A composition with fatigue alleviating and hypoxia tolerance functions and application thereof

Abstract

The present application relates to the field of food biotechnology, in particular to a composition with fatigue relief and hypoxia tolerance function and application thereof. The composition comprises the following components in parts by weight: Gastrodia elata 6 parts, Rhodiola 9 parts, Phyllanthus emblica 3 parts. The composition helps to up-regulate the AMPK / PGC-1 alpha signal pathway, promote mitochondrial biosynthesis and energy metabolism, reduce hippocampal tissue oxidative stress, prolong the time of weight swimming, reduce blood lactic acid and blood urea nitrogen levels, and improve learning and memory ability under fatigue. The present application has high safety, few side effects, and mild effect, and is suitable for early prevention and long-term conditioning of fatigue population caused by high altitude travel, high intensity exercise, long distance driving, brain work and large physical consumption.

Description

Technical Field

[0001] This invention relates to the field of food biotechnology, and in particular to a composition with fatigue-relieving and hypoxia-resistant functions and its applications. Background Technology

[0002] Fatigue is a physiological decline in bodily function that occurs after sustained physical or mental activity, often manifesting as muscle soreness, poor concentration, slow reaction time, dizziness, and headaches. With the fast pace of modern life, scenarios such as high-intensity mental work, long-distance driving, high-altitude travel, and strenuous exercise are becoming increasingly common, making fatigue a significant factor affecting work efficiency and quality of life. If chronic fatigue is not effectively relieved, it can lead to decreased immunity, endocrine disorders, impaired cardiovascular function, and even induce chronic fatigue syndrome.

[0003] The mechanisms of fatigue are complex and mainly include the following aspects: First, energy metabolism disorder, with prolonged high-intensity activity leading to ATP depletion, reduced glycogen reserves, and accumulation of metabolic products such as lactic acid and blood urea nitrogen; second, oxidative stress damage, with large amounts of reactive oxygen species (ROS) produced during strenuous exercise or in hypoxic environments, exceeding the body's antioxidant defense capacity, resulting in lipid peroxidation and cell membrane damage; and third, neuroendocrine disorders, with dysfunction of the hypothalamus-pituitary-adrenal axis and abnormal cortisol secretion, affecting the excitability of the central nervous system.

[0004] Currently, strategies for combating fatigue mainly include rest and sleep, nutritional supplementation, exercise adjustment, and interventions. However, existing methods have significant shortcomings. First, while caffeine-based stimulants can temporarily boost alertness, their effects are short-lived, easily leading to tolerance and dependence, and may cause side effects such as palpitations and insomnia. Second, energy drinks often contain high levels of sugar, posing metabolic risks with long-term consumption. Third, anti-fatigue drugs, such as ginseng preparations, are expensive and not suitable for all populations. Therefore, there is an urgent need for anti-fatigue products that are highly safe, have few side effects, are gentle in effect, and are suitable for long-term use. Summary of the Invention

[0005] To address the aforementioned problems, this invention provides a composition and its application that alleviates fatigue and improves hypoxia tolerance. The composition activates the AMPK / PGC-1α signaling pathway, reduces oxidative stress in brain tissue, prolongs swimming time under load, lowers blood lactate and blood urea nitrogen levels, and improves learning and memory abilities under fatigue. This invention is highly safe, has few side effects, and a mild effect. All raw materials are selected from the "food and medicine homology" and "health food raw materials" lists published by the National Health Commission, making it suitable for early prevention and long-term conditioning of fatigue caused by high-altitude travel, high-intensity exercise, long-distance driving, mental labor, and high physical exertion.

[0006] To achieve the above objectives, the present invention provides the following technical solution: The present invention provides a composition having the functions of relieving fatigue and hypoxia tolerance, comprising the following components in parts by weight: 6 parts of Gastrodia elata, 9 parts of Rhodiola rosea, and 3 parts of Phyllanthus emblica.

[0007] Preferably, the method for preparing the composition includes the following steps: 1) Gastrodia elata, Rhodiola rosea, and Phyllanthus emblica were pulverized and passed through a 60-mesh sieve to obtain powders of each component; 2) Mix the powders of each component obtained in step 1) according to the weight proportions, add 8 times the total mass of pure water, soak for 2 hours, decoct twice, 30 minutes each time, and combine the decoctions; 3) Filter the combined decoction obtained in step 2), concentrate the filtrate to a clear extract with a relative density of 1.15 (60°C), and spray dry to obtain the composition.

[0008] Preferably, the powder pulverized in step 1) is passed through a 60-mesh sieve. Preferably, the soaking time in step 2) is 2 hours, and the decoction is performed twice, 30 minutes each time. Preferably, the inlet air temperature of the spray drying in step 3) is 180°C, and the outlet air temperature is 80°C.

[0009] The present invention also provides a method for preparing the composition described in the above technical solution, comprising the following steps: mixing the Gastrodia elata, Rhodiola rosea and Phyllanthus emblica in parts by weight, adding water to decoct, concentrate and dry to obtain the composition.

[0010] The present invention also provides the application of the composition described above in the preparation of fatigue-relieving products.

[0011] The present invention also provides the application of the composition described in the above technical solution in the preparation of hypoxia-resistant products.

[0012] Preferably, the product is food, medicine, or health product.

[0013] Gastrodia elata, known for its ability to calm liver yang and dispel wind and unblock meridians, can improve symptoms of "failure of clear yang" such as dizziness, heaviness in the head, and vertigo. Its active ingredient, gastrodin, has sedative, anti-vertigo, and neuroprotective effects, providing a foundation for refreshing the mind. Rhodiola rosea has the effects of invigorating qi and blood circulation, and relieving asthma. Its active ingredient, rhodioloside, can significantly enhance the body's tolerance to hypoxia, effectively combating fatigue and restoring energy. It can also activate the AMPK / PGC-1α energy metabolism signaling pathway. Phyllanthus emblica clears heat and generates fluids, and is rich in superoxide dismutase (SOD) and vitamin C, exhibiting significant antioxidant and free radical scavenging effects, which can counteract oxidative stress damage caused by exercise and hypoxia. Through the synergistic effects of calming wind and relieving dizziness, invigorating qi and blood circulation, and clearing heat and resisting oxidation, these three ingredients achieve comprehensive relief of fatigue.

[0014] The beneficial effects of this invention are: The composition provided by this invention can promote mitochondrial biosynthesis and energy metabolism by activating the AMPK / PGC-1α signaling pathway, reduce oxidative stress in brain tissue, significantly prolong the swimming time of mice under load, reduce serum lactate and blood urea nitrogen levels, and improve learning and memory abilities under fatigue. This invention has high safety, few side effects, and a mild effect. All raw materials are food-grade medicinal materials, making it suitable for early prevention and long-term conditioning of fatigue caused by high-altitude travel, high-intensity exercise, long-distance driving, mental labor, and high physical exertion. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0016] Figure 1 shows the effect of the composition on the Morris water maze test in fatigued mice; (A) escape latency in the concealed platform test; (B) number of times the original platform location was crossed in the spatial exploration test; (C) time spent in the original platform quadrant in the spatial exploration test. Figure 2 shows the effect of the composition on the ability of fatigued mice to recognize new objects; (A) total exploration time of mice for new and old objects; (B) resolution index of mice for new objects; Figure 3 shows the effect of the composition on the weight-bearing swimming time of fatigued mice; Figure 4 shows the effects of the composition on serum lactate (LA) and blood urea nitrogen (BUN) levels in fatigued mice; (A) Effect of the composition on serum lactate (LA) levels; (B) Effect of the composition on blood urea nitrogen (BUN) levels. Figure 5 shows the effects of the composition on the activities of SOD, GSH-Px and MDA levels in the hippocampus of fatigued mice; (A) SOD activity; (B) GSH-Px activity; (C) MDA activity; Figure 6 shows the effect of the composition on the expression of AMPK / PGC-1α signaling pathway-related proteins in the hippocampus of fatigued mice; (A) Western blot analysis of the protein expression levels of p-AMPK and AMPK in the hippocampus; (B) Western blot analysis of the protein expression levels of PGC-1α and SIRT1 in the hippocampus; (C) Quantitative analysis results of the corresponding gray values. Detailed Implementation

[0017] The present invention provides a composition having the functions of relieving fatigue and hypoxia tolerance, comprising the following components in parts by weight: 6 parts of Gastrodia elata, 9 parts of Rhodiola rosea, and 3 parts of Phyllanthus emblica.

[0018] In this invention, the preparation method of the composition preferably includes the following steps: 1) Gastrodia elata, Rhodiola rosea, and Phyllanthus emblica were pulverized and passed through a 60-mesh sieve to obtain powders of each component; 2) Mix the powders of each component obtained in step 1) according to the weight proportions, add 8 times the total mass of pure water, soak for 2 hours, decoct twice, 30 minutes each time, and combine the decoctions; 3) Filter the combined decoction obtained in step 2), concentrate the filtrate to a clear extract with a relative density of 1.15 (60°C), and spray dry to obtain the composition.

[0019] In this invention, the preferred method for preparing Gastrodia elata includes the following steps: washing and slicing the Gastrodia elata raw material, drying it at 60°C until the moisture content is below 5%, and pulverizing it through a 60-mesh sieve to obtain Gastrodia elata powder. In this invention, the preferred method for preparing Rhodiola rosea includes the following steps: washing and slicing the Rhodiola rosea rhizome, drying it at 55°C until the moisture content is below 5%, and pulverizing it through a 60-mesh sieve to obtain Rhodiola rosea powder. In this invention, the preferred method for preparing Phyllanthus emblica includes the following steps: washing the fresh Phyllanthus emblica fruit, drying it at 50°C until the moisture content is below 8%, and pulverizing it through a 60-mesh sieve to obtain Phyllanthus emblica powder.

[0020] In this invention, the spray drying conditions preferably include: an inlet air temperature of 180°C and an outlet air temperature of 80°C.

[0021] The present invention also provides a method for preparing the composition described in the above technical solution, comprising the following steps: mixing the Gastrodia elata, Rhodiola rosea and Phyllanthus emblica in parts by weight, adding water to decoct, concentrate and dry to obtain the composition.

[0022] This invention also provides the application of the composition described in the above-described technical solutions in the preparation of fatigue-relieving products. This invention further provides the application of the composition described in the above-described technical solutions in the preparation of hypoxia-resistant products. In this invention, the product is preferably a food, pharmaceutical, or health product.

[0023] To further illustrate the present invention, the following detailed description is provided in conjunction with embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0024] Example 1 A composition with the functions of relieving fatigue and hypoxia tolerance consists of the following components in parts by weight: 6 parts of Gastrodia elata, 9 parts of Rhodiola rosea, and 3 parts of Phyllanthus emblica.

[0025] The preparation method of Gastrodia elata includes the following steps: wash and slice the raw material of Gastrodia elata, dry it at 60°C until the moisture content is less than 5%, and pulverize it through a 60-mesh sieve to obtain Gastrodia elata powder.

[0026] The preparation method of Rhodiola rosea is as follows: wash and slice the rhodiola rosea rhizome, dry it at 55°C until the moisture content is less than 5%, and pulverize it through a 60-mesh sieve to obtain Rhodiola rosea powder.

[0027] The preparation method of Phyllanthus emblica is as follows: wash the fresh Phyllanthus emblica fruit, dry it at 50℃ until the moisture content is less than 8%, pulverize it through a 60-mesh sieve to obtain Phyllanthus emblica powder.

[0028] The preparation method of the composition is as follows: 6 parts of Gastrodia elata powder, 9 parts of Rhodiola rosea powder and 3 parts of Phyllanthus emblica powder are mixed according to the weight ratio, 8 times the total mass of pure water is added, soaked for 2 hours, and then decocted twice, 30 minutes each time. The decoctions are combined, filtered, and the filtrate is concentrated to a clear extract with a relative density of 1.15 (60℃). The extract is then spray-dried (inlet air temperature 180℃, outlet air temperature 80℃) to obtain the composition.

[0029] Example 2 Model preparation: Fifty healthy male ICR mice, weighing 18-22g, were randomly divided into five groups: control group, model group, positive control group, low-dose composition group, and high-dose composition group, with 10 mice in each group. Immediately after grouping, administration of rhodioloside was initiated via gavage: the positive control group received 50 mg / kg / day, the low-dose composition group received 250 mg / kg / day, and the high-dose composition group received 500 mg / kg / day; the control group and model group received an equal volume of physiological saline.

[0030] Simultaneous drug administration and modeling were conducted. Except for the control group, mice in other groups underwent chronic fatigue stress modeling using a combination of forced swimming and sleep deprivation. Mice were placed in a cylindrical bucket with a water depth of 25 cm and a water temperature of 25 ± 1 °C and forced to swim for 15 min, once daily for 14 consecutive days. Starting from the 8th day of modeling, a small platform aquatic environment sleep deprivation treatment was added. The platform had a diameter of 4 cm, and the water surface was about 1 cm below the platform surface. Sleep deprivation was performed for 6 hours daily for 7 consecutive days. Control group mice were fed normally and did not receive modeling stimulation.

[0031] During the experiment, all mice had free access to food and water, and the room temperature was controlled at (24±2)℃. A 12-hour light-dark cycle was set, and animal operations were carried out in accordance with ethical guidelines for laboratory animals. After the modeling period, the weight-bearing swimming time, serum lactate, and blood urea nitrogen levels were measured. The model group showed statistically significant differences compared with the control group, confirming the successful establishment of the chronic fatigue stress model. Subsequently, behavioral tests were carried out. After the behavioral tests were completed, the hippocampus tissue of the mice was isolated for subsequent biochemical index and protein expression detection.

[0032] Indicator Testing: After the drug administration and modeling period for each group of mice was completed, various indicator tests were initiated. The Morris water maze training lasted for 5 days, and all behavioral tests were completed within a 24-hour time window after the last drug administration. After all behavioral tests were completed, a weighted swimming test was conducted. The weighted swimming test was the terminal stress test, and no further behavioral tests were conducted after the swimming test to avoid fatigue stress interfering with the learning and memory behavioral results.

[0033] 1. Morris water maze experiment Twenty-four hours after the last dose, the Morris water maze test was conducted, including a hidden platform learning training test and a spatial exploration memory test. The water maze was a circular pool with a diameter of 0.67 m and a height of 0.25 m. The pool was evenly divided into four quadrants. A circular hidden platform with a diameter of 15 cm was placed in the center of the first quadrant, with the water level 1 cm above the platform surface. The water temperature was kept constant at (25±0.5)℃ throughout the test.

[0034] Concealed Platform Test: For 5 consecutive days prior to the formal test, mice underwent adaptive learning training. Each mouse was trained twice daily, with each training session lasting 60 seconds. The mice were placed into the water pool from different entry points in four quadrants. If a mouse failed to find the platform independently within 60 seconds, it was manually guided to the platform to acclimatize. The daily escape latency for each mouse was recorded.

[0035] Spatial exploration test: The day after the concealed platform training ended, the concealed platform in the pool was removed, and a 60-second spatial exploration test was conducted. The time the mice spent in the first quadrant where the original platform was located, as well as the number of times they crossed the original platform location, were recorded to evaluate the mice's spatial learning and memory abilities.

[0036] Novel object recognition test (NOR) After the water maze experiment was completed, a new object recognition experiment was conducted. The experiment was divided into three phases: environmental adaptation, familiarization training, and testing. Throughout the experiment, all test items and the test chamber were wiped and disinfected with 75% ethanol to eliminate odor interference. Judgment criteria: mice were considered to have explored the object with their face facing it and their mouth and nose ≤ 2 cm away from it. Lying down or leaning against the object was not counted as exploration time.

[0037] ① Environmental adaptation period: 24 hours before the experiment, mice were placed in a cube test box with a side length of 50 cm and allowed to explore freely for 5 minutes to adapt to the test environment and eliminate environmental unfamiliarity stress; several days before the experiment, mice were gently stroked to reduce human operation stress.

[0038] ② Familiarization training period (T1): Place two identical objects A1 and A2 at opposite corners of the test box, and let the mouse explore freely in the box for 5 minutes. Record the total exploration time of the mouse for the two identical objects.

[0039] ③ Test period (T2): After training, the test objects were changed. One original object A was kept and another object B was replaced. The old and new objects were placed diagonally and the mice were allowed to explore freely for 5 minutes. The effective exploration time of the mice for the old and new objects was recorded respectively.

[0040] The discrimination index (DI) was used to evaluate the recognition and memory abilities of mice. The calculation formula is as follows: In the formula: T 新 For the exploration time of new objects, T 旧 The exploration time for old objects; DI is a dimensionless index, ranging from -1 to 1. 3. Weighted swimming test After the new object recognition test, a weighted swimming test was conducted. A lead sheet equivalent to 5% of the mouse's body weight was placed on the base of the mouse's tail. The mouse was then placed in a constant-temperature water pool at a depth of 30 cm and a water temperature of (25±0.5)℃. The swimming time from entry to exhaustion was recorded. Exhaustion was defined as the mouse's head remaining submerged for more than 10 seconds without surfacing or exhibiting any active struggling behavior. This duration was the mouse's weighted swimming exhaustion time.

[0041] 4. Serum lactate (LA) and blood urea nitrogen (BUN) measurements Immediately after the weighted swimming test, blood was collected by enucleation. After standing at room temperature for 30 minutes, the blood was centrifuged at 3000 r / min for 10 minutes to obtain the serum supernatant. The levels of LA and BUN in the serum were measured strictly according to the instructions of the corresponding biochemical reagent kits.

[0042] Detection of SOD, GSH-Px activity and MDA content in hippocampal tissue Mice were euthanized after serum sample collection, and hippocampal tissue was rapidly isolated. A 10% (w / v) hippocampal tissue homogenate was prepared using pre-cooled physiological saline, centrifuged at 3000 r / min for 10 min, and the supernatant was collected for analysis. Using a biochemical colorimetric kit, strictly following the kit instructions, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and the content of malondialdehyde (MDA) in the hippocampal tissue were determined.

[0043] Western blot protein expression detection in hippocampus tissue (1) Sample processing and protein quantification After euthanizing mice, hippocampal tissue was rapidly isolated, minced, and added to RIPA total protein lysis buffer containing protease inhibitors and phosphatase inhibitors. The mixture was thoroughly homogenized in an ice bath and allowed to lyse for 15 min. The mixture was then centrifuged at 12000 r / min for 15 min at low temperature, and the supernatant was collected to obtain total protein samples from the hippocampal tissue. The protein concentration of each sample was determined using a BCA protein quantification kit, and the protein loading amount was standardized.

[0044] (2) Protein denaturation The protein sample was mixed with 5× loading buffer at a volume ratio of 4:1, denatured in a boiling water bath for 15 min, cooled and stored at -20℃ for later use.

[0045] (3) SDS-PAGE electrophoresis Prepare the stacking gel and separating gel, and assemble the electrophoresis apparatus. Perform electrophoresis at a constant voltage of 75 V during the stacking gel stage. After the sample enters the separating gel, adjust the voltage to 120 V. Electrophoresis is terminated when the bromophenol blue migrates to approximately 1 cm from the bottom of the gel.

[0046] (4) Transfer membrane Peel off the gel, and stack filter paper, PVDF membrane and gel in sequence to form a "sandwich" structure. Transfer the membrane at 300mA constant current for 70 min under ice bath conditions to ensure full transfer of large protein molecules.

[0047] (5) Immune incubation After transfer, block with 5% skim milk at room temperature for 1-2 hours. After blocking, discard the blocking solution and incubate with primary antibody at 4°C strictly according to the antibody instructions. After washing, add secondary antibody and incubate at room temperature. Detection indicators include p-AMPK, AMPK, PGC-1α, and SIRT1. p-AMPK is normalized using total AMPK as a reference, while PGC-1α and SIRT1 are normalized using β-actin as an internal control.

[0048] (6) Imaging and Gray Scale Analysis The protein band images were scanned and acquired using an ECL chemiluminescence reagent kit after incubation in the dark. The gray values ​​of each target band and internal control band were detected using Alpha grayscale analysis software, and the relative expression levels of each protein were calculated.

[0049] Results and Analysis: (1) Results of the Morris water maze experiment (I) Results of Covert Platform Tests Table 1 reflects the changes in escape latency of mice in each group during the 5-day concealed platform exploration test. With increasing training days, the escape latency of all groups generally decreased; the control group showed the fastest decrease, while the model group showed the slowest. From day 2 to day 5, compared with the control group, mice in the model group took longer to find the concealed platform, suggesting a significant decline in their learning and memory abilities. Compared with the model group, on day 5 of the concealed platform test, the escape latency of the control group was 26.45±1.15s, the model group was 41.90±3.55s, the positive control group was 31.50±3.25s, the low-dose combination group was 34.85±3.10s, and the high-dose combination group was 29.15±2.80s. The data show that compared with the model group, the escape latency of mice in each treatment group was significantly reduced (P<0.05).

[0050] Table 1. Changes in the 5-day escape latency of mice in each group. Note: ##P<0.01, compared with the control group; ×P<0.05, ××P<0.01, compared with the model group.

[0051] (II) Space Exploration Test Results After the hiding platform was removed, the swimming trajectories of the model group mice became disordered, with most swimming along the pool wall and randomly distributed in the four quadrants. This indicates that the model group mice did not accurately remember the original platform location, and their spatial location memory ability was significantly reduced. The data results are shown in Table 2. The time spent in the original platform quadrant by the control group mice was 25.80±1.75s, which was significantly higher than that of the model group (20.35±4.20s). After the intervention of the composition, the time spent in the original platform quadrant by the positive control group, the low-dose group, and the high-dose group were 25.20±2.45s, 23.85±3.15s, and 28.50±3.20s, respectively, all of which were significantly higher than that of the model group (P<0.05).

[0052] Furthermore, the number of times the model group crossed the original platform was 1.65±0.68, significantly lower than the 2.75±0.25 times in the control group. The positive control group, the low-dose group, and the high-dose group of the composition increased the number of times the mice crossed the original platform, to 2.15±0.72, 2.05±0.70, and 2.65±0.58 times, respectively, significantly higher than the model group (P<0.05). This suggests that the composition intervention can improve the spatial learning and memory ability of fatigue stress model mice.

[0053] Table 2 Results of spatial exploration tests in each group of mice Note: ##P<0.01, compared with the control group; ×P<0.05, ××P<0.01, compared with the model group.

[0054] (2) Results of the new object recognition experiment Novel object recognition memory can be used to evaluate hippocampal-dependent non-spatial learning and memory abilities in mice. Total exploration time reflects the mice's autonomous exploration behavior, and the relative recognition index (DI) is used to evaluate memory level. The total exploration time of the control group mice was 65.20±4.35s, which was significantly higher than that of the model group (49.80±3.95s) (P<0.05). After intervention with the combination or rhodioloside, the total exploration time of mice in all groups for both new and old objects was increased; the low-dose combination group was 58.40±3.50s, the high-dose combination group was 66.85±2.95s, and the positive control group was 63.50±3.10s.

[0055] The relative recognition index (DI) results for each group are shown in Table 3. The DI of the model group was 0.62±0.025, significantly lower than that of the control group (0.85±0.028) (P<0.05). The DI of the low-dose group, high-dose group, and positive control group increased to 0.75±0.024, 0.84±0.027, and 0.80±0.026, respectively, showing significant differences compared with the model group (P<0.05). This suggests that the composition can improve the novel object recognition and memory ability of fatigue-stressed mice.

[0056] Table 3 Results of novel object recognition test for each group of mice Note: ##P<0.01, compared with the control group; ×P<0.05, ××P<0.01, compared with the model group.

[0057] (3) Results of the weighted swimming test Table 4 reflects the changes in weight-bearing swimming time in each group of mice. Compared with the control group, the weight-bearing swimming time of the model group mice was significantly shortened (P < 0.01), indicating that the chronic fatigue stress model was successfully established. Compared with the model group, the weight-bearing swimming time of mice in the positive control group and each group treated with the composition was significantly prolonged (P < 0.05); among them, the swimming time of the high-dose group of the composition was close to that of the control group, suggesting that this composition can improve the exercise endurance of fatigue-stressed mice and exert an anti-fatigue effect.

[0058] Table 4. Swimming time under load for mice in each group Note: ##P<0.01, compared with the control group; ×P<0.05, ××P<0.01, compared with the model group.

[0059] (4) Serum lactate (LA) and blood urea nitrogen (BUN) levels As shown in Table 5, compared with the control group, the serum LA and BUN levels in the model group mice were significantly increased (P < 0.01), indicating the accumulation of metabolites in the body under fatigue. After intervention, the serum LA and BUN levels in the positive control group and each group treated with the composition were significantly decreased (P < 0.05), with the high-dose group of the composition showing a more significant decrease, suggesting that the composition helps promote the clearance of metabolites in the body under fatigue.

[0060] Table 5 Serum LA and BUN levels in mice of each group Note: ##P<0.01, compared with the control group; ×P<0.05, ××P<0.01, compared with the model group.

[0061] (5) Detection of SOD, GSH-Px activity and MDA content in mouse hippocampus tissue The antioxidant effects of the composition on the hippocampus were evaluated by measuring the activities of SOD and GSH-Px and the content of MDA in the hippocampus of mice. As shown in Table 6, compared with the control group, the activities of SOD and GSH-Px in the hippocampus of the model group mice were significantly decreased (P < 0.05), while the MDA level was significantly increased (P < 0.05). After intervention, the activities of SOD and GSH-Px in the hippocampus of the positive control group and each group treated with the composition were significantly restored; in addition, compared with the model group, the MDA content after intervention with the composition and rhodioloside was significantly decreased (P < 0.05), suggesting that the composition can improve the oxidative stress state of the hippocampus in fatigue model mice.

[0062] Table 6. SOD, GSH-Px activities and MDA content in hippocampal tissue of mice in each group Note: ##P<0.01, compared with the control group; ×P<0.05, ××P<0.01, compared with the model group.

[0063] (6) Detection of AMPK / PGC-1α pathway-related protein expression in mouse hippocampus This experiment analyzed the effects of a composition on the AMPK / PGC-1α signaling pathway by detecting the expression levels of p-AMPK, AMPK, PGC-1α, and SIRT1 in the hippocampus of mice. Western blot analysis showed that chronic fatigue significantly inhibited AMPK phosphorylation and PGC-1α protein expression, and the inhibitory effect of fatigue on the AMPK / PGC-1α signaling pathway in the hippocampus was reversed after intervention. The p-AMPK / AMPK ratio, PGC-1α, and SIRT1 protein expression levels in the high-dose group were significantly higher than those in the model group (P < 0.05), and comparable to the positive control group. This suggests that the composition can upregulate the AMPK / PGC-1α signaling pathway, promote mitochondrial biosynthesis and energy metabolism, and improve fatigue-related damage induced by energy depletion.

[0064] In summary, the composition can upregulate the AMPK / PGC-1α signaling pathway, promote mitochondrial biosynthesis and energy metabolism, reduce oxidative stress in hippocampal tissue, prolong weight-bearing swimming time, reduce blood lactate and blood urea nitrogen levels, and improve the learning and memory abilities of mice under fatigue.

[0065] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, not all embodiments. Those skilled in the art can obtain other embodiments based on the present invention without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A composition having the functions of relieving fatigue and improving hypoxia tolerance, characterized in that, The ingredients include the following components in parts by weight: 6 parts Gastrodia elata, 9 parts Rhodiola rosea, and 3 parts Phyllanthus emblica.

2. The composition according to claim 1, characterized in that, The method for preparing the composition includes the following steps: 1) Gastrodia elata, Rhodiola rosea, and Phyllanthus emblica were pulverized and passed through a 60-mesh sieve to obtain powders of each component; 2) Mix the powders of each component obtained in step 1) according to the weight proportions, add 8 times the total mass of pure water, soak for 2 hours, decoct twice, 30 minutes each time, and combine the decoctions; 3) Filter the combined decoction obtained in step 2), concentrate the filtrate to a clear extract with a relative density of 1.15 (60°C), and spray dry to obtain the composition.

3. The composition according to claim 2, characterized in that, Step 1) The pulverized powder is passed through a 60-mesh sieve; Step 2) The soaking time is 2 hours, and the decoction is performed twice, 30 minutes each time; Step 3) The inlet air temperature of the spray dryer is 180℃, and the outlet air temperature is 80℃.

4. The composition according to claim 1, characterized in that, The preparation method of Gastrodia elata includes the following steps: The raw material of Gastrodia elata is washed, sliced, dried at 60℃ until the moisture content is less than 5%, and then pulverized through a 60-mesh sieve to obtain Gastrodia elata powder.

5. The composition according to claim 1, characterized in that, The preparation method of Rhodiola rosea includes the following steps: Wash and slice the rhizome of Rhodiola rosea, dry it at 55°C until the moisture content is less than 5%, and then pulverize it through a 60-mesh sieve to obtain Rhodiola rosea powder.

6. The composition according to claim 1, characterized in that, The preparation method of the amla fruit includes the following steps: Wash the fresh amla fruit, dry it at 50℃ until the moisture content is less than 8%, and then pulverize it through a 60-mesh sieve to obtain amla powder.

7. A method for preparing the composition according to any one of claims 1 to 6, characterized in that, Includes the following steps: The ingredients of Gastrodia elata, Rhodiola rosea and Phyllanthus emblica are mixed in proportions by weight, decocted with water, concentrated and dried to obtain the composition.

8. The use of the composition of claim 1 in the preparation of fatigue-relieving products.

9. The use of the composition of claim 1 in the preparation of hypoxia-tolerant products.

10. The application according to claim 8 or 9, characterized in that, The product in question is food, medicine, or health supplement.

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