Composition for improving alcoholic liver injury and intestinal injury and preparation method thereof

Abstract

The invention discloses a composition for improving alcoholic liver injury and intestinal injury and a preparation method thereof, and relates to the technical field of medicines. The composition is prepared from the following components in parts by weight: 20 to 30 parts of radix puerariae polysaccharide, 10 to 15 parts of betaine, 8 to 14 parts of esculetin and 5 to 12 parts of quercetin. Through drug combination, the levels of ALT and AST in serum are remarkably reduced, pathological injuries of liver and intestinal tissues are improved, oxidative stress and inflammatory response are relieved, and the activity of alcohol metabolic enzyme is increased, so that the drug combination has remarkable liver protection and intestinal repair effects and shows potential drug combination value. Meanwhile, the components of the composition are wide in source, low in cost and easy to obtain, the preparation method is simple, the effect is remarkable, and the composition has a very good market development prospect.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to a composition for improving alcoholic liver damage and intestinal damage and a preparation method thereof. Background Art

[0002] Alcohol-induced liver damage and intestinal damage are common health problems caused by long-term excessive drinking. Alcoholic liver disease (ALD) is one of the main causes of death from liver diseases. It is reported that oxidative stress and inflammatory response play an important role in the development of ALD, and compounds with antioxidant or anti-inflammatory activity can significantly improve ALD. Such as corn germ powder albumin peptides, black bean-derived peptides, soybean meal peptides, coix seed protein hydrolysate, quercetin polyphenols, turmeric polyphenols, kaempferol, lotus root polyphenols, etc. In addition, reactive oxygen species (ROS) are produced during alcohol metabolism in the body. ROS can damage biological macromolecules such as proteins, lipids and nucleic acids, destroying their normal structure and function. ROS may also cause lipid peroxidation and promote liver oxidative stress, trigger cell apoptosis and inflammation, and ultimately lead to liver damage.

[0003] Alcoholic liver disease is an important cause of chronic liver disease, and intestinal barrier damage and intestinal flora imbalance are important pathogenesis. Intestinal epithelial cells are tightly bound together by tight junction proteins, covered with mucus, colonized with intestinal flora, and together with the immune cells of the lamina propria, form an intestinal barrier. After drinking, the normal symbiotic relationship between the intestinal flora and the host is destroyed, the diversity of the flora is reduced, the species and number of the flora change to varying degrees, the integrity of the intestinal epithelial tight junctions is destroyed, and the intestinal permeability increases, causing intestinal damage.

[0004] Currently, the treatment of alcohol-induced liver and intestinal damage mainly includes drug intervention and surgical treatment. However, long-term intake of chemical drugs may increase the burden on the digestive system and liver, and surgical treatment is generally only used for critically ill patients. Therefore, it is urgent to develop natural products or food-derived compounds as compositions to prevent alcohol-induced liver and intestinal damage. Summary of the invention

[0005] In view of the above-mentioned defects of the prior art, the present invention proposes a composition for improving alcoholic liver damage and intestinal damage and a preparation method thereof, so as to solve the problems raised by the above-mentioned background technology.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] A composition for improving alcoholic liver damage and intestinal damage, comprising the following components in parts by weight: 20-30 parts of kudzu root polysaccharide, 10-15 parts of betaine, 8-14 parts of esculenta and 5-12 parts of quercetin.

[0008] Preferably, the composition comprises the following components in parts by weight: 26 parts of pueraria polysaccharide, 13 parts of betaine, 10 parts of esculenta and 7 parts of quercetin.

[0009] Preferably, quercetin is a natural ingredient extracted from the dried root of Gentiana macrophylla Pall., a plant of the Gentianaceae family, and is purchased from Van der Biotech.

[0010] Preferably, the preparation method of Pueraria polysaccharide comprises the following steps:

[0011] (1) Weigh the washed and dried kudzu root and crush it, add water, place it in a 70-90° C. water bath and stir to extract for 2-5 hours, repeat the extraction 2-4 times, filter and combine all the extracts, cool it and concentrate it under reduced pressure, and then wash the precipitate with 75-80% by volume ethanol for 1-3 times to obtain a precipitate;

[0012] (2) drying the precipitate obtained in step (1) in a vacuum drying oven to obtain polysaccharide powder; dissolving the polysaccharide powder in water to obtain a polysaccharide solution for later use; wherein the mass volume ratio of the polysaccharide powder to water is 1 g: (3-6) mL;

[0013] (3) The polysaccharide solution obtained in step (2) is mixed with α-amylase solution, placed in a 40° C. water bath for 15-30 min, and then boiled to inactivate the enzyme. Ethanol with a volume fraction of 85-95% is then added to precipitate the enzyme protein, and the mixture is centrifuged and vacuum dried to obtain Pueraria polysaccharide.

[0014] Preferably, in step (1), the mass volume ratio of Pueraria root to water is 1 g: (10-15) mL; the volume ratio of the extract to ethanol is 1 mL: (3-6) mL.

[0015] Preferably, in step (3), the volume ratio of the polysaccharide solution, the α-amylase solution and the ethanol is 1 mL: (2-3) mL: (5-10) mL.

[0016] Preferably, the α-amylase solution is prepared by dissolving 0.02 g of 5000 u / g amylase powder in 500 ml of pH 4.5 acetate buffer.

[0017] Preferably, the preparation method of aescin comprises the following steps:

[0018] ① Take 150g of Cortex fraxini (the dried branch bark or dry bark of Fraxini fraxini, Fraxinus fraxini, Fraxinus truncatus or Fraxinus truncatus of the Oleaceae family) coarse powder in a Soxhlet extractor, add 400ml of ethanol and reflux for 8-12h to obtain an ethanol extract, and recover the solvent under reduced pressure to an extract;

[0019] ② Add 40 ml of water to the extract and heat to dissolve, extract with an equal volume of chloroform for 30 to 50 minutes, separate the liquid to obtain the aqueous phase, add an equal volume of ethyl acetate to the aqueous phase for extraction to obtain ethyl acetate solution, dehydrate with anhydrous sodium sulfate, and recover the solvent under reduced pressure to dryness to obtain a dry product;

[0020] ③ Add 50-60℃ methanol to the dried product and let it stand for crystallization. Yellow needle-shaped crystals will precipitate. Filter out the crystals to obtain esculetin.

[0021] Preferably, in step ③, the mass volume ratio of the dried product to methanol is 1 g: (5-8) mL.

[0022] Preferably, the composition is in the form of an oral preparation.

[0023] Preferably, a method for preparing a composition for improving alcoholic liver damage and intestinal damage comprises mixing or adding water to uniformly disperse the components.

[0024] Preferably, the composition of the present invention is used in the preparation of products for improving alcoholic liver damage and intestinal damage.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The combined use significantly reduced the levels of ALT and AST in serum, improved the pathological damage of liver and intestinal tissues, reduced oxidative stress and inflammatory response, and increased the activity of alcohol metabolic enzymes, indicating that the combined use has significant liver protection and intestinal repair effects, showing potential combined use value. At the same time, the components of the composition are widely available, cheap and easy to obtain, the preparation method is simple, the effect is significant, and it has a good market development prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is the change chart of serum ALT and AST levels;

[0028] Figure 2 This is the result of HE staining;

[0029] Figure 3 This is the result of Sirius red staining;

[0030] Figure 4 This is the result of colon PAS staining;

[0031] Figure 5This is the immunohistochemical change of colon MUC-2;

[0032] Figure 6 This is the result of ZO-1 staining;

[0033] Figure 7 This is the result of Occludin staining;

[0034] Figure 8 This is a graph showing changes in the activity of alcohol metabolism enzymes ADH and ALDH;

[0035] Fig. 9 This is a graph showing changes in the activity of alcohol-metabolizing enzyme CYP2E1;

[0036] Fig.10 This is a graph showing changes in inflammatory factors; A is IL-6; B is TNF-α; and C is IL-1β. DETAILED DESCRIPTION

[0037] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods.

[0038] Example 1

[0039] A composition for improving alcoholic liver damage and intestinal damage, comprising the following components in parts by weight: 20 parts of kudzu root polysaccharide, 10 parts of betaine, 8 parts of esculenta and 5 parts of quercetin.

[0040] The preparation method of Pueraria polysaccharide comprises the following steps:

[0041] (1) Weigh and crush the washed and dried kudzu root, add water, place in a 70°C water bath, stir and extract for 2 h, repeat the extraction twice, filter and combine all the extracts, cool and concentrate under reduced pressure, and then wash the precipitate once with 75% by volume ethanol to obtain a precipitate; wherein the mass volume ratio of kudzu root to water is 1 g:10 mL; the volume ratio of the extract to ethanol is 1 mL:3 mL;

[0042] (2) drying the precipitate obtained in step (1) in a vacuum drying oven to obtain polysaccharide powder; dissolving the polysaccharide powder in water to obtain a polysaccharide solution for later use; wherein the mass volume ratio of the polysaccharide powder to water is 1 g:3 mL;

[0043] (3) The polysaccharide solution obtained in step (2) is mixed with α-amylase solution, placed in a 40° C. water bath for 15 min, and then boiled to inactivate the enzyme, and then 85% by volume ethanol is added to precipitate the enzyme protein, and centrifuged and vacuum dried to obtain Pueraria polysaccharide; wherein the volume ratio of the polysaccharide solution, α-amylase solution and ethanol is 1 mL:2 mL:5 mL.

[0044] The preparation method of aescin comprises the following steps:

[0045] ① Take 150g of Cortex Fraxini crude powder in a Soxhlet extractor, add 400ml of ethanol and reflux for 8h to obtain an ethanol extract, and recover the solvent under reduced pressure to an extract;

[0046] ② Add 40 ml of water to the extract and heat to dissolve, extract with an equal volume of chloroform for 30 minutes, separate the liquid to obtain the aqueous phase, add an equal volume of ethyl acetate to the aqueous phase for extraction to obtain ethyl acetate solution, dehydrate with anhydrous sodium sulfate, and recover the solvent under reduced pressure to dryness to obtain a dry product;

[0047] ③ Add 60℃ methanol to the dried product and let it stand for crystallization, yellow needle-shaped crystals will precipitate, filter out the crystals, and obtain esculetin. The mass volume ratio of the dried product to methanol is 1g:5mL.

[0048] The dosage form of the composition is an oral preparation.

[0049] A preparation method of a composition for improving alcoholic liver damage and intestinal damage comprises mixing or adding water to disperse the components uniformly.

[0050] Example 2

[0051] A composition for improving alcoholic liver damage and intestinal damage, comprising the following components in parts by weight: 30 parts of kudzu root polysaccharide, 15 parts of betaine, 14 parts of esculent acid and 12 parts of quercetin.

[0052] The preparation method of Pueraria polysaccharide comprises the following steps:

[0053] (1) Weigh and crush the washed and dried kudzu root, add water, place in a 90°C water bath, stir and extract for 5 h, repeat the extraction 4 times, filter and combine all the extracts, cool and concentrate under reduced pressure, and then wash the precipitate 3 times with 80% ethanol by volume to obtain a precipitate; wherein the mass volume ratio of kudzu root to water is 1 g:15 mL; the volume ratio of the extract to ethanol is 1 mL:6 mL;

[0054] (2) drying the precipitate obtained in step (1) in a vacuum drying oven to obtain polysaccharide powder; dissolving the polysaccharide powder in water to obtain a polysaccharide solution for later use; wherein the mass volume ratio of the polysaccharide powder to water is 1 g:6 mL;

[0055] (3) The polysaccharide solution obtained in step (2) is mixed with α-amylase solution, placed in a 40° C. water bath for 30 min, and then boiled to inactivate the enzyme, and then 95% by volume ethanol is added to precipitate the enzyme protein, and centrifuged and vacuum dried to obtain Pueraria polysaccharide; wherein the volume ratio of the polysaccharide solution, α-amylase solution and ethanol is 1 mL:3 mL:10 mL.

[0056] The preparation method of aescin comprises the following steps:

[0057] ① Take 150g of Cortex Fraxini crude powder in a Soxhlet extractor, add 400ml of ethanol and reflux for 12h to obtain an ethanol extract, and recover the solvent under reduced pressure to an extract;

[0058] ② Add 40 ml of water to the extract and heat to dissolve, extract with an equal volume of chloroform for 50 min, separate the liquid to obtain the aqueous phase, add an equal volume of ethyl acetate to the aqueous phase for extraction to obtain ethyl acetate solution, dehydrate with anhydrous sodium sulfate, recover the solvent under reduced pressure to dryness, and obtain a dry product;

[0059] ③ Add 60°C methanol to the dried product and let it stand for crystallization, yellow needle-shaped crystals will precipitate. Filter out the crystals to obtain esculetin, wherein the mass volume ratio of the dried product to methanol is 1g:8mL.

[0060] The dosage form of the composition is an oral preparation.

[0061] A preparation method of a composition for improving alcoholic liver damage and intestinal damage comprises mixing or adding water to disperse the components uniformly.

[0062] Example 3

[0063] A composition for improving alcoholic liver damage and intestinal damage, comprising the following components in parts by weight: 26 parts of kudzu root polysaccharide, 13 parts of betaine, 10 parts of esculent acid and 7 parts of quercetin.

[0064] The preparation method of Pueraria polysaccharide comprises the following steps:

[0065] (1) Weigh and crush the washed and dried kudzu root, add water, place in a water bath at 80°C, stir and extract for 4 h, repeat the extraction 3 times, filter and combine all the extracts, cool and concentrate under reduced pressure, and then wash the precipitate twice with 78% ethanol by volume to obtain a precipitate; wherein the mass volume ratio of kudzu root to water is 1 g:12 mL; the volume ratio of the extract to ethanol is 1 mL:5 mL;

[0066] (2) drying the precipitate obtained in step (1) in a vacuum drying oven to obtain polysaccharide powder; dissolving the polysaccharide powder in water to obtain a polysaccharide solution for later use; wherein the mass volume ratio of the polysaccharide powder to water is 1 g:5 mL;

[0067] (3) The polysaccharide solution obtained in step (2) is mixed with α-amylase solution, placed in a 40° C. water bath for 20 min, and then boiled to inactivate the enzyme, and then 90% by volume ethanol is added to precipitate the enzyme protein, and centrifuged and vacuum dried to obtain Pueraria polysaccharide; wherein the volume ratio of the polysaccharide solution, α-amylase solution and ethanol is 1 mL:3 mL:8 mL.

[0068] The preparation method of aescin comprises the following steps:

[0069] ① Take 150g of Cortex Fraxini crude powder in a Soxhlet extractor, add 400ml of ethanol and reflux for 10h to obtain an ethanol extract, and recover the solvent under reduced pressure to an extract;

[0070] ② Add 40 ml of water to the extract and heat to dissolve, extract with an equal volume of chloroform for 40 min, separate the liquid to obtain the aqueous phase, add an equal volume of ethyl acetate to the aqueous phase for extraction to obtain ethyl acetate solution, dehydrate with anhydrous sodium sulfate, recover the solvent under reduced pressure to dryness, and obtain a dry product;

[0071] ③ Add 55°C methanol to the dried product and allow to stand for crystallization, whereupon yellow needle-shaped crystals are precipitated. The crystals are filtered out to obtain esculetin; the mass volume ratio of the dried product to methanol in step ③ is 1g:7mL.

[0072] The dosage form of the composition is an oral preparation.

[0073] A preparation method of a composition for improving alcoholic liver damage and intestinal damage comprises mixing or adding water to disperse the components uniformly.

[0074] Experimental example

[0075] 1. Experimental Methods

[0076] 1.1 Experimental animals and experimental protocol

[0077] Experimental animals: Six-week-old male C57BL / 6J mice were housed in a standard animal laboratory with a constant room temperature of (22±2)°C, humidity of (50±5)%, 12h light / 12h dark cycle, free access to food and water, and the experiments were conducted after one week of adaptive feeding.

[0078] Experimental plan: The mice were randomly divided into control group (CON), alcohol model group (MOD), Pueraria polysaccharide treatment group (PLP1), betaine treatment group (Betaine), Pueraria polysaccharide + betaine + esculetin + quercetin combined with high-dose treatment group (HD-Combination) and Pueraria polysaccharide + betaine + esculetin + quercetin combined with low-dose treatment group (LD-Combination), with 8 mice in each group, for a total of 48 mice.

[0079] The alcohol model group and each treatment group were given a 16-day liquid diet NIAAA model (refer to Gao Bin et al., Alcoholic liver disease mouse model induced by chronic alcohol feeding plus acute alcohol gavage, [J], 2013, 26(5)). The pueraria polysaccharide treatment group was given pueraria polysaccharide (400 mg / kg) solution, the betaine treatment group was given betaine (200 mg / kg) solution, the combined low-dose treatment group was given pueraria polysaccharide (200 mg / kg) + betaine (100 mg / kg) + esculetin (78 mg / kg) + quercetin (53.8 mg / kg) solution, and the combined high-dose treatment group was given pueraria polysaccharide (400 mg / kg) and betaine (200 mg / kg) + esculetin (156 mg / kg) + quercetin (107.6 mg / kg) solution for 10 consecutive days. The control group was not given any modeling treatment.

[0080] 1.2 Sampling and sample preparation

[0081] After the administration, the mice were fasted overnight but not watered for 12 h, anesthetized with tribromoethanol, and serum samples were collected for biochemical index detection. Liver and intestinal tissues were weighed, some tissues were stored at -80 °C for subsequent analysis, and some tissues were fixed in 4% paraformaldehyde for histological analysis.

[0082] 1.3 Detection indicators

[0083] 1.3.1 Liver function and intestinal permeability testing

[0084] Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured to reflect liver function. Intestinal permeability was measured by tight junction proteins and mucin to evaluate intestinal barrier function.

[0085] 1.3.2 Histopathological observation

[0086] Liver and intestinal tissues were stained with hematoxylin-eosin (HE), liver Sirius red, colon PAS, colon MUC-2 immunohistochemistry, and colon tight junction protein (ZO-1, Occludin) to observe pathological changes in tissues. Optical microscopy was used to take pictures and evaluate liver cell damage, fibrosis, and intestinal barrier damage.

[0087] Liver hematoxylin-eosin (HE) staining: used to evaluate the morphological changes, inflammatory response, fatty degeneration and tissue structural damage of hepatocytes, and is an important tool for studying and diagnosing liver diseases.

[0088] Sirius red staining: used to evaluate the degree of liver fibrosis. Sirius red staining is used to observe the distribution and density of collagen fibers in liver tissue to reflect the progression of fibrosis.

[0089] Colon PAS staining: used to detect the number of goblet cells and mucus secretion in colon tissue. PAS staining can show changes in polysaccharides and glycoproteins in the colon to evaluate the effects of alcohol on the intestinal mucosa.

[0090] Colon MUC-2 immunohistochemistry: used to detect the expression level of MUC-2 protein in colon tissue. MUC-2 is the main component of the intestinal mucus layer and can reflect the function of the intestinal mucus barrier.

[0091] Immunofluorescence staining of colon tight junction proteins (ZO-1, Occludin): used to evaluate the expression of tight junction proteins in the colon epithelium and to evaluate the integrity of the intestinal barrier by observing the expression levels of ZO-1 and Occludin.

[0092] 1.3.3 Determination of alcohol metabolizing enzyme (ADH, ALDH, CYP2E1) activity

[0093] Commercial kits were used to measure the activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) in the liver. Part of the liver tissue was removed and homogenized, and the enzyme activities of ADH and ALDH were measured according to the instructions of the kit to evaluate the effect of combined medication on alcohol metabolism.

[0094] Western Blot analysis was used to evaluate the relative expression of CYP2E1 in the liver of each group of mice.

[0095] 1.3.4 Detection of inflammatory factors (TNF-α, IL-6 and IL-1β)

[0096] The concentrations of inflammatory factors tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in serum were determined by enzyme-linked immunosorbent assay (ELISA). After collecting serum, the contents of TNF-α, IL-6 and IL-1β in the serum of each group of mice were determined according to the operating instructions of the ELISA kit to evaluate the regulatory effect of combined drug use on inflammatory response.

[0097] 1.4 Statistical analysis

[0098] GraphPad 10.0 software was used for analysis. The experimental data were expressed as mean ± standard deviation (X ± SEM), and the differences among the groups were analyzed using one-way ANOVA.

[0099] 2. Experimental results

[0100] 2.1 Effects on liver function

[0101] like Figure 1 As shown, compared with the alcohol model group, the Pueraria polysaccharide treatment group, the betaine treatment group and the combined medication treatment group (high dose and low dose) all significantly reduced the serum ALT and AST levels (p<0.05), among which the combined medication treatment group had the most significant effect, indicating that the Pueraria polysaccharide + betaine + esculenta + quercetin combined medication treatment group has a synergistic effect in alleviating alcohol-induced liver damage.

[0102] 2.2 Effects on liver pathological changes (HE staining)

[0103] like Figure 2 As shown, obvious hepatocyte swelling, necrosis and fatty degeneration were observed in the liver of mice in the alcohol model group, accompanied by a large number of inflammatory cell infiltrations; while the pathological damage of the liver tissues of mice in the pueraria polysaccharide treatment group and the betaine treatment group was alleviated, and the hepatocyte morphology was improved to a certain extent. In particular, in the combined drug treatment group, the swelling and necrosis of hepatocytes were significantly reduced, fatty degeneration was effectively inhibited, and the infiltration of inflammatory cells was also significantly reduced, indicating that the combined drug treatment has a significant effect in protecting the hepatocyte structure and reducing alcohol-induced liver damage.

[0104] 2.3 Effect on liver fibrosis (Sirius red staining)

[0105] like Figure 3 As shown, the collagen fibers in the liver of mice in the alcohol model group increased significantly, and the degree of liver fibrosis increased significantly; while the pueraria polysaccharide treatment group, betaine treatment group and combination treatment group all significantly reduced collagen deposition in the liver, especially the combination group, indicating that pueraria polysaccharide and betaine can effectively inhibit the process of alcohol-induced liver fibrosis. On the basis of the separate use of pueraria polysaccharide and betaine, their combination with esculetin and quercetin had the best effect in inhibiting alcohol-induced liver fibrosis.

[0106] 2.4 Effects on intestinal barrier function (colon PAS staining)

[0107] like Figure 4 As shown, the intestinal permeability of mice in the combined drug treatment group was significantly lower than that in the alcohol model group, the number of goblet cells in the colon of mice in the combined drug treatment group increased significantly, mucus secretion returned to normal, and inflammatory cell infiltration decreased, indicating that Pueraria polysaccharide and betaine alone can effectively improve alcohol-induced intestinal damage, and the combined drug effect is better.

[0108] 2.5 Colon MUC-2 immunohistochemistry

[0109] like Figure 5As shown, the MUC-2 immunohistochemistry results showed that the expression of MUC-2 in the colon of mice in the alcohol model group was significantly reduced, while the expression of MUC-2 in the combined drug treatment group was significantly restored, indicating that the combined drug treatment can improve the intestinal mucus barrier function and protect the integrity of the intestinal epithelium.

[0110] 2.6 Colonic tight junction protein staining

[0111] like Figure 6 and Figure 7 As shown, the ZO-1 and Occludin staining results showed that the expression of tight junction proteins in the colon of mice in the alcohol model group was significantly reduced, and the tight junction structure was destroyed; the combined drug treatment group significantly restored the expression levels of ZO-1 and Occludin, indicating that the combined drug treatment can effectively protect the integrity of the intestinal barrier and reduce the destruction of tight junctions.

[0112] 2.7 Effects on the activity of alcohol metabolizing enzymes (ADH, ALDH, CYP2E1)

[0113] like Figure 8 As shown, compared with the alcohol model group, the ADH and ALDH activities in the liver of mice in the Pueraria polysaccharide treatment group, betaine treatment group and combined drug treatment group were significantly increased (p<0.05), among which the ADH and ALDH activity levels in the combined drug treatment group were the highest, indicating that the combined use of Pueraria polysaccharide + betaine + esculenta lactone + quercetin can significantly enhance the metabolic capacity of alcohol and accelerate its clearance in the body.

[0114] 2.8 CYP2E1 protein expression

[0115] like Fig. 9 As shown, Western Blot results showed that the expression of CYP2E1 in the liver of mice in the alcohol model group was significantly increased (p<0.05), indicating that alcohol induced the upregulation of CYP2E1. The Pueraria polysaccharide treatment group, betaine treatment group and combined drug treatment group all significantly reduced the expression level of CYP2E1, especially the combined drug treatment group, indicating that the combination of Pueraria polysaccharide + betaine + esculetin + quercetin can inhibit alcohol-induced CYP2E1 expression and reduce oxidative stress and liver damage.

[0116] 2.9 Effects on inflammatory factors (TNF-α, IL-6, IL-1β)

[0117] like Fig.10As shown in the figure, the ELISA results showed that the levels of TNF-α, IL-6 and IL-1β in the serum of mice in the alcohol model group were significantly increased (p<0.05), indicating that alcohol induced a significant inflammatory response. The levels of TNF-α, IL-6 and IL-1β in the Pueraria polysaccharide treatment group and the betaine treatment group were significantly reduced, especially in the combined drug treatment group, whose inflammatory factor levels were significantly lower than those in the alcohol model group (p<0.01), indicating that combined drug treatment can effectively inhibit the systemic inflammatory response induced by alcohol.

[0118] 3. Conclusion

[0119] This study shows that the combination of pueraria polysaccharide + betaine + esculetin + quercetin can significantly improve alcohol-induced liver and intestinal damage in mice, and its mechanism may be related to enhancing the activity of alcohol metabolic enzymes, reducing CYP2E1 expression, reducing oxidative stress, reducing inflammatory response, inhibiting liver fibrosis and improving intestinal barrier function. The combined application has potential medicinal value and can be used to prepare a composition for treating and / or preventing alcoholic liver disease and intestinal damage.

[0120] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A composition for improving alcoholic liver damage and intestinal damage, characterized in that: The composition comprises the following components in parts by weight: 20 to 30 parts of kudzu root polysaccharide, 10 to 15 parts of betaine, 8 to 14 parts of esculenta and 5 to 12 parts of quercetin.

2. A composition for improving alcoholic liver damage and intestinal damage according to claim 1, characterized in that: The composition comprises the following components in parts by weight: 26 parts of kudzu root polysaccharide, 13 parts of betaine, 10 parts of esculent acid and 7 parts of quercetin.

3. A composition for improving alcoholic liver damage and intestinal damage according to claim 1, characterized in that: The preparation method of Pueraria polysaccharide comprises the following steps: (1) Weigh the washed and dried kudzu root and crush it, add water, place it in a 70-90° C. water bath and stir to extract for 2-5 hours, repeat the extraction 2-4 times, filter and combine all the extracts, cool it and concentrate it under reduced pressure, and then wash the precipitate with 75-80% by volume ethanol for 1-3 times to obtain a precipitate; (2) drying the precipitate obtained in step (1) in a vacuum drying oven to obtain polysaccharide powder; dissolving the polysaccharide powder in water to obtain a polysaccharide solution for later use; wherein the mass volume ratio of the polysaccharide powder to water is 1 g: (3-6) mL; (3) The polysaccharide solution obtained in step (2) is mixed with α-amylase solution, placed in a 40° C. water bath for 15-30 min, and then boiled to inactivate the enzyme. Ethanol with a volume fraction of 85-95% is then added to precipitate the enzyme protein, and the mixture is centrifuged and vacuum dried to obtain Pueraria polysaccharide.

4. A composition for improving alcoholic liver damage and intestinal damage according to claim 3, characterized in that: In step (1), the mass volume ratio of Pueraria root to water is 1 g: (10-15) mL; the volume ratio of the extract to ethanol is 1 mL: (3-6) mL.

5. A composition for improving alcoholic liver damage and intestinal damage according to claim 3, characterized in that: In step (3), the volume ratio of the polysaccharide solution, the α-amylase solution and the ethanol is 1 mL: (2-3) mL: (5-10) mL.

6. A composition for improving alcoholic liver damage and intestinal damage according to claim 1, characterized in that: The preparation method of esculetin comprises the following steps: ① Take 150g of Cortex Fraxini crude powder in a Soxhlet extractor, add 400ml of ethanol and reflux for 8-12h to obtain ethanol extract, and recover the solvent under reduced pressure to an extract; ② Add 40 ml of water to the extract and heat to dissolve, extract with an equal volume of chloroform for 30 to 50 minutes, separate the liquid to obtain the aqueous phase, add an equal volume of ethyl acetate to the aqueous phase for extraction to obtain ethyl acetate solution, dehydrate with anhydrous sodium sulfate, and recover the solvent under reduced pressure to dryness to obtain a dry product; ③ Add 50-60℃ methanol to the dried product and let it stand for crystallization. Yellow needle-shaped crystals will precipitate. Filter out the crystals to obtain esculetin.

7. A composition for improving alcoholic liver damage and intestinal damage according to claim 6, characterized in that: The mass volume ratio of the dried product and methanol in step ③ is 1g:(5-8)mL.

8. The composition according to claim 1, characterized in that The dosage form of the composition is an oral preparation.

9. The method for preparing a composition for improving alcoholic liver damage and intestinal damage according to claim 1, characterized in that: The method comprises mixing the components or adding water to disperse the components uniformly.

10. Use of the composition according to any one of claims 1 to 8 or the composition prepared by the preparation method according to claim 9 in preparing products for improving alcoholic liver damage and intestinal damage.

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