Brain tissue ethanol metabolism rate evaluation method

Abstract

The invention discloses a brain tissue ethanol metabolism rate evaluation method. The method comprises the following steps: converting the ethanol content of a brain tissue by using serum; and calculating the area (AUC) under the drug-time curve of the brain tissues of the mice in each experimental group, and grading the metabolic capability of the wine body after drinking. According to the present invention, the influence of the alcohol beverage on the ethanol metabolism rate is represented by the ethanol content change of the brain tissue, such that the accumulation degree of the ethanol in the brain can be visually evaluated, the data reference is provided for the research of the relationship between the ethanol concentration and the after-drinking comfort level of the body, and the use of the experimental animal can be reduced through the method;

Description

Technical Field

[0001] The invention relates to the technical field of winemaking, and in particular to a method for evaluating the ethanol metabolic rate of brain tissue after drinking and an application thereof. Background Art

[0002] After drinking, ethanol as a small molecule passes through the blood-brain barrier into the cerebrum and cerebellum and accumulates in the brain, affecting consciousness, memory and body coordination. Studies have shown that there is a significant positive correlation between blood and brain tissue ethanol concentrations and behavior. When the ethanol content is below 0.1%, the drinker's brain nerves are excited, and consciousness and body coordination are retained; when the ethanol content is above 0.1%, ataxia occurs, and consciousness, memory and body coordination are gradually lost, and even death may occur in the drinker. The brain is a high-level nerve center for nerves and behaviors, and monitoring the ethanol concentration in brain tissue is of great significance for the study of comfort after drinking.

[0003] Although it is possible to evaluate the ethanol metabolism capacity of mice by sampling their brain tissue at different times after drinking, this method uses too many animals and is not in line with animal ethics. Summary of the invention

[0004] The purpose of the present invention is to provide a method for evaluating the brain tissue ethanol metabolic rate. The method clarifies the relationship between brain tissue and serum ethanol content, and uses blood to convert brain tissue ethanol content. It not only intuitively reflects the changes in brain tissue ethanol content to lay a foundation for the study of comfort after drinking, but also reduces the use of experimental animals and complies with the ethical requirements of experimental animals.

[0005] The present invention is achieved through the following technical solutions:

[0006] A method for evaluating brain tissue ethanol metabolism rate comprises the following steps:

[0007] (1) Select male BALB / c mice with good adaptability to feeding, fast for 8-16 hours before the experiment, and ensure adequate drinking water;

[0008] (2) Select a variety of wine product samples;

[0009] (3) While ensuring the same alcohol intake, calculate the intragastric volume of different alcoholic products according to the equivalent conversion method, perform intragastric gavage on mice, and record the gavage time;

[0010] (4) Serum samples were collected from each group of mice at different time points and the ethanol content in brain tissue was calculated;

[0011] The calculation formula of brain tissue ethanol content is:

[0012] C 脑 : Brain tissue ethanol content, mg / g;

[0013] C 血 : serum ethanol content, mmol / L;

[0014] M 乙醇 : molar mass of ethanol, g / mol;

[0015] ρ 血清 : BALB / c serum density, g / cm 2 ;

[0016] K: conversion factor, the value is 0-1, preferably 0.2-0.8;

[0017] (5) The statistical data information is analyzed using statistical software to calculate the alcohol metabolism value and grade the wine samples.

[0018] Preferably, in step (2), the concentration of the wine sample is 35-53 degrees, and the intragastric dose of the wine sample is 0.09-0.18 ml / 10 g.bw.

[0019] Preferably, the ethanol detection time after drinking in step (4) is 0-6 hours.

[0020] Preferably, in step (5), ethanol metabolism value = AUC 实验组 / AUC 对照组 ×100%.

[0021] Preferably, in step (4), serum ethanol concentration: brain tissue ethanol concentration = 1:0-1.

[0022] Preferably, in step (5), the alcohol metabolism value of the alcohol solution group is taken as 100 points as the benchmark, and the experimental wine samples are graded respectively: less than 85, special grade; 85-95, excellent grade; 95-105, first grade; 105-115, second grade; greater than 115, third grade.

[0023] Among them, level two and below indicate that the ethanol metabolism of the experimental wine is slow after drinking, and level one and above indicate that the ethanol metabolism of the experimental wine is fast after drinking. The higher the level, the faster the ethanol metabolism.

[0024] Another object of the present invention is to apply the above method in the process of liquor brewing.

[0025] The method provided by the present invention is more intuitive in evaluating the brain burden after drinking alcoholic products directly through the ethanol concentration of brain tissue, laying a foundation for the study of comfort after drinking. DETAILED DESCRIPTION

[0026] In order to facilitate the understanding of the present invention, the present invention will be described more fully below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0027] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the embodiments of the present invention belong. If the definitions set forth in this section are contrary to or otherwise inconsistent with the definitions set forth in the patents, patent applications, published patent applications, and other publications incorporated herein by reference, the definitions listed in this section take precedence over the definitions incorporated herein by reference.

[0029] Unless otherwise specified, the methods used in the following examples are all conventional methods. Unless otherwise specified, the materials, reagents and instruments used are all conventional materials, reagents and instruments in the art, and can be obtained by those skilled in the art through commercial channels.

[0030] When an amount, concentration or other value or parameter is expressed as a range, a preferred range or a range defined by a series of upper preferred values ​​and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any range upper limit or preferred value with any range lower limit or preferred value, regardless of whether the range is disclosed separately. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted as including the range "1 to 4", "1 to 3", "1 to 2", "1 to 2 and 4 to 5", "1 to 3 and 5", etc. When a numerical range is described herein, unless otherwise stated, the range is intended to include its end values ​​and all integers and fractions within the range. In the present specification and claims, range definitions can be combined and / or interchanged, and if not otherwise stated, these ranges include all subranges contained therein.

[0031] The same-flavor liquor raw materials used in the following examples are all original liquors produced by the same batch of Jinpai Distillery.

[0032] The following animal experiment methods were used in the following examples:

[0033] (1) Select male BALB / c mice with good adaptability to feeding, fast for 8-16 hours before the experiment, and ensure adequate drinking water;

[0034] (2) Select a variety of wine product samples;

[0035] (3) While ensuring the same alcohol intake, calculate the intragastric volume of different alcoholic products according to the equivalent conversion method, perform intragastric gavage on mice, and record the gavage time;

[0036] (4) At different time points after drinking, serum and brain tissue samples were collected from each group of mice to detect ethanol content;

[0037] (5) The serum ethanol content is used to convert the brain tissue ethanol content at each time. The calculation formula for the brain tissue ethanol content is:

[0038] C 脑 : Brain tissue ethanol content, mg / g;

[0039] C 血 : serum ethanol content, mmol / L;

[0040] M 乙醇 : molar mass of ethanol, g / mol;

[0041] ρ 血清 : BALB / c serum density, g / cm 2 ;

[0042] K: conversion factor, 0.2-1, preferably 0.2-0.8;

[0043] (6) The statistical data is analyzed using statistical software to calculate the alcohol metabolism value and grade the wine sample. Alcohol metabolism value = AUC 实验组 / AUC 对照组 ×100%.

[0044] Taking the ethanol metabolism value of the alcohol solution group as 100 points as the benchmark, the experimental wine samples were graded as follows: less than 85, special grade; 85-95, excellent grade; 95-105, first grade; 105-115, second grade; greater than 115, third grade.

[0045] Example 1

[0046] The above method was used to verify the accuracy of the brain tissue ethanol content calculated using blood within 6 hours after drinking and the actual measured brain tissue content.

[0047] Table 1 Ethanol content in blood and cerebellum and cerebrum within 6 hours after drinking (mg / g)

[0048]

[0049] The ethanol concentrations in the brain and cerebellum tissues at each time within 6 hours after drinking showed a strict corresponding relationship with the blood, wherein the ethanol contents in the brain and cerebellum tissues were consistent, which is collectively referred to as the brain tissue ethanol concentration in the present invention. The accuracy of converting the brain tissue ethanol concentration using serum can reach as low as 93.3%, indicating that it is feasible to select the blood ethanol concentration to calculate the brain tissue ethanol concentration and infer the degree of ethanol accumulation in the brain tissue.

[0050] Example 2

[0051] This method was applied to evaluate and compare the ethanol metabolism level in brain tissue of the same wine with different alcohol content under the same alcohol intake.

[0052] Table 2 Ethanol metabolism data of mice with different alcohol concentrations after drinking (n=10, intoxicating dose)

[0053]

[0054]

[0055] As shown in Table 2, the ethanol metabolism levels in brain tissue of the same wine sample with different alcohol content are the same, and they all belong to first-grade wine, which is consistent with the facts.

[0056] Example 3

[0057] This method was applied to evaluate and compare the ethanol metabolism levels in brain tissue of wines of the same alcohol content but different alcohol degrees under the same alcohol intake.

[0058] Table 3 Ethanol metabolic dynamics after drinking different alcohol levels in mice (n=10, intoxicating dose)

[0059]

[0060] As shown in the table above, less ethanol accumulated in the brain tissue after drinking samples 3 and 4, which was mainly manifested by less absorption and faster metabolism.

[0061] In summary, the present invention uses blood to calculate the ethanol content in brain tissue, and evaluates how fast alcohol accumulates in the brain after drinking based on the AUC of ethanol in the brain tissue. This not only intuitively reflects the degree of ethanol accumulation in brain tissue and ensures the objectivity of the ethanol metabolism evaluation results, but also saves experimental animals and complies with the 3R principles of experimental animal ethics.

[0062] The above description is only an embodiment of the present application and is not intended to limit the protection scope of the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for evaluating the metabolic rate of ethanol after drinking, characterized in that: The following steps are involved: (1) Select male experimental mice with good adaptability to feeding, fast for 8-16 hours before the experiment, and ensure sufficient drinking water; (2) Select a variety of wine product samples; (3) While ensuring the same alcohol intake, calculate the intragastric volume of different alcoholic products according to the equivalent conversion method, perform intragastric gavage on mice, and record the gavage time; (4) Blood samples were collected at different time points to detect ethanol content, and the ethanol content in brain tissue was calculated based on the serum ethanol content; (5) The statistical data information is analyzed using statistical software to calculate the alcohol metabolism value and grade the wine samples.

2. The method according to claim 1, characterized in that: Alcoholic products include liquor and its blended liquors, with an alcohol content of 35-53 degrees.

3. The method according to claim 1, characterized in that: The gavage volume was 0.09-0.18 ml / 10 g.bw.

4. The method according to claim 1, characterized in that: The ethanol detection time is 0-6h.

5. The method according to claim 1, characterized in that The calculation formula for brain tissue ethanol content is: C 脑 : Brain tissue ethanol content, mg / g; C 血 : serum ethanol content, mmol / L; M 乙醇 : molar mass of ethanol, g / mol; ρ 血清 : BALB / c serum density, g / cm 2 ; K: Conversion coefficient, the value is 0.2-1.

6. The method according to claim 1, characterized in that In step (5), alcohol metabolism value = AUC 实验组 / AUC 对照组 ×100%.

7. The method according to claim 1, characterized in that Taking the ethanol metabolism value of the alcohol solution group as 100 points as the benchmark, the experimental wine samples were graded: less than 85, special grade; 85-95, excellent grade; 95-105, first grade; 105-115, second grade; greater than 115, third grade. Among them, second grade and below indicate that the experimental wine body metabolizes ethanol slowly after drinking, and first grade and above indicate that the experimental wine body metabolizes ethanol quickly after drinking. The higher the grade, the faster the ethanol metabolism.

8. The method according to claim 1, characterized in that In step (4), serum ethanol concentration: brain tissue ethanol concentration = 1: 0-1.

9. Application of any one of the methods described in claims 1 to 8 in liquor brewing.

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