Composition for synergistically alleviating physical fatigue and application thereof

Abstract

The present invention discloses a composition for synergistically relieving physical fatigue and its application. The composition of the present invention can synergistically regulate anti-oxidation, calcium ion metabolism and energy metabolism, synergistically improve the body's ATP content and glycogen level, accelerate the clearance of lactic acid and urea nitrogen, prolong exercise time, and significantly relieve physical fatigue. Its synergistic effect is stronger than that of using red ginseng, black wolfberry and ganoderma lucidum alone. At the same time, the ingredients of the composition of the present invention are simple and all derived from natural plants; at the same time, after scientific demonstration and experimental verification, the mechanism of action of the composition is clear, and the fatigue-relieving interaction relationship between the various ingredients is clear, with the advantages of being green and safe. In addition, the preparation process of the composition of the present invention is simple and easy to operate, and does not require the investment of professional and technical personnel and large-scale instruments and equipment. The production cost is low, which is convenient for industrial development.

Description

Technical Field

[0001] The present invention relates to the fields of medicine and food technology, and in particular to a composition for synergistically alleviating physical fatigue and an application thereof. Background Art

[0002] Fatigue is a common physiological phenomenon in human life, and exercise-induced fatigue refers to the inability to maintain a specific level of function or a predetermined exercise intensity during physiological processes. Some food-derived active ingredients can significantly regulate energy metabolism and metabolite accumulation, alleviating physical fatigue through various pathways, including antioxidant activity, reduced metabolite accumulation, anti-inflammatory effects, and regulation of energy metabolism and the intestinal microbiome.

[0003] In recent years, research in the field of anti-fatigue has made some progress both domestically and internationally, but many unresolved issues remain. For example, while some known active ingredients have some anti-fatigue effects, their effectiveness is limited and they may have certain side effects. Furthermore, the formulation design and preparation processes of existing products have certain limitations, making it difficult to meet market demand. For example, the formulation ingredients are complex, and the selection and proportioning of raw materials lack scientific basis; the mechanism of action of compound formula products is unclear; the vast majority of products are formulated based on practical application experience and lack scientific justification and experimental verification; and the fatigue-relieving interactions between the various ingredients in the formulation are unclear. With the continuous improvement of consumer health awareness, products developed from natural plants, especially traditional edible resources, with minimal side effects and health-promoting functions are more likely to gain public acceptance. Therefore, it is of great significance to develop a scientifically verified, safe, and directly edible product composed of active ingredients from traditional medicinal and edible products or natural plants that has a synergistic effect in relieving physical fatigue.

[0004] In view of this, the present invention is proposed. Summary of the Invention

[0005] The object of the present invention is to provide a composition for synergistically relieving physical fatigue and its application. The composition is composed of at least two of red ginseng, black wolfberry and ganoderma lucidum, which are compounded in a specific proportion so that they can effectively relieve physical fatigue.

[0006] To achieve the above-mentioned purpose, the technical solution adopted by the present invention to solve the technical problem is:

[0007] The invention provides a traditional Chinese medicine composition, the components of which are selected from at least two of red ginseng, black wolfberry and ganoderma lucidum.

[0008] Red Ginseng ( Talinum paniculata(Jacq.) Gaertn. is a perennial herbaceous plant of the genus Panax, Araliaceae. It is a processed product of ginseng, made by removing the lateral and fine roots and then steaming, sun-drying, or oven-drying. It contains a variety of active ingredients, including ginsenosides, polyacetylene compounds, and acidic polysaccharides. Black wolfberry, also known as black wolfberry, belongs to the genus Lycium, Solanaceae. It is sweet and neutral in nature, and enters the liver and kidney meridians. Ganoderma lucidum is a traditional tonic in Traditional Chinese Medicine, and Ganoderma polysaccharides are one of its main active ingredients.

[0009] The present invention screened a number of substances that can relieve physical fatigue from a variety of food-derived ingredients. By analyzing and comparing the effects of various substances on indicators such as the body's ATP content, glycogen level, lactic acid and urea nitrogen clearance capacity, it was found that: red ginseng can exert anti-fatigue effects by regulating mouse serum urea nitrogen and body glycogen content; the main active ingredient in black wolfberry - black wolfberry polysaccharide can increase serum and liver superoxide dismutase activity, enhance the body's antioxidant capacity, and relieve mouse fatigue; Ganoderma lucidum polysaccharide can increase mouse glycogen reserves, reduce lactic acid and urea nitrogen accumulation, and relieve fatigue. Ganoderma lucidum triterpenes have strong antioxidant and anti-inflammatory abilities and can relieve fatigue by regulating body metabolism. Therefore, the three substances of red ginseng, black wolfberry and Ganoderma lucidum are selected for two-by-two compounding or three-by-three compounding. The compounded composition can exert a synergistic effect in terms of body ATP content, glycogen level, lactic acid and urea nitrogen clearance capacity, etc., so that the anti-fatigue effect of the compounded composition is significantly improved.

[0010] Based on this, the composition provided by the present invention can be a composition consisting of two components: red ginseng and black wolfberry, red ginseng and Ganoderma lucidum, or black wolfberry and Ganoderma lucidum; or a composition consisting of three components: red ginseng, black wolfberry and Ganoderma lucidum.

[0011] The forms of the above ingredients include but are not limited to powders, and may also be extracts, extracts, concentrates and slices. Specifically, the extracts may be water extracts or alcohol extracts, and those skilled in the art may compound and adjust them according to actual conditions.

[0012] In some embodiments, when preparing the above composition, red ginseng, black wolfberry and ganoderma lucidum are all processed into powder state for compounding.

[0013] The present invention does not limit the sources, preparation or extraction methods of the above components, and they can be purchased from the market or prepared by conventional processing methods in the art.

[0014] In order to obtain better effects, the present invention also conducts experimental verification on the addition ratios of different components in each combination. The results show that the effects of the compositions at different ratios are significantly different.

[0015] In some embodiments, the composition comprises the following components and their weight parts, measured in parts by weight: 2-8 parts red ginseng powder and 1-9 parts black wolfberry powder. Specifically, the ratio of red ginseng powder to black wolfberry powder can be 2:1, 2:9, 3:1, 3:9, 4:1, 4:9, 5:1, 5:9, 6:1, 6:9, 7:1, 7:9, 8:1, or 8:9, or any other ratio between 2-8:1-9.

[0016] In some embodiments, the composition comprises the following components and their weight parts, measured in parts by weight: 2-8 parts red ginseng powder and 3-10 parts ganoderma lucidum powder. Specifically, the ratio of red ginseng powder to ganoderma lucidum powder can be 2:3, 2:10, 3:3, 3:10, 4:6, 4:10, 5:3, 5:10, 6:3, 6:10, 7:3, 7:10, 8:3, or 8:10, or any other ratio between 2-8:3-10.

[0017] In some embodiments, the composition comprises the following components and their weight parts, measured in parts by weight: 1-9 parts black wolfberry powder and 3-10 parts ganoderma lucidum powder. Specifically, the ratio of black wolfberry powder to ganoderma lucidum powder can be 1:3, 1:10, 2:3, 2:10, 3:3, 3:2, 4:3, 4:10, 5:3, 5:10, 9:3, or 9:10, or any other ratio between 1-9:3-10.

[0018] In some embodiments, the composition comprises the following components and their weight parts, measured in parts by weight: 2-8 parts red ginseng powder, 1-9 parts black wolfberry powder, and 3-10 parts ganoderma lucidum powder. Specifically, the ratio of red ginseng powder, black wolfberry powder, and ganoderma lucidum powder can be 2:1:3, 2:1:10, 2:9:3, 2:9:10, 8:1:3, 8:1:10, 8:9:3, or 8:9:10, or any other ratio between 2-8:1-9:3-10.

[0019] In some embodiments, the composition comprises the following components, measured by weight: 2 parts red ginseng powder, 3 parts black wolfberry powder, and 8 parts ganoderma lucidum powder. In animal experiments, it was found that the composition of the present invention had the best effects on antioxidant activity, calcium ion metabolism, energy metabolism, and glycogen levels when the weight ratio was 2:3:8, and had a lower combined index (CI) value, indicating that the composition is more effective in relieving physical fatigue under these ratios.

[0020] The combination index method is a method for evaluating the combined effects of drugs, particularly suitable for evaluating the interactions between two or more ingredients after compounding. This method is based on the drug's dose-response curve and calculates the ratio of the actual effect to the theoretical effect when the drug is used in combination to determine whether the combination has a synergistic, additive, or antagonistic effect. A combination index (CI) less than 1 indicates a synergistic effect, a CI equal to 1 indicates an additive effect, and a CI greater than 1 indicates an antagonistic effect. Therefore, experimentally screening food-derived ingredients with better fatigue relief effects and combining the combination index method to evaluate the interactions between functional food ingredients is highly beneficial for the development of synergistic functional food formulas.

[0021] The present invention verified the effectiveness of the above-mentioned scheme using a combined index method. The results showed that both the two-component and three-component compositions were able to increase swimming distance, recovery rate, and ATP, reduce lactate content, and increase the expression of the Nrf2, atp2b1, and g6pca genes. Among them, the Nrf2 gene is a gene related to oxidative stress, the atp2b1 gene is a gene related to calcium ion metabolism, and the g6pca gene is a gene related to energy metabolism. By detecting changes in the expression of these three genes, it can be demonstrated that the various components of the composition of the present invention can synergistically regulate the body's antioxidant, calcium ion metabolism, and energy metabolism pathways, thereby alleviating physical fatigue.

[0022] At the same time, the three-component composition of the present invention has also been verified by the standard method of the "Health Food Function Test and Evaluation Method (2023 Edition)". The results show that the composition formed by the three ingredients can prolong swimming time, reduce lactic acid and urea nitrogen levels, and increase muscle glycogen and liver glycogen levels. Mammalian experiments further prove that the three-component composition has anti-fatigue effects.

[0023] Based on the above, the present invention can also provide the use of the above composition in preparing a product for relieving physical fatigue; or, the present invention can also provide the use of the above composition in preparing a product for improving ATP energy metabolism or reducing lactic acid; or, the present invention can also provide the use of the above composition in preparing a product for regulating the expression of Nrf2, atp2b1 and g6pca genes.

[0024] The composition of the present invention has broad application prospects and can be widely used in the fields of food, health care products, medicines, etc. In this regard, the present invention can also provide a food or health care product, the active ingredient of which is the above composition.

[0025] In some embodiments, the compositions of the present invention can be made into food products using techniques known to those skilled in the art.

[0026] In some embodiments, the food or health product prepared from the composition of the present invention may further include food-acceptable excipients, such as solvents, adhesives, fillers, sugars, sweeteners, and the like.

[0027] In some embodiments, the food or health product prepared from the composition of the present invention can be in the following types: tablets, capsules, oral liquids, solid beverages, pills, granules, decoctions, pastes, lotions, pills, syrups, or tea bags.

[0028] In some embodiments, the compositions of the present invention can be formulated into medicaments suitable for topical administration using techniques known to those skilled in the art.

[0029] In some embodiments, the drug prepared from the composition of the present invention may also include pharmaceutically acceptable excipients, such as: buffers, emulsifiers, suspending agents, decomposers, disintegrants, dispersants, binders, excipients, stabilizers, chelating agents, diluents, gelling agents, preservatives, wetting agents, lubricants, absorption delaying agents, liposomes, etc.

[0030] In some embodiments, the medicament prepared from the composition of the present invention can be in the following dosage forms: tablets, oral liquids, hard capsules, soft capsules, granules, powders or pills.

[0031] In the above-mentioned foods and medicines, the addition amount and addition ratio of the composition of the present invention and related auxiliary materials can be adjusted according to actual needs, and the present invention does not limit this.

[0032] The present invention has the following beneficial effects:

[0033] The present invention compounds red ginseng, black wolfberry and ganoderma lucidum to obtain a variety of compositions that can synergistically regulate antioxidant, calcium ion metabolism and energy metabolism, synergistically improve the body's ATP content and glycogen level, accelerate the removal of lactic acid and urea nitrogen, prolong exercise time, and significantly relieve physical fatigue. Its synergistic effect is stronger than that of using red ginseng, black wolfberry and ganoderma lucidum alone. At the same time, the composition of the present invention is simple in ingredients and all derived from natural plants; at the same time, after scientific demonstration and experimental verification, the mechanism of action of the composition is clear, and the fatigue-relieving interaction relationship between the various components is clear, with the advantages of being green and safe. In addition, the preparation process of the composition of the present invention is simple and easy to operate, and does not require the investment of professional and technical personnel and large-scale instruments and equipment. The production cost is low, which is convenient for industrial development. DETAILED DESCRIPTION

[0034] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be purchased commercially.

[0035] The present invention is further described below by way of specific examples. However, it should be understood that these examples are merely provided for more detailed description and are not to be construed as limiting the present invention in any form.

[0036] The red ginseng, black wolfberry and ganoderma lucidum used in the embodiments and comparative examples of the present invention are purchased from the market. The preparation method thereof is as follows: appropriately weighing the qualified red ginseng, black wolfberry and ganoderma lucidum, crushing them into large particles using a grinder, and then placing them in a multifunctional extraction tank, adding water and extracting for 4 hours, filtering the filtrate through 100 mesh, and concentrating the filtered filtrate under reduced pressure to a relative density of 1.06, spray drying to obtain a dry powder, and then crushing the dry powder to pass through an 80-mesh sieve to obtain dry powder of each component for later use.

[0037] Example 1

[0038] This embodiment is a composition, the components of which are as follows: 3 parts of red ginseng powder and 4 parts of black wolfberry powder.

[0039] Example 2

[0040] This embodiment is a composition, the components of which are as follows: 2 parts of red ginseng powder and 9 parts of black wolfberry powder.

[0041] Example 3

[0042] This embodiment is a composition, the components of which are as follows: 8 parts of red ginseng powder and 1 part of black wolfberry powder.

[0043] Example 4

[0044] This embodiment is a composition, the components of which are as follows: 3 parts of red ginseng powder and 4 parts of ganoderma lucidum powder.

[0045] Example 5

[0046] This embodiment is a composition, the components of which are as follows: 1 part of red ginseng powder and 5 parts of ganoderma lucidum powder.

[0047] Example 6

[0048] This embodiment is a composition, the components of which are as follows: 8 parts of red ginseng powder and 3 parts of ganoderma lucidum powder.

[0049] Example 7

[0050] This embodiment is a composition, the components of which are as follows: 3 parts of black wolfberry powder and 4 parts of ganoderma lucidum powder.

[0051] Example 8

[0052] This embodiment is a composition, the components of which are as follows: 1 part of black wolfberry powder and 10 parts of ganoderma lucidum powder.

[0053] Example 9

[0054] This embodiment is a composition, the components of which are as follows: 3 parts of black wolfberry powder and 1 part of ganoderma lucidum powder.

[0055] Example 10

[0056] This embodiment is a composition, the components of which are as follows: 1.5 parts of red ginseng powder, 1.5 parts of black wolfberry powder, and 4 parts of ganoderma lucidum powder.

[0057] Example 11

[0058] This embodiment is a composition, the components of which are as follows: 1.5 parts of red ginseng powder, 1.5 parts of black wolfberry powder, and 10 parts of ganoderma lucidum powder.

[0059] Example 12

[0060] This embodiment is a composition, the components of which are as follows: 3 parts of red ginseng powder, 3 parts of black wolfberry powder, and 1 part of ganoderma lucidum powder.

[0061] Example 13

[0062] This embodiment is a composition, the components of which are as follows: 2 parts of red ginseng powder, 3 parts of black wolfberry powder, and 8 parts of ganoderma lucidum powder.

[0063] Experimental Example 1

[0064] 6 dpf wild-type AB strain zebrafish were selected, and equal amounts of the compositions described in each example were dissolved in water. The zebrafish were then placed in water containing food-derived ingredients overnight at 27°C for 24 hours. The experimental treatments are shown in Table 1. A zebrafish fatigue model was established using sodium sulfite for 30 minutes. The total distance traveled within 30 minutes was analyzed, the fatigue recovery rate was calculated, and lactate and adenosine triphosphate (ATP) levels, as well as the expression of related genes, were measured.

[0065] The formula for calculating the recovery rate of zebrafish larvae is as follows:

[0066] Recovery rate = (swimming distance of treatment group - swimming distance of model group) / (swimming distance of control group - swimming distance of model group) × 100%

[0067] The swimming distance of the control group was 1996.7 ± 263.5 mm. The recovery rate of zebrafish larvae in each group was calculated according to the above formula. The specific results are as follows:

[0068] Table 1 Effects of red ginseng powder and black wolfberry powder on zebrafish fatigue

[0069]

[0070] Note: The addition amount of each experimental group is the same; the symbol " / " indicates no corresponding value; the statistical significance compared with the model group is denoted as * for p < 0.05, ** for p < 0.01, and *** for p < 0.001.

[0071] As shown in Table 1, compared with the control group, the swimming distance of zebrafish in the model group decreased by 50.0%, indicating that sodium sulfite induced zebrafish fatigue; Red Bull, a recognized anti-fatigue health drink, was selected as the positive control group, and the swimming distance of zebrafish increased by 75.6% (p<0.001). The swimming distance was significantly restored compared with the control group, indicating that Red Bull alleviated sodium sulfite-induced zebrafish fatigue and the fatigue model was successfully established.

[0072] When red ginseng powder and black wolfberry powder were used alone, swimming distance increased significantly (p < 0.05), and recovery rates exceeded 50%, indicating that both have significant fatigue-relieving potential. When red ginseng powder and black wolfberry powder were used in a 3:4, 2:9, and 8:1 ratio, swimming distance increased by 69.5%, 79.3%, and 65.8%, respectively, compared with the model group (p < 0.01), and recovery rates were 69.6%, 79.3%, and 65.6%, respectively. The corresponding CI values were 0.53, 0.46, and 0.71, respectively, all less than 1. This indicates that red ginseng powder and black wolfberry powder can synergistically relieve physical fatigue when used in a ratio of 2:9 to 8:1.

[0073] When red ginseng powder was used alone, the ATP content increased (p<0.05), while there was no significant change with black wolfberry powder, indicating that red ginseng powder can increase ATP content and relieve physical fatigue. When red ginseng powder and black wolfberry powder were used in a mixture of 3:4, 2:9 and 8:1, the ATP content increased by 9.6%, 4.7% and 15.2%, respectively (p<0.05), and the corresponding CI values were 0.74, 0.88 and 0.32, respectively, all less than 1, indicating that when the mixture ratio was 2:9~8:1, it could significantly relieve physical fatigue by synergistically increasing ATP content.

[0074] Furthermore, both red ginseng powder and black wolfberry powder reduced lactic acid levels when used alone (p < 0.05), indicating that they can reduce lactic acid levels in the body and alleviate physical fatigue. When red ginseng powder and black wolfberry powder were mixed at 3:4, 2:9, and 8:1, lactic acid levels decreased by 22.5%, 25.0%, and 17.5%, respectively, compared with the model group (p < 0.05). The corresponding CI values were 0.77, 0.63, and 0.86, respectively, all less than 1. This indicates that when mixed at 2:9 to 8:1, red ginseng powder and black wolfberry powder can synergistically reduce lactic acid levels and alleviate physical fatigue.

[0075] When used alone, both red ginseng powder and black wolfberry powder increased Nrf2 gene expression (p < 0.05), suggesting that both can alleviate physical fatigue by enhancing the body's antioxidant capacity. When red ginseng powder and black wolfberry powder were mixed at ratios of 3:4, 2:9, and 8:1, Nrf2 gene expression increased by 4.39-, 2.78-, and 2.21-fold, respectively (p < 0.01), with CI values of 0.32, 0.58, and 0.77, respectively. This suggests that their combination can alleviate physical fatigue by synergistically regulating the body's antioxidant pathways. When used alone, red ginseng powder and black wolfberry powder increased atp2b1 gene expression (p < 0.05). When mixed at 3:4, 2:9, and 8:1, atp2b1 gene expression increased by 1.87-, 1.51-, and 2.10-fold (p < 0.05), with CI values of 0.87, 0.95, and 0.49, respectively. This suggests that the combination alleviates physical fatigue by synergistically regulating calcium metabolism. Red ginseng powder and black wolfberry powder also increased g6pca gene expression when used alone (p < 0.05). When mixed at 3:4, 2:9, and 8:1, g6pca gene expression increased by 1.72-, 1.62-, and 1.97-fold (p < 0.05), with CI values of 0.71, 0.88, and 0.64, respectively. This suggests that the combination can alleviate physical fatigue by synergistically upregulating energy metabolism.

[0076] In summary, when red ginseng powder and black wolfberry powder are mixed in a ratio of 2:9~8:1, they can synergistically regulate the body's antioxidant capacity, calcium ion metabolism and energy metabolism, and synergistically increase the body's ATP content, accelerate lactic acid clearance, thereby increasing the swimming distance of zebrafish, improving the recovery rate, and significantly alleviating physical fatigue. The recovery effect is comparable to that of Red Bull.

[0077] Experimental Example 2

[0078] The experimental method was the same as that of Experimental Example 1. The compound raw materials and proportions are shown in Table 2. The swimming distance of the control group was 2020.4 ± 213.6 mm. The specific results are as follows:

[0079] Table 2 Effects of red ginseng powder and ganoderma lucidum powder on fatigue in zebrafish

[0080]

[0081] Note: The dosage of each experimental group was the same; the symbol “ / ” indicates no corresponding value; the statistical significance compared with the model group is denoted as * for p<0.05, ** for p<0.01, and *** for p<0.001.

[0082] As shown in Table 2, when red ginseng powder and Ganoderma lucidum powder were used alone, they increased the swimming distance of zebrafish (p < 0.05), with recovery rates exceeding 30%, demonstrating their potential to alleviate physical fatigue. When red ginseng powder and Ganoderma lucidum powder were mixed at ratios of 3:4, 1:5, and 8:3, swimming distance increased by 53.4%, 77.8%, and 67.6%, respectively (p < 0.05), with recovery rates of 49.5%, 73.3%, and 63.4%, respectively. The corresponding CI values were 0.79, 0.37, and 0.64, respectively, all less than 1. This indicates that red ginseng powder and Ganoderma lucidum powder can synergistically alleviate physical fatigue when mixed at ratios of 1:5 to 8:3.

[0083] When red ginseng powder and ganoderma lucidum powder were used alone, ATP levels increased (p < 0.05), indicating that both can alleviate fatigue by increasing ATP levels. When red ginseng powder and ganoderma lucidum powder were mixed in ratios of 3:4, 1:5, and 8:3, ATP levels increased by 10.9%, 7.9%, and 17.98%, respectively (p < 0.01). The corresponding CI values were 0.82, 0.91, and 0.28, respectively, all less than 1, indicating that the two powders can synergistically increase ATP levels and alleviate fatigue.

[0084] Furthermore, when used alone, both red ginseng powder and ganoderma lucidum powder reduced lactic acid levels (p < 0.05) and alleviated physical fatigue. When red ginseng powder and ganoderma lucidum powder were combined at ratios of 3:4, 1:5, and 8:3, lactic acid levels decreased by 23.6%, 28.9%, and 18.4%, respectively (p < 0.05), with corresponding CI values of 0.46, 0.44, and 0.89, all less than 1, indicating that the combination synergistically reduced lactic acid levels and alleviated physical fatigue.

[0085] When used alone, both red ginseng powder and Ganoderma lucidum powder increased Nrf2 gene expression (p < 0.05). When mixed at 3:4, 1:5, and 8:3 ratios, Nrf2 gene expression increased by 2.69-, 1.89-, and 2.83-fold (p < 0.01), with CI values of 0.61, 0.88, and 0.44, respectively. This suggests that the combination can synergistically regulate antioxidant pathways and alleviate physical fatigue. Both red ginseng powder and Ganoderma lucidum powder increased atp2b1 gene expression (p < 0.05). When mixed at 3:4, 1:5, and 8:3 ratios, atp2b1 gene expression increased by 2.0-, 1.81-, and 2.1-fold (p < 0.01), with CI values of 0.33, 0.49, and 0.29, respectively. This suggests that the combination of red ginseng powder and Ganoderma lucidum powder can synergistically regulate calcium metabolism and alleviate physical fatigue. When red ginseng powder and Ganoderma lucidum powder were used alone, they also increased the expression of the g6pca gene (p<0.05). After the two were mixed at 3:4, 1:5 and 8:3, the expression of the g6pca gene increased by 1.79, 1.59 and 1.94 times, respectively (p<0.05), with CI values of 0.68, 0.71 and 0.45, respectively. This shows that the combination of red ginseng powder and Ganoderma lucidum powder can synergistically upregulate the body's energy metabolism pathway and relieve physical fatigue.

[0086] In summary, when red ginseng powder and Ganoderma lucidum powder are mixed in a ratio of 1:5~8:3, they can synergistically regulate the body's antioxidant, calcium ion metabolism and energy metabolism, synergistically increase the body's ATP content, accelerate lactic acid clearance, increase the swimming distance of zebrafish and improve the recovery rate, relieve physical fatigue, and the recovery effect is comparable to that of Red Bull.

[0087] Experimental Example 3

[0088] The experimental method was the same as that of Experimental Example 1. The compound raw materials and their proportions are shown in Table 3. The swimming distance of the control group was 1594.8 ± 347.6 mm. The specific results are as follows:

[0089] Table 3 Effects of black wolfberry powder and Ganoderma lucidum powder on zebrafish fatigue

[0090]

[0091] Note: The addition amount of each experimental group is the same; the symbol " / " indicates no corresponding value; the statistical significance compared with the model group is denoted as * for p < 0.05, ** for p < 0.01, and *** for p < 0.001.

[0092] As shown in Table 3, both black wolfberry powder and Ganoderma lucidum powder increased the swimming distance of zebrafish (p < 0.05), and the recovery rates of both were above 30%, indicating that both have the potential to relieve physical fatigue. When black wolfberry powder and Ganoderma lucidum powder were used in a 3:4, 1:10, and 3:1 ratio, the swimming distance increased by 54.49%, 61.0%, and 58.4%, respectively, compared with the model group (p < 0.01), and the recovery rates were 55.7%, 61.0%, and 58.4%, respectively. The corresponding CI values were 0.57, 0.26, and 0.69, respectively, all less than 1. This indicates that when black wolfberry powder and Ganoderma lucidum powder are used in a 1:10 to 3:1 ratio, they can synergistically relieve physical fatigue.

[0093] When black wolfberry powder and Ganoderma lucidum powder were used alone, the ATP content of Ganoderma lucidum powder increased (p<0.05), while there was no significant change in black wolfberry powder. However, after black wolfberry powder and Ganoderma lucidum powder were mixed at ratios of 3:4, 1:10 and 3:1, the ATP content increased by 7.9%, 13.3% and 5.4%, respectively (p<0.05), and the corresponding CI values were 0.44, 0.19 and 0.51, respectively, all less than 1, indicating that when the two are mixed, they can synergistically increase ATP content and relieve physical fatigue.

[0094] In addition, when black wolfberry powder and Ganoderma lucidum powder are used alone, they can reduce lactic acid content (p<0.05) and relieve physical fatigue. When the two are used in a mixture of 3:4, 1:10 and 3:1, the lactic acid content is reduced by 20.5%, 28.2% and 23.0%, respectively (p<0.05), and the corresponding CI values are 0.56, 0.22 and 0.48, respectively, all less than 1, indicating that the combination of black wolfberry powder and Ganoderma lucidum powder can synergistically reduce lactic acid content and relieve physical fatigue.

[0095] Both red ginseng powder and black wolfberry powder can increase the expression of Nrf2 gene when used alone (p<0.05). When the two are mixed at 3:4, 1:10 and 3:1, the expression of Nrf2 gene increases by 3.17, 3.34 and 2.94 times, respectively (p<0.001), and the CI values are 0.54, 0.39 and 0.66, respectively. This shows that the combination of red ginseng powder and black wolfberry powder can relieve physical fatigue by synergistically regulating the body's antioxidant pathways. When black wolfberry powder and Ganoderma lucidum powder were used alone, they both increased the expression of the atp2b1 gene (p<0.05). When the two were mixed at a ratio of 3:4, 1:10 and 3:1, the expression of the atp2b1 gene increased by 2.06, 2.24 and 2.15 times, respectively (p<0.01), with CI values of 0.76, 0.55 and 0.69, indicating that when red ginseng powder and black wolfberry powder are mixed, they can synergistically regulate the body's calcium ion metabolism and relieve physical fatigue. In addition, both black wolfberry powder and Ganoderma lucidum powder increased the expression of the g6pca gene (p<0.05), and after the two were compounded at 3:4, 1:10 and 3:1, the expression levels of the g6pca gene increased by 1.84, 2.0 and 1.64 times, respectively (p<0.01), with CI values of 0.67, 0.43 and 0.77, respectively. This shows that when red ginseng powder and black wolfberry powder are compounded, they can synergistically upregulate the body's energy metabolism pathways and relieve physical fatigue.

[0096] In summary, when red ginseng powder and Ganoderma lucidum powder are mixed in a ratio of 1:10 to 3:1, they can synergistically regulate the body's antioxidant, calcium ion metabolism and energy metabolism, and synergistically increase the body's ATP content, accelerate lactic acid clearance, significantly increase the swimming distance of zebrafish and improve the recovery rate, relieve physical fatigue, and the recovery effect is comparable to that of Red Bull.

[0097] Experimental Example 4

[0098] The experimental method was the same as that of Experimental Example 1. The compound raw materials and their proportions are shown in Table 3. The swimming distance of the control group was 1679.1 ± 251.3 mm. The specific results are as follows:

[0099] Table 4 Effects of red ginseng powder, black wolfberry powder and ganoderma lucidum powder on fatigue in zebrafish

[0100]

[0101] Note: The addition amount of each experimental group is the same; the symbol " / " indicates no corresponding value; the statistical significance compared with the model group is denoted as * for p < 0.05, ** for p < 0.01, and *** for p < 0.001.

[0102] As shown in Table 4, when red ginseng powder, black wolfberry powder, and ganoderma lucidum powder were used alone, they all increased the swimming distance of zebrafish (p < 0.05), with recovery rates of 48.2%, 51.3%, and 38.5%, respectively, indicating that they all have the potential to relieve physical fatigue. When red ginseng powder, black wolfberry powder, and ganoderma lucidum powder were mixed at a ratio of 3:4, 3:10, and 6:1, the swimming distance increased by 77.7%, 73.5%, and 74.3%, respectively, compared with the model group (p < 0.01), with recovery rates of 77.7%, 73.5%, and 74.3%, respectively. The corresponding CI values were 0.21, 0.30, and 0.35, respectively, all less than 1. This indicates that when the ratio range of red ginseng powder, black wolfberry powder, and ganoderma lucidum powder is 3:10 to 6:1, they can significantly and synergistically relieve physical fatigue.

[0103] When red ginseng powder, black wolfberry powder and ganoderma lucidum powder were used alone, they all increased the ATP content (p<0.05). When the three were mixed in ratios of 3:4, 3:10 and 6:1, the ATP content increased by 12.1%, 17.3% and 13.5%, respectively (p<0.001). The corresponding CI values were 0.43, 0.22 and 0.32, respectively, all less than 1. This indicates that the combination of red ginseng powder, black wolfberry powder and ganoderma lucidum powder can synergistically increase ATP content and relieve physical fatigue.

[0104] In addition, red ginseng powder, black wolfberry powder and Ganoderma lucidum powder can all reduce lactic acid content (p<0.05) and relieve physical fatigue. When the three are used in combination, the lactic acid content is reduced by 34.1%, 36.5% and 26.8% respectively compared with the model group (p<0.01). The corresponding CI values are 0.38, 0.33 and 0.29, respectively, all less than 1, indicating that when red ginseng powder, black wolfberry powder and Ganoderma lucidum powder are used in combination at a ratio of 3:10 to 6:1, they can synergistically reduce lactic acid content and relieve physical fatigue.

[0105] When used individually, red ginseng powder, black wolfberry powder, and ganoderma lucidum powder all increased Nrf2 gene expression (p<0.05), suggesting that these three ingredients can alleviate fatigue by regulating the body's antioxidant pathways. When red ginseng powder, black wolfberry powder, and ganoderma lucidum powder were combined at ratios of 3:4, 3:10, and 6:1, Nrf2 gene expression increased by 3.12-, 3.03-, and 2.52-fold, respectively (p<0.001), with CI values of 0.36, 0.46, and 0.55, respectively. This suggests that the three ingredients, when combined, can synergistically regulate the body's antioxidant pathways to alleviate physical fatigue. Red ginseng powder, black wolfberry powder, and ganoderma lucidum powder all increased atp2b1 gene expression (p < 0.05), regulating calcium metabolism and alleviating physical fatigue. Furthermore, when the three were mixed at a ratio of 3:4, 3:10, and 6:1, atp2b1 gene expression increased by 5.17-, 4.29-, and 4.88-fold, respectively (p < 0.001), with CI values of 0.21, 0.41, and 0.33, respectively. This suggests that the combination of red ginseng powder, black wolfberry powder, and ganoderma lucidum powder can synergistically regulate calcium metabolism and alleviate physical fatigue. When used alone, red ginseng powder, black wolfberry powder, and ganoderma lucidum powder also increased g6pca gene expression (p < 0.05), indicating that all three can alleviate physical fatigue by upregulating energy metabolism pathways. When red ginseng powder + black wolfberry powder and Ganoderma lucidum powder were mixed at ratios of 3:4, 3:10 and 6:1, the expression levels of the g6pca gene increased by 2.35, 2.0 and 2.25 times, respectively (p<0.001), and the CI values were 0.33, 0.44 and 0.29, respectively, indicating that the combination of the three can significantly synergistically regulate the body's energy metabolism pathways and relieve physical fatigue.

[0106] In summary, when red ginseng powder + black wolfberry powder and Ganoderma lucidum powder are compounded at a ratio of 3:10~6:1, they can synergistically regulate the body's antioxidant, calcium ion metabolism and energy metabolism, and synergistically increase the body's ATP content, accelerate lactic acid clearance, significantly increase the swimming distance of zebrafish and improve the recovery rate, and effectively relieve physical fatigue.

[0107] Combined with the experimental results of Experimental Examples 1 to 3, it was found that when any two of red ginseng powder, black wolfberry powder and ganoderma lucidum powder are mixed, they can synergistically relieve physical fatigue, and when the three are mixed, they can exert a stronger synergistic effect, and the recovery effect is slightly higher than Red Bull.

[0108] Experimental Example 5

[0109] A compound formula consisting of red ginseng powder, black wolfberry powder, and ganoderma lucidum powder in varying proportions has been shown to significantly relieve physical fatigue in zebrafish experiments. However, zebrafish differ significantly from the human body, so a mouse fatigue model experiment was used to further validate the fatigue-relieving effect of this formula, providing reliable data support for subsequent product development and use. Based on the compound formula dosage combination verified in the zebrafish experiment, and referring to the zebrafish to human dosage conversion method described in patent CN113496071A, the compound formula dosage in the zebrafish experiment was converted to the mouse intake dosage. The fatigue-relieving effect of the compound formula was verified in strict accordance with the standard method of the "Health Food Functional Test and Evaluation Method (2023 Edition)". The specific experimental process is as follows:

[0110] Male C57 / 6J mice weighing approximately 20 ± 2 g were selected and treated with the experimental treatments shown in Table 5. They were gavaged for 30 consecutive days. At the end of the experimental period, 30 minutes after gavage, the mice underwent a weighted swimming test. The time from the start of swimming to the time the head was completely submerged in water and unable to emerge within 5 seconds was recorded as the exhaustive swimming time. Lactate, urea nitrogen, muscle glycogen, and liver glycogen levels were measured. The results are shown below.

[0111] Table 5 Effects of red ginseng powder, black wolfberry powder and ganoderma lucidum powder on fatigue in mice

[0112]

[0113] Note: The addition amount of each experimental group is the same; the statistical significance compared with the model group is denoted as * for p<0.05, ** for p<0.01, and *** for p<0.001.

[0114] As shown in Table 5, using Red Bull, a recognized anti-fatigue health drink, as the positive control group, the mice's exhaustive swimming time was extended by 83.9% (p < 0.01), indicating that Red Bull significantly alleviates physical fatigue in mice. When red ginseng powder, black wolfberry powder, and ganoderma lucidum powder were combined at a ratio of 3:4, 3:10, and 6:1, the exhaustive swimming time was extended by 84.14%, 78.73%, and 66.79%, respectively (p < 0.05), indicating that this formula, at a ratio of 3:10 to 6:1, is effective in relieving physical fatigue in mice.

[0115] When red ginseng powder, black wolfberry powder, and ganoderma lucidum powder were combined in ratios of 3:4, 3:10, and 6:1, lactic acid levels decreased by 36.1%, 35.1%, and 28.4%, respectively (p<0.001), and urea nitrogen levels decreased by 16.1%, 13.8%, and 15.1%, respectively (p<0.05). This suggests that a 3:10-6:1 ratio can significantly alleviate physical fatigue by reducing lactic acid and urea nitrogen levels. Furthermore, when the three ingredients were combined in ratios of 3:4, 3:10, and 6:1, muscle glycogen levels increased by 42.5%, 21.2%, and 18.7%, respectively (p<0.05), and liver glycogen levels increased by 32.2%, 13.7%, and 17.8%, respectively (p<0.05), indicating that a 3:10-6:1 ratio can alleviate physical fatigue by increasing glycogen levels.

[0116] In summary, red ginseng powder, black wolfberry powder, and ganoderma lucidum powder, when combined in a ratio of 3:10 to 6:1, significantly prolonged the mice's exhaustive swimming time by reducing the accumulation of metabolic products and increasing glycogen levels. This indicates that this formula can alleviate physical fatigue in mice, with a fatigue recovery effect slightly superior to that of Red Bull. Thus, the composition of the present invention has been confirmed to have a significant effect on relieving physical fatigue using the standard method of the "Health Food Functional Testing and Evaluation Methods (2023 Edition)."

[0117] Comparative Example 1

[0118] The components in this solution are: wolfberry fruit powder.

[0119] Comparative Example 2

[0120] The components in this solution are: sea buckthorn fruit powder.

[0121] Comparative Example 3

[0122] The components in this scheme are: Polygonatum sibiricum powder.

[0123] Comparative Example 4

[0124] The components in this solution are: 1 part of wolfberry fruit powder and 2 parts of sea buckthorn fruit powder.

[0125] Comparative Example 5

[0126] The components in this solution are: 1 part of wolfberry fruit powder and 3 parts of polygonatum powder.

[0127] Comparative Example 6

[0128] The components in this solution are: 2 parts of sea buckthorn fruit powder and 3 parts of polygonatum powder.

[0129] Comparative Example 7

[0130] The components in this solution are: 1 part of wolfberry fruit powder, 2 parts of sea buckthorn fruit powder and 3 parts of polygonatum powder.

[0131] The above comparative example was subjected to a zebrafish experiment according to the method of the above experimental example, and the test results are as follows:

[0132] Table 6 Effects of control ratios on zebrafish fatigue

[0133]

[0134] The results in the above table show that although wolfberry fruit powder, sea buckthorn fruit powder and polygonatum sibiricum powder each have the potential to relieve physical fatigue, when they are combined, whether in pairs or in combinations of three, the recovery rates do not change much, and the corresponding CI values are 1.78, 1.69, 1.43 and 1.55, respectively, all greater than 1, proving that the various combinations of these three ingredients do not have a synergistic effect. It also shows that food-derived ingredients that have the effect of relieving physical fatigue alone do not necessarily have a synergistic effect when combined with each other.

[0135] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. Use of a traditional Chinese medicine composition in preparing a product for relieving physical fatigue, characterized in that: The composition is composed of red ginseng and black wolfberry, and the weight parts thereof are: 2 parts of red ginseng and 9 parts of black wolfberry; or The composition is composed of red ginseng and ganoderma lucidum, and the weight parts thereof are: 1 part of red ginseng and 5 parts of ganoderma lucidum; or The composition is composed of black wolfberry and ganoderma lucidum, and the weight proportions thereof are: 1 to 9 parts of black wolfberry and 3 to 10 parts of ganoderma lucidum; or The composition consists of red ginseng, black wolfberry and ganoderma lucidum, and the weight proportions thereof are: 2-8 parts of red ginseng, 1-9 parts of black wolfberry and 3-10 parts of ganoderma lucidum.

2. The use according to claim 1, characterized in that The composition consists of the following components in parts by weight: 2 parts of red ginseng, 3 parts of black wolfberry, and 8 parts of ganoderma lucidum.

3. The use according to claim 2, characterized in that The types of products include tablets, oral liquids, capsules, granules or pills.

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