the preparation of products for the prevention and / or treatment of alcoholic liver disease.

[0073] In this invention, the abbreviations are: HMOs for human milk oligosaccharides; 2'-FL for 2'-fucosylated lactose; 3-FL for 3-fucosylated lactose; LNnT for lactose-N-neotetrasaccharide; 3'-SL for 3'-sialic acid lactose; and 6'-SL for 6'-sialic acid lactose.

[0074] In the following examples, all ingredients used are conventional commercially available products, which can be obtained from commercially available products or synthesized using known methods. For example, human milk oligosaccharides were produced by Huangshan Tongxi Biotechnology Co., Ltd.

[0075] The present invention will be described in more detail below through examples. Where specific experimental steps or conditions are not specified in the examples, they should be performed according to conventional practices or conditions in the art.

[0076] Example 1 According to the composition formulations shown in Table 1, weigh out HMOs (2'-FL, 3-FL, LNnT, 3'-SL or 6'-SL), vitamin B1, vitamin D3 and zinc gluconate respectively, mix them and dissolve them in 20 L of physiological saline to prepare different composition solutions.

[0077] Table 1. Composition Formulation

[0078] Example 2 1. Animal model establishment and intervention Thirty 6-week-old male BALB / c mice (weighing 18-20 g) were selected and provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. All mice were specific pathogen-free. Animals were housed following a 12-hour light-dark cycle, at a temperature of 25 ± 2°C and humidity of 50-70%. After acclimatization for 3 days with free access to food and water, they were acclimatized for 7 days with a non-alcoholic Lieber-DeCarli control diet. Subsequently, the mice were randomly divided into 5 groups (n=6 per group): A) Control group: Healthy mice were administered physiological saline by gavage (200 μL / mouse / day); B) Model group: Alcoholic liver disease mouse model + normal saline by gavage (200 μL / mouse / day); C) Composition 1 intervention group: Alcoholic liver disease mouse model + Composition 1 by gavage (200 μL / mouse / day); D) Composition 2 intervention group: Alcoholic liver disease mouse model + Composition 2 by gavage (200 μL / mouse / day); E) Composition 3 intervention group: Alcoholic liver disease mouse model + Composition 3 by gavage (200 μL / mouse / day).

[0079] Except for the control group, the proportion of alcohol in the diet of mice in other groups was gradually increased daily to allow them to adapt to alcohol. Starting on day 7, mice were fed a Lieber-DeCarli alcohol diet containing 5% (v / v) ethanol, while the control group mice were fed an equal-calorie, alcohol-free Lieber-DeCarli control diet. From day 14, mice in the composition intervention group were administered 200 μL of the corresponding composition solution by gavage daily, while mice in the control and model groups were given the same volume of physiological saline. Changes in body weight and food intake were recorded daily for each group until the end of the experiment on day 42.

[0080] 2. Sample Collection On day 42, mice from each group were placed in sterilized, clean metabolic cages, and at least 200 mg of fresh fecal samples were collected and placed in sterile tubes. These samples were then frozen in liquid nitrogen and stored at -80°C. After collecting blood from the orbital fossa, the mice were euthanized, and liver and small intestine tissue samples were collected. Whole blood samples were incubated at room temperature for 2 hours, centrifuged at 2000 r / min for 15 min, and the supernatant serum was collected and frozen at -80°C. Liver samples were weighed and the liver-to-body ratio was calculated before freezing at -80°C.

[0081] 3. Indicator detection and statistical analysis The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected by enzymatic methods; the contents of total triglycerides (TG) and total cholesterol (TC) in liver tissue were detected by colorimetric methods; the expression levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in liver tissue were detected by enzyme-linked immunosorbent assay (ELISA); the levels of lipopolysaccharide (LPS) in serum and feces were detected by ELISA; fecal DNA was extracted using a fecal DNA kit, and the V3-V4 hypervariable region of the 16S rRNA gene of intestinal flora was amplified and high-throughput sequenced using primers 341F (5'-CCTACGGGAGGCAGCAG-3') and 805R (5'-GGACTACVSGGGTATCTAAT-3').

[0082] Data are expressed as mean ± standard deviation. Unpaired t-tests were used to analyze statistical differences between two groups. One-way ANOVA and Tukey's multiple comparison test were used for three or more groups. A p-value less than 0.05 was considered statistically significant. * indicates that compared with the control group, *P<0.05, **P<0.01, ***P<0.001. # indicates that compared with the model group, #P<0.05, ##P<0.01, ###P<0.001.

[0083] 4. Experimental Results 4.1 Effects of the composition on mouse body weight, food intake, and liver index As shown in Table 2, compared with the control group, there was no statistically significant difference in food intake among the groups during the experiment, but the final body weight and liver index of the model group mice were significantly increased (P<0.05). Compared with the model group mice, the final body weight and liver index of the combination intervention group mice were significantly decreased (P<0.05).

[0084] Table 2 Effects of the composition on mouse body weight, food intake, and liver index

[0085] Note: Data are expressed as mean ± standard error (n=6), * indicates P<0.05 compared with the control group, and # indicates P<0.05 compared with the model group.

[0086] 4.2 The composition significantly reduced serum transaminase levels in ALD mice. Alanine aminotransferase (ALT) is mainly distributed in the cytoplasm of hepatocytes. When hepatocytes degenerate and cell membrane permeability increases, ALT is released from the cells into the bloodstream, leading to elevated blood ALT levels. Aspartate aminotransferase (AST) is mainly distributed in the mitochondria of hepatocytes. When hepatocytes are severely damaged or necrotic, AST is released from the mitochondria into the bloodstream, leading to elevated blood AST levels. Therefore, elevations in ALT and AST can reflect the degree of hepatocyte damage to some extent. Figure 1 As shown in A and B, the serum ALT and AST levels of mice in the model group were significantly higher than those in the control group. After the intervention of the composition, the serum ALT and AST levels of mice were significantly lower than those in the model group, especially in the LNnT (composition 2 intervention group) and zinc gluconate (composition 3 intervention group) enhancement groups, where the ALT and AST levels were further reduced, indicating that the composition intervention can effectively improve alcohol-induced hepatocellular damage.

[0087] 4.3 The composition significantly reduced liver lipid levels in ALD mice. Long-term excessive alcohol consumption can also lead to fat accumulation in the liver, causing repeated fatty degeneration, necrosis, and regeneration of liver cells, pushing alcoholic fatty liver further into alcoholic hepatitis, liver fibrosis, or cirrhosis. Figure 2 As shown in Figures A and B, continuous alcohol intake significantly increased the levels of total triglycerides (TG) and total cholesterol (TC) in the liver of mice in the model group compared to the control group. Conversely, the levels of TG and TC in the liver of mice in the three combined intervention groups were significantly lower than those in the model group, with the LNnT and zinc gluconate-enhanced groups showing even more pronounced reductions in TG and TC levels. This indicates that the combined intervention can effectively alleviate alcohol-induced hepatic fat accumulation.

[0088] 4.4 The composition significantly reduced liver inflammation levels in ALD mice. Inflammatory responses are a significant cause of liver damage in patients with alcoholic liver disease. Long-term excessive alcohol consumption induces stress and inflammation in hepatocytes, releasing cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). TNF-α is a crucial pro-inflammatory cytokine that exacerbates liver damage by promoting inflammation, oxidative stress, and cell necrosis. IL-6, on the other hand, is an anti-inflammatory factor with significant cytoprotective effects in alcoholic liver disease, playing a significant role in alleviating acute reactions, reducing liver inflammation, promoting hepatocyte regeneration, and reducing fatty liver. Detecting these two representative pro-inflammatory and anti-inflammatory factors can provide some insight into the level of liver inflammation. Figure 3As shown in A and B, long-term alcohol intake significantly increased the levels of TNF-α and IL-6 in the liver tissue of the model group mice compared to the control group. However, compared to the model group, the intervention of the three compositions could effectively reduce the levels of TNF-α and IL-6 in the liver tissue of mice. In particular, the LNnT enhanced groups (compositions 2 and 3) showed a more significant decrease in the levels of TNF-α and IL-6, indicating that the compositions can effectively reduce the level of inflammation in the liver of ALD mice.

[0089] 4.5 The composition significantly improved intestinal mucosal barrier damage in ALD mice. Lipopolysaccharide (LPS) is a component of the cell wall of Gram-negative bacteria. Alcohol can damage intestinal tight junction proteins, leading to increased intestinal permeability and thus increased entry of enterogenic LPS into the bloodstream. Therefore, serum LPS levels can reflect the extent of damage to the intestinal mucosal barrier. Figure 4 As shown in Figure A, the serum LPS level in the model group mice was significantly higher than that in the control group, while the serum LPS level in the three intervention groups was significantly lower than that in the model group. Composition 2, containing LNnT, was more effective than Composition 1, which did not contain LNnT. Adding zinc gluconate (Composition 3) in addition to LNnT further enhanced its effect on improving intestinal mucosal barrier damage in ALD mice. Continuous alcohol intake also leads to an increase in intestinal LPS levels, such as... Figure 4 As shown in Figure B, the LPS level in the feces of mice in the model group was significantly higher than that in the control group, while the intervention of the composition could reduce the LPS level in the feces to some extent.

[0090] 4.6 The composition significantly increased the abundance of beneficial gut bacteria in ALD mice. Changes in gut microbiota composition were characterized by sequencing the V3-V4 hypervariable region of the 16S rRNA gene of mouse feces. The results are as follows: Figure 5 As shown in A and B, compared with the control group, the Chao 1 index (reflecting the abundance of gut microbiota) and Shannon index (reflecting the diversity of gut microbiota) in the model group were significantly reduced, indicating that long-term alcohol intake reduces the abundance and diversity of gut microbiota. The three combination interventions all improved the abundance and diversity parameters of gut microbiota in mice to varying degrees, with the enhancement of LNnT being more beneficial to improving the diversity of gut microbiota in ALD mice.

[0091] Figure 6 and Figure 7 The changes in the gut microbiota of mice in each group were shown at the phylum and genus levels, respectively. Figure 6 A, B, and C show that, compared to the control group, the model group had a higher incidence of Firmicutes ( ). Firmicutes ) and Proteobacteria ( Proteobacteria The relative abundance of Bacteroidetes (Bacteroidetes) increased significantly. BacteroidetesThe relative abundance of Firmicutes decreased significantly. Excessive proliferation of Proteobacteria is a hallmark of alcoholic liver disease, activating hepatic inflammatory pathways through the release of lipopolysaccharides; this phenomenon was significantly altered by the composition. Although the composition had a smaller effect on the relative abundance of Firmicutes, the combination... Figure 7 Based on the main differences in gut microbiota at the genus level among the various groups of mice, the combined intervention significantly increased beneficial bacteria such as Lactobacillus in Firmicutes. Lactobacillus ), Akkermania ( Akkermansia ) and Prevotella spp. Faecalibacterium prausnitzii The relative abundance of Firmicutes (Bacteria) is likely the main reason why there was no significant change in Firmicutes compared to the model group. At the genus level, long-term alcohol intake increased the abundance of *Desulfovibrio* (Bacteria) compared to the control group. Desulfovibrio The relative abundance of Lactobacillus, Akkermansia, and Prevotella was significantly increased, accompanied by a significant decrease in the relative abundance of Lactobacillus, Akkermansia, and Prevotella. All three combination interventions could significantly reverse the changes in the relative abundance of the above gut microbiota. Comparative analysis showed that adding LNnT was more beneficial in improving alcohol-induced changes in gut microbiota and increasing the abundance of beneficial bacteria.

[0092] It should be understood that the above embodiments are exemplary and are not intended to encompass all possible implementations included in the claims. Various modifications and changes can be made to the above embodiments without departing from the scope of this disclosure. Similarly, the various technical features of the above embodiments can be arbitrarily combined to form other embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments only illustrate several implementations of the present invention and do not limit the scope of protection of this patent.

Claims

1. A composition, characterized in that, The composition comprises: human milk oligosaccharides and vitamins; the human milk oligosaccharides include fucoidyl lactose and sialylated lactose.

2. The composition according to claim 1, characterized in that, The fucoidan is selected from 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), difucosyllactose (DFL), lactose-N-fucopentose I (LNFP I), lactose-N-fucopentose II (LNFP II), lactose-N-fucopentose III (LNFP III), lactose-N-fucopentose V (LNFPV), lactose-N-difucohexasose I (LNDFH I), lactose-N-difucohexasose II (LNDFH II), fucoidan-N-hexasose I (F-LNH I), fucoidan-N-hexasose II (F-LNH II), fucoidan-N-hexasose III (F-LNH III), trifucosyllactose-N-hexasose (TF-LNH), and difucosyllactose-N-hexasose a (DF-LNH-I). a) One or more of the following: df-fucosyllactose-N-hexasaccharide b (DF-LNH-I b), df-fucosyllactose-N-hexasaccharide II (DFLNH II), df-fucosyllactose-N-neohexose I (F-LNnH I), df-fucosyllactose-N-neohexose II (F-LNnH II), df-fucosyllactose-N-neohexose I (DFLNnH I), and df-fucosyllactose-N-neohexose II (DFLNnH II), preferably 2'-fucosyllactose and / or 3-fucosyllactose; Preferably, the sialylated lactose is selected from 3'-sialylated lactose and / or 6'-sialylated lactose; Preferably, the vitamin is selected from one or more of vitamin A, vitamin B, vitamin C, vitamin D, vitamin E, vitamin K, and vitamin P; more preferably, it is selected from one or more of vitamin B and vitamin D; more preferably, it is selected from vitamin B1, vitamin B2, vitamin B6, and vitamin B1. 12 One or more of vitamins D1 and D2; more preferably vitamins B1 and / or vitamin D3.

3. The composition according to claim 2, characterized in that, The human milk oligosaccharides include 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose, and 6'-sialylated lactose; Preferably, the composition comprises: 2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, vitamin B1, and vitamin D3.

4. The composition according to claim 3, characterized in that, The mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose and 6'-sialylated lactose is (25~35):(12~18):(12~18):(12~18), preferably (27~32):(13~16):(13~16):(13~16), further preferably (29~30):(14~15):(14~15):(14~15), and even more preferably 29.1:14.55:14.55:14.55; Preferably, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (25~35):(12~18):(12~18):(12~18):(0.1~3):(1*10) -4 ~1*10 -3 The preferred ratio is (27~32):(13~16):(13~16):(13~16):(1~2):(3*10). -4 ~7*10 -4 ), further preferably (29~30):(14~15):(14~15):(14~15):(1~1.5):(4*10 -4 ~6*10 -4 A further preferred ratio is 29.1:14.55:14.55:14.55:1:5*10. -4 .

5. The composition according to claim 4, characterized in that, The human milk oligosaccharides also include lactose-N-neotetrasaccharide; Preferably, the human milk oligosaccharides include 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, and 6'-sialylated lactose; Preferably, the composition comprises: 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3.

6. The composition according to claim 5, characterized in that, The mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose and 6'-sialylated lactose is (25~35):(12~18):(20~30):(12~18):(12~18), preferably (27~32):(13~16):(22~27):(13~16):(13~16), further preferably (29~30):(14~15):(24~25):(14~15):(14~15), and even more preferably 29.1:14.55:24.25:14.55:14.55; Preferably, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, and vitamin D3 is (25~35):(12~18):(20~30):(12~18):(12~18):(0.1~3):(1*10) -4 ~1*10 -3 The preferred order is (27~32):(13~16):(22~27):(13~16):(13~16):(1~2):(3*10) -4 ~7*10 -4 ), further preferably (29~30):(14~15):(24~25):(14~15):(14~15):(1~1.5):(4*10 -4 ~6*10 -4 A further preferred ratio is 29.1:14.55:24.25:14.55:14.55:1:5*10. -4 .

7. The composition according to claim 6, characterized in that, The composition further includes: zinc element; Preferably, the zinc element is selected from one or more of zinc sulfate, zinc gluconate, zinc glycinate, zinc pyridinecarboxylate, zinc oxide, zinc lactate, zinc citrate, zinc chloride, zinc acetate, zinc carbonate, and zinc citrate trihydrate, with zinc gluconate being the most preferred.

8. The composition according to claim 7, characterized in that, The composition comprises: 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, vitamin D3, and zinc gluconate; Preferably, the mass ratio of 2'-fucosylated lactose, 3-fucosylated lactose, lactose-N-neotetrasaccharide, 3'-sialylated lactose, 6'-sialylated lactose, vitamin B1, vitamin D3, and zinc gluconate is (25~35):(12~18):(20~30):(12~18):(12~18):(0.1~3):(1*10) -4 ~1*10 -3 ): (0.1~6), preferably (27~32): (13~16): (22~27): (13~16): (13~16): (1~2): (3*10 -4 ~7*10 -4 ): (2~4), further preferably (29~30): (14~15): (24~25): (14~15): (14~15): (1~1.5): (4*10 -4 ~6*10 -4 (2~3), and more preferably 29.1:14.55:24.25:14.55:14.55:1:5*10 -4 :

2.

9. An article, characterized in that, The article comprises the composition according to any one of claims 1 to 8.

10. The use of the composition according to any one of claims 1 to 8 or the article according to claim 9 in the preparation of products for the prevention and / or treatment of alcoholic liver disease.

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