Application of ligustilide in preparation of sports fatigue relieving product

Abstract

The invention relates to the field of medicine application, in particular to application of ligustilide in preparation of a product for relieving exercise fatigue. The invention provides an application of ligustilide in preparation of a product for relieving exercise fatigue. The invention provides a natural terpene compound-ligustilide derived from ligusticum wallichii volatile oil. The natural terpene compound-ligustilide has a remarkable effect in the aspect of delaying exercise fatigue. Researches show that ligustilide enhances the oxidative stress resistance and reduces the accumulation of reactive oxygen species (ROS) by inhibiting NSY-1 / SEK-1 / PMK-1 phosphorylation cascade reaction and promoting the expression of GST-4 (glutathione S-transferase), thereby balancing the oxidative stress level and improving the exercise ability. In addition, ligustilide significantly improves the head swing frequency and the body twisting frequency of caenorhabditis elegans, prolongs the service life, and has no negative influence on the growth, development and reproductive capacity of caenorhabditis elegans.

Description

Technical Field

[0001] The invention relates to the field of drug application, and in particular to the application of ligustilide in the preparation of products for relieving sports fatigue. Background Art

[0002] With the accelerated pace of society and unhealthy lifestyles, the harm of physical and mental fatigue to health is becoming more and more obvious. It is reported that exercise is a means to alleviate chronic diseases related to obesity, metabolism, and the immune system, and can effectively enhance human immunity and life span. However, excessive exercise can lead to the massive production of harmful substances in the body, including free radicals, lactic acid, and urea nitrogen. The massive accumulation of these metabolic byproducts can lead to intrinsic metabolic disorders and cause the occurrence of sports fatigue. Therefore, promoting muscle repair, balancing energy metabolism, and inhibiting the production of metabolic byproducts are important ways to relieve sports fatigue.

[0003] Excessive accumulation of free radicals is considered to be one of the important factors of sports fatigue. The massive accumulation of free radicals (such as reactive oxygen, reactive nitrogen and lipid free radicals) can lead to an imbalance between antioxidants and oxidants in the body and cause oxidative stress. The regulation of oxidative stress is related to many human diseases and postoperative treatments, including neurodegenerative diseases, cardiovascular diseases, cancer and diabetes. Once this antioxidant balance is broken, excessive free radicals will react with cells, causing cell membrane damage, DNA damage, and lipid and protein peroxidation, and may even trigger programmed cell death.

[0004] For the inhibition and relief of sports fatigue, the existing technology also has certain treatment methods. For example, a study disclosed a compound preparation based on bioactive ingredients, which can prolong the exercise endurance of experimental animals and effectively regulate multiple physiological indicators such as serum urea nitrogen, lactic acid, creatinine, corticosterone, etc., thereby relieving fatigue symptoms and improving exercise capacity. In addition, there are studies reporting a composition combining specific plant extracts and nutrients, which can promote vasodilation and reduce oxygen consumption, thereby improving exercise performance and energy levels, while exerting antioxidant and anti-inflammatory effects to improve sports fatigue. In addition, some literature also mentions that proteins extracted from animal by-products show certain anti-sports fatigue effects due to their high antioxidant activity, thereby increasing the utilization value of the by-products.

[0005] Although there are certain treatment methods and strategies for delaying and improving sports fatigue, none of them involve the compound of the present invention and the application report in alleviating sports fatigue. Summary of the invention

[0006] In view of this, the present invention provides the use of ligustilide in the preparation of products for relieving sports fatigue. The present invention proposes a natural terpene compound derived from volatile oil of Ligusticum chuanxiong - ligustilide, which has a significant effect in delaying sports fatigue. Studies have shown that ligustilide inhibits the NSY-1 / SEK-1 / PMK-1 phosphorylation cascade reaction and promotes the expression of GST-4 (glutathione S-transferase), thereby enhancing the ability to resist oxidative stress and reducing the accumulation of reactive oxygen species (ROS), thereby balancing the level of oxidative stress and improving exercise capacity. In addition, ligustilide significantly improved the head swing frequency and body twisting frequency of Caenorhabditis elegans, prolonged lifespan, and had no negative impact on its growth and development and reproductive ability.

[0007] Compared with the existing technology, Ligustilide not only covers the functions of anti-oxidation and life extension, but also effectively enhances the anti-oxidative stress response at the molecular level through a multi-dimensional mechanism, thereby improving sports endurance. This multi-level regulatory mechanism enables the present invention to have a more extensive and lasting effect in anti-fatigue and aging delay, which is superior to the common single antioxidant or single sports performance improvement strategies in the existing technology.

[0008] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0009] The invention provides application of ligustilide in preparing a product for relieving sports fatigue.

[0010] In some embodiments of the present invention, in the above uses, ligustilide enhances the ability to resist oxidative stress.

[0011] In some embodiments of the present invention, in the above use, ligustilide promotes the expression of glutathione transferase.

[0012] In some embodiments of the present invention, in the above use, ligustilide inhibits the reaction of NSY-1 / SEK-1 / PMK-1 phosphorylation cascade.

[0013] In some embodiments of the present invention, in the above uses, ligustilide reduces the content of reactive oxygen species.

[0014] In some embodiments of the present invention, in the above uses, ligustilide improves motor performance.

[0015] In some embodiments of the present invention, in the above application, ligustilide is derived from volatile oil of Ligusticum chuanxiong.

[0016] In some embodiments of the present invention, in the above applications, ligustilide is derived from natural plants and / or chemically synthesized.

[0017] In some embodiments of the present invention, in the above applications, the product includes: one or more of: medicines, preparations, health products and cosmetics.

[0018] In some embodiments of the present invention, in the above application, the administration amount of ligustilide is: 0.95-95 μg / day.

[0019] The present invention proposes a natural terpene compound derived from volatile oil of Ligusticum chuanxiong - ligustilide, which has a significant effect in delaying sports fatigue. Studies have shown that ligustilide can enhance the ability to resist oxidative stress and reduce the accumulation of reactive oxygen species (ROS) by inhibiting the NSY-1 / SEK-1 / PMK-1 phosphorylation cascade reaction and promoting the expression of GST-4 (glutathione S-transferase), thereby balancing the level of oxidative stress and improving exercise capacity. In addition, ligustilide significantly improved the head swing frequency and body twisting frequency of Caenorhabditis elegans, prolonged its lifespan, and had no negative impact on its growth and development and reproductive capacity.

[0020] Compared with the existing technology, Ligustilide not only covers the functions of anti-oxidation and life extension, but also effectively enhances the anti-oxidative stress response at the molecular level through a multi-dimensional mechanism, thereby improving sports endurance. This multi-level regulatory mechanism enables the present invention to have a more extensive and lasting effect in anti-fatigue and aging delay, which is superior to the common single antioxidant or single sports performance improvement strategies in the existing technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art are briefly introduced below.

[0022] Figure 1 It shows that ligustilide promotes the movement ability of Caenorhabditis elegans; wherein: A shows the chemical structure of ligustilide; BF shows the effect of ligustilide treatment for 2, 4, 6, 12, and 24 hours on the movement of Caenorhabditis elegans; G shows the dose effect of ligustilide treatment for 12 hours on the movement behavior of Caenorhabditis elegans; H shows the effect of ligustilide on the movement ability of nematodes at different age stages; I shows the body bending frequency of Caenorhabditis elegans after ligustilide treatment; J shows the total number of offspring produced by Caenorhabditis elegans after ligustilide treatment;

[0023] Figure 2 It shows that ligustilide prolongs the life span of nematodes, but does not affect the development of nematodes; A shows representative images of nematodes at different growth and development stages; B shows body length; C shows quantitative analysis of area; D shows the effect of ligustilide treatment on the life span of nematodes;

[0024] Figure 3The figure shows the mechanism of action of ligustilide in alleviating exercise fatigue and anti-oxidative stress; wherein: AD shows the movement frequency of PMK-1 phosphorylation cascade-related mutants nsy-1 (A), sek-1 (B), pmk-1 (C) and gst-4 (D) after ligustilide treatment; EF shows representative images (E) and quantitative analysis (F) of fluorescence expression of gst-4::GFP nematodes of different ages after ligustilide and DMSO treatment; G shows the movement frequency of gst-4; pmk-1 RNAi and wild-type nematodes after ligustilide treatment; HJ shows representative western blot images (H), PMK-1 total protein (I) and phosphorylation levels (J) of wild-type worms. DETAILED DESCRIPTION

[0025] The invention discloses application of ligustilide in preparing a product for relieving sports fatigue.

[0026] It should be understood that the expression "one or more of..." includes each of the items recited after the expression individually and in various different combinations of two or more of the recited items, unless otherwise understood from the context and usage. The expression "and / or" in combination with three or more recited items should be understood to have the same meaning, unless otherwise understood from the context.

[0027] The use of the terms "comprising", "having" or "containing", including their grammatical synonyms, should generally be understood as open and non-restrictive, for example not excluding other unrecited elements or steps, unless otherwise specifically stated or otherwise understood from the context.

[0028] It should be understood that the order of steps or the order in which certain actions are performed is not important as long as the present invention remains operable. In addition, two or more steps or actions may be performed simultaneously.

[0029] The use of any and all examples or exemplary language, such as "for example" or "including", herein is intended only to better illustrate the invention and does not limit the scope of the invention unless otherwise claimed. No language in this specification should be construed as indicating that any non-claimed element is essential to the practice of the invention.

[0030] In addition, the numerical ranges and parameters used to define the present invention are approximate values, and the relevant values ​​in the specific embodiments have been presented as accurately as possible. However, any numerical value inherently inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise expressly stated, it should be understood that all ranges, quantities, values ​​and percentages used in this disclosure are modified by "about". Here, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a specific value or range.

[0031] The present invention provides a terpene compound ligustilide, aiming to provide a compound capable of reducing oxidative stress and delaying sports fatigue.

[0032] The active substance in the Rhizoma Chuanxiong proposed by the present invention is a terpene active small molecule compound ligustilide, which can prolong the life span of nematodes while effectively reducing oxidative stress damage and motor ability decline caused by exercise.

[0033] The active small molecule ligustilide proposed in the present invention is derived from volatile oil of Chuanxiong, and is a natural cyclic terpene compound formed by polymerization of multiple isoprene units. The present invention uses a multicellular model organism Caenorhabditis elegans to conduct experiments, and determines that ligustilide is the main active ingredient of volatile oil of Chuanxiong for delaying sports fatigue. At the same time, it also clarifies the main mechanism of action of ligustilide by increasing the activity of glutathione S-transferase, catalyzing the reduction reaction to neutralize harmful peroxides or active oxygen, reducing oxidative stress and relieving sports fatigue. Therefore, the compound ligustilide has the potential to be developed into a drug for relieving sports fatigue and enhancing antioxidant capacity.

[0034] Compared with previous drug molecules such as resveratrol, green tea polyphenols, and ginsenosides, the ligustilide proposed in the present invention has the common properties of anti-oxidation and anti-aging, and also has the characteristics of significantly enhancing athletic ability and relieving sports fatigue. This makes ligustilide unique in relieving sports fatigue and shows its potential economic value in the field of sports health.

[0035] In Examples 1 to 7 and Effect Examples 1 to 3 of the present invention, all the raw materials and reagents used can be purchased from the market.

[0036] The present invention will be further described below in conjunction with embodiments:

[0037] Example 1 Head swing experiment

[0038] To measure the vitality and health of the worms, the head swing of the worms was evaluated. The worms synchronized to the L4 stage were picked into NGM plates mixed with OP50 bacterial solutions of different concentrations of ligustilide and OP50 bacterial solutions mixed with DMSO, and cultured for 12 hours at 20°C. Then, 20μL of M9 solution was dropped into the center of the NGM plate, and the recording began 30 seconds after the worms moved into the M9. The number of head swings within 30s was set as one cycle. All experiments were carried out in a constant temperature environment of 20°C to ensure that the movement of the worms was not affected by temperature.

[0039] Example 2 Body bending test

[0040] The nematodes synchronized to the L4 stage were placed on the ligustilide plate and the DMSO plate and cultured for 12 hours. Then, the nematodes were moved to a clean NGM plate and after acclimating to the environment for 1 minute, the number of times the nematodes bent within 1 minute was recorded.

[0041] Example 3 Reproductive capacity experiment

[0042] The number of nematodes brooded is an experiment to detect the reproductive ability of nematodes and an important indicator of drug reproductive toxicity. The nematodes synchronized to the L4 period were picked into the ligustilide plate and the DMSO plate respectively, and one L4 period nematode was placed on each NGM plate. The nematodes were transferred every 12 hours and the number of eggs laid was counted until the nematodes no longer laid eggs (usually 4-5 days). Among them, every 10 nematodes were grouped together, and the test was repeated three times.

[0043] Example 4 Life Test

[0044] The life span test of nematodes can effectively reflect the effect of drug treatment on the life span of nematodes. The nematodes synchronized to the L4 period were picked into the ligustilide plate and the DMSO plate respectively, with 50 nematodes on each plate, and the recording time was 0 day. Cultured in a constant temperature incubator at 20°C, the nematodes were transferred to a new NGM plate every 24 hours, and the number of nematodes in each group was recorded. Deaths in the first seven days are considered unnatural deaths and will not be recorded. The death criterion is that if no peristalsis or swallowing movement is observed after lightly touching the head of the nematode, it is recorded as death. Daily survival rate = (total number - cumulative number of deaths) / total number of deaths.

[0045] Example 5 Growth and development experiment

[0046] The determination of the body length and area of ​​nematodes at different stages is an important indicator for evaluating the toxicological effects of drugs on the growth and development of nematodes. The nematodes in the egg-laying period were picked into the ligustilide plate and the DMSO plate respectively, and the eggs produced within 1 hour were obtained through synchronization experiments. Then, the nematodes were continuously cultured in the ligustilide plate and the DMSO plate, and imaging was performed with a multifunctional microscope at each developmental stage of the nematodes (L1, L2, L3, L4). Ten nematodes were photographed in each group at each developmental stage. After obtaining the data, the body length and area of ​​the nematodes at each stage were quantitatively analyzed using Image J software to analyze the differences. The test was repeated three times.

[0047] Example 6 Fluorescence Imaging

[0048] The nematodes synchronized to the L4 stage were picked into the ligustilide plate and DMSO plate respectively and cultured at 20°C for 12h. Weigh 2g agar and 40mL distilled water to prepare a hot agar solution, transfer it to a glass slide and press it into an agar sheet to cool for use. After the nematodes were anesthetized with 25mM sodium azide solution, they were transferred to the agar sheet to make imaging samples. The nematodes were imaged under a multifunctional microscope. In order to quantitatively analyze the experimental results, the exposure time, magnification and other parameters of fluorescence imaging were set as basic parameters, and all nematodes treated in the same batch were used afterwards. The imaging pictures were processed and the fluorescence intensity was analyzed using Image J software, and the bar graph was drawn using GraphpadPrism 8 software.

[0049] Example 7 Protein Molecular Imprinting

[0050] The nematodes synchronized to the L4 stage were picked into the ligustilide plate and the DMSO plate, respectively, and cultured at 20°C for 12 hours. The nematodes were eluted with M9 solution into a 1.5 mL centrifuge tube, centrifuged at 800 rpm for 30 seconds at room temperature, and the supernatant was removed after the nematodes sank to the bottom of the centrifuge tube. Then, 1 mL of M9 solution was added and centrifuged at 800 rpm for 30 seconds, and the solution was removed again. This step was repeated three times to ensure that no OP50 remained.

[0051] The nematode sample containing a small amount of M9 was quickly transferred to liquid nitrogen for quick freezing, and 500 μL of lysis solution containing protease and phosphatase inhibitors was added, and ultrasonic disruption (60 Hz) was performed at 0 ° C for 3 min. Then, it was placed at 4 ° C for full lysis for 1 h, centrifuged at 4 ° C and 10000 rpm for 30 min, and the supernatant was collected. Then the western blot analysis method was used. The total protein of different components was separated by 10% SDS-PAGE gel electrophoresis and electrotransferred to the NC membrane at a constant current of 350 mA. After crossing the membrane, the membrane was blocked and incubated with TBST containing 5% skim milk at room temperature for 30 minutes. Then, appropriate concentrations of primary and secondary antibodies were added to the NC membrane and incubated for 12 h and 1 h, respectively. The membrane was rinsed three times with TBST and then incubated with the prepared ELC solution (ELC A:ELC B=1:1). Finally, the electrophoresis image was obtained using a chemiluminescence imager, and the protein bands of interest were analyzed using ImageJ software.

[0052] Effect Example 1 Ligustilide can relieve nematode exercise fatigue

[0053] Ligustilide (LIG) is a phenolphthalein derivative and a class of lipophilic compounds with multiple biological activities in the volatile oils of Umbelliferae plants, including traditional Chinese herbal medicines such as Chuanxiong and Angelica sinensis. Figure 1 As shown in A, the molecular formula is C 12 H 14 O2 , molecular weight is 190.238 g / mol, CAS number is 81944-09-4. Ligustilide has attracted wide attention due to its diverse biological activities, including anti-inflammatory, anti-tumor, antibacterial and antioxidant effects. Therefore, ligustilide has potential application prospects in delaying sports fatigue.

[0054] Caenorhabditis elegans is a classic model animal with a simple structure. In the past few decades, remarkable results have been achieved in the fields of development, aging, and neurological diseases. This study selected Caenorhabditis elegans as the research object. Caenorhabditis elegans has many advantages in exploring the pharmacological mechanisms of active compounds, including short lifespan, large egg-laying volume, complete cell and developmental lineage, and simple culture. Thrashing experiment is the main method to study movement rate. The study determined the connection between movement and aging by observing the head swing frequency of nematodes of different ages, and studied the changes in the movement rate of nematodes by adding various concentrations of ligustilide. In order to further evaluate the effects of ligustilide on nematodes, we studied the egg-laying rate and body twisting frequency indicators of nematodes exposed to ligustilide.

[0055] We exposed nematodes to ligustilide (10μM, 100μM and 1mM) for different time periods (2h, 4h, 6h, 12h, 24h). Figure 1 As shown in BG, compared with DMSO-treated nematodes, the exposure time was 2h ( Figure 1 The swing frequency of nematodes in B) did not change significantly. However, when the treatment time reached 4h ( Figure 1 C) and 6h( Figure 1 After exposure to the three concentrations of ligustilide, the swing frequency of nematodes was significantly enhanced and increased at 12h( Figure 1 The peak value was reached at E), and the swing frequency of the nematode reached nearly 80 times within 30 seconds. Figure 1 As shown in G, when nematodes were exposed to different concentrations (0.01μM, 0.1μM, 1μM, 10μM, 100μM, 1mM) of ligustilide for 12h, the swing frequency of the nematodes showed a concentration-dependent enhancement as the concentration of ligustilide increased, and was significantly enhanced when the concentration reached 10μM. Therefore, the experiment selected 10μM ligustilide and the exposure time of 12h as the condition to treat nematodes of different ages. The results showed that ligustilide can improve the movement ability of nematodes at different growth stages ( Figure 1 H).

[0056] In addition, the nematode body twisting experiment is also an important indicator for detecting nematode movement and health. In order to further evaluate the effect of ligustilide on nematodes, we measured the effect of ligustilide on the nematode body twisting frequency and egg-laying rate. Figure 1As shown in Figure 1, after exposure to 10 μM ligustilide for 12 hours, the body twisting frequency of nematodes also increased significantly. Compared with the DMSO group, there was no significant change in the egg-laying rate of nematodes exposed to ligustilide ( Figure 1 The results of the study showed that ligustilide can improve the movement ability of nematodes and has no negative impact on the physiological characteristics of nematodes.

[0057] Table 1

[0058]

[0059]

[0060] Table 1 corresponds to Figure 1 B in.

[0061] Table 2

[0062]

[0063]

[0064] Table 2 corresponds to Figure 1 C in.

[0065] Table 3

[0066]

[0067]

[0068] Table 3 corresponds to Figure 1 D in.

[0069] Table 4

[0070]

[0071]

[0072] Table 4 corresponds to Figure 1 E in.

[0073] Table 5

[0074]

[0075]

[0076] Table 5 corresponds to Figure 1 F in.

[0077] Table 6

[0078]

[0079]

[0080] Table 6 corresponds to Figure 1 The G in.

[0081] Table 7

[0082]

[0083]

[0084] Table 7 corresponds to Figure 1 H in.

[0085] Table 8

[0086]

[0087]

[0088] Table 8 corresponds to Figure 1 I in.

[0089] Effect Example 2 Ligustilide can increase the lifespan of nematodes without affecting their growth and development

[0090] Oxidative stress is caused by an imbalance in the generation and neutralization of reactive oxygen species, which leads to the massive generation of multiple reactive oxygen species, which may cause serious oxidative damage to DNA, proteins, and lipids, and cause the occurrence of exercise fatigue. Therefore, oxidative stress is considered to be an important factor leading to exercise fatigue. Aging and growth and development are important indicators of oxidative stress, so we conducted experiments related to lifespan and growth and development on nematodes treated with ligustilide to further study the connection between ligustilide and exercise fatigue.

[0091] Several studies have reported that aging and growth are also important indicators of oxidative stress. To evaluate this possibility, we studied the growth and development of C. elegans exposed to ligustilide ( Figure 2 AC) and life indicators ( Figure 2 D). The experimental results are as follows Figure 2 As shown in AC, no significant changes were found in the body length and area of ​​nematodes at different stages (L1, L2, L3, and L4) exposed to a dose of 10 μM ligustilide. In addition, the effect of ligustilide on the lifespan of nematodes was further studied. Compared with the DMSO group, the average lifespan of nematodes treated with 10 μM ligustilide was significantly increased by 20.67% (p<0.0001). This shows that ligustilide can prolong the lifespan of nematodes, delay the aging of nematodes, and will not affect the normal growth and development process of nematodes. Combined with the effect of ligustilide on improving exercise fatigue in nematodes, these results indicate that the improvement of nematode aging and movement by ligustilide may be associated with oxidative stress.

[0092] Effect Example 3: Ligustilide enhances the antioxidant stress ability of nematodes by increasing the activity of glutathione S-transferase

[0093] PMK-1 is a homologous gene of mammalian MAPK11 and MAPK14, and together with NSY-1 (MAPKKK) and SEK-1 (MAPKK), it forms a phosphorylation cascade reaction, which plays an important role in inflammatory and oxidative stress responses. GST-4 is a glutaminotransferase that participates in the metabolism of the antioxidant molecule glutathione. To clarify the relationship between ligustilide in relieving exercise fatigue and oxidative stress, and to explore the mechanism by which ligustilide enhances exercise ability, fluorescence imaging and Western blotting were used for determination in this study.

[0094] The experimental results showed that the thrashing frequencies of nsy-1, sek-1, pmk-1, and gst-4 mutant nematodes were all significantly decreased compared with those of wild-type nematodes treated with DMSO. At the same time, treatment with ligustilide failed to increase the thrashing frequencies of nsy-1, sek-1, pmk-1, and gst-4 mutant nematodes ( Figure 3 A-D). These results indicate that NSY-1, SEK-1, and PMK-1 are all involved in the mechanism of action of ligustilide, and this mechanism is related to antioxidant stress.

[0095] GST-4 is an important member of the glutathione-S-transferase family, mainly used to promote the synthesis of glutathione to participate in the defense against oxidative stress and neutralize harmful toxins. GST-4 is a highly conserved gene that widely exists in humans, mice, plants, and other invertebrates, and is considered a key indicator for studying the function of antioxidants. Therefore, to further confirm the effect of ligustilide treatment on GST-4, we treated gst-4p::GFP nematodes at different stages with DMSO and 10 μM ligustilide for 12 h respectively, and quantitatively analyzed the fluorescence expression of the nematodes. As Figure 3 shown in E and Figure 3 F, compared with the DMSO group, the fluorescence intensities of nematodes at different stages treated with ligustilide were significantly increased, indicating that ligustilide can effectively improve the antioxidant stress ability of nematodes.

[0096] To further determine the relationship between GST-4 and the PMK-1 signaling pathway in the mechanism of action of ligustilide, we treated gst-4 nematodes after pmk-1 RNAi interference with DMSO and ligustilide. The thrashing frequency phenotypes of WT, gst-4, and pmk-1 RNAi nematodes remained stable. The results showed that the thrashing frequencies of gst-4;pmk-1 RNAi nematodes under DMSO and ligustilide treatments were similar to those of gst-4 nematodes ( Figure 3 G). The results of Western blotting were as shown in Figure 3As shown in HJ, compared with the DMSO group, there was no difference in the expression of total PMK-1 protein in the nematodes treated with ligustilide, while the phosphorylation level of PMK-1 (p-PMK-1) was significantly reduced. These results indicate that GST-4 is a downstream target of PMK-1, and ligustilide enhances the anti-oxidative stress ability of nematodes and delays exercise fatigue by inhibiting the NSY-1 / SEK-1 / PMK-1 phosphorylation cascade reaction process and promoting GST-4 expression.

[0097] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Application of ligustilide in the preparation of products for relieving sports fatigue.

2. The use according to claim 1, characterized in that Ligustilide enhances the ability to resist oxidative stress.

3. The use according to claim 1 or 2, characterized in that: Ligustilide promotes the expression of glutathione S-transferase.

4. The use according to any one of claims 1 to 3, characterized in that Ligustilide inhibits the NSY-1 / SEK-1 / PMK-1 phosphorylation cascade.

5. The use according to any one of claims 1 to 4, characterized in that: Ligustilide reduces the content of reactive oxygen species.

6. The use according to any one of claims 1 to 5, characterized in that Ligustilide improves exercise performance.

7. The use according to any one of claims 1 to 6, characterized in that Ligustilide is derived from the volatile oil of Chuanxiong.

8. The use according to any one of claims 1 to 7, characterized in that Ligustilide is derived from natural plants and / or chemically synthesized.

9. The use according to any one of claims 1 to 8, characterized in that The products include: one or more of medicines, preparations, health products and cosmetics.

10. The use according to any one of claims 1 to 9, characterized in that The dosage of ligustilide is: 0.95~95μg / day.

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