Plant-based compositions, their preparation methods and applications

Abstract

This invention provides a plant-based composition comprising a first component, a second component, and a third component. The first component comprises 1-3 parts by weight of protein melanin and 1-3 parts by weight of phenylethanoid glycoside. The second component comprises 10-14 parts by weight of Polygonatum sibiricum powder, 8-12 parts by weight of licorice powder, 8-12 parts by weight of wolfberry powder, 8-14 parts by weight of longan powder, 8-16 parts by weight of jujube powder, 12-15 parts by weight of Poria cocos powder, and 7-10 parts by weight of red rose powder. The third component comprises 3-5 parts by weight of γ-aminobutyric acid (GABA), 2-6 parts by weight of erythritol, 3-5 parts by weight of citric acid, 2-4 parts by weight of sucralose, and 3-5 parts by weight of ascorbic acid. This invention uses protein melanin and phenylethanoid glycoside as the main active ingredients, supplemented with Polygonatum sibiricum powder, Poria cocos powder, and GABA. Experiments have shown that this composition can prolong the swimming time of mice under load and improve their lymphocyte proliferation capacity. This composition can regulate bodily functions and enhance immunity.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, and in particular to a plant-based composition, its preparation method, and its application. Background Technology

[0002] Protein melanin is a brownish substance, also known as melanoidin, formed by the Maillard reaction between carbonyl compounds (generally sugars, etc.) and nitrogen-containing compounds with free amino groups (such as amino acids, peptides, and proteins). The product undergoes further cyclization, dehydration, reverse condensation, rearrangement, and isomerization condensation. Protein melanin possesses physiological functions such as antioxidation, antibacterial activity, hypoglycemia, and hypotension, and its functionality has been preliminarily verified. Protein melanin is a complex polymer with varying degrees of polymerization. Under different reaction conditions and with different reactants, the composition and structure of protein melanin differ. Currently, researchers are gradually beginning to conduct in-depth studies on the specific composition, molecular structure, mechanism of action, and development and utilization of protein melanin.

[0003] Phenylethanoid glycosides are a class of natural compounds composed of caffeic acid, phenytoin, and glycosyl groups. Their structural characteristics primarily involve a β-glucose core containing ester and oxyglycosidic bonds, with hydroxyl and methoxy groups attached to the phenethyl group. An acetyl or caffeoyl group is often attached to the central glucose group. Modern pharmacological studies have shown that phenylethanoid glycosides play important roles in antibacterial, anti-inflammatory, antioxidant, antiviral, and immune-enhancing effects. Currently, research on their development and utilization is gradually underway.

[0004] Currently, most immune-boosting substances are chemical drugs, and traditional substances are banned in some countries due to their potential carcinogenicity. Immunity-boosting compositions extracted from natural plant-derived products may be one of the safer alternatives. Summary of the Invention

[0005] In view of this, the present invention provides a plant-based composition that has the effects of enhancing immunity and anti-fatigue.

[0006] This invention provides a plant-based composition comprising a first component, a second component, and a third component;

[0007] The first component raw materials include 1-3 parts by weight of protein melanin and 1-3 parts by weight of phenylethanol glycoside;

[0008] The second component of raw materials includes 10-14 parts by weight of Polygonatum sibiricum powder, 8-12 parts by weight of Glycyrrhiza uralensis powder, 8-12 parts by weight of Lycium barbarum powder, 8-14 parts by weight of Longan powder, 8-16 parts by weight of Jujube powder, 12-15 parts by weight of Poria cocos powder, and 7-10 parts by weight of Rose powder.

[0009] The third component includes 3-5 parts by weight of γ-aminobutyric acid, 2-6 parts by weight of erythritol, 3-5 parts by weight of citric acid, 2-4 parts by weight of sucralose, and 3-5 parts by weight of ascorbic acid.

[0010] Preferably, it is characterized in that,

[0011] The first component raw material includes 2 parts by mass of protein melanin and 2 parts by mass of phenylethanol glycoside;

[0012] The second component of raw materials includes 12 parts by weight of Polygonatum sibiricum powder, 10 parts by weight of Glycyrrhiza uralensis powder, 8 parts by weight of Lycium barbarum powder, 8 parts by weight of Longan powder, 16 parts by weight of Jujube powder, 15 parts by weight of Poria cocos powder, and 8 parts by weight of Rose powder.

[0013] The third component comprises 5 parts by mass of γ-aminobutyric acid, 4 parts by mass of erythritol, 4 parts by mass of citric acid, 3 parts by mass of sucralose, and 3 parts by mass of ascorbic acid.

[0014] Preferably, the mass ratio of the first component, the second component, and the third component is 4:77:19.

[0015] This invention provides a method for preparing the plant-based composition according to any one of the above technical solutions, comprising:

[0016] A) The first component is obtained by mixing protein melanin and phenylethanol glycoside;

[0017] Mix the Polygonatum powder, licorice powder, wolfberry powder, longan powder, jujube powder, Poria powder and red rose powder to obtain the second component;

[0018] The third component is obtained by mixing γ-aminobutyric acid, erythritol, citric acid, sucralose and ascorbic acid;

[0019] B) Mix the first, second and third components, sterilize, fill, and sterilize again to obtain the final product.

[0020] Preferably, the sterilization temperature is 105°C and the time is 5 minutes;

[0021] The secondary sterilization temperature is 115℃ and the time is 5 minutes.

[0022] This invention provides the use of the plant-based composition described in any one of the above claims in the preparation of anti-fatigue products.

[0023] Preferably, the anti-fatigue measures include extending the duration of weight-bearing swimming, increasing glycogen reserves, reducing serum lactate (BLA) levels, and increasing lactate dehydrogenase (LDH) activity.

[0024] This invention provides the use of the plant-based compositions described in any one of the above-mentioned claims in the preparation of products that enhance immunity.

[0025] Preferably, the enhanced immunity includes increasing the number of antibody-producing cells, the half-hemolysis value, carbon clearance capacity, the phagocytic rate and phagocytic index of macrophages phagocytizing chicken erythrocytes, the lymphocyte proliferation capacity, and reducing foot swelling.

[0026] This invention provides a product for enhancing immunity / anti-fatigue, comprising the plant-based composition described in any one of the above technical solutions.

[0027] Compared with existing technologies, this invention provides a plant-based composition comprising a first component, a second component, and a third component. The first component comprises 1-3 parts by weight of protein melanin and 1-3 parts by weight of phenylethanoid glycoside. The second component comprises 10-14 parts by weight of Polygonatum sibiricum powder, 8-12 parts by weight of licorice powder, 8-12 parts by weight of wolfberry powder, 8-14 parts by weight of longan powder, 8-16 parts by weight of jujube powder, 12-15 parts by weight of Poria cocos powder, and 7-10 parts by weight of rose powder. The third component comprises 3-5 parts by weight of γ-aminobutyric acid (GABA), 2-6 parts by weight of erythritol, 3-5 parts by weight of citric acid, 2-4 parts by weight of sucralose, and 3-5 parts by weight of ascorbic acid. This invention uses protein melanin and phenylethanoid glycoside as the main active ingredients, supplemented with Polygonatum sibiricum powder, licorice powder, wolfberry powder, longan powder, jujube powder, Poria cocos powder, and rose powder; as well as GABA, erythritol, citric acid, sucralose, and ascorbic acid. By adding various food-grade medicinal ingredients, and through swimming tests on mice to assess fatigue resistance and immunity, the optimal composition was determined. Results showed that this composition can better regulate bodily functions and enhance immunity, and can be applied in beverages, meal replacements, and special dietary products. Attached Figure Description

[0028] Figure 1 This is a flowchart of the present invention. Detailed Implementation

[0029] This invention provides a plant-based composition, its preparation method, and its application. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and fall within the scope of protection of this invention. The method and application of this invention have been described through preferred embodiments. Those skilled in the art can clearly modify or appropriately change and combine the method and application described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0030] This invention provides a plant-based composition comprising a first component, a second component, and a third component;

[0031] The first component raw materials include 1-3 parts by weight of protein melanin and 1-3 parts by weight of phenylethanol glycoside;

[0032] The second component of raw materials includes 10-14 parts by weight of Polygonatum sibiricum powder, 8-12 parts by weight of Glycyrrhiza uralensis powder, 8-12 parts by weight of Lycium barbarum powder, 8-14 parts by weight of Longan powder, 8-16 parts by weight of Jujube powder, 12-15 parts by weight of Poria cocos powder, and 7-10 parts by weight of Rose powder.

[0033] The third component includes 3-5 parts by weight of γ-aminobutyric acid, 2-6 parts by weight of erythritol, 3-5 parts by weight of citric acid, 2-4 parts by weight of sucralose, and 3-5 parts by weight of ascorbic acid.

[0034] The plant-based composition provided by the present invention includes a first component.

[0035] The first component of the present invention comprises 1-3 parts by weight of protein melanin and 1-3 parts by weight of phenylethanol glycoside.

[0036] In one embodiment of the present invention, the first component raw material includes 2 parts by mass of protein melanin and 2 parts by mass of phenylethanol glycoside.

[0037] In one embodiment of the present invention, the first component raw material includes 1 part by mass of protein melanin and 3 parts by mass of phenylethanol glycoside.

[0038] In one embodiment of the present invention, the first component raw material includes 3 parts by mass of protein melanin and 1 part by mass of phenylethanol glycoside.

[0039] The inventors have discovered that when the ratio of protein melanin to phenylethanol glycoside in the first component is 1:1, the composition exhibits the best anti-fatigue effect and synergistic immune-enhancing effect in mice.

[0040] Protein melanin is a brown substance formed by the Maillard reaction between carbonyl compounds (generally sugars, etc.) and nitrogen-containing compounds with free amino groups (such as amino acids, peptides, and proteins, etc.). The product is further processed through cyclization, dehydration, reverse condensation, rearrangement, isomerization, and condensation. It is also called melanoidin.

[0041] Phenylethanoid glycosides are a class of natural compounds composed of caffeic acid, phenytoin, and glycosyl groups. Their structural features are mainly β-glucose as the parent nucleus, containing ester bonds and oxyglycosidic bonds, with hydroxyl and methoxy groups attached to the phenethyl group, and acetyl and caffeoyl groups often attached to the central glucose group.

[0042] The plant-based composition provided by this invention includes a second component.

[0043] The second component consists of 10-14 parts by weight of Polygonatum sibiricum powder, 8-12 parts by weight of Glycyrrhiza uralensis powder, 8-12 parts by weight of Lycium barbarum powder, 8-14 parts by weight of Longan powder, 8-16 parts by weight of Jujube powder, 12-15 parts by weight of Poria cocos powder, and 7-10 parts by weight of Rose powder.

[0044] This invention does not limit the source of the above-mentioned raw materials; as long as they are commercially available, meet food and drug standards, and are safe and of high quality, they are acceptable.

[0045] In one embodiment of the present invention, the second component raw materials include 12 parts by weight of Polygonatum sibiricum powder, 10 parts by weight of Glycyrrhiza uralensis powder, 8 parts by weight of Lycium barbarum powder, 8 parts by weight of Longan powder, 16 parts by weight of Jujube powder, 15 parts by weight of Poria cocos powder, and 8 parts by weight of Rose powder.

[0046] In one embodiment of the present invention, the second component raw materials include 10 parts by weight of Polygonatum sibiricum powder, 8 parts by weight of licorice powder, 12 parts by weight of wolfberry powder, 12 parts by weight of longan powder, 12 parts by weight of jujube powder, 13 parts by weight of Poria cocos powder and 10 parts by weight of red rose powder.

[0047] In one embodiment of the present invention, the second component raw materials include 14 parts by weight of Polygonatum sibiricum powder, 12 parts by weight of Glycyrrhiza uralensis powder, 10 parts by weight of Lycium barbarum powder, 14 parts by weight of Longan powder, 8 parts by weight of Jujube powder, 12 parts by weight of Poria cocos powder, and 7 parts by weight of Rose powder.

[0048] The plant-based composition provided by this invention includes a third component.

[0049] The third component of this invention comprises 3-5 parts by weight of γ-aminobutyric acid, 2-6 parts by weight of erythritol, 3-5 parts by weight of citric acid, 2-4 parts by weight of sucralose, and 3-5 parts by weight of ascorbic acid.

[0050] This invention does not limit the source of the above-mentioned raw materials; as long as they are commercially available, meet food and drug standards, and are safe and of high quality, they are acceptable.

[0051] In one embodiment of the present invention, the third component comprises 5 parts by mass of γ-aminobutyric acid, 4 parts by mass of erythritol, 4 parts by mass of citric acid, 3 parts by mass of sucralose, and 3 parts by mass of ascorbic acid.

[0052] In one embodiment of the present invention, the third component comprises 4 parts by mass of γ-aminobutyric acid, 6 parts by mass of erythritol, 3 parts by mass of citric acid, 2 parts by mass of sucralose, and 4 parts by mass of ascorbic acid.

[0053] In one embodiment of the present invention, the third component comprises 3 parts by mass of γ-aminobutyric acid, 2 parts by mass of erythritol, 5 parts by mass of citric acid, 4 parts by mass of sucralose, and 5 parts by mass of ascorbic acid.

[0054] In the most preferred embodiment of the present invention, the mass ratio of the first component, the second component, and the third component is 4:77:19.

[0055] This invention provides a method for preparing the plant-based composition according to any one of the above technical solutions, comprising:

[0056] A) The first component is obtained by mixing protein melanin and phenylethanol glycoside;

[0057] Mix the Polygonatum powder, licorice powder, wolfberry powder, longan powder, jujube powder, Poria powder and red rose powder to obtain the second component;

[0058] The third component is obtained by mixing γ-aminobutyric acid, erythritol, citric acid, sucralose and ascorbic acid;

[0059] B) Mix the first, second and third components, sterilize, fill, and sterilize again to obtain the final product.

[0060] The method for preparing the plant-based composition provided by this invention first prepares protein melanin and phenylethanol glycoside according to the above method. The preparation method described above has already been clearly described in this invention and will not be repeated here.

[0061] The present invention has already clearly described the above components and proportions, and will not repeat them here.

[0062] The first component is obtained by mixing protein melanin and phenylethanoid glycoside; the second component is obtained by mixing Polygonatum sibiricum powder, licorice powder, wolfberry powder, longan powder, jujube powder, Poria cocos powder, and red rose powder evenly; the third component is obtained by mixing γ-aminobutyric acid, erythritol, citric acid, sucralose, and ascorbic acid evenly. This invention does not limit the mixing method; any method known to those skilled in the art is acceptable.

[0063] The preferred method for mixing the first component, the second component, and the third component is as follows:

[0064] Add the first component to water, stir until dissolved, then add the second component, continue stirring until completely dissolved, then add the third component according to the formula and stir until completely dissolved.

[0065] The product is then sterilized, filled, and sterilized a second time. The sterilization temperature is 105℃ for 5 minutes; the second sterilization temperature is 115℃ for 5 minutes.

[0066] This invention provides the use of the plant-based composition described in any one of the above claims in the preparation of anti-fatigue products.

[0067] The anti-fatigue measures described in this invention include extending the duration of weight-bearing swimming, increasing glycogen reserves, reducing BLA levels, and increasing LDH activity.

[0068] This invention provides the use of the plant-based compositions described in any one of the above-mentioned claims in the preparation of products that enhance immunity.

[0069] The immune enhancement described in this invention includes increasing the number of antibody-producing cells, the half-hemolysis value, carbon clearance capacity, the phagocytic rate of macrophages phagocytizing chicken erythrocytes, the phagocytic index, the lymphocyte proliferation capacity, and reducing foot swelling.

[0070] This invention provides a product for enhancing immunity / anti-fatigue, comprising the plant-based composition described in any one of the above technical solutions.

[0071] This invention provides a plant-based composition comprising a first component, a second component, and a third component. The first component comprises 1-3 parts by weight of protein melanin and 1-3 parts by weight of phenylethanoid glycoside. The second component comprises 10-14 parts by weight of Polygonatum sibiricum powder, 8-12 parts by weight of licorice powder, 8-12 parts by weight of wolfberry powder, 8-14 parts by weight of longan powder, 8-16 parts by weight of jujube powder, 12-15 parts by weight of Poria cocos powder, and 7-10 parts by weight of rose powder. The third component comprises 3-5 parts by weight of γ-aminobutyric acid (GABA), 2-6 parts by weight of erythritol, 3-5 parts by weight of citric acid, 2-4 parts by weight of sucralose, and 3-5 parts by weight of ascorbic acid. This invention uses protein melanin and phenylethanoid glycoside as the main active ingredients, supplemented with Polygonatum sibiricum powder, licorice powder, wolfberry powder, longan powder, jujube powder, Poria cocos powder, and rose powder; as well as GABA, erythritol, citric acid, sucralose, and ascorbic acid. By adding various food-grade medicinal ingredients, and using mouse swimming tests to assess fatigue resistance and immunity, the optimal composition was selected. Results showed that this composition can better regulate bodily functions and improve immunity, and can be applied in beverages, meal replacements, and special dietary products.

[0072] To further illustrate the present invention, the following detailed description of a plant-based composition, its preparation method, and its application, in conjunction with embodiments, is provided by the present invention.

[0073] The raw materials used in this invention are all commercially available products that meet food and drug standards and are safe and of high quality.

[0074] All embodiments and comparative examples of this invention have the same total amount, only the proportions are different.

[0075] Example 1:

[0076] The plant-based composition comprises component A, component B, and component C, and by weight, includes the following components: component A consists of 2 parts of dietary protein melanin and 2 parts of phenylethanoid glycoside; component B consists of 12 parts of Polygonatum sibiricum powder, 10 parts of licorice powder, 8 parts of wolfberry powder, 8 parts of longan powder, 16 parts of jujube powder, 15 parts of Poria cocos powder, and 8 parts of red rose powder; component C consists of 5 parts of γ-aminobutyric acid, 4 parts of erythritol, 4 parts of citric acid, 3 parts of sucralose, and 3 parts of ascorbic acid.

[0077] (1) Preparation method, including the following steps: ① Take the amount of component A raw material in the formula and mix it evenly to obtain mixture A, which is set aside; ② Take the amount of component B raw material in the formula and mix it evenly to obtain mixture B, which is set aside; ③ Take the amount of component C raw material in the formula and mix it evenly to obtain mixture C, which is set aside.

[0078] Add mixture A to water, stir until dissolved, then add mixture B, continue stirring until completely dissolved, then add the required amount of mixture B in sequence, stir until completely dissolved, sterilize, fill, and sterilize a second time to obtain the plant-based composition, and dry for later use.

[0079] (2) Fatigue resistance test

[0080] 1) Laboratory animals

[0081] Male Kunming mice, 6–8 weeks old, 18–20g.

[0082] 2) Method

[0083] ① Animal grouping and treatment

[0084] Eighty healthy male Kunming mice were randomly divided into three dosage groups according to their body weight: a blank control group, a low-dose group (S1-L), a medium-dose group (S1-M), and a high-dose group (S1-H) of the plant-based composition from Example 1. Each group consisted of 20 mice with free access to food and water. The dosages for the treatment groups were 200, 400, and 800 mg / kg, respectively. The blank control group received an equal volume of physiological saline. The medication was administered once daily by gavage for 14 consecutive days. The mice were given the medication at regular intervals according to their body weight (0.1 mL / 10 g) at a temperature of 22 ± 2℃ and a humidity of 60% ± 5%.

[0085] ② Mouse weight-bearing swimming experiment: 30 minutes after the last administration, 10 mice were taken from each of the blank control group, S1-L group, S1-M group and S1-H group. The mice were placed in a swimming tank with a lead weight of 5% of their body weight on their tails and placed in water at a depth of 40 cm and a water temperature of 25±1℃. The weight-bearing swimming time of the mice was recorded (the time from the start of swimming until they sank below the surface for 10 seconds and could not float back up).

[0086] ③ Establishment of mouse fatigue model: 30 min after the last administration, 10 mice were taken from each of the blank control group, S1-L group, S1-M group and S1-H group. The mice were made to swim without load for 60 min in a swimming tank with a water depth of 40 cm and a water temperature of 25±1℃ to induce fatigue. After resting for 10 min, serum, muscle and liver samples were collected from the mice to measure muscle glycogen (MG), liver glycogen (LG), serum lactate (BLA) and LDH and other indicators.

[0087] ④ Determination of MG and LG content: After mice were euthanized by cervical dislocation, fresh muscle and liver samples were immediately taken, rinsed in physiological saline at 4°C, blotted dry with filter paper, and the MG and LG content were determined according to the instructions of the glycogen content kit.

[0088] ⑤ Determination of BLA content and LDH activity in serum: Whole blood was collected from mice after enucleation. The blood was incubated at room temperature (approximately 37°C) for 30 minutes, then centrifuged at 3000 rpm for 5 minutes to obtain the supernatant, which was then the mouse serum. BLA content and LDH activity were determined according to the kit instructions.

[0089] Statistical methods: Experimental data were statistically analyzed using SPSS 17.0 software. Results are expressed as mean ± standard deviation. One-way ANOVA and Tukey's test were used for comparisons among multiple groups. P < 0.05 was considered statistically significant.

[0090] 4) Results

[0091] ① Example 1 Plant-based composition

[0092] The weight-bearing swimming time in mice is often used as an important indicator of fatigue level. Improved exercise endurance is the most direct manifestation of anti-fatigue effect; the longer the weight-bearing swimming time in mice, the higher the exercise endurance and the stronger the anti-fatigue effect. As shown in Table 1-1, the weight-bearing swimming time of mice in the low, medium, and high dose groups of the plant-based composition in Example 1 was significantly prolonged compared to the blank control group, with statistically significant differences (P<0.05 or P<0.01), extending the time by 27.88%, 95.17%, and 67.91%, respectively. The medium dose group (400 mg / kg) of the plant-based composition in Example 1 showed the best effect. The results indicate that the extract of Example 1 has the effect of prolonging the weight-bearing swimming time in mice.

[0093] Table 1-1 Swimming time of mice under load ( n=10)

[0094]

[0095] Note: *P vs. blank control group, **P<0.01 vs. blank control group.

[0096] ② Effect of plant-based compositions on glycogen content

[0097] MG and LG levels can be used to assess the degree of fatigue. As shown in Tables 1-2, compared with the blank control group, the MG and LG reserves of mice in the low, medium, and high dose groups of the plant-based composition were all increased after swimming, with statistically significant differences (P<0.05 or P<0.01). The medium dose group (400 mg / kg) of the plant-based composition showed the highest MG and LG reserves. These results indicate that the plant-based extract can increase glycogen reserves and alleviate fatigue.

[0098] Table 1-2 MG and LG content in mice after swimming ( n=10)

[0099]

[0100] Note: *P vs. blank control group, **P<0.01 vs. blank control group.

[0101] ③ Effect of plant-based compositions on BLA content

[0102] BLA levels can serve as an important indicator for assessing post-exercise fatigue. As shown in Tables 1-3, compared with the blank control group, mice in the medium and high dose groups of the plant-based composition showed decreased BLA levels after swimming, with statistically significant differences (P<0.05 or P<0.01). The medium dose group (400 mg / kg) of the plant-based composition showed the most significant inhibitory effect on BLA accumulation. These results indicate that the plant-based composition may inhibit BLA production or accelerate BLA metabolism after exercise, reducing BLA accumulation and delaying the onset of fatigue.

[0103] ④ Effect of plant-based compositions on LDH activity

[0104] LDH activity can serve as an important indicator for assessing the degree of fatigue after exercise. As shown in Table 3, the LDH activity of mice in the low, medium, and high dose groups of the plant-based composition after swimming was significantly higher than that in the blank control group, with statistically significant differences (P<0.01). These results indicate that the plant-based extract can significantly increase LDH activity, accelerate BLA metabolism, reduce BLA accumulation, and delay the onset of fatigue.

[0105] Table 1-3 Blood lactate levels and LDH activity in mice after swimming ( n=10)

[0106]

[0107] Note: *P vs. blank control group, **P<0.01 vs. blank control group.

[0108] 5) Discussion

[0109] Fatigue is a common symptom of physical discomfort, often caused by prolonged work or exercise, leading to a decline in physical function or poor mental state, resulting in decreased work efficiency. It can also trigger sudden illnesses, exacerbate existing conditions, or even threaten life. Studies often use weight-bearing swimming tests and biochemical indicators such as MG, LG, BLA, and LDH to evaluate fatigue levels and anti-fatigue effects. The weight-bearing swimming test is the most commonly used exercise endurance test. Mice in the plant-based composition group showed a significantly longer weight-bearing swimming time compared to the control group, indicating that the plant-based composition can improve exercise endurance and has a significant anti-fatigue effect. The higher the body's MG and LG reserves, the stronger the fatigue-delaying effect. The anti-fatigue effect of the plant-based composition may be related to its improvement of energy metabolism and increase of MG and LG reserves. The effects of the plant-based composition on BLA content and LDH activity showed that by increasing LDH activity, inhibiting BLA production or accelerating BLA metabolism, and reducing BLA accumulation, fatigue onset is delayed. The mechanism may be related to the presence of protein melanin and phenylethyl glycosides in the plant-based composition, thus achieving the fatigue-delaying effect. In summary, the plant-based composition prepared in Example 1 of this invention can prolong the swimming time of mice under load, increase the reserves of MG and LG after exercise, improve blood LDH activity, inhibit BLA production or accelerate BLA metabolism, and produce a significant anti-fatigue effect. Moreover, it has a certain dose-response relationship with the dosage and ratio of the plant-based composition, especially the 400mg / kg dosage group has the best anti-fatigue effect.

[0110] (3) Immunity testing experiment

[0111] 1) Laboratory animals and test samples

[0112] 192 female SPF-grade KM mice, weighing 18–20g, were selected.

[0113] 2) Experimental methods

[0114] After a period of acclimatization and observation, the mice were randomly divided into 16 groups of 12 mice each, with 4 groups constituting one experimental batch, for a total of 4 experimental batches. Experimental batch one underwent organ / body weight measurements, delayed-type hypersensitivity tests, hemolysis half-maximum (HMI) values, and antibody-producing cell counts. Experimental batch two underwent carbon clearance tests. Experimental batch three underwent ConA-induced mouse lymphocyte transformation and natural killer cell activity assays. Experimental batch four underwent mouse peritoneal macrophage phagocytosis of chicken erythrocytes experiments. Each group was further divided into three dosage groups based on the recommended daily oral dose of 1.6g for adults (adult body weight based on 60kg): 5 times, 10 times, and 30 times the recommended dose, i.e., 0.14g / kg BW, 0.27g / kg BW, and 0.81g / kg BW daily for low, medium, and high dose groups, respectively. For these groups, 0.70g, 1.35g, and 4.05g of the plant-based composition sample were respectively added to sterile water to 50mL, and the gavage volume for mice was 10mL / kg BW. The test substance was administered orally once daily for 33 days, after which various indicators were measured. A blank control group (0.00g / kg BW) was also established, using sterile water instead of the test substance, with the same daily gavage volume as the test substance groups. Each dosage group was given a maintenance diet at regular intervals each day. After 33 days, relevant immune function indicators were measured.

[0115] 3) Statistical methods

[0116] Data with This indicates that SPSS was used to perform analysis of variance on the data, and t-tests were used to compare data between groups.

[0117] 4) Experimental Results

[0118] ① Effect of the test substance on mouse body weight

[0119] Four batches of mice, 12 mice in each group, had initial body weights that were not statistically significantly different from the blank control group. Mice were orally administered different doses of the test substance (0.14 g / kg BW, 0.27 g / kg BW, and 0.81 g / kg BW). After 33 days, there were no statistically significant differences in body weight between the four batches of mice in different dose groups and the blank control group. See Tables 1-4 and 1-5.

[0120] Table 1-4 Initial body weight of mice in each group ( g)

[0121]

[0122] Table 1-5 Effects of different doses of the test substance on the body weight of mice in each group ( g)

[0123]

[0124]

[0125] ② Effect of the test substance on the organ / body weight ratio of experimental mice

[0126] Twelve mice in each group were orally administered different doses of the test substance (0.14 g / kg BW, 0.27 g / kg BW, and 0.81 g / kg BW) for 33 days. There were no statistically significant differences in spleen / body weight and thymus / body weight values ​​between the dose groups and the blank control group. (See Tables 1-6).

[0127] Table 1-6 Effects of different doses of the test substance on spleen / body weight and thymus / body weight values

[0128]

[0129] ③ Effects of the test substance on mouse cellular immune function

[0130] After oral administration of high, medium, and low doses of the test substance to 12 mice in each group for 33 days, the high-dose group (0.81 g / kg BW) showed statistically significant differences in paw edema and lymphocyte proliferation compared to the blank control group (P < 0.05). See Tables 1-7.

[0131] Table 1-7 Effects of different oral doses of the test substance on delayed-type hypersensitivity and lymphocyte transformation assays

[0132]

[0133] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0134] ④ Effects of the test substance on humoral immunity in mice

[0135] After oral administration of high, medium, and low doses of the test substance to mice for 33 days, the number of hemolytic plaques and the half-maximal hemolysis value in each dose group were significantly different from those in the blank control group (P < 0.05 or P < 0.01). See Table 1-8.

[0136] Table 1-8 Effects of different oral doses of the test substance on antibody-producing cell count and half-hemolytic value.

[0137]

[0138]

[0139] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0140] ⑤ Effect of the test substance on the phagocytic function of mouse mononuclear macrophages

[0141] After oral administration of high, medium, and low doses of the test substance to mice for 33 days, the phagocytic index of mice in the medium-dose and high-dose groups increased significantly compared with the blank control group (P < 0.05). See Table 1-9. The phagocytic rate and phagocytic index of macrophages phagocytosing chicken erythrocytes were also increased in the medium-dose and high-dose groups, with statistically significant differences compared with the blank control group (P < 0.05 or P < 0.01). See Table 1-10.

[0142] Table 1-9 Effects of different oral doses of the test substance on carbon clearance capacity

[0143]

[0144] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0145] Table 1-10 Effects of different oral doses of the test substance on the phagocytic rate and phagocytic index of macrophages phagocytosis of chicken erythrocytes.

[0146]

[0147] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0148] ⑥ Effect of the test substance on the activity of mouse natural killer cells

[0149] Mice were orally administered high, medium, and low doses of the test substance. After 33 days, the natural killer cell activity of mice at each dose was compared with that of the blank control group. The natural killer cell activity of the high-dose group (0.81 g / kg BW) was increased, but the difference was not statistically significant. See Table 1-11.

[0150] Table 1-11 Effects of different oral doses of the test substance on natural killer cell activity

[0151]

[0152]

[0153] 5) Discussion

[0154] The immune-enhancing mechanism of plant-based compositions does not follow a single biological model, but rather involves the comprehensive regulation of the body's immune function through multiple immune pathways and antibody targets, thereby significantly improving the organism's immunity. Studies have shown that many plant-based compositions, such as the protein melanin and phenylethyl glycosides in [the text abruptly ends here, so the translation stops here as well.]

[0155] The results of this study show that after oral administration of different doses of the test substance to mice for 33 days, the test substance had no adverse effect on the weight gain of the mice.

[0156] Compared with the blank control group, the high, medium and low dose groups of the test substance increased the number of antibody-producing cells and the half-hemolysis value in mice, with the high dose group showing a significant increase. The medium and high dose groups improved the carbon clearance ability of mice, with the high dose group showing a significant increase. Both the medium and high dose groups significantly increased the phagocytic rate and phagocytic index of mouse macrophages phagocytosis of chicken erythrocytes. The high dose group improved the proliferation ability of mouse lymphocytes and reduced the swelling of mouse paws.

[0157] Example 1 showed positive results in both cellular and humoral immune function tests of the plant-based composition, suggesting that the product enhances immunity. This study provides strong evidence for the development and widespread application of related products, and offers a scientific basis for further clinical applications and resource development.

[0158] Example 2:

[0159] The plant-based composition comprises component A, component B, and component C, and by weight, includes the following components: component A consists of 1 part of dietary protein melanin and 3 parts of phenylethanoid glycoside; component B consists of 10 parts of Polygonatum sibiricum powder, 8 parts of licorice powder, 12 parts of wolfberry powder, 12 parts of longan powder, 12 parts of jujube powder, 13 parts of Poria cocos powder, and 10 parts of red rose powder; component C consists of 4 parts of γ-aminobutyric acid, 6 parts of erythritol, 3 parts of citric acid, 2 parts of sucralose, and 4 parts of ascorbic acid.

[0160] (1) Preparation method, including the following steps: ① Take the amount of component A raw material in the formula and mix it evenly to obtain mixture A, which is set aside; ② Take the amount of component B raw material in the formula and mix it evenly to obtain mixture B, which is set aside; ③ Take the amount of component C raw material in the formula and mix it evenly to obtain mixture C, which is set aside.

[0161] Add mixture A to water, stir until dissolved, then add mixture B, continue stirring until completely dissolved, then add the required amount of mixture B in sequence, stir until completely dissolved, sterilize, fill, and sterilize a second time to obtain the plant-based composition, and dry for later use.

[0162] (2) Experimental results of anti-fatigue effect in Example 2

[0163] The experimental method in Example 2 of this invention is the same as that in Example 1.

[0164] ① Example 2: Mouse weighted swimming experiment using plant-based composition

[0165] The weight-bearing swimming time in mice is often used as an important indicator of fatigue level. Improved exercise endurance is the most direct manifestation of anti-fatigue effect; the longer the weight-bearing swimming time in mice, the higher the exercise endurance and the stronger the anti-fatigue effect. As shown in Table 2-1, the weight-bearing swimming time of mice in the low, medium, and high dose groups of the plant-based composition in Example 2 was significantly prolonged compared to the blank control group, with statistically significant differences (P<0.05 or P<0.01), extending the time by 15.85%, 65.16%, and 54.09%, respectively. The medium dose group (400 mg / kg) of the plant-based composition in Example 2 showed the best effect. The results indicate that the extract in Example 2 has the effect of prolonging the weight-bearing swimming time in mice.

[0166] Table 2-1 Swimming time of mice under load ( n=10)

[0167]

[0168] Note: *P vs. blank control group, **P<0.01 vs. blank control group.

[0169] ② Effect of plant-based compositions on glycogen content

[0170] MG and LG levels can be used to assess the degree of fatigue. As shown in Table 2-2, compared with the blank control group, the MG and LG reserves of mice in the low, medium, and high dose groups of the plant-based composition were all increased after swimming, with statistically significant differences (P<0.05 or P<0.01). The medium dose group (400 mg / kg) of the plant-based composition had the highest MG and LG reserves. The results indicate that the plant-based composition can increase glycogen reserves and alleviate fatigue.

[0171] Table 2-2 MG and LG content in mice after swimming ( n=10)

[0172]

[0173] Note: *P vs. blank control group, **P<0.01 vs. blank control group.

[0174] ③ Effect of plant-based compositions on BLA content

[0175] BLA levels can serve as an important indicator for assessing post-exercise fatigue. Table 3 shows that, compared to the blank control group, mice in the medium and high dose groups of the plant-based composition exhibited significantly lower BLA levels after swimming (P<0.05 or P<0.01), with the medium dose group (400 mg / kg) showing the most significant inhibitory effect on BLA accumulation. These results indicate that the plant-based composition may inhibit BLA production or accelerate BLA metabolism after exercise, reducing BLA accumulation and delaying the onset of fatigue.

[0176] ④ Effect of plant-based compositions on LDH activity

[0177] LDH activity can serve as an important indicator for assessing the degree of fatigue after exercise. As shown in Tables 2-3, the LDH activity of mice in the low, medium, and high dose groups of the plant-based composition after swimming was significantly higher than that in the blank control group, with statistically significant differences (P<0.01). These results indicate that the plant-based composition can significantly increase LDH activity, accelerate BLA metabolism, reduce BLA accumulation, and delay the onset of fatigue.

[0178] Table 2-3 Blood lactate levels and LDH activity in mice after swimming ( n=10)

[0179]

[0180] Note: *P vs. blank control group, **P<0.01 vs. blank control group.

[0181] 4) Discussion

[0182] In summary, the plant-based composition prepared in Example 2 of this invention can prolong the swimming time of mice under load, increase the reserves of MG and LG after exercise, improve blood LDH activity, inhibit BLA production or accelerate BLA metabolism, and produce a significant anti-fatigue effect. Moreover, it has a certain dose-response relationship with the dosage and ratio of the plant-based composition, especially the 400mg / kg dosage group has the best anti-fatigue effect.

[0183] (3) Results of Immunity Test

[0184] 1) Laboratory animals and test samples

[0185] 192 female SPF-grade KM mice, weighing 18–20g, were selected.

[0186] 2) Experimental methods

[0187] After a period of acclimatization and observation, the mice were randomly divided into 16 groups of 12 mice each, with 4 groups constituting one experimental batch, for a total of 4 experimental batches. Experimental batch one underwent organ / body weight measurements, delayed-type hypersensitivity tests, hemolysis half-maximum (HMI) values, and antibody-producing cell counts. Experimental batch two underwent carbon clearance tests. Experimental batch three underwent ConA-induced mouse lymphocyte transformation and natural killer cell activity assays. Experimental batch four underwent mouse peritoneal macrophage phagocytosis of chicken erythrocytes experiments. Each group was further divided into three dosage groups based on the recommended daily oral dose of 1.6g for adults (adult body weight based on 60kg): 5 times, 10 times, and 30 times the recommended dose, i.e., 0.14g / kg BW, 0.27g / kg BW, and 0.81g / kg BW daily for low, medium, and high dose groups, respectively. For these groups, 0.70g, 1.35g, and 4.05g of the plant-based composition sample were respectively added to sterile water to 50mL, and the gavage volume for mice was 10mL / kg BW. The test substance was administered orally once daily for 33 days, after which various indicators were measured. A blank control group (0.00g / kg BW) was also established, using sterile water instead of the test substance, with the same daily gavage volume as the test substance groups. Each dosage group was given a maintenance diet at regular intervals each day. After 33 days, relevant immune function indicators were measured.

[0188] 3) Statistical methods

[0189] Data with This indicates that SPSS was used to perform analysis of variance on the data, and t-tests were used to compare data between groups.

[0190] 4) Experimental Results

[0191] ① Effect of the test substance on mouse body weight

[0192] Four batches of mice, 12 mice in each group, had initial body weights that were not statistically significantly different from the blank control group. Mice were orally administered different doses of the test substance (0.14 g / kg BW, 0.27 g / kg BW, and 0.81 g / kg BW). After 33 days, there were no statistically significant differences in body weight between the four batches of mice in different dose groups and the blank control group. See Tables 2-4 and 2-5.

[0193] Table 2-4 Initial body weight of mice in each group ( g)

[0194]

[0195] Table 2-5 Effects of different doses of the test substance on the body weight of mice in each group ( g)

[0196]

[0197] ② Effect of the test substance on the organ / body weight ratio of experimental mice

[0198] Twelve mice in each group were orally administered different doses of the test substance (0.14 g / kg BW, 0.27 g / kg BW, and 0.81 g / kg BW) for 33 days. There were no statistically significant differences in spleen / body weight and thymus / body weight ratios between the dose groups and the blank control group. (See Table 2-6).

[0199] Table 2-6 Effects of different doses of the test substance on spleen / body weight and thymus / body weight values

[0200]

[0201] ③ Effects of the test substance on mouse cellular immune function: After oral administration of high, medium, and low doses of the test substance to 12 mice in each group for 33 days, the high-dose group (0.81 g / kg BW) showed statistically significant differences in paw edema and lymphocyte proliferation compared to the blank control group (P < 0.05). See Table 2-7.

[0202] Table 2-7 Effects of different oral doses of the test substance on delayed-type hypersensitivity and lymphocyte transformation assays

[0203]

[0204]

[0205] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0206] ④ Effects of the test substance on humoral immunity in mice: After oral administration of high, medium, and low doses of the test substance to mice for 33 days, the number of hemolytic plaques and the half-maximal hemolytic value in each dose group were significantly different from those in the blank control group (P < 0.05 or P < 0.01). See Table 2-8.

[0207] Table 2-8 Effects of different oral doses of the test substance on antibody-producing cell count and half-hemolytic value

[0208]

[0209] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0210] ⑤ Effect of the test substance on the phagocytic function of mouse mononuclear macrophages

[0211] After oral administration of high, medium, and low doses of the test substance to mice for 33 days, the phagocytic index of mice in the medium-dose and high-dose groups increased significantly compared with the blank control group (P < 0.05). See Table 2-9. The phagocytic rate and phagocytic index of macrophages phagocytosing chicken erythrocytes were also increased in the medium-dose and high-dose groups, with statistically significant differences compared with the blank control group (P < 0.05 or P < 0.01). See Table 2-10.

[0212] Table 2-9 Effects of different oral doses of the test substance on carbon clearance capacity

[0213]

[0214] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0215] Table 2-10 Effects of different oral doses of the test substance on the phagocytic rate and phagocytic index of macrophages phagocytosis of chicken erythrocytes.

[0216]

[0217]

[0218] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0219] ⑥ Effect of the test substance on the activity of mouse natural killer cells

[0220] Mice were orally administered high, medium, and low doses of the test substance. After 33 days, the natural killer cell activity of mice at each dose was compared with that of the blank control group. The natural killer cell activity of the high-dose group (0.81 g / kg BW) was increased, but the difference was not statistically significant. See Table 2-11.

[0221] Table 2-11 Effects of different oral doses of the test substance on the activity of natural killer cells

[0222]

[0223] 5) Discussion

[0224] The results of this Example 2 study show that after oral administration of different doses of the test substance to mice for 33 days, the test substance had no adverse effect on the weight gain of the mice.

[0225] Compared with the blank control group, the high, medium and low dose groups of the test substance increased the number of antibody-producing cells and the half-hemolysis value in mice, with the high dose group showing a significant increase. The medium and high dose groups improved the carbon clearance ability of mice, with the high dose group showing a significant increase. Both the medium and high dose groups significantly increased the phagocytic rate and phagocytic index of mouse macrophages phagocytosis of chicken erythrocytes. The high dose group effectively reduced the paw edema of mice and enhanced the proliferation ability of mouse lymphocytes.

[0226] Example 2 showed positive results in both cellular and humoral immune function tests of the plant-based composition, suggesting that the product enhances immunity. This study provides strong evidence for the development and widespread application of related products, and offers a scientific basis for further clinical applications and resource development.

[0227] Example 3:

[0228] The plant-based composition comprises component A, component B, and component C, and by weight, includes the following components: component A consists of 3 parts of dietary protein melanin and 1 part of phenylethanoid glycoside; component B consists of 14 parts of Polygonatum sibiricum powder, 12 parts of licorice powder, 10 parts of wolfberry powder, 14 parts of longan powder, 8 parts of jujube powder, 12 parts of Poria cocos powder, and 7 parts of red rose powder; component C consists of 3 parts of γ-aminobutyric acid, 2 parts of erythritol, 5 parts of citric acid, 4 parts of sucralose, and 5 parts of ascorbic acid.

[0229] (1) Preparation method, including the following steps: ① Take the amount of component A raw material in the formula and mix it evenly to obtain mixture A, which is set aside; ② Take the amount of component B raw material in the formula and mix it evenly to obtain mixture B, which is set aside; ③ Take the amount of component C raw material in the formula and mix it evenly to obtain mixture C, which is set aside.

[0230] Add mixture A to water, stir until dissolved, then add mixture B, continue stirring until completely dissolved, then add the required amount of mixture B in sequence, stir until completely dissolved, sterilize, fill, and sterilize a second time to obtain the plant-based composition, and dry for later use.

[0231] (2) Example 3: Experiment on the anti-fatigue effect of plant-based composition

[0232] The experimental method described above is the same as in Example 1.

[0233] 1) Example 3: Mouse weight-bearing swimming experiment with plant-based composition

[0234] Swimming time under load in mice is often used as an important indicator of fatigue level. Improved exercise endurance is the most direct manifestation of anti-fatigue effect; the longer the weight-bearing swimming time, the higher the exercise endurance and the stronger the anti-fatigue effect. As shown in Table 3-1, the low, medium, and high dose groups of the plant-based composition in Example 1 significantly prolonged the weight-bearing swimming time of mice compared to the blank control group, with statistically significant differences (P<0.05 or P<0.01), extending it by 21.52%, 90.55%, and 79.13%, respectively. The medium dose group (400 mg / kg) of the plant-based composition in Example 1 showed the best effect. The results indicate that the extract in Example 1 has the effect of prolonging the weight-bearing swimming time of mice.

[0235] Table 3-1 Swimming time of mice under load ( n=10)

[0236]

[0237] Note: *P vs. blank control group, **P<0.01 vs. blank control group.

[0238] 2) Effect of plant-based compositions on glycogen content

[0239] MG and LG levels can be used to assess the degree of fatigue. As shown in Table 3-2, compared with the blank control group, the MG and LG reserves of mice in the low, medium, and high dose groups of the plant-based composition were all increased after swimming, with statistically significant differences (P<0.05 or P<0.01). The medium dose group (400 mg / kg) of the plant-based composition had the highest MG and LG reserves. The results indicate that the plant-based composition can increase glycogen reserves and alleviate fatigue.

[0240] Table 3-2 MG and LG content in mice after swimming ( n=10)

[0241]

[0242] Note: *P vs. blank control group, **P<0.01 vs. blank control group.

[0243] 3) Effect of plant-based compositions on BLA content

[0244] BLA levels can serve as an important indicator for assessing post-exercise fatigue. Table 3 shows that, compared to the blank control group, mice in the medium and high dose groups of the plant-based composition exhibited significantly lower BLA levels after swimming (P<0.05 or P<0.01), with the medium dose group (400 mg / kg) showing the most significant inhibitory effect on BLA accumulation. These results indicate that the plant-based composition may inhibit BLA production or accelerate BLA metabolism after exercise, reducing BLA accumulation and delaying the onset of fatigue.

[0245] 4) Effect of plant-based compositions on LDH activity

[0246] LDH activity can serve as an important indicator for assessing the degree of fatigue after exercise. As shown in Table 3-3, the LDH activity of mice in the low, medium, and high dose groups of the plant-based composition after swimming was significantly higher than that in the blank control group, with statistically significant differences (P<0.01). These results indicate that the plant-based composition can significantly increase LDH activity, accelerate BLA metabolism, reduce BLA accumulation, and delay the onset of fatigue.

[0247] Table 3-3 Blood lactate content and LDH activity in mice after swimming ( n=10)

[0248]

[0249] Note: *P vs. blank control group, **P<0.01 vs. blank control group.

[0250] Example 3 showed positive results in both cellular and humoral immune function tests of the plant-based composition, suggesting that the product enhances immunity. This study provides strong evidence for the development and widespread application of related products, and offers a scientific basis for further clinical applications and resource development.

[0251] (3) Results of Immunity Test in Example 3

[0252] 1) Laboratory animals and test samples

[0253] 192 female SPF-grade KM mice, weighing 18–20g, were selected.

[0254] 2) Experimental methods

[0255] After a period of acclimatization and observation, the mice were randomly divided into 16 groups of 12 mice each, with 4 groups constituting one experimental batch, for a total of 4 experimental batches. Experimental batch one underwent organ / body weight measurements, delayed-type hypersensitivity tests, hemolysis half-maximum (HMI) values, and antibody-producing cell counts. Experimental batch two underwent carbon clearance tests. Experimental batch three underwent ConA-induced mouse lymphocyte transformation and natural killer cell activity assays. Experimental batch four underwent mouse peritoneal macrophage phagocytosis of chicken erythrocytes experiments. Each group was further divided into three dosage groups based on the recommended daily oral dose of 1.6g for adults (adult body weight based on 60kg): 5 times, 10 times, and 30 times the recommended dose, i.e., 0.14g / kg BW, 0.27g / kg BW, and 0.81g / kg BW daily for low, medium, and high dose groups, respectively. For these groups, 0.70g, 1.35g, and 4.05g of the plant-based composition sample were respectively added to sterile water to 50mL, and the gavage volume for mice was 10mL / kg BW. The test substance was administered orally once daily for 33 days, after which various indicators were measured. A blank control group (0.00g / kg BW) was also established, using sterile water instead of the test substance, with the same daily gavage volume as the test substance groups. Each dosage group was given a maintenance diet at regular intervals each day. After 33 days, relevant immune function indicators were measured.

[0256] 3) Statistical methods

[0257] Data with This indicates that SPSS was used to perform analysis of variance on the data, and t-tests were used to compare data between groups.

[0258] ① Effect of the test substance on mouse body weight

[0259] Four batches of mice, 12 mice in each group, had initial body weights that were not statistically significantly different from the blank control group. Mice were orally administered different doses of the test substance (0.14 g / kg BW, 0.27 g / kg BW, and 0.81 g / kg BW). After 33 days, there were no statistically significant differences in body weight between the four batches of mice in different dose groups and the blank control group. See Tables 3-4 and 3-5.

[0260] Table 3-4 Initial body weight of mice in each group ( g)

[0261]

[0262] Table 3-5 Effects of different doses of the test substance on the body weight of mice in each group ( g)

[0263]

[0264] ② Effect of the test substance on the organ / body weight ratio of experimental mice

[0265] Twelve mice in each group were orally administered different doses of the test substance (0.14 g / kg BW, 0.27 g / kg BW, and 0.81 g / kg BW) for 33 days. There were no statistically significant differences in spleen / body weight and thymus / body weight ratios between the control groups and the control group. (See Table 3-6).

[0266] Table 3-6 Effects of different doses of the test substance on spleen / body weight and thymus / body weight values

[0267]

[0268] ③ Effects of the test substance on mouse cellular immune function

[0269] After oral administration of high, medium, and low doses of the test substance to 12 mice in each group for 33 days, the high-dose group (0.81 g / kg BW) showed statistically significant differences in paw edema and lymphocyte proliferation compared to the blank control group (P < 0.05). See Table 3-7.

[0270] Table 3-7 Effects of different oral doses of the test substance on delayed-type hypersensitivity and lymphocyte transformation assays

[0271]

[0272] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0273] ④ Effects of the test substance on humoral immunity in mice

[0274] After oral administration of high, medium, and low doses of the test substance to mice for 33 days, the number of hemolytic plaques and the half-maximal hemolysis value in each dose group were significantly different from those in the blank control group (P < 0.05 or P < 0.01). See Table 3-8.

[0275] Table 3-8 Effects of different oral doses of the test substance on antibody-producing cell count and half-hemolytic value.

[0276]

[0277] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0278] ⑤ Effect of the test substance on the phagocytic function of mouse mononuclear macrophages

[0279] After oral administration of high, medium, and low doses of the test substance to mice for 33 days, the phagocytic index of mice in the medium-dose and high-dose groups increased significantly compared with the blank control group (P < 0.05). See Table 3-9. The phagocytic rate and phagocytic index of macrophages phagocytosing chicken erythrocytes were also increased in the medium-dose and high-dose groups, with statistically significant differences compared with the blank control group (P < 0.05 or P < 0.01). See Table 3-10.

[0280] Table 3-9 Effects of different oral doses of the test substance on carbon clearance capacity

[0281]

[0282]

[0283] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0284] Table 3-10 Effects of different oral doses of the test substance on the phagocytic rate and phagocytic index of macrophages phagocytosis of chicken erythrocytes.

[0285]

[0286] Note: * indicates that the difference is statistically significant compared with the 0.00 g / kg BW group.

[0287] ⑥ Effect of the test substance on the activity of mouse natural killer cells

[0288] Mice were orally administered high, medium, and low doses of the test substance. After 33 days, the natural killer cell activity of mice at each dose was compared with that of the blank control group. The natural killer cell activity of the high-dose group (0.81 g / kg BW) was increased, but the difference was not statistically significant. See Table 3-11.

[0289] Table 3-11 Effects of different oral doses of the test substance on natural killer cell activity

[0290]

[0291] Example 3 showed positive results in both cellular and humoral immune function tests of the plant-based composition, suggesting that the product enhances immunity. This study provides strong evidence for the development and widespread application of related products, and offers a scientific basis for further clinical applications and resource development.

[0292] discuss:

[0293] In summary, the plant-based composition prepared in Example 3 of this invention can prolong the swimming time of mice under load, increase the reserves of MG and LG after exercise, improve blood LDH activity, inhibit BLA production or accelerate BLA metabolism, and produce a significant anti-fatigue effect. Moreover, it has a certain dose-response relationship with the dosage and ratio of the plant-based composition, especially the 400mg / kg dosage group has the best anti-fatigue effect.

[0294] The immune-enhancing mechanism of plant-based compositions does not follow a single biological model, but rather involves the comprehensive regulation of the body's immune function through multiple immune pathways and antibody targets, thereby significantly improving the organism's immunity. Studies have shown that many plant-based compositions, such as protein melanin and phenylethanoid glycosides, have immune-enhancing effects and are novel natural immunomodulators. They can activate immune cells such as macrophages, T / B lymphocytes, and natural killer cells, promote antibody production, promote the release of various cytokines, and activate the complement system.

[0295] The results of this study show that after oral administration of different doses of the test substance to mice for 33 days, the test substance had no adverse effect on the weight gain of the mice.

[0296] Compared with the blank control group, the high, medium and low dose groups of the test substance increased the number of antibody-producing cells and the half-hemolysis value in mice, with the high dose group showing a significant increase. The medium and high dose groups improved the carbon clearance ability of mice, with the high dose group showing a significant increase. Both the medium and high dose groups significantly increased the phagocytic rate and phagocytic index of mouse macrophages phagocytosis of chicken erythrocytes. The high dose group effectively reduced the paw edema of mice and improved the proliferation ability of mouse lymphocytes.

[0297] When the ratio of component A, component B, and component C is controlled at 4:77:19, and the ratio of protein melanin to phenylethanol glycoside in component A is 1:1, the composition exhibits the best anti-fatigue effect and synergistic immune-enhancing effect in mice.

[0298] Example 1 differs from Examples 2 and 3 in that the mass ratio of melanin protein to phenylethyl glycoside in component A is 1:1, and the distribution ratios of each component in components B and C are different. The study found that the mass ratio of melanin protein to phenylethyl glycoside has the greatest impact on the anti-fatigue and immune-enhancing effects of the composition.

[0299] Comparative Example 1 (lacking protein melanin compared to Example 1)

[0300] The plant-based composition comprises component A, component B, and component C, and by weight, includes the following components: component A contains no dietary protein melanin and only 2 parts of phenylethanoid glycoside; component B includes 12 parts of Polygonatum sibiricum powder, 10 parts of licorice powder, 8 parts of wolfberry powder, 8 parts of longan powder, 16 parts of jujube powder, 15 parts of Poria cocos powder, and 8 parts of red rose powder; component C includes 5 parts of γ-aminobutyric acid, 4 parts of erythritol, 4 parts of citric acid, 3 parts of sucralose, and 3 parts of ascorbic acid.

[0301] (1) Preparation method, including the following steps: ① Take the amount of component A raw material in the formula and mix it evenly to obtain mixture A, which is set aside; ② Take the amount of component B raw material in the formula and mix it evenly to obtain mixture B, which is set aside; ③ Take the amount of component C raw material in the formula and mix it evenly to obtain mixture C, which is set aside.

[0302] Add mixture A to water, stir until dissolved, then add mixture B, continue stirring until completely dissolved, then add the required amount of mixture B in sequence, stir until completely dissolved, sterilize, fill, and sterilize a second time to obtain the plant-based composition, and dry for later use.

[0303] (2) Fatigue resistance test

[0304] 1) Laboratory animals

[0305] Male Kunming mice, 6–8 weeks old, 18–20g.

[0306] 2) Method

[0307] ① Animal grouping and treatment

[0308] Eighty healthy male Kunming mice were randomly divided into three dosage groups according to their body weight: a blank control group, a low-dose group (S1-L), a medium-dose group (S1-M), and a high-dose group (S1-H) of the plant-based composition from Example 1. Each group consisted of 20 mice with free access to food and water. The dosages for the treatment groups were 200, 400, and 800 mg / kg, respectively. The blank control group received an equal volume of physiological saline. The medication was administered once daily by gavage for 14 consecutive days. The mice were given the medication at regular intervals according to their body weight (0.1 mL / 10 g) at a temperature of 22 ± 2℃ and a humidity of 60% ± 5%.

[0309] ② Mouse weight-bearing swimming experiment: 30 minutes after the last administration, 10 mice were taken from each of the blank control group, S1-L group, S1-M group and S1-H group. The mice were placed in a swimming tank with a lead weight of 5% of their body weight on their tails and placed in water at a depth of 40 cm and a water temperature of 25±1℃. The weight-bearing swimming time of the mice was recorded (the time from the start of swimming until they sank below the surface for 10 seconds and could not float back up).

[0310] ③ Establishment of mouse fatigue model: 30 min after the last administration, 10 mice were taken from each of the blank control group, S1-L group, S1-M group and S1-H group. The mice were made to swim without load for 60 min in a swimming tank with a water depth of 40 cm and a water temperature of 25±1℃ to induce fatigue. After resting for 10 min, serum, muscle and liver samples were collected from the mice to measure muscle glycogen (MG), liver glycogen (LG), serum lactate (BLA) and LDH and other indicators.

[0311] 4) Results

[0312] ①Comparative Example 1: Plant-based Composition

[0313] The weight-bearing swimming time of mice is often used as an important indicator of fatigue level. Improved exercise endurance is the most direct manifestation of anti-fatigue effect. The longer the weight-bearing swimming time of mice, the higher the exercise endurance and the stronger the anti-fatigue effect. As shown in Table 1-Comparative Example 1, the weight-bearing swimming time of mice in the low, medium, and high dose groups of the plant-based composition in Comparative Example 1 was slightly longer than that of the blank control group, but the difference was not significant. The increases were 10.64%, 29.65%, and 18.75%, respectively. The medium dose group (400 mg / kg) of the plant-based composition in Comparative Example 1 showed the best effect.

[0314] Table 4-1(1) Swimming time of mice under load (±s, n=10)

[0315]

[0316] ② Effect of plant-based compositions on glycogen content

[0317] MG and LG levels can be used to evaluate the degree of fatigue. As shown in Table 1-2(1), compared with the blank control group, the MG and LG reserves of mice in the low, medium, and high dose groups of the plant-based composition were slightly increased after swimming, but the differences were not statistically significant. Among them, the medium dose group (400 mg / kg) of the plant-based composition had the highest MG and LG reserves. The results indicate that the plant-based extract can increase glycogen reserves and reduce fatigue.

[0318] Table 4-2(1) MG and LG content in mice after swimming (±s, n=10)

[0319]

[0320] (3) Results of Immunity Test

[0321] ① Effects of the test substance on humoral immunity in mice

[0322] After oral administration of high, medium, and low doses of the test substance to mice for 33 days, the number of hemolytic plaques and the half-maximal hemolysis value in each dose group were significantly different from those in the blank control group (P < 0.05 or P < 0.01). See Table 1-3 (1).

[0323] Table 4-3(1) Effects of different oral doses of the test substance on the number of antibody-producing cells and the half-hemolytic value (±s)

[0324]

[0325] ② Effect of the test substance on the activity of mouse natural killer cells

[0326] Mice were orally administered high, medium, and low doses of the test substance. After 33 days, the natural killer cell activity of mice at each dose was compared with that of the blank control group. The natural killer cell activity of the high dose group (0.81 g / kg BW) was increased, but the difference was not statistically significant. See Table 1-4(1).

[0327] Table 4-4(1) Effects of different oral doses of the test substance on the activity of natural killer cells

[0328]

[0329] Comparative Example 2 (replacing erythritol with xylitol in contrast to Example 2)

[0330] The plant-based composition comprises component A, component B, and component C, and by weight, includes the following components: component A contains 2 parts of dietary protein melanin and 2 parts of phenylethanoid glycoside only; component B includes 12 parts of Polygonatum sibiricum powder, 10 parts of licorice powder, 8 parts of wolfberry powder, 8 parts of longan powder, 16 parts of jujube powder, 15 parts of Poria cocos powder, and 8 parts of red rose powder; component C includes 5 parts of γ-aminobutyric acid, 4 parts of xylitol, 4 parts of citric acid, 3 parts of sucralose, and 3 parts of ascorbic acid.

[0331] According to the results of anti-fatigue and immunity experiments, in Comparative Example 2, after replacing 4 parts of erythritol with 4 parts of xylitol, the swimming time of mice under load and the ability to increase lymphocyte proliferation were slightly reduced compared with the Example.

[0332] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A plant-based composition, characterized in that, It consists of a first component, a second component, and a third component; The first component consists of 1-3 parts by weight of protein melanin and 1-3 parts by weight of phenylethanol glycoside. The second component consists of 10-14 parts by weight of Polygonatum sibiricum powder, 8-12 parts by weight of Glycyrrhiza uralensis powder, 8-12 parts by weight of Lycium barbarum powder, 8-14 parts by weight of Longan powder, 8-16 parts by weight of Jujube powder, 12-15 parts by weight of Poria cocos powder, and 7-10 parts by weight of Rose powder. The third component consists of 3-5 parts by weight of γ-aminobutyric acid, 2-6 parts by weight of erythritol, 3-5 parts by weight of citric acid, 2-4 parts by weight of sucralose, and 3-5 parts by weight of ascorbic acid.

2. The plant-based composition according to claim 1, characterized in that, The first component raw material is 2 parts by mass of protein melanin and 2 parts by mass of phenylethanol glycoside; The second component consists of 12 parts by weight of Polygonatum sibiricum powder, 10 parts by weight of Glycyrrhiza uralensis powder, 8 parts by weight of Lycium barbarum powder, 8 parts by weight of Longan powder, 16 parts by weight of Jujube powder, 15 parts by weight of Poria cocos powder, and 8 parts by weight of Rose powder. The third component consists of 5 parts by mass of γ-aminobutyric acid, 4 parts by mass of erythritol, 4 parts by mass of citric acid, 3 parts by mass of sucralose, and 3 parts by mass of ascorbic acid.

3. The plant-based composition according to claim 1 or 2, characterized in that, The mass ratio of the first component, the second component, and the third component is 4:77:

19.

4. The method for preparing a plant-based composition according to any one of claims 1 to 3, characterized in that, include: A) The first component is obtained by mixing protein melanin and phenylethanol glycoside; Mix the Polygonatum powder, licorice powder, wolfberry powder, longan powder, jujube powder, Poria powder and red rose powder to obtain the second component; The third component is obtained by mixing γ-aminobutyric acid, erythritol, citric acid, sucralose and ascorbic acid; B) Mix the first component, the second component, and the third component, sterilize, fill, and sterilize again to obtain the final product.

5. The preparation method according to claim 4, characterized in that, The sterilization temperature is 105 ℃ and the sterilization time is 5 min; The secondary sterilization was performed at a temperature of 115 °C for 5 minutes.

6. The use of the plant-based composition according to any one of claims 1 to 3 in the preparation of anti-fatigue products.

7. Use according to claim 6, characterized in that, The anti-fatigue measures include extending the duration of weight-bearing swimming, increasing glycogen reserves, reducing serum lactate levels, and increasing lactate dehydrogenase activity.

8. The use of the plant-based composition according to any one of claims 1 to 3 in the preparation of products that enhance immunity.

9. Use according to claim 8, characterized in that, The enhanced immunity includes increasing the number of antibody-producing cells, the half-hemolysis value, carbon clearance capacity, macrophage phagocytosis rate and phagocytic index of chicken red blood cells, lymphocyte proliferation capacity, and reducing foot swelling.

10. A product for enhancing immunity / anti-fatigue, characterized by, Includes the plant-based composition according to any one of claims 1 to 3.

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