Application of astragaloside compound-containing composition in preparation of plateau fatigue-resistant medicine

Abstract

The invention discloses an application of a compound composition containing astragaloside IV in preparation of a medicine for resisting plateau fatigue, and the compound composition containing astragaloside IV is prepared from the following raw materials in parts by weight: 500 to 2000 parts of astragaloside IV, 5000 to 20000 parts of ginsenoside Rg1, 2 to 5 parts of vitamin D3 and 30000 to 150000 parts of puerarin. Animal experiments prove that the compound composition can relieve plateau exercise fatigue, improve the hypoxia tolerance, exercise fatigue resistance and oxidative stress resistance of a body and improve the comprehensive exercise performance of the body. According to the invention, the defect of single-component efficacy is overcome, a compound preparation is designed in a targeted manner, an innovative and feasible compound combination is provided, and a new scheme is provided for promoting athletic performance under plateau hypoxia.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of functional compound compositions, and particularly relates to application of a composition containing a compound of astragaloside in preparation of anti-high-altitude fatigue medicine. BACKGROUND

[0002] The plateau environment is a special natural environment, and its main features are low pressure, low oxygen, high cold, strong ultraviolet radiation and dryness. With the increase of altitude, the atmospheric pressure and oxygen partial pressure decrease, and this low oxygen environment has a profound impact on human physiological functions, especially the interference with the energy metabolism system is particularly significant.

[0003] Under the plateau hypoxic environment, the body's energy metabolism process changes significantly. Under normal oxygen partial pressure, cells mainly produce ATP through aerobic oxidation, while under hypoxic environment, they have to rely more on anaerobic glycolysis to provide energy. This metabolic conversion leads to a significant reduction in energy production efficiency, and an increase in lactic acid accumulation. Studies have shown that under the plateau hypoxic environment, protein, carbohydrate and lipid metabolism are negatively affected, leading to fatigue, hyperlipidemia and hindered body repair during high-altitude work. The plateau hypoxic environment also causes a series of physiological stress responses, including increased heart rate, elevated blood pressure, rapid breathing, and compensatory increase in red blood cells. Long-term exposure to this environment may cause serious conditions such as altitude sickness, high-altitude pulmonary edema, high-altitude cerebral edema, and even irreversible damage to multiple organ systems. In recent years, with the increase of activities such as high-altitude tourism, scientific exploration, military operations and sports training, the prevention and treatment of high-altitude exercise fatigue has become a research hotspot in the fields of sports science, high-altitude medicine and military medicine.

[0004] Although existing research has found that many components have the potential to resist high-altitude exercise fatigue, there are still the following main problems: (1) Single component limitation. Current research focuses on the anti-high-altitude fatigue effects of single components, and although these components have some effect to some extent, they are difficult to comprehensively address the multi-factor, multi-mechanism complex pathophysiological process of high-altitude exercise fatigue.

[0005] (2) Lack of targeted compound preparation design. At present, there are relatively few compound preparations in the research of anti-high-altitude exercise fatigue drugs, but they have shown good application prospects. The advantage of compound preparations is that they can simultaneously target multiple aspects of high-altitude exercise fatigue through the synergistic effect of multiple components, thereby improving the overall effect. SUMMARY

[0006] In view of the above problems existing in the prior art, the application provides application of a compound composition containing astragaloside in preparation of anti-high altitude fatigue medicine.

[0007] The technical scheme of the application is as follows: The first object of the application is to provide application of a compound composition containing astragaloside in preparation of anti-high altitude fatigue medicine, wherein the compound composition containing astragaloside is composed of 500-2000 parts of astragaloside IV, 5000-20000 parts of ginsenoside Rg1, and 2-5 parts of vitamin D3.

[0008] In an embodiment of the application, the purity of the astragaloside IV is greater than or equal to 98%.

[0009] In an embodiment of the application, the purity of the ginsenoside Rg1 is greater than or equal to 50%.

[0010] In an embodiment of the application, the purity of the ginsenoside Rg1 is greater than or equal to 50%.

[0011] In an embodiment of the application, the compound composition containing astragaloside is composed of 2000 parts of astragaloside IV, 20000 parts of ginsenoside Rg1, 100000 parts of puerarin, and 5 parts of vitamin D3.

[0012] In an embodiment of the application, the medicine further comprises a drug carrier and / or a pharmaceutical excipient.

[0013] In an embodiment of the application, the drug carrier comprises microcapsules, microspheres, nanoparticles and / or liposomes.

[0014] In an embodiment of the application, the pharmaceutical excipient comprises excipients and / or additional agents.

[0015] In an embodiment of the application, the excipients comprise anti-adhesion agents, penetration promoters, buffering agents, plasticizers, surfactants, antifoaming agents, thickening agents, inclusion agents, absorbents, humectants, solvents, propellants, solubilizers, cosolvents, emulsifiers, coloring agents, pH regulators, adhesives, disintegrants, fillers, lubricants, wetting agents, integrating agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, foaming agents, suspending agents, coating materials, fragrances, diluents, flocculants and deflocculants, filtration aids and / or release retardants; and the additional agents comprise microcrystalline cellulose, hydroxypropyl methylcellulose and / or refined lecithin.

[0016] In an embodiment of the present application, the anti-high altitude fatigue medicine is an oral medicine preparation, and the dosage form is one of a tablet, a granule, a capsule, a mixture, and an oral liquid.

[0017] In an embodiment of the present application, the anti-high altitude fatigue is to improve the exercise performance of a hypoxic animal.

[0018] In an embodiment of the present application, the effect of improving the exercise performance in a hypoxic environment includes relieving adverse reactions in a high altitude, improving the body's anti-exercise fatigue ability and anti-oxidative stress ability, and enhancing the overall exercise performance of the body.

[0019] In an embodiment of the present application, the effect of the anti-high altitude fatigue includes at least one of (a)-(g): (a) reducing muscle damage of a high altitude individual: reducing blood creatinine content; (b) relieving exercise fatigue of a high altitude individual: reducing lactic acid accumulation or improving lactic acid clearance rate; (c) relieving exercise fatigue of a high altitude individual: improving muscle glycogen reserves or accelerating muscle glycogen recovery; (d) improving the anti-oxidative stress ability of a high altitude individual: reducing MDA level; (e) improving the anti-oxidative stress ability of a high altitude individual: increasing SOD level; (f) improving the exercise performance of a high altitude individual: increasing exhaustive swimming time; (g) improving the exercise performance of a high altitude individual: increasing grip strength.

[0020] The present application has the following beneficial technical effects: The present application verifies through animal experiments that the compound combination of Astragaloside IV, Ginsenoside Rg1, Puerarin, and Vitamin D3 has the effect of anti-high altitude exercise fatigue, which is embodied in the following aspects: (1) The compound combination significantly reduces the blood creatinine content of a hypoxic mouse; (2) The compound combination significantly reduces the lactic acid accumulation in the blood of a hypoxic mouse; (3) The compound combination significantly improves the muscle glycogen reserves of a hypoxic mouse; (4) The compound combination significantly reduces the MDA level in the serum of a hypoxic mouse; (5) The compound combination significantly improves the SOD level in the serum of a hypoxic mouse; (6) The compound combination significantly increases the exhaustive swimming time of a hypoxic mouse; (7) The compound combination significantly improves the grip strength of a hypoxic mouse.

[0021] Therefore, the compound combination has great application prospects in relieving high altitude exercise fatigue, improving the body's anti-exercise fatigue ability and anti-oxidative stress ability, and improving the overall exercise performance of the body. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 Effect of compound combination on blood creatinine content of hypoxic mice after exercise; Figure 2 Effect of compound combination on serum lactic acid content of hypoxic mice after exercise; Figure 3 Effect of compound combination on muscle glycogen level of hypoxic mice; Figure 4 Effect of compound combination on MDA level in serum of hypoxic mice; Figure 5 Effect of compound combination on SOD level in serum of hypoxic mice; Figure 6 Effect of compound combination on exhaustive swimming time of hypoxic mice; Figure 7 Effect of compound combination on grip strength of hypoxic mice. DETAILED DESCRIPTION

[0023] The present application will be described in detail below in combination with the drawings and examples.

[0024] The acetazolamide involved in the following examples is purchased from Suzhou Greeter Pharmaceutical Technology Co., Ltd.

[0025] The compound components involved in the following examples are: Astragalus membranaceus glycoside, ginsenoside Rg1, puerarin, all purchased from Bofei Meike Company, and vitamin D3 is purchased from China National Pharmaceutical Group Star Shark Pharmaceutical Co., Ltd.

[0026] The configuration method of the compound combination involved in the following examples is as follows: Acetazolamide solution: a certain mass of acetazolamide powder is weighed and dissolved in sterile normal saline to prepare a 3 mg / mL acetazolamide solution.

[0027] Compound combination solution: 0.9 mg of Astragalus membranaceus glycoside IV, 9 mg of ginsenoside Rg1, 45 mg of puerarin, and 2.25 μg of vitamin D3 are dissolved in 3 ml of sterile normal saline to prepare a compound combination solution.

[0028] Example 1: Simulation of hypoxic modeling results of high altitude The specific steps are as follows: 24 SPF level male ICR mice (6 weeks old, 20 g ~ 25 g) were randomly divided into 4 groups, 6 in each group, respectively blank group, model group, Yangshen group (acetazolamide), and compound group (compound combination). The mice were raised in the experimental animal center of Jiangnan University, with constant temperature 21 26℃, humidity 40 70%, noise level less than or equal to 60 dB, animal illumination 15 20LX (All animal experimental procedures were reviewed and approved by the Animal Welfare and Ethics Management Committee of Jiangnan University).

[0029] The experiment lasted for 5 weeks: animal experimental grouping information is shown in Table 1. Weeks 0-1 were the mouse acclimatization period; weeks 1-5 were the mouse intervention period. During weeks 1-5, mice were administered 200 μL of sterile saline via gavage daily. Mice in the blank control group and model group were administered 200 μL of 3 mg / mL acetazolamide solution via gavage daily, while mice in the compound group were administered 200 μL of the compound combination solution via gavage daily. Weeks 3-5 were the mouse hypoxia exposure period, during which mice underwent simulated high-altitude hypoxia modeling combined with exercise modeling.

[0030] Table 1

[0031] Modeling Procedure: Hypoxia Modeling: Mice in the model group, Yangshen group, and chemical combination group were placed in a hypobaric chamber for hypoxia exposure for 3-5 weeks, simulating an altitude of 5000 m for 2 weeks. Specifically, the altitude was gradually increased to 5000 m at a rate of 500 m / min and maintained for 2 weeks. During intervention, the altitude was reduced to normal at a rate of 1000 m / min. Mice were then removed for gavage intervention, supplemental feeding, and water bottle replacement. After the intervention, the altitude was increased to 5000 m again at a rate of 500 m / min. Exercise Modeling: Mice were trained to swim for 20 minutes daily for 3-5 weeks.

[0032] After the experiment, the mice were subjected to exhaustive swimming. Immediately after exhaustion, the mice were euthanized, and blood was collected from their orbital fossa. Two to three drops of whole blood were collected in EDTA-K2 anticoagulant tubes, and the whole blood was analyzed using an automated blood cell analyzer to determine the levels of erythrocytes, hemoglobin, hematocrit, and mean corpuscular volume. The results of the whole blood analysis of hypoxic mice are shown in Table 2.

[0033] Table 2

[0034] Note: Compared with the blank group, This indicates that P < 0.05. This indicates that P < 0.01. This indicates that P < 0.001. This indicates that P < 0.0001. The results show: Compared with the control group, the model group showed significantly increased levels of erythrocytes, hemoglobin, and hematocrit, indicating the successful establishment of the high-altitude hypoxia mouse model. Furthermore, under hypobaric and hypoxic conditions, the use of acetazolamide as a stimulant did not reduce the compensatory effects of the hematopoietic system (the compensatory effects of hypoxia promoting increased erythrocyte production and reduced oxygen consumption to reduce damage caused by insufficient oxygen) in the compound group. In contrast, the levels of erythrocytes, hemoglobin, and hematocrit in the compound group were close to those in the control group, suggesting that the compound group could reduce the compensatory effects caused by hypoxia, and the compound combination may have effectively inhibited excessive erythrocyte proliferation.

[0035] Example 2: Effect of compound combination on serum creatinine levels in hypoxic mice after exercise The specific method is as follows: After the experiment, mice were subjected to exhaustive swimming. Immediately after exhaustion, the mice were euthanized, and blood was collected from their orbital fossa. The blood was placed in 1.5 mL enzyme-free EP tubes and allowed to stand for at least 2 hours. Serum was obtained by centrifugation at 4°C and 3000 r / min for 15 min. The serum was carefully aspirated into enzyme-free PCR tubes, aliquoted, and stored at -80°C. Serum creatinine levels were measured using an automated biochemical analyzer. The results are shown below. Figure 1 .

[0036] The results show: The serum creatinine levels in the control group mice were 5.164±0.5049 μmol / L, in the model group mice were 5.600±0.7550 μmol / L, in the Yangshen group mice were 2.700±0.3000 μmol / L, and in the combined group mice were 2.300±0.9165 μmol / L. Compared with the control group, the serum creatinine level in the model group increased by 8.44%, the serum creatinine level in the Yangshen group decreased by 51.79%, and the serum creatinine level in the combined group decreased by 58.93%. Changes in serum creatinine levels are used to assess exercise intensity and muscle metabolic load, reflecting transient changes in renal function: serum creatinine levels increase after exhaustive exercise. Serum creatinine levels primarily reflect the body's muscle metabolic load. The greater the exercise intensity, the greater the muscle load, and the more rapidly serum creatinine levels will rise. Compared with the model group, both the Yangshen group and the chemical combination group showed significant decreases, indicating that the muscle load had not reached its peak, and the chemical combination group showed the best effect.

[0037] Example 3: Effect of compound combination on serum lactate levels in hypoxic mice after exercise The specific method is as follows: The specific implementation method is the same as in Example 2. The lactate level in the serum of hypoxic mice was detected according to the instructions of the lactate biochemistry kit (Shanghai ELISA kit). The results are as follows: Figure 2 As shown.

[0038] The results show: The blood lactate levels in the control group mice were 4.691±0.9644 μmol / mL, in the model group mice were 6.753±0.6663 μmol / mL, in the *Gynostemma pentaphyllum* group mice were 5.527±0.7816 μmol / mL, and in the compound group mice were 3.778±0.8853 μmol / mL. Compared with the control group, the blood lactate content in the model group was significantly increased, increasing by 43.96%, indicating significant lactate accumulation. Compared with the model group, the blood lactate content in the *Gynostemma pentaphyllum* group decreased by 18.15%, and the blood lactate content in the compound group decreased by 44.05%, indicating a significant effect of the compound group in clearing lactate accumulation. These experimental results indicate that exercise under hypoxic conditions exacerbates lactate accumulation, leading to more severe exercise fatigue. The compound combination can significantly clear lactate accumulation and alleviate exercise fatigue, with a better effect than acetazolamide.

[0039] Example 4: Effects of compound combinations on muscle glycogen levels in hypoxic mice The specific method is as follows: The specific implementation method is the same as in Example 1. After the mice were sacrificed, the gastrocnemius muscle of the hind leg was removed and placed in liquid nitrogen, stored at -80°C. The glycogen level in the gastrocnemius muscle of hypoxic mice was detected according to the instructions of the glycogenochemistry kit (Shanghai ELISA kit). The results are as follows. Figure 3 As shown.

[0040] The results show: The muscle glycogen levels in the control group were 0.7566±0.2107 mg / g, in the model group were 0.5165±0.1436 mg / g, in the Yangshen group were 0.6777±0.09109 mg / g, and in the chemical combination group were 0.9303±0.08386 mg / g. Compared with the control group, the muscle glycogen content in the model group decreased by 31.73%, indicating that exercise in hypoxic mice accelerates the consumption of muscle glycogen. Compared with the model group, the muscle glycogen content in the Yangshen group increased by 31.21%, and the muscle glycogen content in the chemical combination group increased by 80.12%. The chemical combination group significantly increased muscle glycogen reserves and reduced muscle glycogen loss. The above experimental results indicate that exhaustive swimming is a prolonged endurance exercise. After exercise, the muscle glycogen level of mice will decrease significantly. Muscles are the main energy-consuming organs during exercise, and muscle glycogen is the most direct and rapid energy source for muscle contraction (especially in high-intensity exercise). The combination of compounds can significantly restore muscle glycogen content, and the effect is better than that of acetazolamide.

[0041] Example 5: Effect of compound combinations on serum MDA levels in hypoxic mice The specific method is as follows: The specific implementation method is the same as in Example 2. The malondialdehyde (MDA) level in mouse serum was detected according to the instructions of the serum malondialdehyde (MDA) level ELISA kit. The results are as follows. Figure 4 As shown.

[0042] The results show: The serum MDA levels in the control group were 3.089±0.3635 nmol / mL, in the model group were 3.956±0.5619 nmol / mL, in the *Gynostemma pentaphyllum* group were 3.523±0.4425 nmol / mL, and in the compound group were 3.319±0.5586 nmol / mL. Compared with the control group, the MDA level in the model group was significantly increased by 28.07%, indicating that exercise in high-altitude hypoxic mice significantly increased the level of oxidative stress, which may have caused a certain degree of oxidative damage. Compared with the model group, the MDA level in the *Gynostemma pentaphyllum* group decreased by 10.95%, and the MDA level in the compound group decreased by 16.10%. These experimental results indicate that high-altitude hypoxic exercise aggravates the oxidative stress state of the body, causing oxidative stress damage. The compound combination can reduce the body's MDA level and alleviate oxidative stress damage, and the anti-oxidative stress effect of the compound combination is better than that of acetazolamide.

[0043] Example 6: Effect of compound combinations on serum SOD levels in hypoxic mice The specific method is as follows: The specific implementation method is the same as in Example 2. The superoxide dismutase (SOD) level in mouse serum was detected according to the instructions of the serum superoxide dismutase (SOD) level ELISA kit. The results are as follows: Figure 5 As shown.

[0044] The results show: The serum SOD level in the control group was 21.55±5.852 U / mL, the serum SOD level in the model group was 8.125±2.863 U / mL, the serum SOD level in the *Gynostemma pentaphyllum* group was 20.76±8.854 U / mL, and the serum SOD level in the chemical combination group was 32.36±2.513 U / mL. Compared with the control group, the serum SOD level in the model group was significantly reduced, decreasing by 62.29%, indicating that exercise under high-altitude hypoxia caused severe oxidative stress in mice, resulting in reduced SOD levels and significantly decreased antioxidant capacity. Compared with the model group, the serum SOD level in the *Gynostemma pentaphyllum* group increased by 155.51%, and the SOD level in the chemical combination group increased significantly by 298.3%, indicating that the chemical combination group can maximize the increase of SOD level in the body and enhance the body's antioxidant capacity. The above experimental results show that the combination of compounds can significantly increase the body's SOD level and improve the body's antioxidant stress capacity, with the best effect, which is superior to acetazolamide.

[0045] Example 7: Effect of compound combinations on exhaustive swimming time in hypoxic mice The specific method is as follows: During the animal experiments, the model group, the *Gynostemma pentaphyllum* group, and the compound group underwent exercise training for 3-5 weeks to establish the mouse model. Mice were trained to swim for 20 minutes daily. On the day before the end of the experiment, after the last gavage and 30 minutes of rest, the time to exhaustion during weight-bearing swimming was measured. A 5% weight was applied to the base of the mouse's tail, and the temperature was controlled at 24-26℃. Exhaustion was defined as the mouse's hind limb movement becoming sluggish, the swimming range decreasing, and the head sinking to the surface within 3 seconds. Results are shown in […]. Figure 6 .

[0046] The results show: The average exhaustive swimming time for mice in the control group was 25.61 min, the average exhaustive swimming time for mice in the model group was 18.14 min, the average exhaustive swimming time for mice in the *Gynostemma pentaphyllum* group was 21.95 min, and the average exhaustive swimming time for mice in the compound group was 26.83 min. Compared with the control group, the exhaustive swimming time in the model group decreased by 29.17%, indicating that the high-altitude hypoxic environment greatly limits individual athletic performance. Compared with the model group, the exhaustive swimming time in the *Gynostemma pentaphyllum* group increased by 21.00%, and the exhaustive swimming time in the compound group increased by 47.91%. The exhaustive swimming time of mice in the compound group was significantly prolonged, and the effect was better than that in the *Gynostemma pentaphyllum* group. The above experimental results show that the compound combination can better and significantly improve the exhaustive swimming time of mice in high-altitude hypoxic environments and enhance the athletic performance of hypoxic mice.

[0047] Example 8: Effect of compound combinations on grip strength in hypoxic mice The specific method is as follows: During the animal experiments, on the last day of the experiment, the gripping force of the mice's limbs was tested using a gripping force meter (Jinan Yiyan Technology Co., Ltd.). The mice's limbs were placed horizontally on the gripping net, their tails were grasped, and the mice were slowly pulled horizontally. This operation was repeated 5 times, and the maximum value was recorded. The results are shown in […]. Figure 7 .

[0048] The results show: The average grip strength of mice in the control group was 243.6 gf, the average grip strength of mice in the model group was 215.0 gf, the average grip strength of mice in the *Gynostemma pentaphyllum* group was 241.8 gf, and the average grip strength of mice in the compound group was 247.3 gf. Compared with the control group, the grip strength of mice in the model group decreased by 11.74%, indicating that the high-altitude hypoxic environment limited the limb strength of individuals. Compared with the model group, the grip strength of the *Gynostemma pentaphyllum* group increased by 12.47%, and the grip strength of the compound group increased by 15.02%. The grip strength of mice in the compound group was significantly improved, and the effect was better than that of the *Gynostemma pentaphyllum* group. The above experimental results show that the compound combination can significantly improve the grip strength of mice and alleviate the fatigue state of mice in high-altitude hypoxic environments, and the effect is better than that of acetazolamide.

[0049] The embodiments provided above are not intended to limit the scope of the invention, nor are the described steps intended to limit the order of execution. Any obvious modifications made to the invention by those skilled in the art based on existing common knowledge also fall within the scope of protection defined by the claims.

Claims

1. The application of a composition containing astragaloside A in the preparation of drugs for combating altitude fatigue, characterized in that, The composition of the astragaloside IV compound, by weight, is as follows: astragaloside IV 500-2000 parts, ginsenoside Rg 15000-20000 parts, vitamin D3 2-5 parts, and puerarin 30000-150000 parts.

2. The application according to claim 1, characterized in that, The purity of astragaloside IV is ≥98%.

3. The application according to claim 1, characterized in that, The purity of ginsenoside Rg1 is ≥50%.

4. The application according to claim 1, characterized in that, Puerarin purity ≥ 98%.

5. The application according to claim 1, characterized in that, The composition of the astragaloside IV compound, by weight, is as follows: 2000 parts of astragaloside IV, 20000 parts of ginsenoside Rg1, 100000 parts of puerarin, and 5 parts of vitamin D3.

6. The application according to claim 1, characterized in that, Pharmaceuticals also include drug carriers and / or pharmaceutical excipients.

7. The application according to claim 6, characterized in that, Drug carriers include microcapsules, microspheres, nanoparticles, and / or liposomes.

8. The application according to claim 6, characterized in that, Pharmaceutical excipients include excipients and / or additives.

9. The application according to claim 8, characterized in that, The excipients include anti-adhesives, penetration enhancers, buffers, plasticizers, surfactants, defoamers, thickeners, encapsulating agents, absorbents, humectants, solvents, propellants, solubilizers, co-solvents, emulsifiers, colorants, pH adjusters, binders, disintegrants, fillers, lubricants, wetting agents, binding agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, foaming agents, suspending agents, coating materials, fragrances, diluents, flocculants and anti-flocculation agents, filter aids and / or release inhibitors; the additives include microcrystalline cellulose, hydroxypropyl methylcellulose and / or refined lecithin.

10. The application according to claim 1, characterized in that, Anti-altitude fatigue is used to improve the athletic performance of hypoxic animals.

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