Determination method for hangover-alleviating and liver-protecting effects of hangover-alleviating and liver-protecting drink on mice with alcoholic liver injury

Abstract

The invention relates to the technical field of prevention and treatment of alcoholic liver injury, and discloses a method for determining the hangover alleviating and liver protecting effects of a hangover alleviating and liver protecting drink on a mouse with alcoholic liver injury, and the method comprises the following steps: S1, preparing an extract; s2, establishing an animal model; s3, performing a drug experiment; s4, detecting experimental indexes; according to the alcohol effect dispelling and liver protecting drink, through compound compatibility of the radix puerariae, the hovenia dulcis thunb and the dendrobium officinale, key links of alcohol metabolism are comprehensively regulated and controlled, accumulation of liver tissue lipid peroxidation products (such as MDA) is remarkably reduced, triglyceride metabolic disorder can be improved, and the alcohol effect dispelling and liver protecting drink has the effects of dispelling the effects of alcohol and protecting liver. Liver cell injury is relieved through oxidative injury and lipid accumulation, the treatment effect is enhanced, and compared with a single-component medicine, the compound shows a more comprehensive substantive liver protection effect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of alcoholic liver injury prevention and treatment, in particular to a method for determining the alcoholism and liver protection effects of Jiujiuhuan'gan drink on alcoholic liver injury mice. BACKGROUND

[0002] Alcoholic liver injury (ALD) is a liver disease caused by long-term or excessive alcohol intake, and its pathogenesis is complex, involving oxidative stress, lipid metabolism disorder, inflammatory response and hepatocyte damage. Modern pharmacological research often uses animal models (such as alcohol-induced mouse liver injury model) to evaluate drug effects, and analyzes the mechanism of action of drugs by detecting drunk time, liver weight ratio, malondialdehyde (MDA), alkaline phosphatase (ALP) and other indicators.

[0003] At present, the treatment of alcoholic liver injury has disadvantages: the effect of single target treatment is limited, and the current research on alcoholic liver injury mainly focuses on single component or single mechanism (such as antioxidant), and the commonly used drugs such as silybin have certain liver protection effect, but there are problems such as single action target and unstable efficacy, and the liver damage caused by alcohol metabolism is the result of the joint action of multiple pathological links, and existing drugs often cannot comprehensively cover key links such as lipid peroxidation and triglyceride accumulation in the alcohol metabolism process, resulting in limited treatment effect; traditional alcoholism drugs mainly focus on relieving drunk symptoms, and the effect is limited, and lack of protection for liver parenchymal damage, which is difficult to meet the health needs of long-term drinkers. Therefore, a method for determining the alcoholism and liver protection effects of Jiujiuhuan'gan drink on alcoholic liver injury mice is proposed. SUMMARY

[0004] The purpose of the present application is to provide a method for determining the alcoholism and liver protection effects of Jiujiuhuan'gan drink on alcoholic liver injury mice, to solve the problems raised in the background art.

[0005] To achieve the above purpose, the present application provides the following technical scheme: a method for determining the alcoholism and liver protection effects of Jiujiuhuan'gan drink on alcoholic liver injury mice, comprising the following steps:

[0006] S1, preparing an extract;

[0007] S2, establishing an animal model;

[0008] S3, drug experiment;

[0009] S4, detecting experimental indicators;

[0010] S5, data analysis.

[0011] As a preferred, in S1, the extract is made of Pueraria lobata, Hovenia dulcis and Dendrobium officinale, and the extract content in each 100g product is 8.84g.

[0012] As preferred, in S2, the above comprises the following sub-steps:

[0013] S201, select 48 SPF KM mice, after adaptive feeding for 5 days, fast before the experiment;

[0014] S202, 48 mice are randomly divided into 6 groups, 8 in each group, which are normal group, model group, positive drug group, Jiujiuhelan drink high dose group, Jiujiuhelan drink medium dose group and Jiujiuhelan drink low dose group.

[0015] As preferred, in S202, the normal group includes a blank control group, the model group includes an alcohol group, the positive drug group includes an EtOH+shikimic acid group, and the doses of the Jiujiuhelan drink high / medium / low dose groups are obtained by converting the body surface area method.

[0016] As preferred, in S3, the above comprises the following sub-steps:

[0017] S301, administration period (1-21 days):

[0018] Normal group: intragastrically administered with normal saline;

[0019] Model group: intragastrically administered with normal saline;

[0020] Positive drug group: intragastrically administered with a set dose of shikimic acid;

[0021] Jiujiuhelan drink high dose group: intragastrically administered with an equal amount of extract containing 321.75 mg / 20 g of Jiujiuhelan drink;

[0022] Jiujiuhelan drink medium dose group: intragastrically administered with an equal amount of extract containing 160.8 mg / 20 g of Jiujiuhelan drink;

[0023] Jiujiuhelan drink low dose group: intragastrically administered with an equal amount of extract containing 80.4 mg / 20 g of Jiujiuhelan drink;

[0024] S302, 30 min after the last administration, all groups of mice except the normal group were intragastrically administered with 55% alcohol (0.32 mL / 20 g) to establish the alcohol model;

[0025] S303, perform the righting experiment.

[0026] As preferred, in S303, the specific operation of the righting experiment comprises:

[0027] S3031, place the alcohol model mice in a supine position;

[0028] S3032, keep the mouse in a supine position for 30 seconds as the judgment standard for the disappearance of righting reflex;

[0029] S3033, record the drunk time;

[0030] S3034, record the sober time.

[0031] As preferred, the drunk time is the time interval from the beginning of alcohol gavage to the disappearance of the mouse righting reflex, and the sober time is the time interval from the beginning of alcohol gavage to the recovery of the mouse righting reflex.

[0032] As preferred, during the drug experiment, the mice in each group are fed with ordinary feed and drink water freely, and there is no difference between each factor in each group.

[0033] As preferred, in S4, the following sub-steps are included:

[0034] S401, after 16h of alcohol modeling, the mice are anesthetized with diethyl ether and sacrificed;

[0035] S402, collect blood samples and liver tissue samples of the dead mice, and weigh the liver tissue samples;

[0036] S403, detect the related biochemical indexes of the mice in each group.

[0037] As preferred, in S5, the experimental data of statistical analysis is represented by x±s, and is calculated and processed by Microsoft Excel and GraphPad prism 6.0 software, the statistical method is one-way analysis of variance, and P<0.05 is statistically significant.

[0038] Compared with the prior art, the above technical scheme has the following technical effects:

[0039] Firstly, the present application proposes a multi-target synergistic liver protection effect: the Jiuji Hugan drink is compounded by Pueraria lobata, Hovenia dulcis and Dendrobium officinale, which comprehensively regulates the key link of alcohol metabolism, not only significantly reduces the accumulation of lipid peroxidation products (such as MDA) in liver tissue, but also improves the disorder of triglyceride metabolism, reduces liver cell damage from oxidative damage and lipid accumulation, and enhances the treatment effect, compared with single-component drugs, the compound shows more comprehensive and substantial liver protection effect.

[0040] Secondly, the alcohol-eliminating and liver-protecting drink can effectively prolong the drunkenness latency of mice, can play an anti-intoxication role by regulating the activity of alcohol metabolism enzymes or delaying the absorption of the gastrointestinal tract, and the sober-up time is shortened after the intervention of the compound, which indicates that it can accelerate the decomposition and metabolism of alcohol, relieve the drunken state, and overcome the limitation of traditional alcohol-eliminating drugs which only target the symptoms, the high, medium and low dose groups of the compound show different effects, among which the low dose has advantages in delaying the drunken time and the medium and high doses have advantages in improving oxidative stress, which provides a scientific basis for clinical dose selection, in addition, the liver weight ratio and other indicators show that it has no negative impact on the liver index and has good safety, and is suitable for preventive intervention of long-term drinkers. BRIEF DESCRIPTION OF DRAWINGS

[0041] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description are only some embodiments of the present application, and for those skilled in the art, other drawings can be obtained without creative labor on the basis of these drawings.

[0042] Figure 1 The flowchart of the present application. DETAILED DESCRIPTION

[0043] The technical solutions in the embodiments of the present application will be described clearly and completely in the following with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some embodiments of the present application, not all embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0044] EMBODIMENT

[0045] Please refer to Figure 1 The present application provides a technical solution: a method for determining the alcohol-eliminating and liver-protecting effect of the alcohol-eliminating and liver-protecting drink on alcoholic liver injury mice, comprising the following steps:

[0046] The method for determining the alcohol-eliminating and liver-protecting effect of the alcohol-eliminating and liver-protecting drink on alcoholic liver injury mice comprises the following steps:

[0047] S1, preparing an extract, the extract is prepared from pueraria, hovenia dulcis and dendrobium officinale, and the extract content in 100g of the product is 8.84g;

[0048] S2, establishing an animal model;

[0049] Comprising the following sub-steps:

[0050] S201, selecting 48 SPF KM mice, and after adaptive feeding for 5 days, fasting but not water fasting before the experiment;

[0051] S202, 48 mice were randomly divided into 6 groups, 8 in each group, which were normal group, model group, positive drug group, high dose group of Jiejiuhelan drink, medium dose group of Jiejiuhelan drink and low dose group of Jiejiuhelan drink. The normal group included the blank control group, the model group included the alcohol group, the positive drug group included the EtOH+shikimic acid pinus group, and the doses of the high / medium / low dose groups of Jiejiuhelan drink were obtained by the body surface area method: the content of the high dose group of Jiejiuhelan drink (EtOH+J-H group) was 321.75 mg / 20 g, according to the calculation of human daily consumption of 300 g of the product, the mouse dose was equivalent to 4 times the human clinical dose; the content of the medium dose group of Jiejiuhelan drink (EtOH+J-M group) was 160.8 mg / 20 g, according to the calculation of human daily consumption of 300 g of the product, the mouse dose was equivalent to 2 times the human clinical dose; the content of the low dose group of Jiejiuhelan drink (EtOH+J-L group) was 80.4 mg / 20 g, according to the calculation of human daily consumption of 300 g of the product, the mouse dose was equivalent to the human clinical dose;

[0052] S3, drug experiment: during the drug experiment, the mice in each group were fed with ordinary feed and free water, and there was no difference between each factor in each group;

[0053] including the following sub-steps:

[0054] S301, administration period (1-21 days):

[0055] Normal group: intragastrically administered with normal saline;

[0056] Model group: intragastrically administered with normal saline;

[0057] Positive drug group: intragastrically administered with a set dose of shikimic acid pinus;

[0058] High dose group of Jiejiuhelan drink: intragastrically administered with an equal amount of extract containing 321.75 mg / 20 g of Jiejiuhelan drink;

[0059] Medium dose group of Jiejiuhelan drink: intragastrically administered with an equal amount of extract containing 160.8 mg / 20 g of Jiejiuhelan drink;

[0060] Low dose group of Jiejiuhelan drink: intragastrically administered with an equal amount of extract containing 80.4 mg / 20 g of Jiejiuhelan drink;

[0061] S302, 30 min after the last administration, the mice in each group except the normal group were intragastrically administered with 55% alcohol (0.32 mL / 20 g) to perform alcohol modeling;

[0062] S303, the righting experiment was performed;

[0063] The specific operation of the righting experiment includes:

[0064] S3031, the alcohol-modeled mice were placed in a supine position;

[0065] S3032, the mouse keeps the supine position for 30 seconds as the judgment standard for the disappearance of the righting reflex;

[0066] S3033, record the drunk time: the time interval from the start of alcohol gavage to the disappearance of the righting reflex of the mouse;

[0067] S3034, record the sober-up time: the time interval from the start of alcohol gavage to the recovery of the righting reflex of the mouse;

[0068] S4, detection of experimental indexes;

[0069] Including the following sub-steps:

[0070] S401, after 16 hours of alcohol modeling, the mice were anesthetized with ether and sacrificed;

[0071] S402, collect blood samples and liver tissue samples from dead mice, and weigh the liver tissue samples;

[0072] S403, detect the related biochemical indexes of each group of mice, including:

[0073] Determination of serum ALT and AST content: the mice were anesthetized with ether, and blood was taken from the orbit and placed at 4℃. Centrifuge at 14000r / min for 15min, take the supernatant to get serum, according to the kit instructions to detect each biochemical index;

[0074] Determination of ALT, MDA and TG content in liver tissue: after the mice were anesthetized and blood was taken from the orbit, the mice were dissected, the liver was taken out, and after weighing, the supernatant was extracted according to the kit instructions, and the protein concentration in the tissue homogenate was quantified by BCA method. According to the kit instructions to determine the content of ALT, MDA and TG in liver tissue;

[0075] S5, data analysis: the experimental data of statistical analysis is expressed as x±s, and Microsoft Excel and GraphPad prism 6.0 software are used for calculation and processing, the statistical method is one-way analysis of variance, P<0.05 is statistically significant.

[0076] Results and analysis:

[0077] I. Effect of Jiuji Hugan Drink on the drunk and sober-up time of ALD (alcoholic liver injury) mice:

[0078] The determination of drunken time and sober time can show whether Jiejiuhuganyin has the effect of relieving alcoholism. After the alcoholism model of mice is established, the righting reflex experiment is carried out, and the righting reflex disappearance time of each group of mice is recorded as the drunken time, and the righting reflex recovery time of mice is the sober time of mice. The experimental results (as shown in Table 1) show that the drunken time of mice is compared: model group > high-dose Jiejiuhuganyin group > positive drug group > medium-dose Jiejiuhuganyin group > low-dose Jiejiuhuganyin group, and the sober time of mice is compared: medium-dose Jiejiuhuganyin group < low-dose Jiejiuhuganyin group < high-dose Jiejiuhuganyin group < positive drug group < model group, but compared with the model group, the drunken time and sober time of the high-dose, medium-dose and low-dose Jiejiuhuganyin groups have no statistical difference.

[0079] Table 1 Effect of Jiejiuhuganyin on righting reflect time in micewith ALD( n=7)

[0080] Table 1 Effect of Jiejiuhuganyin on righting reflect time in micewith ALD( n=7)

[0081]

[0082] Note: compared with the normal group, 1) p<0.05, 2) p<0.01; compared with the control group 3) p<0.05, 4) p<0.01 (Table 2-5 same). The index calculation standard of each group: the drunken mouse percentage = the number of drunken mice / the total number of mice given alcoholism, wherein the total number of mice includes the number of mice that are drunk within 10 min after being given alcohol and the number of mice that are not drunk after 2 h; the drunken time: the time from when the mice are given alcohol to when the righting reflex of the mice disappears, and the sober time: the time from when the mice are given alcohol to when the righting reflex of the mice recovers.

[0083] II. Effect of Jiejiuhuganyin on liver weight and liver weight ratio of ALD mice:

[0084] The mice were treated 16 h after modeling, the liver of the mice was taken out, the liver weight was weighed, and the liver weight ratio of the mice was investigated. The liver weight can reflect whether the liver of the mice is enlarged, and the liver weight ratio can evaluate the liver index of the mice. The experimental results (as shown in Table 2) show that there is no significant difference in the liver weight and liver weight ratio of the six groups of mice.

[0085] Table 2 Effect of Jiejiuhuganyin on liver weight and liver weight ratio of ALD mice

[0086] Table 2 Effect of Jiejiuhuganyin on liver weight and liver / body weight in mice with ALD

[0087]

[0088] Three, the effect of Jiejiuhuganyin on serum transaminase levels in ALD mice:

[0089] Excessive alcohol intake can cause severe liver damage in mice, and transaminase (ALT, AST) is a sensitive marker of acute hepatocyte damage. Therefore, detecting the levels of ALT and AST in serum can reflect whether liver damage has occurred. The experimental results (Table 3) show that compared with the normal group, the serum ALT and AST levels of the model group were significantly increased (p<0.01), indicating that the modeling was successful; administration of high and low doses of Jiejiuhuganyin and silybin could significantly reduce the levels of ALT and AST in the serum of mice (p<0.01, p<0.05), indicating that Jiejiuhuganyin and silybin have a protective effect on the liver of ALD mice.

[0090] Table 3 Effect of Jiejiuhuganyin on the levels of ALT,AST in serum of mice with ALD

[0091] Table 3 Effect of Jiejiuhuganyin on the levels of ALT,AST in serum of mice with ALD

[0092]

[0093] Four, the effect of Jiejiuhuganyin on MDA, ALP and TG levels in the liver of ALD mice:

[0094] Alcohol can cause intrahepatic cholestasis, leading to elevated alkaline phosphatase (ALP). Both animal and human experiments have shown that acute or chronic alcoholism damages the structure and function of various organelles and enzymes of hepatocytes, and affects the beta oxidation of fatty acids in mitochondria, causing lipid peroxidation, malondialdehyde (MDA) is the end product of lipid peroxidation, which affects the structure and function of hepatocytes through free radical damage, while the damage of hepatocyte membrane caused by lipid peroxidation causes the output of triglyceride (TG) from hepatocytes to be impaired, leading to the accumulation of triglyceride in the liver. To verify whether the effect of Jiejiuhuganyin on inhibiting alcohol-induced acute liver damage is achieved by improving oxidative stress, the levels of MDA, ALP and TG in the liver tissue of mice were detected. As shown in Table 4, the MDA content of the model group was significantly higher than that of the normal group (p<0.01), while Jiejiuhuganyin could significantly reduce the MDA content in the tissue (p<0.01); but the ALP content determination results showed that compared with the normal group, the ALP level of the model group did not change significantly, while the ALP content of the liver tissue of mice in the high-dose group of Jiejiuhuganyin increased significantly; at the same time, the TG content determination results in liver tissue showed that compared with the model group, the TG level in the liver tissue of mice in the high / medium / low dose group of Jiejiuhuganyin increased significantly (p<0.05, p(0.01). The above experimental results show that Jiejiuhuganyin cannot play a protective role on ALD by improving oxidative stress in mice.

[0095] Table 4 Effect of Jiejiuhuganyin on the levels ofALP,MDA and TG inhepatic tissue ofmice withALD( n=6)

[0096] Table 4 Effect of Jiejiuhuganyin on the levels ofALP,MDA and TG inhepatic tissue ofmice withALD( n=6)

[0097]

[0098] Summary:

[0099] 1) The results of Jiejiuhuganyin anti-drunk experiment showed that low-dose Jiejiuhuganyin could delay the drunk time of mice; medium-dose Jiejiuhuganyin could shorten the sober-up time of mice;

[0100] 2) The liver protection experiment of Jiejiuhuganyin showed that Jiejiuhuganyin could reduce the serum transaminase level and had a protective effect on the liver of ALD mice; among them, the effect of reducing transaminase of mice in the high-dose and low-dose groups of Jiejiuhuganyin was more significant than that in the medium-dose group;

[0101] 3) The effect of Jiejiuhugan drink in inhibiting alcohol-induced acute liver injury is achieved by improving partial oxidative stress; wherein, compared with the model group, the high / middle / low dose of Jiejiuhugan drink can significantly reduce the MDA level and can slow down the lipid peroxidation to a certain extent;

[0102] 4) The maximum amount of Jiejiuhugan drink taken by an adult each time is not more than 26.52g of the low-dose group (corresponding to 80.4mg of the extract of mice), that is, 300g of Jiejiuhugan drink product.

[0103] In summary, the Jiejiuhugan drink of the present application, through the compound compatibility of Pueraria lobata, Hovenia dulcis and Dendrobium officinale, comprehensively regulates the key link of alcohol metabolism, not only significantly reduces the accumulation of liver tissue lipid peroxidation products (such as MDA), but also improves the disorder of triglyceride metabolism, reduces liver cell damage from both oxidative damage and lipid accumulation. Compared with single-component drugs, the compound shows a more comprehensive liver protection effect; experiments have proved that Jiejiuhugan drink can effectively prolong the drunkenness latency of mice, especially in the low-dose group, suggesting that it can play an anti-drunken role by regulating the activity of alcohol metabolism enzymes or delaying gastrointestinal absorption. At the same time, the sober-up time is shortened after the intervention of the compound, indicating that it can accelerate the decomposition and metabolism of alcohol, relieve the drunken state, and make up for the limitations of traditional alcohol-curing drugs that only target symptoms; the high / middle / low dose groups of the compound show differential effects, among which the low dose has advantages in delaying drunkenness time, and the middle and high doses have advantages in improving oxidative stress, providing a scientific basis for clinical dose selection. In addition, indicators such as liver weight ratio show that it has no negative impact on liver index and has good safety, and is suitable for preventive intervention of long-term drinkers.

[0104] Those skilled in the art can understand that the features described in various embodiments and / or claims of the present application can be combined or / and combined, even if such combinations or combinations are not explicitly described in the present application. In particular, the features described in various embodiments and / or claims of the present application can be combined and / or combined in various combinations without departing from the spirit and teachings of the present application. All these combinations and / or combinations fall within the scope of the present application.

Claims

1. A method for determining the alcoholism-preventing and liver-protecting effects of Jiujiuhelan drink on alcoholic liver injury mice, characterized in that, Comprise the following steps: S1, preparation of extract; S2, the establishment of animal models; S3, drug experiment; S4, experimental index detection; S5, data analysis.

2. The method for determining the alcohol elimination and liver protection effects of the alcohol elimination and liver protection drink according to claim 1 on alcoholic liver injury mice, characterized in that: In S1, the extract is made of Pueraria lobata, Hovenia dulcis and Dendrobium officinale, and the extract content in 100g product is 8.84g.

3. The method for determining the alcohol elimination and liver protection effects of the alcohol elimination and liver protection drink according to claim 2 on alcoholic liver injury mice, characterized in that: In S2, it comprises the following sub-steps: S201, select 48 SPF KM mice, adaptively feed for 5 days, and fast before the experiment; S202, 48 mice are randomly divided into 6 groups, 8 in each group, which are normal group, model group, positive drug group, high dose of Jiujiuhelan drink group, medium dose of Jiujiuhelan drink group and low dose of Jiujiuhelan drink group.

4. The method for determining the alcohol elimination and liver protection effects of the alcohol elimination and liver protection drink according to claim 3 on alcoholic liver injury mice, characterized in that: In S202, the normal group includes a blank control group, the model group includes an alcohol group, the positive drug group includes an EtOH+shikimic acid group, and the doses of the high / medium / low dose of Jiujiuhelan drink group are obtained by body surface area method.

5. The method for determining the anti-alcoholic and liver-protecting effects of the anti-alcoholic and liver-protecting drink according to claim 1 on alcoholic liver injury mice, characterized in that: In S3, it comprises the following sub-steps: S301, dosing period (1-21 days): Normal group: intragastrically administered with normal saline; Model group: intragastrically administered with normal saline; Positive drug group: intragastrically administered with a set dose of shikimic acid; High dose of Jiujiuhelan drink group: intragastrically administered with an equivalent amount of extract content of 321.75mg / 20g of Jiujiuhelan drink; Medium dose of Jiujiuhelan drink group: intragastrically administered with an equivalent amount of extract content of 160.8mg / 20g of Jiujiuhelan drink; Low dose of Jiujiuhelan drink group: intragastrically administered with an equivalent amount of extract content of 80.4mg / 20g of Jiujiuhelan drink; S302, 30min after the last administration, the mice in each group except the normal group were intragastrically administered with 55% alcohol (0.32mL / 20g) to perform alcohol modeling; S303, perform the righting experiment.

6. The method for determining the alcohol elimination and liver protection effects of the alcohol elimination and liver protection drink according to claim 5 on alcoholic liver injury mice, characterized in that: In S303, the specific operation of the righting experiment comprises: S3031, place the alcohol modeled mice in a supine position; S3032, take the mouse maintaining the supine position for 30 seconds as the judgment standard for the disappearance of the righting reflex; S3033, record the drunken time; S3034, record the sober-up time.

7. The method for determining the anti-alcoholic and liver-protecting effects of the anti-alcoholic and liver-protecting drink according to claim 6 on alcoholic liver injury mice, characterized in that: The drunken time is the time interval from the start of alcohol intragastrical administration to the disappearance of the righting reflex of the mouse, and the sober-up time is the time interval from the start of alcohol intragastrical administration to the recovery of the righting reflex of the mouse.

8. The method for determining the anti-alcoholic and liver-protecting effects of the anti-alcoholic and liver-protecting drink according to claim 5 on alcoholic liver injury mice, characterized in that: During the drug experiment, each group of mice is fed with ordinary feed and free water, and there is no difference between each factor in each group.

9. The method for determining the anti-alcoholic and liver-protecting effects of the anti-alcoholic and liver-protecting drink according to claim 1 on alcoholic liver injury mice, characterized in that: In S4, it comprises the following sub-steps: S401, after 16h of alcohol modeling, the mice are euthanized under ether anesthesia; S402, collect the blood samples and liver tissue samples of the dead mice, and weigh the liver tissue samples; S403, detect the related biochemical indexes of each group of mice.

10. The method for determining the anti-alcoholic and liver-protecting effects of the anti-alcoholic and liver-protecting drink according to claim 7 on alcoholic liver injury mice, characterized in that: In S5, the experimental data of statistical analysis is represented by x±s, and is calculated and processed by Microsoft Excel and GraphPad prism 6.0 software, the statistical method is one-way analysis of variance, and P<0.05 is statistically significant.

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