Liquor with anti-fatigue and liver-protecting effects and preparation method thereof
Patent Information
- Application Number
- CN202610242293.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-02-28
- Publication Date
- 2026-06-16
Smart Images
Figure CN122214110A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of liqueur production technology, and in particular to a liqueur with anti-fatigue and liver-protecting effects and its preparation method. Background Technology
[0002] With societal development, the pace of life is accelerating, and work pressure is increasing. Long hours of work, study, and various daily chores lead to widespread physical and mental fatigue, making the search for effective anti-fatigue methods an urgent need.
[0003] my country has a rich and traditional drinking culture, and alcohol is an indispensable beverage in daily life. However, excessive drinking can severely damage health, especially liver function. When ethanol enters the body, 10% is excreted by the kidneys, while 90% is metabolized and broken down in the liver. In the liver, alcohol dehydrogenase oxidizes ethanol into acetaldehyde, which is then oxidized by acetaldehyde dehydrogenase into acetic acid, entering the tricarboxylic acid cycle, and ultimately metabolized into carbon dioxide and water. Excessive drinking can lead to alcohol poisoning, and long-term alcoholism can cause alcohol dependence. In addition, it can induce cirrhosis and liver cancer, as well as obesity, hyperlipidemia, and arteriosclerosis.
[0004] Penicillium cicadae ( Paecilomyces cicadae *Penicillium cicadae* is a common entomopathogenic fungus in southern my country. It possesses anti-cancer, analgesic, metabolic-regulating, kidney-enhancing, nerve-protecting, and sedative effects.
[0005] Patent application CN105219608 A discloses a Cordyceps militaris maca wine and its preparation method. This Cordyceps militaris maca wine comprises the following components by weight: 35-50 parts maca extract, 2-10 parts Cordyceps militaris extract, 5-10 parts crushed wolfberry, 5-10 parts crushed red dates, 2-8 parts rock sugar, 300-450 parts white wine, and 100-200 parts water. This Cordyceps militaris maca wine is said to have beneficial effects such as tonifying the kidneys and boosting yang, and improving immunity. However, this Cordyceps militaris maca wine uses Cordyceps militaris fruiting bodies as its raw material, requires the addition of active dry yeast, has a long cultivation period and is expensive, and does not have anti-fatigue or liver-protective effects.
[0006] Therefore, liqueurs with health benefits are favored by consumers, and there is an urgent need for a type of liqueur that can be consumed long-term to combat fatigue and protect the liver, has a short aging period, and is inexpensive. Summary of the Invention
[0007] This invention provides a liqueur with anti-fatigue and liver-protecting effects and its preparation method, in order to solve the defects of existing liqueurs that do not have anti-fatigue and liver-protecting effects, have long cultivation cycles and are expensive, so as to provide a liqueur that has anti-fatigue and liver-protecting effects for long-term consumption, has a short cultivation cycle and is inexpensive.
[0008] In a first aspect, the present invention provides a liquor with anti-fatigue and liver-protecting effects, comprising base liquor, and by weight, further comprising 3-12 parts wheat oligopeptide, 1-3 parts collagen tripeptide, 5-15 parts corn oligopeptide, 2-14 parts sea cucumber peptide, 5-10 parts fermentation liquid, and 400-600 parts base liquor. The fermentation broth was prepared by fermenting golden chrysanthemum, chrysanthemum and wolfberry with Penicillium cicadae.
[0009] The liqueur with anti-fatigue and liver-protecting effects provided by the present invention comprises 3-12 parts of wheat oligopeptides, for example, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, or 12 parts, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0010] The liqueur with anti-fatigue and liver-protecting effects provided by this invention includes 1-3 parts of collagen tripeptide, for example, 1 part, 1.5 parts, 2 parts, 2.5 parts, or 3 parts, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0011] The liqueur with anti-fatigue and liver-protecting effects provided by this invention comprises 5-15 parts of corn oligopeptide powder, for example, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, or 15 parts, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0012] The liqueur with anti-fatigue and liver-protecting effects provided by this invention includes 2-14 parts of sea cucumber peptide, for example, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, or 14 parts, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0013] The liqueur with anti-fatigue and liver-protecting effects provided by this invention comprises 5-10 parts of fermentation liquid, for example, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, or 10 parts, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0014] The liqueur with anti-fatigue and liver-protecting effects provided by this invention comprises 400-600 parts of wine, for example, 400 parts, 410 parts, 420 parts, 430 parts, 440 parts, 450 parts, 460 parts, 470 parts, 480 parts, 490 parts, 500 parts, 510 parts, 520 parts, 530 parts, 540 parts, 550 parts, 560 parts, 570 parts, 580 parts, 590 parts, and 600 parts, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0015] This invention provides a liquor with anti-fatigue and liver-protecting effects. The liquor contains a fermentation liquid of medicinal and edible Chinese herbal medicines with Penicillium cicadae as the strain and a polypeptide composition. Through a specific ratio, the liquor can promote the relief of fatigue and reduce liver damage caused by alcohol.
[0016] In some implementation schemes, *Penicillium cicadae* ( Paecilomyces cicadae The accession number of the specimen is CGMCC No. 3453, and it is deposited at the China General Microbiological Culture Collection Center of the China Committee on the Preservation and Management of Microbial Cultures.
[0017] In some implementations, the liqueur with anti-fatigue and liver-protecting effects comprises, by weight, 3-6 parts wheat oligopeptides, 1-2 parts collagen tripeptides, 5-8 parts corn oligopeptides, 2-6 parts sea cucumber peptides, 5-8 parts fermentation liquid, and 400-600 parts base liquor.
[0018] In some embodiments, the fermentation broth is prepared by fermentation of 1-3 parts by weight of goldenrod, 1-3 parts by weight of chrysanthemum and 3-12 parts by weight of wolfberry using Penicillium cicadae.
[0019] In some embodiments, the fermentation broth is prepared by the following method: S1. Mix 1-3 parts of golden flower, 1-3 parts of chrysanthemum and 3-12 parts of wolfberry with water and extract at 80℃~100℃ to obtain an extract; S2. Increase the concentration of live bacteria to OD. 600 The extract was inoculated with 0.5-0.6% Penicillium cicadae seed solution at an inoculation rate of 3.5%-5% (w / w), and fermentation was carried out to obtain the fermentation broth.
[0020] In some implementations, the amount of gold in step S1 is 1-3 parts, such as 1 part, 1.5 parts, 2 parts, 2.5 parts, or 3 parts, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0021] In some implementations, the amount of chrysanthemum in step S1 is 1-3 parts, such as 1 part, 1.5 parts, 2 parts, 2.5 parts, or 3 parts, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0022] In some implementations, the amount of goji berries in step S1 is 3-12 parts, for example, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, or 12 parts, but not limited to the listed values. Other unlisted values within the range are also applicable.
[0023] In some implementations, the temperature extracted in step S1 is 80°C to 100°C, for example, 80°C, 85°C, 90°C, 95°C, or 100°C, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0024] In some implementations, the viable bacterial concentration value (OD) of the *Penicillium cicadae* seed solution in step S2 is... 600 The range is 0.5-0.6, for example, it can be 0.5, 0.51, 0.52, 0.53, 0.54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.6, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0025] In some implementations, the inoculation amount of *Penicillium cicadae* seed solution is 3.5% to 5% (w / w), for example, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0026] In some implementation schemes, step S1 specifically involves: mixing 1-3 parts of goldenrod, 1-3 parts of chrysanthemum, and 3-12 parts of wolfberry with water at a material-to-liquid ratio of 1 g:(10-15) mL, and extracting at 80℃-100℃ for 1-3 h to obtain a first filtrate; mixing the filtered residue with water at a material-to-liquid ratio of 1 g:(10-15), and extracting at 80℃-100℃ for 1-3 h to obtain a second filtrate; combining the first filtrate and the second filtrate and concentrating to 1 / 2-1 / 3 of the original volume to obtain an extract.
[0027] In some implementations, the material-to-liquid ratio for extracting the first filtrate and the second filtrate is 1 g: (10~15) mL, for example, it can be 1 g: 10 mL, 1 g: 11 mL, 1 g: 12 mL, 1 g: 13 mL, 1 g: 14 mL, 1 g: 15 mL, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0028] In some implementations, the extraction time for the first and second filtrates is 1-3 hours, for example, 1 hour, 1.5 hours, 2 hours, 2.5 hours, or 3 hours, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0029] In some embodiments, in step S2, the *Penicillium cicadae* seed solution is prepared by the following method: S21. Slant culture expansion of Penicillium cicadae strain: Inoculate Penicillium cicadae strain into slant culture medium and culture at 25~30℃ for 5-9 days; S22. Preparation of Penicillium ciliata seed culture: Select sporulating Penicillium ciliata fungi and inoculate them into seed culture medium. Culture them in a shaker at 25-30℃ and 110-130 r / min for 4-7 days.
[0030] In some embodiments, the slant culture medium is a solid PDA medium, and the seed culture medium is a liquid PDA medium.
[0031] In some implementations, the culture temperature in step S21 is 25~30℃, for example, it can be 25℃, 26℃, 27℃, 28℃, 29℃, or 30℃, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0032] In some implementations, the culture time in step S21 is 5-9 days, for example, 5 days, 6 days, 7 days, 8 days, or 9 days, but not limited to the listed values. Other unlisted values within the range are also applicable.
[0033] In some implementations, the shaking incubation temperature in step S22 is 25~30℃, for example, it can be 25℃, 26℃, 27℃, 28℃, 29℃, or 30℃, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0034] In some implementations, the shaking speed in step S22 is 110-130 r / min, for example, it can be 110 r / min, 115 r / min, 120 r / min, 125 r / min, or 130 r / min, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0035] In some implementations, the shaker culture time in step S22 is 4-7 days, for example, 4 days, 5 days, 6 days, or 7 days, but not limited to the listed values. Other unlisted values within the range are also applicable.
[0036] In some implementations, in step S2, the fermentation temperature is 25~30℃, for example, it can be 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, but is not limited to the listed values, and other unlisted values within the range are also applicable; in step S2, the fermentation time is 5-9 days, for example, it can be 5 days, 6 days, 7 days, 8 days, 9 days, but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0037] In some embodiments, the fermentation broth is prepared by the following method: S1. Mix 1-3 parts of goldenrod, 1-3 parts of chrysanthemum, and 3-12 parts of wolfberry with water at a material-to-liquid ratio of 1 g:(10-15) mL, and extract at 80℃-100℃ for 1-3 h to obtain the first filtrate; mix the filtered residue with water at a material-to-liquid ratio of 1 g:(10-15), and extract at 80℃-100℃ for 1-3 h to obtain the second filtrate; combine the first filtrate and the second filtrate and concentrate to 1 / 2-1 / 3 of the original volume to obtain the extract; S2. Increase the concentration of live bacteria to OD. 600 The extract was inoculated with 0.5-0.6% Penicillium cicadae seed solution at an inoculation rate of 3.5%-5% (w / w), and fermented at 25-30℃ for 5-9 days to obtain the fermentation broth. In step S2, the Penicillium cicadae seed liquid is prepared by the following method: S21. Slant culture expansion of Penicillium cicadae strain: Inoculate Penicillium cicadae strain into slant culture medium and culture at 25~30℃ for 5-9 days; S22. Preparation of Penicillium ciliata seed culture: Select sporulating Penicillium ciliata fungi and inoculate them into seed culture medium. Culture them in a shaker at 25-30℃ and 110-130 r / min for 4-7 days.
[0038] This scheme controls the fermentation broth to be prepared by the specific method described above, which can fully convert the active ingredients in the raw materials into small molecules that are more easily absorbed by the human body, while generating a variety of prebiotics and antioxidant active ingredients to help relieve physical fatigue.
[0039] In some implementations, the base liquor has an alcohol content of 50-55 degrees. For example, it can be 50 degrees, 51 degrees, 52 degrees, 53 degrees, 54 degrees, or 55 degrees, but is not limited to the listed values. Other unlisted values within the range are also applicable.
[0040] This formula controls the alcohol content of the base liquor to 50-55 degrees. While improving the taste of the liqueur, it also allows the active ingredients in the fermentation liquid of medicinal and edible herbs with Penicillium cicadae as the strain and the polypeptide composition system to fully exert their effects in the liquor at this alcohol content, thereby playing a role in anti-fatigue and liver protection.
[0041] A second aspect of the present invention provides a method for preparing a liqueur with anti-fatigue and liver-protective effects as described in the first aspect, comprising: The wheat oligopeptide, the collagen tripeptide, the corn oligopeptide powder, and the sea cucumber peptide are added to the mixture of the fermentation liquid and the base wine, and mixed evenly to obtain the liqueur with anti-fatigue and liver-protecting effects.
[0042] Compared with the prior art, the beneficial technical effects of the present invention are as follows: This invention achieves a synergistic enhancement of anti-fatigue and liver-protective effects through the scientific combination of fermentation of medicinal and edible strains and polypeptide components. During fermentation, the medicinal and edible strain (Penicillium cicadae strain) transforms the active ingredients in the raw materials into smaller molecules that are more easily absorbed by the human body, while simultaneously generating various prebiotics and antioxidant active ingredients to help alleviate fatigue. Meanwhile, the exogenously added polypeptides can rapidly replenish the amino acid substrates needed for energy metabolism, reduce muscle protein breakdown, and further enhance the anti-fatigue effect. The combined effect of these two components protects liver cells from damage by regulating the activity of antioxidant enzymes in the liver and inhibiting lipid peroxidation, while also accelerating the clearance of fatigue metabolites such as lactic acid and urea nitrogen. Compared to liqueurs with a single active ingredient, this invention offers a longer duration of anti-fatigue effects and enhanced liver-protective activity.
[0043] (2) This invention uses Penicillium cicadae strain to ferment the ingredients that are both food and medicine. On the one hand, it can reduce the anti-nutritional factors or irritating components that may exist in the raw materials through microbial metabolism, reduce the metabolic burden on the liver, and conform to the safety concept of "food and medicine are of the same origin". On the other hand, the flavor substances such as esters and organic acids produced during the fermentation process can effectively neutralize the fishy smell of polypeptides, improve the taste and flavor of the liqueur, solve the problem that the efficacy and palatability of traditional functional liqueurs are difficult to balance, improve the acceptance of the product, and are suitable for long-term daily health care scenarios.
[0044] (3) The product of this invention has both anti-fatigue and liver protection functions. It can meet the anti-fatigue needs of people with high-intensity work and sports enthusiasts, and also meet the liver health needs of people who drink alcohol for a long time and stay up late. It breaks through the bottleneck of traditional functional liquors that have "single function and limited target population". It can be widely used in daily health care, business banquets, post-exercise recovery and other scenarios, and enhance the market competitiveness and application value of the product. Attached Figure Description
[0045] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.
[0046] Figure 1 The graph shows the effects of the blank control, base wine group, and liqueur group on the weight-bearing swimming time of mice in Example 2.
[0047] Figure 2 The graph shows the effects of the blank control, base wine group, and liqueur group on plasma urea nitrogen content in Example 2.
[0048] Figure 3 The graph shows the effects of the blank control, base wine group, and liqueur group on liver glycogen and muscle glycogen in Example 2.
[0049] Figure 4 The figure shows the effects of the blank control, base wine group, and liqueur group on ALT and AST in mice in Example 3.
[0050] Figure 5 The figure shows the effects of the blank control, base wine group, and liqueur group on ADH and ALDH in mice in Example 3.
[0051] Figure 6 The graph shows the effects of the blank control, base wine group, and liqueur group on serum SOD, GSH, and MDA in mice in Example 3. Detailed Implementation
[0052] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.
[0053] Unless otherwise specified, the techniques or conditions described in the examples are as described in the literature in this field, or as per the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.
[0054] Example 1 Microbial culture 1. PDA culture medium Liquid PDA medium: 200 g / L potato, 20 g / L glucose, 5 g / L peptone, 3 g / L KH2PO4, 1.5 g / L MgSO4·7H2O, sterilized at 121℃ for 20 min.
[0055] Solid PDA medium: 200 g / L potato, 20 g / L glucose, 5 g / L peptone, 3 g / L KH2PO4, 1.5 g / L MgSO4·7H2O, 20 g / L agar, sterilized at 121℃ for 20 min.
[0056] 2. Slant culture expansion cicada-like Penicillium ( Paecilomyces cicadaeThe strain (CGMCC No. 3453, deposited at the China General Microbiological Culture Collection Center) was used as the strain source. After preparing the slant solid medium (solid PDA medium), it was sterilized in an autoclave and then inoculated on a clean bench. The spore powder of *Penicillium ciliata* was picked up with an inoculation loop and inoculated onto the slant medium (solid PDA medium), and then placed in a constant temperature incubator at 28°C for 7 days.
[0057] 3. Preparation of Penicillium cicadae seed liquid Select *Penicillium cicadae* fungi that showed good growth and were confirmed to produce sporulation after 7 days of incubation in an incubator. Inject 5 mL of Tween 80 solution into the slant culture medium. Use an inoculation loop to scrape the *Penicillium cicadae* fungi from the culture medium and inoculate them into seed culture medium (liquid PDA medium). Place the incubator in a constant temperature shaker and incubate at 28℃ and 120 r / min for 5 days, until the viable cell concentration reaches OD500. 600 The seed solution of Penicillium cicadae was 0.55.
[0058] 4. Preparation of fermentation broth for food and medicine homology Weigh out 2.5 parts by weight of golden flower, 2.5 parts by weight of chrysanthemum and 7 parts by weight of wolfberry, mix them together, add water (material-to-liquid ratio 1 g:10 mL) and soak for 6 hours, decoct at 85℃ for 2 hours, filter and set aside the filtrate; add water (material-to-liquid ratio 1 g:10 mL) to the filtered residue and continue to decoct for 2 hours, filter and set aside the filtrate. The filtrates obtained above are combined, concentrated by rotation to half of the original volume, centrifuged, and the supernatant is collected and sterilized to obtain the extract.
[0059] OD concentration of live bacteria 600 The seed culture of *Penicillium ciliata* was inoculated into the extract at a concentration of 0.55% (w / w), and fermented at 28°C for 5 days to obtain the fermentation broth.
[0060] 5. Preparation of liqueurs with anti-fatigue and liver-protecting effects Weigh out 4 parts wheat oligopeptide, 1.5 parts collagen tripeptide, 6 parts corn oligopeptide powder, and 4 parts sea cucumber peptide (commercially available) and add them to a mixture of 8 parts fermentation liquid and 500 parts base liquor (baijiu, 52 degrees). Stir well and dissolve completely. Filter at 4°C to remove impurities. After blending, you will get 52-degree liqueur.
[0061] The prepared liqueur was subjected to experimental research. The following is the implementation process and the results obtained: All experimental data in this study are presented as mean ± standard deviation (Mean ± SD). SPSS 26.0 statistical software was used for data analysis. Before comparing between groups, the normality of the data was verified by the Shapiro-Wilk test, and the homogeneity of variance was assessed using the Levene test. For data that conformed to a normal distribution and had homogeneous variance, one-way ANOVA was used to compare differences among multiple groups. The statistical significance criteria were as follows: * P <0.05 indicates that the difference is statistically significant. ** P <0.01 indicates that the difference is statistically significant.
[0062] Example 2 Anti-fatigue experimental study 1.1 Materials Analytical balance; weighing scale; thermometer; lead sheet; exercise timer; swimming tank (50 cm × 50 cm × 40 cm); blood urea nitrogen (BUN) test kit (urease method); liver / muscle glycogen assay kit (colorimetric method): all purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd.; full-wavelength microplate reader (Mutiskan SkyHigh type): Thermo Fisher Scientific; refrigerated centrifuge (TGL-20MB type): Changsha Xiangzhi Centrifuge Instrument Co., Ltd.
[0063] 1.2 Animals Specific pathogen-free (SPF) grade male Kunming mice: 4-5 weeks old, weighing (22±2) g, 45 mice. The animals were housed in an environment with a temperature of (23±2) ℃, humidity of 55%±5%, and a light-dark cycle of 12 h, with free access to food and water.
[0064] 1.3 Animal grouping and model establishment Forty-five male Kunming mice were used and, after a 7-day acclimatization period, were randomly divided into three groups: a blank control group (no oral administration of alcohol), a base liquor group (52-degree baijiu), and a spirit group (52-degree spirit prepared in Example 1), with 15 mice in each group. Each group was administered the corresponding test substance by gavage at a dose of 0.2 mL / 20 g body weight (determined according to pharmacological dosage conversion), once daily for 30 consecutive days.
[0065] 1.4 Determination of the swimming time of mice under load Kunming mice in each group were administered the medication by gavage for 30 days. Half an hour after the last gavage, six mice from each group were randomly selected and placed in a swimming tank at a water temperature of 25℃±0.5℃. The mice were loaded with lead weights equal to 5% of their body weight at the base of their tails. The time from the start of swimming to exhaustion was recorded. The criterion for exhaustion was that the mouse's head submerged in the water and it could not return to the surface within 10 seconds. The calculation method for the weighted swimming time was the time from the start of swimming to exhaustion.
[0066] 1.5 Effects on plasma urea nitrogen content and muscle / liver glycogen content in mice Thirty minutes after the last administration of the test sample, the remaining mice swam in water at 30°C without load for 90 minutes. After resting for 60 minutes, blood was collected, centrifuged, and plasma was used to measure blood urea nitrogen. 100 mg of liver and 100 mg of leg muscle were collected to measure liver / muscle glycogen content.
[0067] 2. Results 2.1 Mouse weight-bearing swimming experiment Figure 1 This is a graph showing the effects of the blank control, base wine group, and liqueur group on the weight-bearing swimming time of mice in Example 2. Figure 1 It was found that the weight-bearing swimming time of mice in the base wine group and the liqueur group was significantly increased compared with that in the blank control group, with the liqueur group showing the best performance in weight-bearing swimming. The results indicate that liqueur can effectively prolong the weight-bearing swimming time of mice, and its active ingredients significantly enhance the anti-fatigue effect.
[0068] 2.2 Effect on plasma urea nitrogen content The body's primary energy source is blood glucose. After prolonged strenuous exercise, a significant amount of energy is consumed, leading to a drop in blood glucose levels. When energy supply is insufficient, the body breaks down protein to meet its energy needs. Protein breakdown produces ammonia, which is metabolized into urea. The level of blood urea nitrogen (BUN) in plasma increases with increasing exercise load; the stronger the body's adaptation to exercise, the lower the BUN level. Blood urea nitrogen is an important indicator for evaluating protein catabolism.
[0069] Figure 2 The graph shows the effects of the blank control, base wine group, and liqueur group on plasma urea nitrogen content in Example 2 (compared to the blank group). ### P <0.001; compared with the base wine group, ** P <0.01; n=9). (From...) Figure 2 It can be seen that the plasma urea nitrogen level of mice in the base wine group was significantly higher than that in the blank control group. P<0.001); Compared with the base wine group, the plasma urea nitrogen level in mice was significantly lower in the liqueur group. This indicates that the production of plasma urea nitrogen in mice after exercise is slower or the clearance of plasma urea nitrogen is faster, which improves the mice's exercise adaptability and delays the onset of fatigue after exercise.
[0070] 2.3 Effects on liver glycogen and muscle glycogen The primary function of liver glycogen is to maintain stable blood glucose levels. When blood glucose levels in mice decrease, liver glycogen is rapidly broken down into glucose, which is released into the bloodstream to maintain blood glucose concentration within a normal range, providing energy for vital organs such as the brain and red blood cells. After glucose enters mouse muscle cells, it initiates the synthesis of muscle glycogen under the catalysis of enzymes such as hexokinase. Similar to liver glycogen synthesis, this process involves a series of reactions, including phosphorylation, ultimately forming muscle glycogen stored in muscle cells. Muscle glycogen provides energy for muscle contraction. During exercise in mice, muscle glycogen is broken down into glucose-6-phosphate, which then produces ATP through glycolysis and other pathways, providing a direct energy source for muscle contraction. Studying the effects of exhaustive exercise on liver and muscle glycogen levels in mice can help understand the mechanisms of exercise fatigue.
[0071] Figure 3 The graph shows the effects of the blank control, base wine group, and liqueur group on liver glycogen and muscle glycogen in Example 2 (compared to the blank group). ## P <0.01, ### P <0.001; compared with the base wine group, * P <0.05, ** P <0.01, *** P <0.001, n=9). From Figure 3 It can be seen that, compared with the blank control group, the liver glycogen and muscle glycogen content in the base liquor group was significantly reduced. P <0.001 or P <0.01), compared with the base wine group, the liver glycogen and muscle glycogen content in the liqueur group was significantly increased ( P <0.001 or P <0.01). The results show that liqueur can effectively reduce the consumption of liver glycogen and muscle glycogen compared with base wine.
[0072] Example 3 Liver protection experimental study 1.1 Materials Alanine aminotransferase (ALT / GPT) test kit, aspartate aminotransferase (AST / GOT) test kit, total superoxide dismutase (T-SOD) assay kit, reduced glutathione (GSH) assay kit, malondialdehyde (MDA) content assay kit, triglyceride (TG) assay kit, alcohol dehydrogenase (ADH) activity assay kit, aldehyde dehydrogenase (ALDH) activity assay kit, 4% paraformaldehyde, BCA protein quantification kit. All reagents were purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd.; Multiskan SkyHigh microplate reader: Thermo Fisher Scientific; TGL-20MB refrigerated centrifuge: Changsha Xiangzhi Centrifuge Instrument Co., Ltd.; analytical balance, weighing scale.
[0073] 1.2 Animals Specific pathogen-free (SPF) grade male Kunming mice: 4-5 weeks old, weighing (22±2) g, 45 mice. The animals were housed in an environment with a temperature of (23±2) ℃, humidity of 55%±5%, and a light-dark cycle of 12 h, with free access to food and water.
[0074] 1.3 Animal grouping and model establishment Forty-five male Kunming mice were used and, after a 7-day acclimatization period, were randomly divided into three groups: a blank control group (no oral administration of alcohol), a base alcohol group, and a spirits group, with ten mice in each group. Each group was administered the corresponding test substance by gavage at a dose of 0.2 mL / 20 g body weight (determined according to pharmacological dosage conversion), once daily for 10 consecutive days.
[0075] 1.4 Sample Collection and Processing Two hours after the last gavage, mice in each group were anesthetized with isoflurane, and blood was collected by enucleation (centrifugation at 3000 r / min for 15 min at 4℃, and the upper serum was collected and stored in a -80℃ refrigerator). The mice were dissected and the livers were removed. The liver tissue was repeatedly rinsed in physiological saline at 4℃ and then cut off. The liver tissue was stored in a -80℃ refrigerator for the detection of liver biochemical indicators.
[0076] 1.5 Determination of Biochemical Indicators Pre-cooled physiological saline was added to the liver tissue, and homogenized under ice bath conditions to prepare a 10% (w / w) liver tissue homogenate. The homogenate was centrifuged (3000 r / min, 4℃, 20 min), and the supernatant was collected. The activity of SOD, GSH, and MDA in mouse liver, as well as the activities of ethanol metabolism-related enzymes ADH and ALDH, were measured according to the kit instructions. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in mouse serum were measured according to the kit instructions.
[0077] 2. Results 2.1 Effects on ALT and AST in mice Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are important indicators for evaluating liver damage. When hepatocytes are abnormal, they can cause inflammatory response and hepatocyte necrosis. Increased cell membrane permeability allows ALT and AST from hepatocytes to enter the bloodstream.
[0078] Figure 4 The graph shows the effects of the blank control, base wine group, and liqueur group on ALT and AST in mice in Example 3 (compared to the blank control group). ## P <0.01, ### P <0.001; compared with the base wine group, * P <0.05, ** P <0.01, *** P <0.001, n=15). From Figure 4 It can be seen that, compared with the blank control group, the ALT and AST levels in the base wine group mice were significantly increased. P <0.001); compared with the base wine group, the ALT and AST levels in the liqueur group mice were significantly reduced ( P <0.01 or P <0.001). The results showed that the liver damage in the base wine group was significantly greater than that in the blank control group, and the liqueur could effectively reduce the damage of alcohol to the liver.
[0079] 2.2 Effects on ADH and ALDH in mice ADH (alcohol dehydrogenase) and ALDH (acetaldehyde dehydrogenase) are two key enzymes in alcohol metabolism. After alcohol (ethanol) enters the body, ADH oxidizes ethanol to acetaldehyde, which is the first step in alcohol metabolism. This catalytic process relies on the coenzyme nicotinamide adenine dinucleotide (NAD+). ADH dehydrogenates the hydroxyl group of ethanol and simultaneously reduces NAD+ to reduced nicotinamide adenine dinucleotide (NADH). Acetaldehyde produced by ADH catalysis is toxic and can cause various adverse reactions in the human body, such as facial flushing and rapid heartbeat. ALDH further oxidizes acetaldehyde to acetic acid, which is ultimately broken down into carbon dioxide and water and excreted from the body, thus completing the alcohol metabolism process. The activity levels of ADH and ALDH directly affect an individual's ability to metabolize alcohol.
[0080] Figure 5 The graph shows the effects of the blank control, base wine group, and liqueur group on ADH and ALDH in mice in Example 3 (compared to the blank group). ## P <0.01, ###P <0.001; compared with the base wine group, * P <0.05, ** P <0.01, *** P <0.001, n=15). From Figure 5 It can be seen that, compared with the blank control group, the activities of ADH and ALDH in the base wine group were significantly reduced. P <0.01 or P <0.001); compared with the base wine group, the activities of ADH and ALDH were significantly increased in the liqueur group ( P <0.001). The results indicate that liqueur can increase the activity of ADH and ALDH and accelerate alcohol metabolism.
[0081] 2.3 Effects on serum SOD, GSH and MDA in mice Superoxide dismutase (SOD), glutathione peroxidase (GSH), and malondialdehyde (MDA) are important biochemical indicators for assessing liver health and oxidative stress. SOD is an important antioxidant enzyme that catalyzes the oxidation of superoxide anion radicals (O2). - Superoxide anion (SOD) is converted into hydrogen peroxide (H2O2) and oxygen, thereby reducing oxidative damage to cells caused by free radicals. Free radicals are produced during liver metabolism, and the level of superoxide anion increases, especially in cases of liver damage, inflammation, or other pathological states. Decreased SOD levels generally indicate increased oxidative stress in the body and may be associated with liver diseases such as hepatitis, cirrhosis, and fatty liver. Increased SOD levels may reflect the liver's adaptive response to combat free radical damage.
[0082] Glutathione-dependent glutathione (GSH) catalyzes the reduction of hydrogen peroxide (H₂O₂) and organic peroxides, protecting cells from oxidative damage. It is primarily found in the liver and other important antioxidant defense systems. GSH levels are commonly used to assess the liver's antioxidant capacity. Low GSH levels may indicate impaired liver antioxidant defenses, increased oxidative stress, and increased susceptibility to liver cell damage.
[0083] MDA is a product of lipid peroxidation and is commonly used to measure the degree of lipid peroxidation in the body. Lipid peroxidation is part of the oxidative stress response, and MDA production is usually associated with cell membrane damage, especially in hepatocytes. Elevated MDA levels generally indicate that the liver has undergone lipid peroxidation under oxidative stress, resulting in cell membrane damage. MDA is used as a marker of oxidative damage, and its increase can reflect the degree of liver damage; for example, elevated MDA levels may be seen in diseases such as hepatitis, cirrhosis, and alcoholic liver disease.
[0084] Figure 6 The graph shows the effects of the blank control, base wine group, and liqueur group on serum SOD, GSH, and MDA in mice in Example 3 (compared to the blank group). ## P <0.01, ### P <0.001; compared with the base wine group, * P <0.05, ** P <0.01, *** P <0.001, n=15). From Figure 6 It can be seen that, compared with the blank control group, the levels of SOD and GSH in the base wine group mice were significantly reduced. P <0.01 or P <0.001), MDA levels increased significantly ( P <0.001); Compared with the base wine group, the levels of SOD and GSH in the liqueur group were significantly increased ( P <0.01 or P <0.001), MDA levels decreased significantly ( P <0.001). The results indicate that liqueur can enhance the body's antioxidant capacity and reduce oxidative stress in the liver.
[0085] In summary, the analysis shows that this liqueur has a certain protective effect on relieving fatigue and protecting the liver.
[0086] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims
1. A liqueur with anti-fatigue and liver-protecting effects, comprising a base spirit, characterized in that, By weight, it also includes 3-12 parts wheat oligopeptides, 1-3 parts collagen tripeptides, 5-15 parts corn oligopeptides, 2-14 parts sea cucumber peptides, and 5-10 parts fermentation liquid. The fermentation broth was prepared by fermenting golden chrysanthemum, chrysanthemum and wolfberry with Penicillium cicadae.
2. The liqueur with anti-fatigue and liver-protecting effects according to claim 1, characterized in that, By weight, the liqueur comprises 3-6 parts wheat oligopeptides, 1-2 parts collagen tripeptides, 5-8 parts corn oligopeptides, 2-6 parts sea cucumber peptides, 5-8 parts fermentation liquid, and 400-600 parts base liquor.
3. The liqueur with anti-fatigue and liver-protecting effects according to claim 1 or 2, characterized in that, The fermentation broth, by weight, is prepared by fermenting 1-3 parts of goldenrod, 1-3 parts of chrysanthemum and 3-12 parts of wolfberry with Penicillium cicadae.
4. The liqueur with anti-fatigue and liver-protecting effects according to claim 1 or 3, characterized in that, The fermentation broth was prepared by the following method: S1. Mix 1-3 parts of golden flower, 1-3 parts of chrysanthemum and 3-12 parts of wolfberry with water and extract at 80℃~100℃ to obtain an extract; S2. Increase the concentration of live bacteria to OD. 600 The extract was inoculated with 0.5-0.6% Penicillium cicadae seed solution at an inoculation rate of 3.5%-5% (w / w), and fermentation was carried out to obtain the fermentation broth.
5. The liqueur with anti-fatigue and liver-protecting effects according to claim 4, characterized in that, Step S1 is as follows: Mix 1-3 parts of golden flower, 1-3 parts of chrysanthemum and 3-12 parts of wolfberry with water at a material-to-liquid ratio of 1 g: (10-15) mL and extract at 80℃-100℃ for 1-3 h to obtain the first filtrate; mix the filtered residue with water at a material-to-liquid ratio of 1 g: (10-15) and extract at 80℃-100℃ for 1-3 h to obtain the second filtrate; combine the first filtrate and the second filtrate and concentrate to 1 / 2-1 / 3 of the original volume to obtain the extract.
6. The liqueur with anti-fatigue and liver-protecting effects according to claim 4 or 5, characterized in that, In step S2, the Penicillium cicadae seed liquid is prepared by the following method: S21. Slant culture expansion of Penicillium cicadae strain: Inoculate Penicillium cicadae strain into slant culture medium and culture at 25~30℃ for 5-9 days; S22. Preparation of Penicillium ciliata seed culture: Select sporulating Penicillium ciliata fungi and inoculate them into seed culture medium. Culture them in a shaker at 25-30℃ and 110-130 r / min for 4-7 days.
7. The liqueur with anti-fatigue and liver-protecting effects according to claim 6, characterized in that, The slant culture medium is a solid PDA medium, and the seed culture medium is a liquid PDA medium.
8. The liqueur with anti-fatigue and liver-protecting effects according to any one of claims 4-7, characterized in that, In step S2, the fermentation temperature is 25~30℃, and the fermentation time is 5-9 days.
9. The liqueur with anti-fatigue and liver-protecting effects according to claim 1, characterized in that, The base liquor has an alcohol content of 50-55%.
10. A method for preparing a liqueur with anti-fatigue and liver-protecting effects as described in any one of claims 1 to 9, characterized in that, include: The wheat oligopeptide, the collagen tripeptide, the corn oligopeptide powder, and the sea cucumber peptide are added to the mixture of the fermentation liquid and the base wine, and mixed evenly to obtain the liqueur with anti-fatigue and liver-protecting effects.
Citation Information
Patent Citations
Isaria cicadae miq lepidium meyenii wine and brewing method thereof
CN105219608A