Application of compound radix astragali seu hedysari-essence oral liquid in anti-fatigue effect of mice

Abstract

The invention relates to the technical field of application of radix astragali seu hedysari oral liquid, and particularly discloses application of a compound radix astragali seu hedysari oral liquid in the anti-fatigue effect of mice, which comprises the following steps: S1, preparing an intragastric solution, S2, carrying out grouped administration on the mice, S3, carrying out a weight-bearing exhaustive swimming test on the mice, S4, determining related indexes, and S5, carrying out statistical analysis. The compound radix astragali seu hedysari oral liquid can reduce glycogen consumption and BUN accumulation of exercise mice and prolong the exercise time of the mice, so that the compound radix astragali seu hedysari oral liquid has a certain anti-fatigue effect, and the compound radix astragali seu hedysari oral liquid consisting of the three traditional Chinese medicinal materials can be used as an antioxidant and a supplement for improving exercise tolerance, so that the anti-fatigue function is achieved; different doses of compound radix astragali seu hedysari oral liquid can reduce the accumulation of malonaldehyde, reduce the release of lactic dehydrogenase and creatine kinase into blood, increase the accumulation of superoxide dismutase and glutathione peroxidase, and relieve the muscle injury of mice with excessive movement.

Description

Technical Field

[0001] The invention belongs to the technical field of application of Qijing oral liquid, and specifically relates to application of a compound Qijing oral liquid in anti-fatigue effect in mice. Background Art

[0002] Sports fatigue is a form of fatigue. When the body is in high-intensity exercise, it will cause energy consumption, metabolite accumulation, internal environment imbalance, oxidative damage, low immune function, and increase in inflammatory cytokines related to muscle contraction. The typical manifestation is the increase in the production of lactic acid and urea nitrogen, which further causes muscle damage and oxidative stress response, and has an adverse effect on the recovery of normal body functions after exercise.

[0003] In order to relieve the damage caused by sports fatigue to the body, the development of Chinese medicines such as Astragalus membranaceus as medicine and food has been increased. By consulting relevant literature and expert discussions, the prescription of compound Qijing oral liquid was formed, which is composed of Astragalus membranaceus, Polygonatum sibiricum and Cistanche deserticola. The main components of Astragalus membranaceus include polysaccharides, saponins, flavonoids, etc., which have the effects of anti-fatigue, regulating immunity, anti-oxidation, and protecting cardiovascular and cerebrovascular vessels. The main components of Polygonatum sibiricum include polysaccharides, flavonoids, alkaloids, etc., which have the effects of regulating immunity, anti-aging, anti-fatigue, and protecting myocardial cells. The main components of Cistanche deserticola include phenylethanol glycosides, cycloterpenes, lignin, etc., which have the effects of improving immunity, anti-oxidation, anti-aging, and anti-fatigue. In the compound, Polygonatum sibiricum is a commonly used Chinese medicine for relieving fatigue. Astragalus membranaceus replenishes qi and nourishes yin, and Cistanche deserticola replenishes the middle qi. The whole prescription is light and mild, replenishing but not harsh. The three Chinese medicines for both medicine and food play the effects of strengthening the body, replenishing qi, and resisting fatigue. Relevant studies have shown that single Chinese medicines such as Astragalus, Polygonatum, and Cistanche have anti-fatigue effects, but there are no reports on the anti-fatigue effects of the three-drug combination. Therefore, based on the development of Astragalus Oral Liquid, this study explored the improvement effect of Astragalus Oral Liquid on the exercise endurance of mice, and determined its improvement effect on exercise fatigue by measuring relevant biochemical indicators, providing a basis for the development and research of this compound and related preparations.

[0004] In view of this, the inventors proposed the use of a compound Qijing oral liquid in the anti-fatigue effect of mice to solve the above problems. Summary of the invention

[0005] The purpose of the present invention is to provide an application of compound Qijing oral liquid in the anti-fatigue effect of mice, so as to solve the problems raised in the above background technology.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] An application of compound Qijing oral liquid in the anti-fatigue effect of mice comprises the following steps:

[0008] S1, preparing a gavage solution, preparing Qijing oral liquid according to the prescription ratio, extracting and concentrating to obtain a gavage solution;

[0009] S2, grouping and administering drugs to mice: the mice were randomly grouped and fed with different doses of the gavage solution obtained in step S1 in sequence, and a Red Bull energy drink control group and a normal control group were set up;

[0010] S3, weight-loaded exhaustive swimming test for mice, the mice grouped in step S2 are subjected to swimming training, and the time is recorded;

[0011] S4, measuring relevant indicators, sampling and measuring the remaining mice in step S2;

[0012] S5. Statistical analysis: SPSS25.0 software was used to analyze the experimental data to determine whether they conformed to the normal distribution, and then one-way analysis of variance and post hoc multiple comparisons (LSD) were performed. The results were expressed as "mean ± standard deviation".

[0013] Preferably, the method for preparing the intragastric solution in step S1 is: weighing the Cistanche deserticola, Astragalus, and Polygonatum sibiricum slices, adding purified water, soaking, reflux extraction, adding water to make up for weight loss, filtering, combining the filtrate, and concentrating to obtain the intragastric solution.

[0014] Preferably, the ratio of Cistanche deserticola, Astragalus membranaceus and Polygonatum sibiricum slices is 1:1:1, a total of 45-60 g, 10 times the amount of purified water is added, the soaking time is 1.5-2 h, reflux extraction is performed 3 times, each time for 2-3 h, water is added to make up for the weight loss, filtered, the filtrates of 3 times are combined, and concentrated to 0.5 g / mL to obtain the intragastric solution.

[0015] Preferably, in step S2, the mice are randomly divided into 5 groups: a normal control group, a Red Bull functional drink control group, a low-dose Qijing oral liquid group, a medium-dose Qijing oral liquid group, and a high-dose Qijing oral liquid group, and gavage is performed, and the body weights of the mice before and after the experiment are recorded.

[0016] Preferably, in step S2, the mice are adaptively fed for 3 days, during which they are free to eat and drink water. There are 18 mice in each group. The low, medium and high dose groups are gavaged with 1.8 g / kg·BW·d, 3.6 g / kg·BW·d and 7.2 g / kg·BW·d, respectively, which are 5, 10 and 20 times the human dose, respectively. The Red Bull functional drink group and the normal control group are gavaged with equal volumes of Red Bull drink and normal saline, respectively, once a day, 0.1 mL / 10 g each time, for 10 consecutive days. The body weight of the mice is recorded before and after the experiment.

[0017] Preferably, in step S3, the mice are gavaged for 30 minutes every day for 5 days before sampling and then subjected to unweighted swimming training, the training is 40 minutes on the first day, and the training is increased by 10 minutes every day thereafter, and 30 minutes after the end of gavage on the 11th day, a number of mice are randomly selected from each group, and a lead sheet weighing 3% to 5% of their body weight is tied to the tail for a weighted swimming test, and the time from the start of swimming to the death of the mice is recorded.

[0018] Preferably, 30 minutes after gavage on the 11th day in step S4, the remaining mice are allowed to swim without weight bearing, and blood is collected from the eyeballs after being taken out and dried, and placed in a 1.5 mL centrifuge tube. After the mice are killed, the liver and gastrocnemius muscles of both hind legs are taken, rinsed with physiological saline to remove blood stains, and divided into centrifuge tubes. The blood is centrifuged 30 minutes after sampling, and the upper serum is taken and divided into 1.5 mL centrifuge tubes. The serum and tissues are stored at -80°C; according to the method in the kit instructions, multiple indicators in the serum and liver of each group of mice are measured.

[0019] Preferably, the multiple indicators measured in mouse serum and liver include blood lactate LA, urea nitrogen BUN, superoxide dismutase SOD, malondialdehyde MDA, lactate dehydrogenase LDH, glutathione peroxidase GSH-Px in serum; and glycogen, SOD, MDA, and GSH-Px in liver.

[0020] Preferably, the blood is centrifuged at 4°C and 3500-4000 r / min for 10-15 min.

[0021] Preferably, the left gastrocnemius muscle of both hind legs of the mouse is fixed with 10% formaldehyde fixative, embedded in paraffin, cut into 4 μm sections, stained with hematoxylin-eosin, and the morphology of the muscle tissue is observed under an optical microscope.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] (1) The compound Qijing oral liquid of the present invention can reduce the glycogen consumption and BUN accumulation of exercise mice and prolong the exercise time of mice, indicating that the compound Qijing oral liquid has a certain anti-fatigue effect. The compound Qijing oral liquid composed of three kinds of Chinese medicinal materials can be used as an antioxidant and a supplement to improve exercise endurance, thereby achieving anti-fatigue function.

[0024] (2) Different doses of the compound Qijing oral liquid of the present invention all reduce the accumulation of malondialdehyde, reduce the release of lactate dehydrogenase and creatine kinase into the blood, increase the accumulation of superoxide dismutase and glutathione peroxidase, and alleviate muscle damage in mice subjected to excessive exercise. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a comparison diagram of the effect of the compound Qijing oral liquid of the present invention on glycogen in mouse tissues;

[0026] Figure 2 This is a graph showing the effect of the compound Qijing oral liquid of the present invention on the BUN index of mice;

[0027] Figure 3 This is a graph showing the effect of the compound Qijing oral liquid of the present invention on the LA index of mice;

[0028] Figure 4 This is a graph showing the effect of the compound Qijing oral liquid of the present invention on the LDH index of mice;

[0029] Figure 5 This is a graph showing the effect of the compound Qijing oral liquid of the present invention on the CK index of mice;

[0030] Figure 6 This is a graph showing the effect of the compound Qijing oral liquid of the present invention on the MDA index of mice;

[0031] Figure 7 This is a graph showing the effect of the compound Qijing oral liquid of the present invention on the SOD index of mice;

[0032] Figure 8 This is a graph showing the effect of the compound Qijing oral liquid of the present invention on the GSH-Px index of mice;

[0033] Fig. 9 The diagrams show the effects of the compound Qijing oral liquid of the present invention on the gastrocnemius muscle structure of mice, (a) NC Group, (b) HN Group, (c) LG Group, (d) MG Group, (e) HG Group;

[0034] Fig.10 The present invention is a flowchart of the application of the compound Qijing oral liquid in the anti-fatigue effect on mice. DETAILED DESCRIPTION

[0035] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0036] Embodiment 1:

[0037] See also Figures 1 to 6 As shown, the use of a compound Qijing oral liquid in the anti-fatigue effect of mice comprises the following steps:

[0038] S1. Prepare intragastric solution. Weigh 45 g of Cistanche deserticola, Astragalus membranaceus and Polygonatum sibiricum slices in a ratio of 1:1:1, add 10 times the amount of purified water, soak for 1.5 h, reflux extraction 3 times, each time for 2 h, add water to make up for weight loss, filter, combine the 3 filtrates, and concentrate to 0.5 g / mL to obtain intragastric solution.

[0039] S2. Mice were divided into groups for drug administration. They were adaptively fed for 3 days from the time of purchase, and were free to eat and drink water during the period. The mice were randomly divided into 5 groups: normal control group, Red Bull functional drink control group, Qijing oral liquid low-dose group, medium-dose group, and high-dose group, with 18 mice in each group. According to the results of the preliminary experiment, the low, medium, and high-dose groups were gavaged at 1.8g / (kg·BW·d), 3.6g / (kg·BW·d), and 7.2g / (kg·BW·d) (5, 10, and 20 times the human dose, respectively), and the Red Bull functional drink group and the normal control group (gavage with an equal volume of Red Bull drink and normal saline) were gavaged once a day, 0.1mL / 10g each time, for 10 consecutive days. The weight of the mice was recorded before and after the experiment.

[0040] S3. Weight-exhausted swimming test for mice. For 5 days before sampling, mice were gavaged for 30 minutes every day and then subjected to unweighted swimming training. All mice were placed in a swimming box with a water depth of 30 cm and a water temperature of 25°C. During the period, mice were driven away with wooden sticks to ensure that the mice were in a state of continuous swimming. The training was conducted for 40 minutes on the first day, and then increased by 10 minutes every day. 30 minutes after the gavage on the 11th day, 9 mice were randomly selected from each group, and a lead sheet of 5% of their body weight was tied to the tail for a weighted swimming test. The time from the beginning of swimming to death of the mice was recorded.

[0041] S4. Determination of relevant indicators: 30 minutes after gavage on the 11th day, the remaining 45 mice were allowed to swim for 90 minutes without weight bearing, and blood was collected from the eyeballs after being taken out and dried, and placed in 1.5mL centrifuge tubes. After the mice were killed, the liver and gastrocnemius muscles of both hind legs were taken, rinsed with physiological saline to remove blood stains, and divided into centrifuge tubes. 30 minutes after sampling, the blood was centrifuged at 4℃ (3500r / min, 10min), and the upper serum was divided into 1.5mL centrifuge tubes. The serum and tissues were stored at -80℃.

[0042] According to the instructions of the kit, LA (blood lactate), BUN (urea nitrogen), SOD (superoxide dismutase), MDA (malondialdehyde), LDH (lactate dehydrogenase), and GSH-Px (glutathione peroxidase) were measured in the serum of each group of mice; and glycogen, SOD, MDA, and GSH-Px were measured in the liver.

[0043] Morphological observation of gastrocnemius muscle tissue: The left posterior gastrocnemius muscles were fixed with 10% formaldehyde fixative, embedded in paraffin, cut into 4 μm sections, and stained with hematoxylin-eosin. The morphology of muscle tissue was observed under an optical microscope.

[0044] S5. Statistical analysis: SPSS25.0 software was used to analyze the experimental data to determine whether they conformed to the normal distribution, and then one-way analysis of variance and post hoc multiple comparisons (LSD) were performed. The results were expressed as "mean ± standard deviation". P < 0.05 indicated that the differences between the groups were statistically significant.

[0045] The experimental instruments, materials and animal information involved in this invention are as follows:

[0046] Test equipment

[0047] ME204 electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd.); Ecopure ultrapure water machine (Chongqing Yiyang Enterprise Development Co., Ltd.); 5424R refrigerated centrifuge (Eppendorf, Germany); DE06082 microplate reader (Molecular Instrument (Shanghai) Co., Ltd., USA); U410 ultra-low temperature refrigerator (Eppendorf, Germany); swimming box (Shanghai Yuyan Scientific Instrument Co., Ltd.); lead sheets and handmade ceramic mortars were purchased from local wholesale markets.

[0048] Test reagents

[0049] Blood urea nitrogen (BUN) assay kit, blood lactate (LA) assay kit, liver glycogen (LG) assay kit, and muscle glycogen (MG) assay kit were all purchased from Shanghai ELISA Biotechnology Co., Ltd.; lactate dehydrogenase (LDH) assay kit, superoxide dismutase (SOD) assay kit, malondialdehyde (MDA) test kit, creatine kinase (CK) test kit, and glutathione reductase (GSH-Px) assay reagent were all purchased from Shanghai Yuanju Biotechnology Center; water was homemade purified water.

[0050] Test materials

[0051] Cistanche decoction pieces (batch number 20180711) and Astragalus decoction pieces (batch number 20180711) were purchased from Anning Xinglin Medicinal Materials Wholesale Department, Lanzhou City, Gansu Province; Polygonatum sibiricum decoction pieces (batch number 20190127) were purchased from Longnan Huikang Chinese Medicinal Materials Co., Ltd. The above decoction pieces were inspected and identified as authentic by Shi Xiaofeng, chief pharmacist of Gansu Academy of Medical Sciences, and they comply with the relevant provisions of the 2020 edition of the Pharmacopoeia of the People's Republic of China (Volume 1) under each medicinal material decoction piece.

[0052] animal

[0053] 90 SPF male Balb / C mice, 6-8 weeks old, weighing 20±2g. They were purchased from Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, with the animal production license number: SCXK(Gan)-2023-0046. The mouse breeding environment: temperature (25±2)℃, humidity (50±5)%, day and night light and dark alternation time of 12 / 12h. The mice were fed for 3 days before the experiment. The experimental plan was approved by the Experimental Animal Ethics Review Committee of Lanzhou Food and Drug Inspection and Testing Institute, and was carried out in strict accordance with relevant regulations, with the ethics review approval number: 2022-002.

[0054] Effect of compound Qijing oral liquid on body weight of mice:

[0055] The amount of gavage administered to mice varied with their body weight. Table 1 reflects the changes in body weight of mice before and after the experiment.

[0056] Table 1 Body weight changes of mice before and after the experiment ( n=9)

[0057]

[0058] Table 1

[0059] The results showed that different intragastric doses of normal saline, Red Bull functional drink, and compound Qijing oral liquid had no significant effect on the body weight of mice.

[0060] Embodiment 2:

[0061] Effect of compound Qijing oral liquid on weight-loaded exhaustive swimming time in mice:

[0062] As shown in Table 2, compared with the normal control group gavaged with normal saline, the low-dose (P<0.05), medium-dose (P<0.01), and high-dose (P<0.01) compound Qijing oral liquid group and Red Bull functional drink group (P<0.01) gavaged with low-dose (P<0.05), medium-dose (P<0.01), and high-dose (P<0.01) compound Qijing oral liquid group and Red Bull functional drink group (P<0.01) can prolong the weight-bearing exhaustive swimming time of mice, and the medium-dose and high-dose groups prolong the swimming time of mice more significantly than the low-dose group. This indicates that compound Qijing oral liquid and Red Bull functional drink can enhance the exercise endurance of mice; compared with the Red Bull functional drink control group, the high-dose group can significantly prolong the weight-bearing exhaustive swimming time of mice (P<0.01); and the high-dose group is better than the medium and low-dose groups. This indicates that the high-dose compound Qijing oral liquid has a better anti-fatigue effect than Red Bull functional drink, and different doses of compound Qijing oral liquid can significantly improve the endurance of mice.

[0063] Table 2 Effect of compound Qijing oral liquid on the duration of weight-bearing exhaustive swimming in mice ( n=9)

[0064]

[0065] Table 2

[0066] Note: *P<0.05 indicates a significant difference compared with the blank control group, **P<0.01 indicates an extremely significant difference compared with the blank control group; #P<0.05 indicates a significant difference compared with the Red Bull energy drink control group, ##P<0.01 indicates an extremely significant difference compared with the Red Bull energy drink control group.

[0067] Embodiment three:

[0068] Effects of compound Qijing oral liquid on glycogen in liver and muscle of mice:

[0069] From Table 3 and Figure 1 It can be seen that compared with the normal control group, the liver glycogen content in different doses of oral liquid groups and the Red Bull functional drink control group was significantly increased (P<0.01), and the muscle glycogen and liver glycogen content in the medium dose group (P<0.05), high dose group (P<0.01), and Red Bull functional drink control group (P<0.01) were significantly increased; compared with the Red Bull functional drink control group, the muscle glycogen content in the high dose group increased but there was no significant difference, and the liver glycogen content in the medium and high dose groups was significantly increased (P<0.01).

[0070] Table 3 Effect of compound Qijing oral liquid on the content of liver glycogen and muscle glycogen in mouse tissues ( n=9)

[0071]

[0072] Table 3

[0073] Note: *P<0.05 indicates a significant difference compared with the blank control group, **P<0.01 indicates an extremely significant difference compared with the blank control group; #P<0.05 indicates a significant difference compared with the Red Bull energy drink control group, ##P<0.01 indicates an extremely significant difference compared with the Red Bull energy drink control group.

[0074] It can be seen from this that both Compound Qijing Oral Liquid and Red Bull Energy Drink can increase the replenishment of glycogen in mice after exercise.

[0075] Embodiment 4:

[0076] Effects of compound Qijing oral liquid on biochemical parameters in mouse serum:

[0077] From Table 4 and Figure 2 - Figure 5It can be seen that compared with the normal control group, the serum BUN level in the low-dose group (P<0.05), medium-dose group, high-dose group and Red Bull functional drink control group was significantly reduced (P<0.01); compared with the Red Bull functional drink control group, the serum BUN level in the high-dose group (P<0.05) was significantly reduced, but there was no significant difference between the low and medium dose groups (P>0.05).

[0078] Compared with the normal control group, the serum LA levels in the different dose gavage groups and the Red Bull energy drink control group were significantly decreased (P<0.01); compared with the Red Bull energy drink control group, the serum LA level in the high dose group (P<0.01) was significantly decreased, and the serum LA level in the medium dose group was also decreased but with no significant difference (P>0.05).

[0079] Compared with the normal control group, the serum LDH levels in the medium-dose group, high-dose group and Red Bull functional drink group were significantly decreased (P<0.01); compared with the Red Bull functional drink control group, the serum LDH levels in the medium-dose group and high-dose group were decreased but there was no significant difference (P>0.05).

[0080] Compared with the normal control group, the serum CK levels in the medium and high dose groups and the Red Bull energy drink control group were significantly reduced (P<0.01), while there was no significant difference in the low dose group (P>0.05); compared with the Red Bull energy drink control group, the serum CK levels in the medium and high dose gavage groups were significantly reduced (P<0.01).

[0081] This indicates that compound Qijing oral liquid can promote energy metabolism in exercising mice, reduce the accumulation of metabolites, and thus play an anti-fatigue role.

[0082] Table 4 Effects of compound Qijing oral liquid on biochemical indexes of mice ( n=9)

[0083]

[0084] Table 4

[0085] Note: *P<0.05 indicates a significant difference compared with the blank control group, **P<0.01 indicates an extremely significant difference compared with the blank control group; #P<0.05 indicates a significant difference compared with the Red Bull energy drink control group, ##P<0.01 indicates an extremely significant difference compared with the Red Bull energy drink control group.

[0086] Embodiment five:

[0087] Effects of compound Qijing oral liquid on antioxidant indexes in mice:

[0088] Table 5 Effects of compound Qijing oral liquid on antioxidant indexes in mice ( n=9)

[0089]

[0090]

[0091] Table 5

[0092] Note: *P<0.05 indicates a significant difference compared with the blank control group, **P<0.01 indicates an extremely significant difference compared with the blank control group; #P<0.05 indicates a significant difference compared with the Red Bull energy drink control group, ##P<0.01 indicates an extremely significant difference compared with the Red Bull energy drink control group

[0093] From Table 5 and Figure 6 - Figure 8 It can be seen that compared with the normal control group, the MDA content in serum and liver of the oral compound Qijing liquid group and the Red Bull functional drink control group was significantly reduced (P<0.01); compared with the Red Bull functional drink control group, the MDA content in serum and liver of the high-dose group was significantly reduced (P<0.01). The MDA activity in serum and liver increased with the increase of oral dose.

[0094] Compared with the normal control group, the SOD activities in serum and liver of the oral compound Qijing liquid group and the Red Bull functional drink control group were significantly increased after intragastric administration (P<0.01); compared with the Red Bull functional drink control group, the SOD activities in serum and liver of the high-dose group were significantly increased (P<0.01). The SOD activities in serum and liver increased with the increase of intragastric administration dose.

[0095] Compared with the normal control group, the GSH-Px activity in the liver of the oral compound Qijing liquid group and the Red Bull functional drink control group was significantly enhanced (P<0.01). In the Red Bull functional drink control group, the serum GSH-Px activity in the medium and high dose groups was significantly enhanced (P<0.01), and the serum GSH-Px activity in the low dose group was enhanced but there was no significant difference. Compared with the Red Bull functional drink control group, the liver and serum GSH-Px activity in the high dose group was significantly enhanced (P<0.01). The serum and liver GSH-Px activity increased with the increase of the oral dose.

[0096] The activities of SOD and GSH-Px in the different doses of Compound Qijing Oral Liquid groups were significantly enhanced, and the MDA level was significantly reduced, indicating that Compound Qijing Oral Liquid can reduce antioxidant damage in exercised mice, alleviate lipid peroxidation and increase the activity of antioxidant enzymes, and the antioxidant stress capacity is dose-dependent with the dose of Compound Qijing Oral Liquid.

[0097] Embodiment six:

[0098] As a modern health product integrating the wisdom of traditional Chinese medicine, the compound Qijing oral liquid designed in this paper has shown significant anti-fatigue effects in mouse experiments, indicating that it has broad and real application prospects in promoting human physical recovery, improving sports endurance and alleviating sports fatigue.

[0099] Furthermore, the effect of compound Qijing oral liquid on the gastrocnemius muscle structure of mice:

[0100] HE staining was used to observe the morphological changes of the gastrocnemius muscle structure in mice. Fig. 9 As shown in the figure, in the normal control group, the muscle fibers were arranged in a disordered manner, with uneven sizes and thicknesses, wide gaps, aggregated nuclei, and local congestion. In comparison, the muscle fibers in the low-dose group were arranged slightly neatly, with smaller gaps, but the nuclei were still obviously aggregated, and there was still local congestion; the muscle fibers in the medium-dose group were arranged neatly, but the nuclei were relatively evenly distributed, without congestion and inflammatory cell infiltration; the muscle fibers in the high-dose group were arranged neatly, with small gaps, evenly distributed nuclei, and no congestion and inflammatory cell infiltration; the muscle fibers in the Red Bull functional drink control group were arranged neatly, with slightly larger gaps than the high-dose group, evenly distributed nuclei, and no congestion and inflammatory cell infiltration. Compared with the normal control group, the histological observation results of the gastrocnemius in the five groups were significantly different. This shows that high-intensity exercise causes skeletal muscle damage in mice, and compound Qijing oral liquid can reduce muscle damage, and the degree of damage is positively correlated with the dose.

[0101] As can be seen from the above, the present invention measures the relevant biochemical indexes, antioxidant indexes, and skeletal muscle tissue morphology of mice after swimming training and fatigue mice after a certain period of swimming. The generation of fatigue is closely related to factors such as energy material consumption, metabolite accumulation, and increased oxidative stress levels. Weight-bearing exhaustive swimming is one of the important indicators for evaluating anti-fatigue effects, and the swimming duration can directly reflect the body's exercise endurance. Mice perform aerobic exercise by weighted swimming in a short period of time, and the glycogen in the body is consumed and converted into glucose to provide the first energy source. Therefore, the changes in LG and MG content are important indicators for judging fatigue status. When the body is in a long-term high-intensity exercise process, the energy generated by aerobic respiration is not enough to meet the needs of the organization, and muscle movement will be converted into anaerobic exercise, and the protein and amino acids in the skeletal muscle will be decomposed to produce too much lactic acid, and the metabolite BUN content will increase accordingly. The present invention's research shows that the compound Qijing oral liquid can reduce the glycogen consumption and BUN accumulation of exercise mice, and prolong the exercise time of mice, indicating that the compound Qijing oral liquid has a certain anti-fatigue effect.

[0102] Muscle damage and excessive stretching of muscle fibers caused by high-intensity exercise cause changes in serum CK levels. The role of LDH is to remove lactic acid produced by the body. LDH and CK leak into the blood, increasing the CK content in the serum. At the same time, strenuous exercise will cause the human body to consume a lot of energy, produce too much reactive oxygen, lead to the generation of free radicals, cause oxidative stress, increase the level of lipid peroxide MDA in the serum, and increase antioxidant enzymes such as SOD and GSH-Px to remove free radicals and prevent lipid peroxidation. The present invention shows that different doses of compound Qijing oral liquid reduce the accumulation of malondialdehyde; reduce the release of lactate dehydrogenase and creatine kinase into the blood; increase the accumulation of superoxide dismutase and glutathione peroxidase; and reduce muscle damage in mice with excessive exercise. The reason may be related to the effective ingredients in the compound Qijing oral liquid, proving that the total polysaccharides in Astragalus, Polygonatum, and Cistanche have anti-fatigue effects. It shows that the compound Qijing oral liquid composed of three kinds of Chinese medicinal materials can be used as an antioxidant and a supplement to improve exercise endurance, thereby achieving anti-fatigue function. The invention can provide a theoretical basis for the subsequent development of Qijing health products.

[0103] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An application of compound Qijing oral liquid in the anti-fatigue effect of mice, characterized in that: The following steps are involved: S1. preparing a gavage solution, preparing Qijing oral liquid according to the prescription ratio, extracting and concentrating to obtain a gavage solution; S2, grouping and administering drugs to mice: the mice were randomly grouped and fed with different doses of the gavage solution obtained in step S1 in sequence, and a Red Bull energy drink control group and a normal control group were set up; S3, weight-loaded exhaustive swimming test for mice, the mice grouped in step S2 are subjected to swimming training, and the time is recorded; S4, measuring relevant indicators, sampling and measuring the remaining mice in step S2; S5. Statistical analysis: SPSS25.0 software was used to analyze the experimental data to determine whether they conformed to the normal distribution, and then one-way analysis of variance and post hoc multiple comparisons (LSD) were performed. The results were expressed as "mean ± standard deviation".

2. The use of a compound Qijing oral liquid in the anti-fatigue effect of mice according to claim 1, characterized in that: The method for preparing the intragastric solution in step S1 is: weighing the Cistanche deserticola, Astragalus, and Polygonatum sibiricum slices, adding purified water, soaking, reflux extraction, adding water to make up for weight loss, filtering, combining the filtrate, and concentrating to obtain the intragastric solution.

3. The use of a compound Qijing oral liquid in the anti-fatigue effect of mice according to claim 2, characterized in that: Weigh 45-60 g of Cistanche deserticola, Astragalus membranaceus, and Polygonatum sibiricum slices in a ratio of 1:1:1, add 10 times the amount of purified water, soak for 1.5-2 h, reflux extraction 3 times, each time for 2-3 h, add water to make up for the weight loss, filter, combine the 3 filtrates, and concentrate to 0.5 g / mL to obtain the intragastric solution.

4. The use of a compound Qijing oral liquid in the anti-fatigue effect of mice according to claim 1, characterized in that: In step S2, the mice were randomly divided into five groups: a normal control group, a Red Bull functional drink control group, a low-dose Qijing oral liquid group, a medium-dose Qijing oral liquid group, and a high-dose Qijing oral liquid group. The mice were gavaged and their body weights were recorded before and after the experiment.

5. The use of a compound Qijing oral liquid in the anti-fatigue effect of mice according to claim 4, characterized in that: In step S2, the mice were adaptively fed for 3 days, during which they were free to eat and drink water. There were 18 mice in each group. The low, medium and high dose groups were gavaged with 1.8 g / kg·BW·d, 3.6 g / kg·BW·d and 7.2 g / kg·BW·d, respectively, which are 5, 10 and 20 times the human dose, respectively. The Red Bull functional drink group and the normal control group were gavaged with equal volumes of Red Bull drink and normal saline, respectively, once a day, 0.1 mL / 10 g each time, for 10 consecutive days. The body weights of the mice were recorded before and after the experiment.

6. The use of a compound Qijing oral liquid in the anti-fatigue effect on mice according to claim 1, characterized in that: In step S3, the mice were gavaged for 30 minutes every day for 5 days before sampling, and then subjected to unweighted swimming training. The training was conducted for 40 minutes on the first day, and then increased by 10 minutes every day. 30 minutes after the end of gavage on the 11th day, several mice were randomly selected from each group, and lead sheets weighing 3% to 5% of their body weight were tied to their tails for a weighted swimming test. The time from the start of swimming to the death of the mice was recorded.

7. The use of a compound Qijing oral liquid in the anti-fatigue effect on mice according to claim 1, characterized in that: 30 minutes after gavage on the 11th day in step S4, the remaining mice were allowed to swim without weight bearing, and blood was collected from the eyeballs after being taken out and dried, and placed in 1.5 mL centrifuge tubes. After the mice were killed, the liver and gastrocnemius muscles of both hind legs were taken, rinsed with physiological saline to remove blood stains, and divided into centrifuge tubes. The blood was centrifuged 30 minutes after the sample was taken, and the upper serum was taken and divided into 1.5 mL centrifuge tubes. The serum and tissues were stored at -80°C; according to the method in the kit instructions, multiple indicators in the serum and liver of each group of mice were measured.

8. The use of a compound Qijing oral liquid in the anti-fatigue effect on mice according to claim 7, characterized in that: Multiple indicators measured in mouse serum and liver include blood lactate LA, urea nitrogen BUN, superoxide dismutase SOD, malondialdehyde MDA, lactate dehydrogenase LDH, glutathione peroxidase GSH-Px in serum; and glycogen, SOD, MDA, and GSH-Px in the liver.

9. The use of a compound Qijing oral liquid in the anti-fatigue effect on mice according to claim 7, characterized in that: The conditions for blood centrifugation are 4°C, 3500-4000 r / min for 10-15 min.

10. The use of a compound Qijing oral liquid in the anti-fatigue effect on mice according to claim 7, characterized in that: The left gastrocnemius muscle of both hind legs of mice was fixed with 10% formaldehyde fixative, embedded in paraffin, cut into 4 μm sections, stained with hematoxylin-eosin, and the morphology of the muscle tissue was observed under an optical microscope.

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