A comprehensive evaluation method for the comfort level after drinking Chinese liquor at the animal level

Abstract

The present invention discloses a comprehensive animal-level evaluation method for the comfort level after drinking baijiu. This method uses the enzyme activity of aldehyde dehydrogenase in the cerebellar tissue of hangover mice, the content of 5-hydroxytryptamine in the hippocampus of hangover mice, the number of microglial cells in the brain tissue of hangover mice, the content of FGF21 protein in the serum of hangover mice, the enzyme activity of catalase in the liver tissue of hangover mice, the content of malondialdehyde in the liver tissue, the content of TNF-α in the liver tissue, the enzyme activity of alanine aminotransferase in the liver tissue, and the enzyme activity of aldehyde dehydrogenase in the liver tissue as indicators. The present invention establishes an evaluation method for the comfort level after drinking baijiu at the animal mouse level. By using appropriate statistical analysis methods to obtain a comprehensive score of comfort level after drinking to evaluate the product comfort, a set of relatively scientific, comprehensive, stable, and reliable comprehensive animal-level evaluation method for the comfort level after drinking baijiu is formed.

Description

Technical Field

[0001] The present invention belongs to the technical field of food and health products, and particularly relates to a comprehensive evaluation method for the comfort level after drinking liquor at the animal level and a liquor beverage. Background Art

[0002] Chinese liquor culture has a long history and is profound, embodying cultural and spiritual requirements. With the enhancement of people's health awareness, the drinking comfort and health of liquor have attracted more and more attention from consumers. Drinking liquor should not only satisfy the wonderful enjoyment but also not affect work and health after drinking. The drinking comfort of liquor is generally divided into three levels: comfort before drinking, comfort during drinking, and comfort after drinking. Among them, comfort after drinking refers to a series of physiological reactions after drinking, especially the impact on the nervous system and digestive system. A liquor with good comfort is specifically manifested as no headache, no dizziness, no dry mouth, and quick sobering up after drinking. At present, the evaluation of drinking comfort before and during drinking is highly subjective and greatly affected by individual differences. For the evaluation of comfort after drinking, since drinkers will have a series of physiological reactions, the evaluation indicators are more objective and are also closely related to the drinking health of liquor. Therefore, the comfort level after drinking liquor can more accurately and reliably reflect the quality of liquor.

[0003] For the evaluation of drinking comfort at home and abroad, in the early stage, questionnaires were mostly used to comprehensively judge after scoring a series of physical signs of drinking comfort / hangover (abroad, "hangover / hangover" is mostly used to represent the drinking experience). In recent years, with the continuous development of life science research methods, the animal behavior evaluation method is expected to become a feasible and relatively objective evaluation method for the comfort after drinking liquor. However, the existing animal behavior evaluation methods are not comprehensive and systematic enough, do not accurately grasp the change rules of key parameters in the body after animals drink, and fail to find the key physiological and biochemical indicators that can characterize comfort and their analysis and detection methods; they do not comprehensively evaluate the comfort level after drinking liquor from multiple dimensions and multiple indicators based on the action mechanism of comfort after drinking liquor (headache, dry mouth) and combining animal behavior characteristics. Therefore, there is an urgent need to construct a scientific, objective, and quantifiable evaluation method for comprehensively evaluating the comfort level after drinking liquor at the animal level. Summary of the Invention

[0004] The purpose of this part is to outline some aspects of the embodiments of the present invention and briefly introduce some preferred embodiments. Simplifications or omissions may be made in this part, as well as in the abstract and title of the present application, to avoid obscuring the purpose of this part, the abstract, and the title, and such simplifications or omissions shall not be used to limit the scope of the present invention.

[0005] In view of the above and / or problems existing in the prior art, the present invention is proposed.

[0006] One of the objectives of the present invention is to provide a comprehensive evaluation method for the comfort level after consuming baijiu at the animal level. By using appropriate statistical analysis methods to obtain a comprehensive comfort score after consumption to evaluate the comfort level of the product, a relatively scientific, comprehensive, stable, and reliable comprehensive evaluation method for the comfort level after consuming baijiu at the animal level is formed.

[0007] To solve the above technical problems, the present invention provides the following technical solution: A comprehensive evaluation method for the comfort level after consuming baijiu at the animal level, which uses the enzyme activity of aldehyde dehydrogenase in the cerebellar tissue of hangover mice, the content of 5-hydroxytryptamine in the hippocampus of hangover mice, the number of microglial cells in the brain tissue of hangover mice, the content of FGF21 protein in the serum of hangover mice, the enzyme activity of catalase in the liver tissue of hangover mice, the content of malondialdehyde in the liver tissue, the content of TNF-α in the liver tissue, the enzyme activity of alanine aminotransferase in the liver tissue, and the enzyme activity of aldehyde dehydrogenase in the liver tissue as indicators.

[0008] As a preferred embodiment of the comprehensive evaluation method for the comfort level after consuming baijiu at the animal level of the present invention, it includes the steps of measuring the enzyme activity of aldehyde dehydrogenase in the cerebellar tissue of hangover mice, the content of 5-hydroxytryptamine in the hippocampus of hangover mice, the number of microglial cells in the brain tissue of hangover mice, the content of FGF21 protein in the serum of hangover mice, the enzyme activity of catalase in the liver tissue of hangover mice, the content of malondialdehyde in the liver tissue of hangover mice, the content of TNF-α in the liver tissue of hangover mice, the enzyme activity of alanine aminotransferase in the liver tissue of hangover mice, and the enzyme activity of aldehyde dehydrogenase in the liver tissue of hangover mice.

[0009] As a preferred embodiment of the comprehensive evaluation method for the comfort level after consuming baijiu at the animal level of the present invention, for the measurement of the enzyme activity of aldehyde dehydrogenase in the cerebellar tissue of hangover mice, the cerebellum of hangover mice is separated and homogenized, and the ALDH enzyme activity is measured according to the ALDH detection kit.

[0010] For the measurement of the content of 5-hydroxytryptamine in the hippocampus of hangover mice, the hippocampus of the brain tissue of hangover mice is collected and homogenized, and the 5-HT content is measured according to the 5-hydroxytryptamine ELISA kit.

[0011] For the measurement of the number of microglial cells in the brain tissue of hangover mice, pathological sections of the brain tissue of hangover mice are made, and the number of microglial cells is calculated by IbA.

[0012] As a preferred embodiment of the comprehensive evaluation method for the comfort level after consuming baijiu at the animal level of the present invention, for the measurement of the content of FGF21 protein in the serum of hangover mice, the serum of hangover mice is collected, and the FGF21 content is measured according to the FGF21 ELISA kit.

[0013] As a preferred embodiment of the comprehensive animal-level evaluation method for the comfort level after drinking the white liquor of the present invention, wherein: the enzyme activity of aldehyde dehydrogenase in the liver tissue of the hangover mice is measured. The liver of the drunken mice is taken, homogenized, and the protein is extracted, and the ALDH enzyme activity is measured according to the ALDH detection kit;

[0014] The enzyme activity of alanine aminotransferase in the liver tissue of the hangover mice is measured. The liver of the drunken mice is taken, homogenized, and the protein is extracted, and the alanine aminotransferase enzyme activity is measured according to the alanine aminotransferase detection kit;

[0015] The enzyme activity of catalase in the liver tissue of the hangover mice is measured. The liver of the drunken mice is taken, homogenized, and the protein is extracted, and the CAT enzyme activity is measured according to the CAT detection kit;

[0016] The content of malondialdehyde in the liver tissue of the hangover mice is measured. The liver of the drunken mice is taken, homogenized, and the protein is extracted, and the MDA content is measured according to the MDA detection kit;

[0017] The content of TNF-α in the liver tissue of the hangover mice is measured. The liver tissue of the drunken mice is collected, homogenized, and the protein is extracted, and the expression level of the inflammatory factor TNF-α is measured according to the ELISA detection kit.

[0018] As a preferred embodiment of the comprehensive animal-level evaluation method for the comfort level after drinking the white liquor of the present invention, wherein: the enzyme activity of aldehyde dehydrogenase in the cerebellar tissue of the hangover mice, the content of 5-hydroxytryptamine in the hippocampus of the hangover mice, the number of microglial cells in the brain tissue of the hangover mice, the content of FGF21 protein in the serum of the hangover mice, the enzyme activity of catalase in the liver tissue of the hangover mice, the content of malondialdehyde in the liver tissue of the hangover mice, the content of TNF-α in the liver tissue of the hangover mice, the enzyme activity of alanine aminotransferase in the liver tissue of the hangover mice, and the enzyme activity of aldehyde dehydrogenase in the liver tissue of the hangover mice are statistically analyzed. The higher the comprehensive score, the better the comfort level after drinking the liquor.

[0019] As a preferred embodiment of the comprehensive animal-level evaluation method for the comfort level after drinking the white liquor of the present invention, wherein: the statistical analysis includes,

[0020] Performing principal component analysis on the data of each index;

[0021] Using the principal component analysis method to calculate the scores of each principal component, and the calculation formula is:

[0022]

[0023] F i =W i1 X 1 +W i2 X 2 +…W in X n(2)

[0024] In the formula, W in is the coefficient in the linear combination corresponding to the i-th component of the n-th index, and θ n represents the component matrix value corresponding to the i-th component of the n-th index, represents the square root of the eigenvalue of the i-th component; X n represents the value of the n-th index after data standardization; where, i = 1, 2, and n = 1, 2, 3, 4, 5, 6, 7;

[0025] Calculate the comprehensive score, and the calculation formula is:

[0026] F = α 1 F 1 + α 2 F 2 + … α i F i (3)

[0027] In the formula, α i represents the variance percentage of the i-th principal component.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] The present invention solves the problem that there is currently no mechanism for the effects of headache and dry mouth after drinking liquor, and there is a lack of a method for comprehensively evaluating the comfort level of liquor after drinking from multiple dimensions and multiple indicators at the animal level. The present invention establishes an evaluation method for the comfort level of liquor after drinking at the animal mouse level. By using an appropriate statistical analysis method to obtain the comprehensive score of the comfort level after drinking to evaluate the product comfort level, a relatively scientific, comprehensive, stable, and reliable comprehensive evaluation method for the comfort level of liquor after drinking at the animal level is formed. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings. Among them:

[0031] Figure 1 is the result of the mouse righting reflex experiment in Example 2 of the present invention;

[0032] Figure 2 is the result of the mouse rotarod test in Example 2 of the present invention;

[0033] Figure 3 is the result of the mouse drinking experiment in Example 2 of the present invention;

[0034] Figure 4 It is the experimental result of the "upper head" situation investigation in Example 3 of the present invention;

[0035] Figure 5 It is the determination result of the content of the "dry mouth" biomarker in Example 4 of the present invention;

[0036] Figure 6 It is the experimental result of inflammatory damage in Example 5 of the present invention;

[0037] Figure 7 It is the experimental result of oxidative damage in Example 5 of the present invention;

[0038] Figure 8 It is the experimental result of the metabolic rate of ethanol in mice in Example 5 of the present invention;

[0039] Figure 9 It is the comparison chart among Comprehensive Scores 1, 2, 3, and 4 in Example 6 of the present invention. Detailed implementation manners

[0040] To make the above objects, features, and advantages of the present invention more obvious and understandable, the following will describe the detailed implementation manners of the present invention in conjunction with the embodiments of the specification.

[0041] In the following description, many specific details are set forth to fully understand the present invention. However, the present invention can also be implemented in other ways different from those described herein. Those skilled in the art can make similar generalizations without departing from the connotation of the present invention. Therefore, the present invention is not limited by the specific embodiments disclosed below.

[0042] Secondly, the so-called "one embodiment" or "embodiment" herein refers to a specific feature, structure, or characteristic that can be included in at least one implementation manner of the present invention. The phrase "in one embodiment" appearing in different places in this specification does not necessarily refer to the same embodiment, nor is it a separate or alternative embodiment that is mutually exclusive with other embodiments.

[0043] Unless otherwise specified, the raw materials used in the embodiments are all commercially purchased.

[0044] According to the sensory evaluation of the population in Yanghe Distillery, there is the following relationship between the comfort level after drinking the M3 series of Baijiu and the production year: the longer the year, the higher the comfort level. Five wine samples are respectively marked as M3-1 (7 years), M3-2 (5 years), M3-3 (3 years), M3-4 (1 year), and M3-5 (0 year) according to the different production times.

[0045] Example 1 Determine the conditions such as the dose and time for the sample to cause intoxication in animal mice

[0046] Wine samples: M3 Yanghe wines from different years were respectively labeled as M3-1 (7 years), M3-2 (5 years), M3-3 (3 years), M3-4 (1 year), and M3-5 (0 year) according to the different storage times.

[0047] Thirty-two healthy clean-grade female Balb / c mice (6 - 8 weeks old, weighing 18 - 22 g, provided by Vital River Laboratory Animal Technology Co., Ltd.) were raised at 21 ± 2 °C, fed with normal feed, allowed free access to food and water, and raised under a 12-h day-night cycle. The groups were set as a normal control group; an alcohol group; and an experimental group.

[0048] The mice were administered different amounts of wine samples by gavage. The mice at the lowest gavage dose would show a slightly drunk state, while the mice at the highest gavage dose would show a completely drunk state. The standard dose was set as the third gavage dose, at which the mice would not be completely drunk to death. According to the drunkenness evaluation model established in the literature (Zhang Mengyan, Zheng Lu, Dang Hao, et al. Establishment of an animal model for evaluating the drunkenness degree of Luzhou-flavor liquor [J]. Modern Preventive Medicine, 2010, 37(19): 3.), the mice were tested for the righting reflex, running on a rolling drum, and swimming, and the drunkenness degree was calculated. The results showed that the mice in the alcohol group were gavaged with 0.13 ml / 10 g of 40.8% (v / v) alcohol, the mice in the experimental group were gavaged with samples of the same alcohol content, and the mice in the normal group were gavaged with the corresponding volume of PBS.

[0049] Example 2 Investigation of the behavior of mice after drinking M3 wines from different years

[0050] (1) Righting reflex

[0051] Twenty-four SPF-grade male Balb / c mice, provided by Vital River Laboratory Animal Technology Co., Ltd., weighing 18 - 22 g, 6 - 8 weeks old. They were randomly divided into a normal group, an alcohol group, and a wine sample group, with 8 mice in each group. The mice in the alcohol group were gavaged with 0.13 ml / 10 g of 40.8% (v / v) alcohol, the mice in the experimental group were gavaged with samples of the same alcohol content, and the mice in the normal group were gavaged with the corresponding volume of PBS. The righting reflex of the mice was observed, and the time for the righting reflex to recover was recorded. The drunkenness time or sobering-up time of the mice does not represent the drunkenness degree of the mice. Therefore, we calculated the drunkenness degree of the mice according to the formula in the above-mentioned literature. As Figure 1 shown, the drunkenness degree of the liquor group was significantly lower than that of the ethanol group, and the order of the drunkenness degree of the liquor group was M3-1 < M3-2 < M3-3 < M3-4 < M3-5.

[0052] (2) Rotarod test

[0053] After the righting reflex experiment, the rotation of the mice on the rod was observed, and the effective climbing time of the mice was recorded.

[0054] As Figure 2As shown, the time of the mice in the ethanol group on the rotating shaft was 186.25 ± 40 s, the time of the mice in the M3-1 group on the rotating shaft was 229 ± 25.7 s, which was 1.23 times that of the ethanol group. The time of the mice in the M3-2, M3-3, and M3-4 groups on the rotating shaft was comparable to that of the ethanol group. The time of the mice in the M3-5 group on the rotating shaft was even lower than that of the ethanol group. The falling time order of the liquor group was M3-1 > M3-2 > M3-3 > M3-4 > M3-5.

[0055] (3) Drinking water experiment

[0056] After the righting reflex of the mice recovered, they were given water to drink. The water intake of the mice was detected every 1 h.

[0057] As Figure 3 shown, 6 h after the mice were drunk, the water intakes of the M3-5 and ethanol groups of mice were the highest, and the water intake of the M3-1 group of mice was the lowest. The water intake order of the liquor group was M3-1 < M3-2 < M3-3 < M3-4 < M3-5.

[0058] It can be seen from the above experiments that the longer the age of the liquor, the lower the drunkenness degree of the mice, the faster the sobering up, the less interference with the motor coordination ability, and the less likely to have dry mouth.

[0059] Example 3 Investigation of the "hangover" situation of mice after drinking M3 liquor samples of different years

[0060] The mice were sacrificed 6 h later, the cerebellums of the experimental mice were separated and homogenized, and ALDH was determined according to the operation procedure of the ALDH detection kit; the hippocampi of the experimental mice were collected and homogenized, and the 5-hydroxytryptamine (5-HT) content was determined according to the operation procedure of the 5-HT ELISA kit, and the γ-GABA content was determined according to the operation procedure of the γ-GABA ELISA kit.

[0061] As Figure 4 shown, after drinking, the ALDH enzyme activity and γ-GABA content in the cerebellum of the mice would increase, and the 5-HT content would decrease. In the liquor group, the order of ALDH enzyme activity and γ-GABA content was: M3-1 < M3-2 < M3-3 < M3-4 < M3-5; the order of 5-HT content was: M3-1 > M3-2 > M3-3 > M3-4 > M3-5.

[0062] Example 4 Determination of the content of "dry mouth" biomarkers after drinking liquor

[0063] The mice were sacrificed 6 h later, and the hypothalami of each group of mice were taken on ice. They were ground by adding RIPA protein lysate at a ratio of 1:9, centrifuged at 5000 rpm for 10 min, and the supernatant was taken. The FGF21 content in the hypothalami of each group of mice was detected using the FGF21 kit.

[0064] AsFigure 5 As shown, the content of FGF21 in the serum of mice in the ethanol group was significantly higher than that in the control group. The order of the FGF21 content in the liquor group was: M3-1 < M3-2 < M3-3 < M3-4 < M3-5.

[0065] Determination of main physiological and biochemical indexes such as inflammatory injury and oxidative injury in the blood, liver tissue, brain tissue, etc. of experimental mice after drinking M3 of different years in Example 5

[0066] Inflammatory injury: Mice were sacrificed after 6 h, and pathological sections of the brain tissue of experimental mice were made. The number of microglial cells was calculated by double staining with IbA and CD68 to evaluate the brain inflammation.

[0067] Figure 6 Experimental methods for IL-1β, TNF-α, and iNOS mRNA: After grinding the liver tissue, centrifuge at 3000 rpm at 4 °C for 10 min, and aspirate the supernatant. The contents of IL-1β and TNF-α in the liver tissue of mice were detected according to the ELISA detection kit respectively; grind and lyse the brain tissue, extract mRNA, reverse transcribe to obtain cDNA, perform PCR with iNOS primers, take pictures in a nucleic acid imager, and perform gray analysis on the iNOS band to obtain its relative mRNA expression level.

[0068] As Figure 6 shown, by detecting the inflammatory injury of the brain tissue and liver tissue of mice, we can obtain that the order of the inflammatory injury of mice in the liquor group is: M3-1 < M3-2 < M3-3 < M3-4 < M3-5.

[0069] Oxidative injury: Mice were sacrificed after 6 h, the liver tissue of experimental mice was taken and made into homogenate, and the expression / activity of metabolic enzymes (ALT, AST, CAT, MDA, GSH, SOD) was measured as required according to the instructions of different detection kits.

[0070] As Figure 7 shown, by detecting the liver injury markers and liver antioxidant enzymes of mice, we can obtain that the order of the degree of liver oxidative injury of mice in the liquor group is: M3-1 < M3-2 < M3-3 < M3-4 < M3-5.

[0071] Detection of ADH / ALDH / CYP2E1 enzyme activity: Take the liver of drunken mice, homogenize and extract proteins, and test according to the instructions of the ADH, ALDH, and CYP2E1 enzyme activity detection kits.

[0072] As Figure 8As shown in the figure, by detecting the enzyme activities of the key ethanol-metabolizing enzymes ADH and ALDH in the livers of mice, we can obtain the following order of ethanol metabolism rates in the mice of the Chinese liquor group: M3-1 > M3-2 > M3-3 > M3-4 > M3-5.

[0073] By detecting the CYP2E1 mRNA expression of the key ethanol-metabolizing enzyme in the livers of mice, we can obtain the following order of ethanol metabolism rates in the mice of the Chinese liquor group: M3-3 > M3-4 > M3-5 > M3-2 > M3-1.

[0074] Statistical analysis of various evaluation indicators of the comfort level after drinking the samples in Example 6 and calculation of the comprehensive score

[0075] Through statistical analysis of the data of various evaluation indicators of the samples using SPSS software and the least squares method, the comprehensive score of the comfort level after drinking the samples to be tested is obtained.

[0076] ① According to the results in Examples 3 to 5, comprehensive analysis is performed on the five groups of different types of experimental data.

[0077] ② PCA analysis is performed on the data of each index of the sample, and 2 common factors are extracted, which can reflect 91.035% of the variance of the original variables.

[0078] ③ Use the principal component analysis method to calculate the scores of each principal component. The calculation formula is:

[0079] F i =W i1 X 1 +W i2 X 2 +…W in X n (1)

[0080] In formula (1), represents the coefficient in the linear combination corresponding to the nth index and the ith component, θ n represents the component matrix value corresponding to the nth index and the ith component, represents the square root of the eigenvalue of the ith component, X n represents the value of the nth index after data standardization; i = 1, 2, n = 1, 2, 3, 4, 5.

[0081] ④ Calculate the comprehensive score. The calculation formula is:

[0082] F=α 1 F 1 +α 2 F 2 +…α n F n (4)

[0083] ⑤ Standardize the data of various evaluation indexes of the samples of Yanghe Dream 3 liquor (M3-1, M3-2, M3-3, M3-4, M3-5) used in the examples in different years, and then substitute them into formulas (2), (3), and (5) for statistical analysis.

[0084] First, calculate the SPSS scores for all the results in Examples 3 to 5 to obtain the comprehensive score 1.

[0085] Remove the two indexes (iNOS mRNA expression and CYP2E1 mRNA expression in mouse brain tissue) that are inconsistent with the law of comprehensive score 1 to obtain the comprehensive score 2.

[0086] Respectively select the ALDH enzyme activity in mouse cerebellum and the GABA content in mouse hippocampus in the "getting drunk" index, the serum FGF21 content in the "dry mouth" index, CAT and GSH in the oxidation index, the number of microglial cells in mouse brain tissue and the liver TNF-α content in the inflammation index, the liver injury index ALT, and the liver ALDH enzyme activity in the ethanol metabolism index, and conduct statistical analysis to obtain the comprehensive score 3.

[0087] Respectively select the ALDH enzyme activity in mouse cerebellum and the 5-HT content in mouse hippocampus in the "getting drunk" index, the serum FGF21 content in the "dry mouth" index, CAT and MDA in the oxidation index, the number of microglial cells in mouse brain tissue and the liver TNF-α content in the inflammation index, the liver injury index ALT, and the liver ALDH enzyme activity in the ethanol metabolism index, and conduct statistical analysis to obtain the comprehensive score 4.

[0088] The statistical results are shown in Table 1.

[0089] Table 1

[0090]

[0091]

[0092] The results show that the order of the post-drinking comfort of the Dream 3 series of Baijiu evaluated by the above 4 sets of methods is: M3-1 > M3-2 > M3-3 > M3-4 > M3-5. This result is consistent with the sensory evaluation results of the population, indicating that the established animal-level evaluation method for the post-drinking comfort of Baijiu is feasible. Calculate the differences between the comprehensive scores of EtOH and the 5 liquor samples and the Ctrl group respectively, and then define the difference of the EtOH group as 1, and draw a bar chart of the relative differences, as Figure 9 shown. The results show that the comprehensive score 4 is both simple and can reflect the relationship between the aged liquor and the post-drinking comfort in a relatively reasonable and discriminatory form.

[0093] Verification of the evaluation method in Example 7

[0094] In the previous research of this laboratory, it was found that oleuropein is the preferred natural active substance for improving the comfort after drinking alcohol. Oleuropein was selected to verify the reliability of the evaluation method.

[0095] According to the methods of Examples 3 to 5, mice were randomly divided into a normal saline control group, an alcohol group, and an oleuropein group, each with 8 mice, and isoamyl alcohol, which is recognized to reduce the comfort after drinking liquor, was used as a control. The oleuropein group was gavaged with a 50 mg / kg oleuropein solution 30 minutes in advance, and the isoamyl alcohol group was gavaged with a 0.0065% isoamyl alcohol solution 30 minutes in advance, and the remaining groups were given normal saline. The alcohol group, the oleuropein group, and the isoamyl alcohol group were all gavaged with 42% (v / v) alcohol 0.12 mL / 10 g.

[0096] The comprehensive scores of the saline control group, alcohol group, oleuropein group, and isopentanol group were calculated according to the comprehensive score 4 standard. The calculation results are shown in Table 2.

[0097] Table 2

[0098]

[0099] It can be seen that the scores of mice in the alcohol group were much lower than those in the saline control group, proving that the comfort level after drinking liquor was reduced; and isopentanol, which is generally recognized to be detrimental to the comfort level after drinking liquor, had a lower comprehensive score than pure alcohol; compared with the alcohol group, oleuropein can effectively alleviate the comfort level after drinking liquor. The experimental results further illustrate the reliability or effectiveness of the evaluation method of the comprehensive score 4.

[0100] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A comprehensive animal-level evaluation method for the comfort level after drinking Chinese liquor, characterized in that: This method statistically analyzes the enzyme activity of aldehyde dehydrogenase in the cerebellar tissue of hungover mice, the content of 5-hydroxytryptamine in the hippocampus of hungover mice, the number of microglial cells in the brain tissue of hungover mice, the content of FGF21 protein in the serum of hungover mice, the enzyme activity of catalase in the liver tissue of hungover mice, the content of malondialdehyde in the liver tissue of hungover mice, the content of TNF-α in the liver tissue of hungover mice, the enzyme activity of alanine aminotransferase in the liver tissue of hungover mice, and the enzyme activity of aldehyde dehydrogenase in the liver tissue of hungover mice. The higher the comprehensive score, the better the comfort level after drinking the liquor; The said statistical analysis includes, Performing principal component analysis on the data of each index; Using the principal component analysis method to calculate the scores of each principal component, and the calculation formula is: F i = W i1 X 1 + W i2 X 2 + … W in X n (2) Where, W in is the coefficient in the linear combination corresponding to the i-th component of the n-th index, and θ n represents the component matrix value corresponding to the i-th component of the n-th index, represents the square root of the eigenvalue of the i-th component; X n represents the value after data standardization of the n-th index; where, i = 1, 2, and n = 1, 2, 3, 4, 5, 6, 7; Calculating the comprehensive score, and the calculation formula is: F = α 1 F 1 + α 2 F 2 + …α i F i (3) where α i represents the variance percentage of the i-th principal component.

2. The method according to claim 1, characterized in that: For the determination of the enzyme activity of aldehyde dehydrogenase in the cerebellar tissue of hungover mice, isolate the cerebellum of hungover mice and make it into a homogenate, and determine the ALDH enzyme activity according to the ALDH detection kit; For the determination of the number of microglial cells in the brain tissue of hungover mice, make a pathological section of the brain tissue of hungover mice, and calculate the number of microglial cells by double staining with IbA and CD68.

3. The method according to claim 1, characterized in that: For the determination of the content of 5-hydroxytryptamine in the hippocampus of hungover mice, collect the hippocampus of the brain tissue of hungover mice and make it into a homogenate, and determine the 5-HT content according to the 5-hydroxytryptamine ELISA kit.

4. The method according to claim 1, characterized in that: For the determination of the content of FGF21 protein in the serum of hungover mice, collect the serum of hungover mice, and determine the FGF21 content according to the FGF21 ELISA kit.

5. The method according to claim 1, characterized in that: For the determination of the enzyme activity of aldehyde dehydrogenase in the liver tissue of hungover mice, take the liver of drunk mice, homogenize and extract proteins, and determine the ALDH enzyme activity according to the ALDH detection kit; For the determination of the enzyme activity of alanine aminotransferase in the liver tissue of hungover mice, take the liver of drunk mice, homogenize and extract proteins, and determine the alanine aminotransferase enzyme activity according to the alanine aminotransferase detection kit; For the determination of the enzyme activity of catalase in the liver tissue of hungover mice, take the liver of drunk mice, homogenize and extract proteins, and determine the CAT enzyme activity according to the CAT detection kit.

6. The method according to claim 1, characterized in that: For the determination of the content of malondialdehyde in the liver tissue of hungover mice, take the liver of drunk mice, homogenize and extract proteins, and determine the MDA content according to the MDA detection kit.

7. The method according to claim 1, characterized in that: For the determination of the content of TNF-α in the liver tissue of hungover mice, collect the liver tissue of drunk mice, homogenize and extract proteins, and determine the expression level of the inflammatory factor TNF-α according to the ELISA detection kit.

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