Application of perilla frutescens extract in preparation of sports fatigue resisting product

Abstract

The invention belongs to the technical field of biology, and particularly relates to application of a perilla frutescens extract in preparation of an anti-fatigue product. The perillene volatile oil is obtained by adopting a steam distillation method, and the perillene accounts for more than 10% of the total volatile oil. Further verification shows that the extracted perillene type volatile oil is safe and non-toxic, can improve exercise tolerance, improve muscle state and body physiological indexes, and has a good anti-fatigue effect. The perillene volatile oil can be used for developing anti-fatigue drugs, health care products or food products, and has a good application prospect.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and particularly relates to the application of perilla extract in the preparation of anti-motion fatigue products. Background Art

[0002] Fatigue is a state of tiredness and exhaustion caused by the decline of human body functions and mental state, and it is a special physiological state reaction of the body (Yu Yihao. Research on the anti-fatigue effect and mechanism of sea cucumber peptides [D]. Jiangnan University, 2021.). With the increasingly fierce competition in today's society, in order to adapt to the current pace of life, people's work and life pressures are constantly increasing, and the incidence of fatigue symptoms is also increasing day by day (Gong Yun, Chen Chen. Analysis of the research progress of domestic exercise-induced fatigue based on knowledge graph [J]. Journal of Northwest Adult Education College, 2022, (01): 67-68.). Under the influence of the fast pace of today's society, problems such as lack of exercise, irregular life, habitual staying up late, and high work pressure are becoming more prominent, making the incidence of fatigue increase year by year and has become one of the major health problems of the human body. Research shows that fatigue is a common feature of many physical and neurological diseases, and about 20% of the global population is suffering from fatigue (Abd-Elfattah, H.M., Abdelazeim, F.H. and Elshennawy, S., Physical and cognitive consequences of fatigue: A review. J Adv Res, 2015. 6(3): 351-8).

[0003] ROGER M. ENOKA classifies fatigue into two types. One is the limitation of physical motor function, manifested as peripheral physical fatigue (PF), and the other is central fatigue (CF) with limited cognitive function (Enoka, R.M. and Duchateau, J., Translating Fatigue to Human Performance. Med Sci Sports Exerc, 2016. 48(11):2228-2238.). Physical fatigue usually results from changes in the muscle action potential transmission mechanism caused by excessive exercise, neuromuscular junction transmission disorders, and decreased muscle fiber contractility, thus hindering normal physiological activities (Tornero-Aguilera, J.F., Jimenez-Morcillo, J., Rubio-Zarapuz, A., et al., Central and Peripheral Fatigue in Physical Exercise Explained: A Narrative Review. Int J Environ Res Public Health, 2022. 19(7)). Central fatigue is the weakening of the ability of the central nervous system to receive, send, and transmit nerve impulses under physiological and pathological conditions, manifested as persistent fatigue, cognitive dysfunction, and decreased thinking activity (Chaudhuri, A. and Behan, P.O., Fatigue in neurological disorders. Lancet, 2004. 363(9413):978-88). Fatigue belongs to sub-healthy diseases and affects multiple organs of the body. Long-term fatigue will cause a series of physiological and psychological changes in the body, accompanied by mental tension and irritability. In severe cases, it will lead to neurasthenia, palpitations, shortness of breath, and decreased immunity, and even induce serious diseases (Han Jianan, Liu Zhuorui, Zeng Peiyong, et al. Anti-fatigue effect and mechanism of Wujiashengmaiyin on mice [J]. Journal of Jilin University (Medicine Edition), 2024, 50(3):689-696.).

[0004] The mechanisms underlying physical fatigue mainly involve energy depletion, metabolite accumulation, and oxidative stress (Coqueiro, A.Y., Rogero, M.M. and Tirapegui, J., Glutamine as an Anti-Fatigue Amino Acid in Sports Nutrition. Nutrients, 2019. 11(4).). Research has shown that insufficient energy supply caused by muscle movement is mainly related to glycogen consumption, leading to the accumulation of blood lactate and serum urea nitrogen, disrupting the homeostasis of the internal environment, and thus triggering physical fatigue (Mohamed, M.E., Younis, N.S., El-Beltagi, H.S., et al., The Synergistic Hepatoprotective Activity of Rosemary Essential Oil and Curcumin: The Role of the MEK / ERK Pathway. Molecules, 2022. 27(24).). In addition, reactive oxygen species (ROS) generated after exercise can damage cell membranes, leading to oxidative stress, which in turn affects various tissues such as skeletal muscle (Chung, Y.H., Park, T.K., Yim, S.H., et al., Polysaccharide-Rich Extract of Phragmites rhizome Attenuates Water Immersion Stress and Forced Swimming Fatigue in Rodent Animal Model. J Med Food, 2019. 22(4):355-364.). Excessive accumulation of ROS can promote the occurrence of inflammatory processes, thus triggering a series of pathophysiological processes. In addition, long-term excessive exercise may lead to dysregulation of the homeostasis of the central nervous system, making it difficult to effectively regulate the activities of motor neurons. Eventually, it leads to a decline in muscle motor function. Exercise-induced central fatigue is mainly caused by long-term engagement in repetitive, monotonous, and high-intensity physical work or training. Long-term excessive exercise can also lead to dysregulation of the homeostasis within the central nervous system, unable to control motor neurons, resulting in a decline in muscle motor function, which is common among athletes; central fatigue is not only caused by excessive exercise but also related to long-term mental stress and sleep deprivation, leading to changes in the body's cognitive function and emotions. Severe cases may suffer from central nervous system diseases such as depression and anxiety (Ridouh, I. and Hackshaw, K.V., Essential Oils and Neuropathic Pain. Plants (Basel), 2022. 11(14).).Factors leading to central fatigue include work intensity and duration, mental load, work methods and environment, as well as an individual's physical health status. When external stimuli are received by the body's sensory organs, relevant information is transmitted through nerves into the central nervous system of the brain for comprehensive processing, and then the corresponding effector organs are stimulated through efferent nerves. However, if these stimuli repeatedly occur inappropriately, it will cause problems in the operation of the central nervous system of the brain, overload the psychological regulation function, and thus induce central fatigue.

[0005] Currently, the methods of drug intervention for relieving fatigue are mainly divided into two categories. One is chemically synthesized drugs, such as amphetamine, caffeine, sympathomimetic stimulants, etc.; the other is natural plant active ingredients, such as traditional Chinese medicine compounds, single traditional Chinese medicines, main active ingredients, etc. Chemical drugs with anti-exercise fatigue effects are mostly cerebral cortex excitatory drugs, and their mechanism of action is mostly to excite the central nervous system, relieve the fatigue state caused by central inhibition, eliminate drowsiness, and enable the taker to maintain a long-term or high-intensity working state. However, this does not solve the problems caused by the depletion of energy substances and the accumulation of metabolites. Therefore, chemical drugs only create an "illusion" of fatigue relief, rather than truly relieve fatigue (Lü Yaobo. Anti-fatigue effect and its mechanism of wild ginseng ultramicro powder [D]. Jilin: Jilin University, 2023.). In addition, many adverse reactions have also occurred during the application of chemical drugs, such as addiction, liver and kidney toxicity, and central toxicity, etc. (DE SOUZA J G, DEL COSO J, FONSECA F S, et al. Risk or benefit? Side effects of caffeine supplementation in sport: a systematic review [J]. Eur J Nutr, 2022, 61(8): 3823-3834.). Natural drugs exert special curative effects due to their unique structures. Screening anti-exercise fatigue from safe natural drugs has become a research hotspot.

[0006] Perilla frutescens is an annual upright herbaceous plant of the genus Perilla in the Lamiaceae family. It is widely distributed and is a traditional medicinal and edible plant in China. Perilla porridge, Perilla tea, etc. are popular among the people for their beauty and health care effects. Perilla leaves are consumed in large quantities in China, Japan and South Korea as vegetables and spices. Perilla seeds can not only be used as condiments, but the pressed Perilla oil is rich in α-linolenic acid and has good health care effects. In addition, as a commonly used medicinal material for multi-site medication in humans, the Pharmacopoeia of the People's Republic of China has included three medicinal materials: Perilla leaf, Perilla stem and Perilla seed (Qian Jinxiu, Meng Wuwei, Liu Huihui, et al. Textual research on Perilla medicinal materials in classical famous prescriptions [J]. Chinese Journal of Experimental Traditional Medical Formulae, 2022, 28(10): 55-67.). The 2020 edition of the Chinese Pharmacopoeia records that Perilla has the effects of relieving exterior cold, regulating qi and harmonizing the stomach, and is used for wind-cold colds, cough and nausea, pregnancy vomiting and fish and crab poisoning. Leaves are the commonly consumed part of Perilla. Modern research shows that it contains various bioactive components such as flavonoids, volatile oils, phenolic acids, triterpenoid sterols, glycosides, etc., and has been proven to have anti-inflammatory, antibacterial, antioxidant, antidepressant, liver-protecting and other effects (LI HZ, REN Z Q, REDDY N V, et al. In silico evaluation of antimicrobial, antihyaluronidase and bioavailability parameters of rosmarinic acid in Perilla frutescens leaf extracts [J]. SN Applied Sciences, 2020, 2(9): 2397-2404; Zhang Liangqi, Li Wenjiao, Xiao Meifeng. Research progress on the comparison of active components and pharmacological effects of different parts of Perilla [J]. China Journal of Chinese Materia Medica, 2023, 48(24): 6551-6571.). Perilla leaf volatile oil is a volatile aromatic oily liquid obtained by a specific extraction method from Perilla leaves and is the main active component of Perilla leaves. In recent years, the development and application research of Perilla leaf volatile oil in various fields has been in the ascendant. In the food industry, for food preservation, antibacterial and color enhancement, in the pharmaceutical field, for the research and development of special drugs and the promotion of clinical medicine, and in chemical production, for the supply of industrial raw materials such as cosmetics and varnishes, Perilla leaf volatile oil has brought considerable economic benefits and has great scientific research value (Xue Shan. Study on the composition and antioxidant efficacy of Perilla leaf essential oil under different extraction methods [J]. Science and Technology of Food Industry, 2016, 19(37): 67-74.). Summary of the Invention

[0007] The present invention discovers that Perilla extract has the effect of anti-exercise fatigue and can improve the body's fatigue tolerance ability, and has great market value for developing products related to relieving physical fatigue. Based on this, the present invention is completed.

[0008] In a first aspect, the present invention provides an application of a perilla extract in the preparation of an anti-fatigue product, wherein the perilla extract is perillene-type volatile oil; the perillene-type volatile oil can improve the exercise endurance of the body, and improve the muscle state and physiological indexes of the body.

[0009] Furthermore, the fatigue is exercise fatigue.

[0010] Furthermore, the content of the perillene-type volatile oil is 1% - 100%.

[0011] Even further, the content of the perillene-type volatile oil is 10% - 100%.

[0012] Furthermore, the perillene-type volatile oil can be used independently or in combination with anti-fatigue components other than the perillene-type volatile oil.

[0013] Furthermore, the anti-exercise fatigue product includes food, health products or food additives.

[0014] Even further, the health products include various forms such as oral liquids, tablets or capsules.

[0015] In a second aspect, the present invention provides an application of a perilla extract in the preparation of an anti-fatigue drug, wherein the perilla extract is perillene-type volatile oil.

[0016] Furthermore, the fatigue is exercise fatigue.

[0017] Furthermore, the content of the perillene-type volatile oil is 1% - 100%.

[0018] Even further, the content of the perillene-type volatile oil is 10% - 100%.

[0019] Furthermore, the perillene-type volatile oil can be used independently or in combination with anti-fatigue components other than the perillene-type volatile oil.

[0020] Furthermore, one or more pharmaceutically acceptable carriers can also be added to the drug.

[0021] Furthermore, the drug can be made into various dosage forms, including but not limited to one or more of tablets, capsules, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents and / or suppositories.

[0022] Furthermore, the preparation can be one or more of ordinary preparations, sustained-release preparations and / or controlled-release preparations.

[0023] Furthermore, if necessary, coloring agents, preservatives, fragrances, flavoring agents, sweeteners or other materials can also be added to the drug preparation for various preparations.

[0024] Furthermore, the drug can be administered by injection, via body cavities, or through the respiratory tract.

[0025] Furthermore, the injection administration includes subcutaneous injection, intravenous injection, intramuscular injection, and intracavitary injection, etc.; the administration via body cavities includes rectal or vaginal administration; and the administration through the respiratory tract includes nasal administration.

[0026] Beneficial effects

[0027] In the present invention, a perillene-type volatile oil is obtained by steam distillation, and perillene accounts for more than 10% of the total volatile oil content. Through experimental verification, the perillene-type volatile oil has the following advantages:

[0028] 1) Improving the body's exercise endurance and relieving exercise fatigue;

[0029] 2) Improving muscle atrophy caused by exercise fatigue and restoring the atrophy, deformation and other adverse states of animal skeletal muscle fibers;

[0030] 3) Reducing the activity of animal lactate dehydrogenase, decreasing lactate accumulation, and relieving muscle soreness;

[0031] 4) Enhancing the activities of SOD and CAT, scavenging free radicals in the animal body, and improving the antioxidant capacity;

[0032] 5) Reducing the hydrogen peroxide content in the serum of mice and improving the oxidative stress of the mouse body;

[0033] 6) Increasing the glycogen content in the skeletal muscle of animals and improving exercise endurance and anti-exercise fatigue ability;

[0034] 7) Having no negative impact on the phenotypes, viscera, etc. of biological organisms, and being safe and non-toxic.

[0035] In summary, the perillene-type volatile oil extracted in the present invention can improve exercise endurance, improve muscle status and physiological indexes of the body, and has a good anti-exercise fatigue effect. The perillene-type volatile oil can be used to develop products for anti-exercise fatigue drugs, health products or foods, and has good application prospects. Brief description of the drawings

[0036] Figure 1 It is a schematic diagram of the technical process of the present invention.

[0037] Figure 2 It shows the evaluation results of the ATP content in muscle cells after administration of the perillene-type volatile oil in Example 1 of the present invention.

[0038] Figure 3 It is the evaluation result of the 10% load swimming of mice after administration of the perillene-type volatile oil in Example 2 of the present invention.

[0039] Figure 4 It is the evaluation results of the organ indices of mice after administration of the perillene-type volatile oil in Example 3 of the present invention.

[0040] Figure 5 It is the evaluation results of the serum physiological indices of mice after administration of the perillene-type volatile oil in Example 4 of the present invention.

[0041] Figure 6 It is the evaluation results of the glycogen content in the skeletal muscles of mice after administration of the perillene-type volatile oil in Example 5 of the present invention.

[0042] Figure 7 It is the HE staining results of the liver of mice after administration of the perillene-type volatile oil in Example 6 of the present invention.

[0043] Figure 8 It is the HE staining results of the cross-section of the skeletal muscles of mice after administration of the perillene-type volatile oil in Example 6 of the present invention.

[0044] Figure 9 It is the results of the muscle fiber area of the skeletal muscles of mice after administration of the perillene-type volatile oil in Example 6 of the present invention. Detailed implementation manners

[0045] The following further describes the detailed implementation manners of the present invention. It should be noted here that the descriptions of these implementation manners are for helping to understand the present invention, but do not constitute a limitation to the present invention. In addition, the technical features involved in the following described implementation manners can be combined with each other as long as they do not conflict with each other.

[0046] The experimental methods in the following examples are all conventional methods unless otherwise specified, and the test materials used in the following examples are all commercially available through conventional channels unless otherwise specified.

[0047] The model animals used in the present invention to verify the efficacy of the perillene-type volatile oil are 8-week-old male ICR mice. The genome of the mice contains about 90% homologous genes in the human genome and has been widely used in the screening of gene functions and substance functions of human diseases.

[0048] D-galactose is a physiological nutrient component that can be converted into glucose in normal body metabolism and participate in glucose metabolism. However, excessive supply will lead to metabolic disorders. Continuously injecting a large dose of D-galactose into mice within a certain period of time will cause an overly strong oxidative stress response in the mice, generating a large amount of reactive oxygen species and free radicals, resulting in cell tissue damage, affecting the normal functions of the body, and further causing the mice to show an obvious fatigue state in terms of motor ability.

[0049] Taurine is a sulfur-containing non-protein amino acid and a conditional essential amino acid for the human body. A large number of studies have shown that taurine has the effects of improving exercise ability and resisting exercise fatigue. Therefore, in the application experiment of the present invention, taurine is used as a control group to ensure the authenticity and certainty of the experiment of the present invention.

[0050] Example 1 Extraction, Identification of Perillene-Type Volatile Oil and Preliminary Evaluation of Its Anti-Exercise Fatigue Effect

[0051] Referring to the "Chinese Pharmacopoeia", with a material-liquid ratio of 1:6, add an appropriate amount of water and several grains of zeolite, soak at a constant temperature for a certain time and then shake and mix. Connect the volatile oil detector to the reflux condenser. After heating to boiling, keep slightly boiling for 6 h for extraction. Collect the volatile oil and add an appropriate amount of anhydrous sodium sulfate for dehydration. After extraction, the main substances of the extracted perilla leaf volatile oil are identified by GC-MS method. The main parameters of GC are: RXT-5MS quartz capillary column (30 m * 0.25 μm * 0.25 μm); the pre-column pressure is 53.5 kPa; the split ratio is 50:1; the injection volume is 1 μL; the injection port temperature is 250 °C; the carrier gas is He.

[0052] Skeletal muscle cells play a key role in energy metabolism. Starvation can lead to insufficient energy supply to cells. Simulating the physiological response of the body under long-term exercise or energy deprivation is a commonly used method to induce cell fatigue. First, culture C2C12 muscle cells with complete medium. When subculturing, divide the C2C12 muscle cells into a normal group (cultured with complete medium), a model group (cultured with serum-free and sugar-free medium), and a drug administration group. The model group and the drug administration group are changed to serum-free and sugar-free DMEM medium for culture for 24 h. The drug administration group is respectively added with perillene-type volatile oil containing 10%, 35%, 51%, 68%, and 80%. The normal group and the fatigue model group are added with an equal volume of serum-free and sugar-free DMEM medium and continue to culture for 24 h. Collect the cells, lyse the cells with cell lysate, and operate according to the instructions of the ATP detection kit. Use an enzyme-labeled instrument to measure the content of ATP in the cells.

[0053] The results are as Figure 2 shown. Compared with the normal group, the content of ATP in the cells of the model group decreased, indicating that a starvation-induced muscle cell fatigue model was successfully established. Compared with the model group, the content of ATP in the cells of the drug administration group increased, suggesting that the volatile oil containing 10%-80% perillene all has the effect of resisting exercise fatigue, and the perillene-type volatile oil with a perillene content of 68% has the best effect. This content is selected for subsequent effect verification.

[0054] Example 2 Evaluation of the Weight-Bearing Swimming of Perillene-Type Volatile Oil in Mice

[0055] Twenty-five 8-week-old male SPF-grade ICR mice were selected and divided into 5 groups of 5 mice each. The breeding temperature was 20-25 °C. All mice were allowed to eat freely. After three days of adaptive breeding, subsequent experiments were carried out.

[0056] Grouping, modeling and drug administration:

[0057] Normal group (NC): Intraperitoneally injected with normal saline for 6 weeks, and then gavaged with water for 3 weeks simultaneously. During this period, the mice ate freely;

[0058] Model group (MC): Intraperitoneally injected with a D-galactose solution at a dose of 500 mg / kg, and then gavaged with water for 3 weeks simultaneously. During this period, the mice ate freely;

[0059] Positive control group (PC): Intraperitoneally injected with a D-galactose solution at a dose of 500 mg / kg, and then gavaged with a taurine solution at a dose of 100 mg / kg for 3 weeks simultaneously. During this period, the mice ate freely;

[0060] Low-dose administration group (XL): Intraperitoneally injected with a D-galactose solution at a dose of 500 mg / kg, and then gavaged with perillene-type volatile oil at a dose of 0.01 mL / kg for 3 weeks simultaneously. During this period, the mice ate freely;

[0061] Medium-dose administration group (XM): Intraperitoneally injected with a D-galactose solution at a dose of 500 mg / kg, and then gavaged with perillene-type volatile oil at a dose of 0.05 mL / kg for 3 weeks simultaneously. During this period, the mice ate freely;

[0062] High-dose administration group (XH): Intraperitoneally injected with a D-galactose solution at a dose of 500 mg / kg, and then gavaged with perillene-type volatile oil at a dose of 0.1 mL / kg for 3 weeks simultaneously. During this period, the mice ate freely.

[0063] One hour after the above grouping and drug administration, lead sheets equivalent to 10% of the body weight were wound around the middle of the tails of all mice. In water at a depth of 50 cm and 25 ± 2 °C, the exhaustion standard was that the mouse's nose tip could not float after being immersed in water for 10 s. The exhaustion time of the mice was recorded.

[0064] The results were as Figure 3 shown. The weight-bearing swimming duration of the model group was significantly shorter than that of the normal group. The weight-bearing swimming durations of the positive control group and the administration groups were significantly longer than that of the model group. At the same time, the durations of the administration groups were longer than that of the normal group, and the effect of the administration groups was better than that of the positive control group, indicating that perillene-type volatile oil could significantly prolong the weight-bearing swimming time, improve the body's exercise endurance and relieve exercise fatigue.

[0065] Example 3 Toxicity of perillene-type volatile oil to the main organs of mice

[0066] One hour after the weight-bearing swimming of each group of mice, the mice were anesthetized with 1% sodium pentobarbital, and then blood was collected by enucleating the eyeballs. The mice were sacrificed by cervical dislocation, and the livers, spleens, kidneys and skeletal muscles of the mice were taken out.

[0067] Weigh the livers, spleens, and kidneys of the mice and record the weights.

[0068] The organ index formulas for the mice are as follows:

[0069] (1) Liver index (mg / g) = liver weight / body weight;

[0070] (2) Spleen index (mg / g) = spleen weight / body weight;

[0071] (3) Kidney index (mg / g) = kidney weight / body weight

[0072] The results are as Figure 4 shown. There were no significant differences in the liver index, spleen index, and kidney index of the drug-administered group compared with the normal group. That is, the drug-administered group had no significant negative impact on the metabolic functions of the livers, spleens, and kidneys of the mice, indicating that perillene-type volatile oil has no obvious toxicity to mice.

[0073] Example 4 Effects of Perillene-Type Volatile Oil on the Activities of LDH, SOD, and CAT in Mouse Serum

[0074] Take the mouse serum obtained from the dissection in Example 3 and measure the activities of LDH, SOD, CAT, and the hydrogen peroxide content with reference to the instruction manual of the Nanjing Jiancheng kit.

[0075] Lactate dehydrogenase (LDH) can catalyze the conversion of pyruvate into lactate, causing muscle soreness, and it is one of the evaluation indicators for anti-exercise fatigue effects. Measure the activity of LDH in the serum. The results are as Figure 5 shown. Compared with the model group, the activities of lactate dehydrogenase in the sera of the normal group and the drug-administered group of mice were lower, indicating that perillene-type volatile oil can reduce the activity of lactate dehydrogenase in mice after exhaustive swimming, reduce the accumulation of lactate in mice, relieve the muscle soreness of mice, and thus play an anti-exercise fatigue role.

[0076] When the body is in a state of oxidative stress for a long time, the energy metabolism in the body will be affected, resulting in a sense of fatigue. SOD and CAT are antioxidant enzymes that catalyze the decomposition of reactive oxygen species widely present in the body and are commonly used to evaluate the antioxidant activity in the body. Measure the antioxidant indices SOD and CAT in the serum. The results are as Figure 5 shown. The drug-administered group can restore the activities of SOD and CAT to normal levels, and the SOD activity of the control group is higher than that of the normal group. Moreover, after the administration of perillene-type volatile oil, its SOD activity is almost the same as that of the control group. At the same time, the CAT activity of the perillene-type volatile oil-administered group increases with the concentration gradient, indicating that perillene-type volatile oil can enhance the activities of SOD and CAT, scavenge free radicals in mice, and improve the antioxidant ability of mice.

[0077] Imbalance of redox homeostasis in the body can lead to an increase in hydrogen peroxide content, causing the body to be in a state of peroxidation and fatigue. The hydrogen peroxide content in serum was measured, and the results are as Figure 5 shown. Compared with the normal group, the hydrogen peroxide content in the model group was significantly increased, indicating that D-galactose injection can cause peroxidation in the body. After administration, the hydrogen peroxide content decreased. The hydrogen peroxide content in the serum of the control group and the high-dose perillene-type volatile oil group returned to the normal group level, indicating that the perillene-type volatile oil can reduce the hydrogen peroxide content in the serum of mice and improve the oxidative stress of the mouse body.

[0078] Example 5 Effect of perillene-type volatile oil on glycogen storage in mouse muscle tissue

[0079] The skeletal muscle of the mouse obtained by dissection in Example 3 was taken, and its glycogen content was measured with reference to the instruction manual of the Nanjing Jiancheng kit.

[0080] Skeletal muscle is the main organ of movement. The energy metabolism of skeletal muscle is closely related to the body's exercise ability and exercise endurance. The increase in glycogen content is the main reason for the improvement of exercise endurance, and skeletal muscle is one of the main tissues for glycogen storage. The glycogen content in skeletal muscle was measured, and the results are as Figure 6 shown. The glycogen content in the normal group was significantly higher than that in the model group; compared with the model group, the glycogen content in the administration group was improved, indicating that the perillene-type volatile oil can increase the glycogen content in the skeletal muscle of mice, improve the exercise endurance and anti-exercise fatigue ability of mice.

[0081] Example 6 Effect of perillene-type volatile oil on the pathology of mouse liver and skeletal muscle tissues

[0082] The liver and skeletal muscle of the mouse obtained by dissection in Example 3 were taken, and an appropriate amount of 4% paraformaldehyde was added to fix the tissues. The tissues were dehydrated, infiltrated with wax, embedded and sectioned. The sections were placed in hematoxylin staining solution and rinsed with running water, then dehydrated in 95% ethanol, and then stained with eosin staining solution and dehydrated and sealed to obtain HE-stained tissues. After microscopic examination, image acquisition and analysis were carried out.

[0083] Pathological analysis results of liver tissue Figure 7 shown: The hepatic lobule structure of the liver tissues in each group was clear and complete, and the hepatic cell cords were arranged neatly, indicating again that the perillene-type volatile oil is non-toxic to the mouse liver.

[0084] The pathological analysis results of skeletal muscle tissue are as Figure 8 、 Figure 9 shown: The muscle tissue structure of the normal group mice was clear, and the muscle fibers were arranged tightly. In the model group mice, the muscle fibers were arranged loosely, and the morphology of multiple muscle fibers was atrophied, deformed and necrotic; compared with the model group, the muscle fibers in the administration group were arranged more tightly and had a good morphology. As the drug dose increased, the gap between muscle fibers became smaller, and the area of muscle fibers gradually increased to the normal level, and the muscle bundle was flat.

Claims

1. An application of a perilla extract in the preparation of an anti-fatigue product, wherein the perilla extract is a perilla ene-type volatile oil; the perilla ene-type volatile oil can improve the body's exercise endurance, muscle state and body physiological indicators.

2. The use according to claim 1, wherein the content of the perillene-type volatile oil is 1%-100%.

3. The use as claimed in claim 1, wherein the content of the perillene-type volatile oil is 10%-100%.

4. The use as claimed in claim 1, wherein the perillene-type volatile oil can be used independently or in combination with an anti-fatigue component other than the perillene-type volatile oil.

5. The use according to claim 1, wherein the anti-fatigue product comprises food, health care product or food additive.

6. Use of a perilla extract in the preparation of an anti-fatigue drug, wherein the perilla extract is a perilla ene-type volatile oil.

7. The use according to claim 6, wherein the content of the perillene-type volatile oil is 1%-100%.

8. The use according to any one of claims 6 or 7, wherein the perillene-type volatile oil can be used independently or in combination with an anti-fatigue component other than the perillene-type volatile oil.

9. The use according to any one of claims 6 to 8, wherein the drug can be prepared into a variety of dosage forms, including but not limited to one or more of tablets, capsules, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents and / or suppositories.

10. The use according to any one of claims 6 to 9, wherein the drug can be administered by injection, cavity administration or respiratory tract administration; the injection administration includes subcutaneous injection, intravenous injection, intramuscular injection and intracavitary injection, etc., the cavity administration includes rectal or vaginal administration, and the respiratory tract administration includes nasal administration.

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