CN114224978B

CN114224978B - Traditional Chinese medicine compound composition with anti-fatigue function and application thereof

Info

Publication number: CN114224978B
Application number: CN202210092781.6A
Authority: CN
Inventors: 邹圣灿; 李鑫; 董海燕; 王杰; 李莉; 王尚龙; 宗建成
Original assignee: Qingdao Chenlan Health Industry Group Co ltd; Chenlan Usa Nutritional Products Co ltd
Current assignee: Qingdao Chenlan Health Industry Group Co ltd; Chenlan Usa Nutritional Products Co ltd
Priority date: 2022-01-26
Filing date: 2022-01-26
Publication date: 2023-02-03
Anticipated expiration: 2042-01-26
Also published as: CN114224978A

Abstract

The invention discloses a Chinese medicinal compound composition with an anti-fatigue function and application thereof.

Description

Traditional Chinese medicine compound composition with anti-fatigue function and application thereof

Technical Field

The invention belongs to the technical field of natural medicines, in particular relates to a formula of a traditional Chinese medicine compound composition and application thereof, and particularly relates to a traditional Chinese medicine compound composition with an anti-fatigue function and application thereof.

Background

Fatigue is a complex physiological phenomenon which exists widely, serious pathological reactions such as cardiovascular dysfunction and mental disorder can be induced by long-term fatigue, the harm to human health is great, if the fatigue cannot be eliminated in time, chronic fatigue syndrome can be caused, and the life and health of people are seriously influenced. At present, the efficacy of chemical drugs for relieving fatigue is limited, so that natural anti-fatigue drugs have small toxic and side effects, wide raw material sources and almost no addiction and become research and development hotspots.

Furthermore, although the understanding of the pathogenesis of exercise-induced fatigue by Chinese medical researchers is different, the treatment is based on qi, blood, yin and yang and the whole zang-fu organs. In the market for providing energy by resisting fatigue, popular raw materials such as maca roots, L-carnitine, theanine, B vitamins and the like are mainly used, and few plants are extracted to obtain a compound; in the energy supplement raw materials, the plant extract compound is a blank.

Therefore, how to develop a natural Chinese herbal compound composition with an anti-fatigue function is a problem that needs to be solved urgently by the technical personnel in the field.

Disclosure of Invention

In view of this, the present invention aims to solve the problems in the prior art, and provides a traditional Chinese medicine compound composition with an anti-fatigue function and an application thereof in the preparation of health care products.

The traditional Chinese medicine compound composition with the anti-fatigue function disclosed by the invention is developed to resist fatigue and relieve physical discomfort and muscle weakness caused by fatigue aiming at wide demands of anti-fatigue products in the market. The system is suitable for the juvenile group with repeated examination of learning tasks; workers who have high working pressure and are prone to long-term fatigue; the driver who is easy to be tired in sight during long-term driving, the driver who likes body-building can reach the person, and the like, so that the main symptoms related to the CFS (chronic fatigue syndrome) are reduced. The natural antifatigue medicine has little toxic and side effect, wide raw material source and no caffeine, can be used as a caffeine substitute to enhance the exercise capacity and improve the physical and mental performance, and has almost no addiction.

In order to achieve the purpose, the invention adopts the following technical scheme:

a traditional Chinese medicine compound composition with an anti-fatigue function is mainly prepared by separately extracting rhodiola rosea, rhizoma polygonati and hairyvein agrimony and mixing, and comprises the following components in percentage by weight:

10 to 40 percent of rhodiola rosea extract;

40 to 70 percent of rhizoma polygonati extract;

10 to 40 percent of agrimony extract.

Preferably, the traditional Chinese medicine compound composition comprises the following components in percentage by weight:

25% of rhodiola rosea extract;

50% of polygonatum extract;

25% of hairyvein agrimonia herb and bud extract.

Further, the traditional Chinese medicine compound composition also comprises a cassia twig extract, and the traditional Chinese medicine compound composition comprises the following components in percentage by weight:

40 to 70 percent of rhizoma polygonati extract; 10 to 40 percent of rhodiola rosea extract; 10 to 40 percent of hairyvein agrimony extract and 10 to 40 percent of cassia twig extract.

40% of polygonatum extract; 15% of rhodiola rosea extract; 31 percent of hairyvein agrimony extract and 14 percent of cassia twig extract.

Further, the traditional Chinese medicine compound composition also comprises a ginger extract, and the traditional Chinese medicine compound composition comprises the following components in percentage by weight:

40 to 70 percent of rhizoma polygonati extract; 10 to 40 percent of rhodiola rosea extract; 10 to 40 percent of hairyvein agrimony extract and 10 to 40 percent of ginger juice.

46% of polygonatum sibiricum extract; rhodiola rosea extract 9%; 19% of hairyvein agrimonia herb and bud extract and 26% of ginger juice.

Further, the compound traditional Chinese medicine composition also comprises a broadleaf holly leaf extract, and the compound traditional Chinese medicine composition comprises the following components in percentage by weight:

40 to 70 percent of sealwort extract; 10 to 40 percent of rhodiola rosea extract; 10 to 40 percent of hairyvein agrimony extract and 10 to 40 percent of broadleaf holly leaf extract.

40 percent of polygonatum extract; 10% of rhodiola rosea extract; 20% of hairyvein agrimony extract and 30% of broadleaf holly leaf extract.

The invention discloses a Chinese medicinal compound composition with an anti-fatigue function, which is prepared from rhodiola rosea, rhizoma polygonati, hairyvein agrimony, ginger, cassia twig and broadleaf holly leaf according to a certain weight percentage.

In addition, the current research on natural anti-fatigue drugs is as follows:

1) Insect research with anti-fatigue effect

Insects contain abundant amino acids and polypeptide fragments, and the content of vitamins, minerals and bioactive components is also very high, so the insect-resistant active substance is a key research field of natural anti-fatigue substances. The main mechanisms are as follows: (1) improving the oxidation resistance of the organism; (2) influencing the storage and mobilization of energetic materials in the body; (3) reducing the accumulation of metabolites; (4) it is also possible to exert its antifatigue effect by enhancing the body's immune function.

2) Study on fungi with anti-fatigue effect

Various kinds of various anti-fatigue edible fungi are contained in nature, some of which are widely developed and utilized, such as shiitake, ganoderma lucidum, cordyceps sinensis, poria cocos and the like, and some of which are in urgent need to be developed. Many fungi contain active polysaccharide, alkaloid, trace elements, etc., and most of the substances have the functions of improving immunity, regulating neuroendocrine, protecting cardiovascular system, etc., and have the function of resisting fatigue.

3) Marine organism research with anti-fatigue effect

The sea is the most abundant area of the largest species on the earth, and contains a plurality of animal and plant resources with novel structures and specific functions, such as oysters, stichopus japonicus, spirulina and the like. Abundant marine organisms contain a large amount of components with biological activity, and the compound preparation is an important treasury for anti-fatigue research.

4) Research on Chinese herbal medicine with anti-fatigue effect

In long-term experimental practice, traditional Chinese medicine summarizes a batch of edible Chinese herbal medicines with anti-fatigue effect, wherein a lot of Chinese herbal medicines with clear anti-fatigue function are approved by Ministry of health, and are summarized as follows: rhizomes of plants (radix Panacis Quinquefolii, ginseng radix, radix Codonopsis, polygoni Multiflori radix, radix Puerariae, rhizoma Dioscoreae, radix Ophiopogonis, etc.); fruits of plants (hawthorn, sea buckthorn, mulberry, wolfberry, etc.); the flower and leaf of plant (radix Rhodiolae, bulbus Lilii, pollen, folium Ginkgo, aloe, etc.). In recent years, most of researches have focused on the verification of the anti-fatigue effect of the Chinese herbal medicines and the discussion of the anti-fatigue mechanism. The research on the anti-fatigue effect of the Chinese herbal medicines, whether in quantity or content form, has rich results and has prominent anti-fatigue effect, so that the Chinese herbal medicines are an important field of anti-fatigue research.

In addition, the traditional Chinese medicinal materials used in the traditional Chinese medicine extract disclosed by the invention are screened according to scientific basis, organically combined and prepared according to the traditional Chinese medicine theory, rather than simply superposed according to the efficacy of a single traditional Chinese medicine;

and the invention searches and summarizes the traditional Chinese medicine with the anti-fatigue effect through the safety and the effectiveness and the selection principles of referring to American DS, ODI and the like, and preferably selects the compatibility of the sealwort, the agrimony, the rhodiola rosea, the cassia twig, the ginger and the broadleaf holly leaf under the guidance of the traditional Chinese medicine theory.

Specifically, the efficacy of each Chinese medicinal material is as follows:

rhizoma polygonati is sweet, mild and nontoxic in taste, and has the effects of tonifying middle-jiao and Qi, eliminating wind and dampness, calming the five internal organs, reducing weight after being taken for a long time, and prolonging life without hunger. Modern researches show that rhizoma polygonati contains various chemical components such as polysaccharide, oligosaccharide, flavone, steroid saponin, alkaloid, lignan, amino acid, inorganic elements, volatile oil and the like. The sealwort polysaccharide has various pharmacological activities such as oxidation resistance, has good total reducing capability and capacity of removing DPPH and ABTS free radicals, shows good oxidation resistance, regulates immunity, can remarkably relieve spleen hypoimmunity caused by forced exercise and restore the spleen hypoimmunity to a normal level, and can enhance erythrocyte immune function of children suffering from nephrotic syndrome. Research shows that the polygonatum polysaccharide can obviously prolong the weight swimming time of mice, increase the content of glycogen of the liver of the mice and have obvious anti-fatigue effect.

The herba et Gemma Agrimoniae contains flavonoids, triterpenes, volatile oil, tannin, phenols, etc., and contains trace elements such as magnesium, potassium, phosphorus, iron, sulfur, etc. The pharmacological activity research mainly has antioxidant activity, antitumor effect, anti-inflammatory effect, etc. Research shows that the myocardial MDA content of the rats with the exercise-induced fatigue is obviously increased, the activities of SOD, GSH-Px and T-AOC are obviously reduced, and the alcohol extract of hairyvein agrimony can increase the exercise time of the rats with the exercise-induced fatigue, reduce the myocardial MDA content, increase the activities of SOD, GSH-Px and T-AOC, so that the alcohol extract of hairyvein agrimony can relieve the myocardial oxidative stress injury of the exercise-induced fatigue by improving the myocardial oxidation resistance and reducing the lipid peroxidation of the rats with the exercise-induced fatigue. The hypoactive vitamin has the advantages of low LDH content and high LA content in blood serum of sports fatigue rats, which shows that the anaerobic metabolism capability of an organism is weakened, and simultaneously, the free radical MDA content and the SOD activity of the rat are improved, which shows that the scavenging capability of the organism to free radicals is reduced and the oxidation resistance is weakened. After the agrimony extract is administrated for 14 days by gastric lavage, the LA and MDA contents in the serum of a rat are reduced, and the activities of SOD and LDH are enhanced, which indicates that the agrimony has a certain anti-fatigue effect.

Rhodiola rosea contains glucoside, flavone, alkaloid, phenols, volatile oil, coumarins, steroids, organic acid, trace inorganic elements and the like. Research shows that rhodiola can obviously prolong the swimming time of mice, increase the content of liver glycogen, reduce the content of serum BUN and BLA, and has good anti-fatigue activity. In vitro electrical stimulation of isolated toad gastrocnemius muscle is used as a muscle contraction fatigue model to observe the influence of rhodiola rosea polysaccharide on the motion of the toad gastrocnemius muscle. The results show that rhodiola polysaccharide can eliminate free radical and increase Na ⁺ -K ⁺ -ATPase and Ca ²⁺ -Mg ²⁺ -ATPase activity. Researches show that the rhodiola rosea can obviously improve exercise endurance of mice with large-intensity exercise amount, improve the levels of mitochondrion division related genes Drp-1 and mitochondrion autophagy initiation factors in skeletal muscles of the mice and reduce the levels of mitochondrial fusion related genes Mfn-1, mfn-2 and Opa-1. Salidroside intervention is carried out on a fatigue model rat, then the change of central neurotransmitter of the rat is observed, and the result shows that rhodiola rosea is subjected toThe glycoside can reduce 5-HT content in rat brain, and increase DA and NE content and ratio of DA and 5-HT. The rhodiola root extract SHR-5 (a stabilized extract by Swedish Herbal Institute) was found to inhibit the increase of cortisol level. The researches show that the rhodiola rosea polysaccharide and the salidroside can relieve peripheral fatigue by increasing the storage of energy substances, reducing the accumulation of metabolites and improving the mitophagy and mitochondrion fusion-division of skeletal muscle mitochondria, delay central fatigue by improving the content of central neurotransmitter and also relieve pathological fatigue by regulating the HPA axis.

Kuding tea is a famous national medicine in Yangtze river basin and south area thereof, is bitter and sweet in taste, is cool in nature, enters liver, lung and stomach, and has the effects of clearing summer heat, detoxifying and promoting fluid production. Modern pharmacological research shows that the ilex kudingcha has the activities of reducing fat and losing weight, reducing blood sugar, reducing blood pressure, resisting atherosclerosis, resisting oxidation, resisting bacteria, resisting viruses and the like. There are documents in fatigue resistance tests: the experimental mice are weighed and divided into two groups, wherein the group A is used for drinking normal water, and the group B is subjected to intragastric administration of the broadleaf holly leaf liquid for 0.4 ml/mouse (divided into two times of administration) for 7 days, and then the experiment is carried out. After 7 days, the mice were put into constant temperature water (28 Shi 1) deg.C for swimming test, and the swimming time of each mouse was recorded by using the inability of the mice to float and sink to the water bottom as an observation index. The experimental result also proves that the broadleaf holly leaf tea has the functions of improving the hypoxia tolerance and low temperature resistance of the mice and improving the stress of the mice when the mice move.

In addition, animal behavior experiments such as swimming exhaustion experiments, running table experiments, hypoxia tolerance experiments and the like and biochemical indexes are preferably selected for carrying out curative effect verification so as to research the vigor improving effect of the traditional Chinese medicine compound composition, and research results show that the traditional Chinese medicine compound composition has an obvious vigor improving effect.

In addition, in the prior art, although the common dosage of each traditional Chinese medicine is known, the prescription is an organic whole prepared by combining a plurality of medicines according to diseases, the drug effect of the prescription is not equal to the simple superposition of each medicine on the basis of the common dosage, the dosage of each traditional Chinese medicine in the prescription can not be determined according to the common dosage of each traditional Chinese medicine, the dosage proportion of the medicines is determined by various considerations such as the characteristics of the medicines, monarch, minister, assistant and guide compatibility and the like, and the prescription can not be obtained by simple experimental means such as a comparison method, an orthogonal method and the like.

Further, the preparation method of the traditional Chinese medicine compound composition with the anti-fatigue function comprises the following steps:

(1) The traditional Chinese medicine extract can be prepared by a heating reflux extraction method, the rhodiola root medicinal material is added with 6 to 12 times of aqueous solution, the heating reflux extraction is carried out for 2 to 3 times, each time lasts for 1 to 2.0 hours, the filter liquor is merged, the filtration and the centrifugation are carried out, the supernatant fluid is concentrated to the relative density of about 1.02 to 1.05, 80 to 95 percent of ethanol is added for stirring until the alcohol content is 60 to 70 percent, the mixture is kept stand for 12 to 24 hours, the precipitate is filtered, the supernatant fluid is concentrated and then is dried under reduced pressure to obtain the rhodiola root extract A;

(2) Cutting rhizoma Polygonati into pieces, reflux-extracting with 6-8 times of water for 2-3 times each for 1.5-2.0 hr, mixing the filtrates, concentrating to relative density of 1.10-1.20, precipitating with 70-80% ethanol, magnetically stirring for 1 hr, standing for more than 24 hr, and spin-drying the precipitate to obtain B;

(3) Extracting herba et Gemma Agrimoniae with 4-6 times of 60-80% ethanol for 2-3 times, each for 1.5-2.0 hr, concentrating the extractive solution until no ethanol exists, adding water, concentrating, standing, centrifuging to remove precipitate, collecting supernatant, concentrating, and drying to obtain C;

(4) Extracting ramulus Cinnamomi with 4-6 times of 70-80% ethanol for 2-3 times (each for 1.5-2.0 hr), concentrating the extractive solution until there is no ethanol, adding water, concentrating, and drying under reduced pressure to obtain D;

(5) Cutting rhizoma Zingiberis recens into pieces, adding 10-12 times of water, squeezing, filtering, and drying under reduced pressure to obtain E;

(6) 4-6 times of broadleaf holly leaf medicinal material, extracting with 70% -80% ethanol for 2-3 times, each time for 1.5-2.0 hours, concentrating the extract until no ethanol exists, adding water, continuing to concentrate, and drying under reduced pressure to obtain F;

(7) Extracting Folum Ilicis with 10-12 times of water solution for 2-3 times (each for 1.5-2.0 hr), concentrating the extractive solution, and drying under reduced pressure to obtain G;

(8) The six extracts comprise 0.1-3% of salidroside as the main component of the extract A; (ii) a The extract B mainly comprises 10-20% of polygonatum polysaccharide; the main component of the extract D is 0.1% -3.0% of cinnamic acid; the main components of the extracts F and G are chlorogenic acid content of 1% -5.0%;

(9) Mixing 1-7 kinds of the above extracts at a certain proportion to obtain Chinese medicinal composition, and preparing into different dosage forms according to requirement.

The compound prepared by the method can be prepared into capsules, tablets, granules and other dosage forms, and the application direction is to improve energy.

Specifically, the invention also aims to provide application of the traditional Chinese medicine compound composition with the function of resisting fatigue in preparing health-care products.

The compound composition disclosed by the invention can be used as a dietary supplement or a health food raw material to solve the problem of easy fatigue.

According to the technical scheme, compared with the prior art, the traditional Chinese medicine compound composition with the anti-fatigue function and the application thereof provided by the invention have the following excellent effects:

the intervention of each compound composition medicine reduces the accumulation of lactic acid, enhances the endurance capacity and improves the fatigue state; can obviously provide energy for the organism, enhance the motion ability, reduce the fatigue feeling, contribute to the enhancement of skeletal muscles and enhance the motion ability; and the reserve of muscle glycogen and the compensatory quantity of blood sugar of the body are increased to resist fatigue, the generation of ATP is promoted, the motor capacity of the body is further enhanced, the fatigue is reduced, and the synthesis of cell factors is promoted to act on various tissues and organs, maintain the biological steady state of the body, play an active protection role and resist the pathophysiological injury caused by movement.

The rhodiola rosea, the rhizoma polygonati, the agrimony, the ginger, the cassia twig and the broadleaf holly leaf are prepared into the traditional Chinese medicine compound composition with the anti-fatigue function according to a certain weight percentage, and the traditional Chinese medicine compound composition has an obvious effect on improving energy and is a safe and effective dietary supplement.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.

FIG. 1 shows the effect of the drug to be tested on the first sinking time of the mouse weight-bearing swimming experimentp < 0.05）。

FIG. 2 shows the effect of the drug to be tested on the experimental total swimming time of weight swimming in micep<0.05）。

FIG. 3 shows the effect of the tested drugs on the hypoxia tolerance time of the mouse in the normal pressure hypoxia tolerance experimentp < 0.05）。

Detailed Description

The technical solutions disclosed in the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.

The embodiment of the invention discloses a traditional Chinese medicine compound composition which has an anti-fatigue function and can be used as a dietary supplement or a health food raw material for improving energy.

The present invention will be further specifically illustrated by the following examples for better understanding, but the present invention is not to be construed as being limited thereto, and certain insubstantial modifications and adaptations of the invention by those skilled in the art based on the foregoing disclosure are intended to be included within the scope of the invention.

The technical solution disclosed in the present invention will be further described with reference to the following specific examples.

Example 1

A preparation method of a traditional Chinese medicine compound composition with an anti-fatigue function comprises the following specific preparation steps:

(1) Weighing 300g of rhodiola rosea, rhizoma polygonati and hairyvein agrimony respectively;

(2) Adding 6, 6 and 6 times of water solution into rhodiola rosea medicinal material, heating and refluxing for extraction for 3 times, wherein each time is 2.0 hours, combining filtrates, filtering, centrifuging, concentrating supernate to relative density of about 1.02-1.05, adding 95% ethanol, stirring until the alcohol content is 70%, standing for 12 hours, filtering out precipitates, concentrating supernate, and drying under reduced pressure to obtain rhodiola rosea extract with the rhodiola rosea glycoside content of 3%;

(3) Cutting rhizoma Polygonati into pieces, reflux-extracting with 6 and 8 times of water for 2 times (each time for 1.5 hr), mixing the filtrates, concentrating to relative density of 1.10, precipitating with 70% ethanol, magnetically stirring for 1 hr, standing for more than 24 hr, and spin-drying the precipitate to obtain rhizoma Polygonati extract with rhizoma Polygonati polysaccharide content of 20%;

(4) Extracting herba et Gemma Agrimoniae with 4, 5, 6 times of 70% ethanol for 3 times, each for 1.5 hr, concentrating the extractive solution until there is no ethanol, adding water, concentrating, standing, centrifuging to remove precipitate, collecting supernatant, concentrating, and drying to obtain herba et Gemma Agrimoniae extract;

taking the three extract powders, wherein the weight ratio of the rhodiola rosea extract, the rhizoma polygonati extract, the hairyvein agrimony extract and the three formula compositions is 25%:50%:25 percent, thus obtaining the product.

Example 2

(1) Weighing 300g of rhodiola rosea, rhizoma polygonati, hairyvein agrimony, cassia twig, ginger and broadleaf holly leaf respectively;

(5) 4 and 6 times of cassia twig medicinal material, extracting with 70% ethanol for 2 times, each time for 2.0 hours, concentrating the extracting solution until no ethanol exists, adding water, continuously concentrating, and drying under reduced pressure to obtain a cassia twig extract containing 2% of cinnamic acid;

(6) Cutting rhizoma Zingiberis recens into pieces, adding 10-12 times of water, squeezing, filtering, and drying under reduced pressure to obtain rhizoma Zingiberis recens extract;

(7) Extracting Folum Ilicis with 4, 6 times of 70% ethanol for 2 times, each for 2.0 hr, concentrating the extractive solution until there is no ethanol, adding water, concentrating, and drying under reduced pressure to obtain Folum Ilicis extract containing chlorogenic acid 3%;

taking the six kinds of extract powder, and mixing the six kinds of extract powder according to the weight ratio of 10% of four formula compositions of rhodiola rosea extract, rhizoma polygonati extract, hairyvein agrimony extract, cassia twig extract, ginger extract and broadleaf holly leaf extract: 30%:10%:10%:10%:30 percent.

Example 3

A preparation method of a traditional Chinese medicine composition with an anti-fatigue function comprises the following specific preparation steps:

(1) Weighing 300g of rhodiola rosea, rhizoma polygonati, hairyvein agrimony and broadleaf holly leaf respectively;

(5) Folium Ilicis extract containing chlorogenic acid 3% is prepared by extracting folium Ilicis with 4, 6 times of 70% ethanol for 2 times, each for 2.0 hr, concentrating the extractive solution until no ethanol exists, adding water, concentrating, and drying under reduced pressure.

Taking the four extract powders, wherein the weight ratio of the four formula compositions of the rhodiola rosea extract, the sealwort extract, the hairyvein agrimonia herb and bud tea extract is 10 percent: 40%:20%: and (5) 30 percent.

Example 4

(1) Weighing 300g of rhodiola rosea, rhizoma polygonati, hairyvein agrimony and cassia twig respectively;

(5) The cassia twig is extracted by 4 and 6 times of ethanol with 70 percent of ethanol for 2 times, each time lasts for 2.0 hours, the extracting solution is concentrated to be free of alcohol, water is added for continuous concentration, and the cassia twig extract containing 2 percent of cinnamic acid is obtained after reduced pressure drying.

Taking the four extract powders, wherein the weight ratio of the four formula compositions of the rhodiola rosea extract, the sealwort extract, the hairyvein agrimony extract and the cassia twig extract is 15 percent: 40%:31%: and (5) 14 percent, thus obtaining the product.

Example 5

(1) Weighing 300g of rhodiola rosea, rhizoma polygonati, hairyvein agrimony and ginger respectively;

(3) Cutting rhizoma Polygonati medicinal materials into blocks, reflux-extracting with 6 and 8 times of water respectively for 2 times, each time for 1.5 hr, mixing the two filtrates, concentrating to relative density of 1.10, adding 70% ethanol for precipitating, magnetically stirring for 1 hr, standing for more than 24 hr, and spin-drying the precipitate to obtain rhizoma Polygonati extract with rhizoma Polygonati polysaccharide content of 20%;

(5) Cutting rhizoma Zingiberis recens into pieces, adding 10-12 times of water, squeezing, filtering, and drying under reduced pressure to obtain rhizoma Zingiberis recens extract.

Taking the four extract powders, wherein the weight ratio of the four formula compositions of rhodiola rosea extract, rhizoma polygonati extract, hairyvein agrimony extract and broadleaf holly leaf extract is 9%:46%:19%:26 percent.

Example 6: capsule preparation

(1) 30kg of each of six medicinal materials of rhodiola rosea, rhizoma polygonati, hairyvein agrimony, cassia twig, ginger and broadleaf holly leaf, adding 6, 6 and 6 times of aqueous solution into the rhodiola rosea, heating and refluxing for 3 times, extracting for 2.0 hours each time, merging filtrate, filtering, centrifuging, concentrating supernatant till the relative density is about 1.02-1.05, adding 95% ethanol, stirring till the alcohol content is 70%, standing for 12 hours, filtering out precipitate, concentrating supernatant, and drying under reduced pressure to obtain rhodiola rosea extract, wherein the content of salidroside is 3%;

(2) Cutting rhizoma Polygonati into pieces, reflux-extracting with 6 and 8 times of water for 2 times (each time for 1.5 hr), mixing the filtrates, concentrating to relative density of 1.10, precipitating with 70% ethanol, magnetically stirring for 1 hr, standing for more than 24 hr, and spin-drying the precipitate to obtain rhizoma Polygonati extract with rhizoma Polygonati polysaccharide content of 20%;

(3) Extracting herba et Gemma Agrimoniae with 4, 5, 6 times of 70% ethanol for 3 times, each for 1.5 hr, concentrating the extractive solution until there is no ethanol, adding water, concentrating, standing, centrifuging to remove precipitate, collecting supernatant, concentrating, and drying to obtain herba et Gemma Agrimoniae extract.

(4) The cassia twig is extracted by 4 and 6 times of ethanol with 70 percent of ethanol for 2 times, each time lasts for 2.0 hours, the extracting solution is concentrated to be free of alcohol, water is added for continuous concentration, and the cassia twig extract containing 2 percent of cinnamic acid is obtained after reduced pressure drying.

(6) The broadleaf holly leaf extract is extracted by 4 and 6 times of 70 percent ethanol for 2 times, each time lasts for 2.0 hours, the extract is concentrated to be free of alcohol, water is added for continuous concentration, and the broadleaf holly leaf extract containing 3 percent chlorogenic acid is obtained by decompression and drying.

Taking the six extract powders, and mixing the six formula compositions according to the weight ratio of 10% of rhodiola rosea extract, rhizoma polygonati extract, hairyvein agrimony extract, cassia twig extract, ginger extract and broadleaf holly leaf extract: 30%:10%:10%:10%:30 percent.

(7) Continuously concentrating the prepared extract until the relative density is 1.08 to 1.10, uniformly mixing the extract according to the weight ratio after spray drying, sieving the medicinal powder with a 40-mesh sieve, adding pregelatinized starch and talcum powder and magnesium stearate, uniformly mixing, and encapsulating to prepare capsules.

Example 7: tablet preparation

(1) 30kg of rhodiola rosea, rhizoma polygonati, hairyvein agrimony, cassia twig, ginger and broadleaf holly leaf, wherein the rhodiola rosea is added with 6, 6 and 6 times of water solution, heated and refluxed for extraction for 3 times, each time lasts for 2.0 hours, filtrate is combined, filtered and centrifuged, supernatant is concentrated to the relative density of about 1.02-1.05, 95 percent ethanol is added for stirring until the alcohol content is 70 percent, the mixture is kept stand for 12 hours, precipitate is filtered out, supernatant is concentrated and then is dried under reduced pressure to obtain rhodiola rosea extract, and the content of salidroside is 3 percent;

Taking the six extract powders, wherein the weight ratio of six formula compositions of rhodiola rosea extract, rhizoma polygonati extract, hairyvein agrimony extract, cassia twig extract, ginger extract and broadleaf holly leaf extract is 10 percent: 30%:10%:10%:10%:30 percent.

(7) Continuously concentrating the prepared extract until the relative density is 1.08 to 1.10, uniformly mixing the extract according to the weight ratio after spray drying, adding starch and magnesium stearate, uniformly mixing, pressing into tablets, coating film coatings and preparing into tablets.

Comparative example 1

300g of rhodiola rosea medicinal material, adding 6, 6 and 6 times of water solution into the rhodiola rosea medicinal material, heating, refluxing and extracting for 3 times, each time for 2.0 hours, combining filter liquor, filtering, centrifuging, concentrating supernate to the relative density of about 1.02-1.05, adding 95% ethanol, stirring until the alcohol content is 70%, standing for 12 hours, filtering out precipitates, concentrating supernate, and drying under reduced pressure to obtain the rhodiola rosea extract, wherein the content of salidroside is 3%. Cutting 300g of rhizoma polygonati medicinal material into blocks, respectively carrying out reflux extraction for 2 times by using 6 and 8 times of water as raw materials, carrying out 1.5 hours each time, combining the two filtrates, concentrating until the relative density is 1.10, adding 70% ethanol for alcohol precipitation, carrying out magnetic stirring for 1 hour, standing for more than 24 hours, respectively taking the precipitate and carrying out spin drying to obtain the rhizoma polygonati extract, wherein the rhizoma polygonati polysaccharide content is 20%; uniformly mixing the rhodiola rosea extract and the polygonatum extract according to the ratio of 1.

Comparative example 2

300g of rhodiola rosea medicinal material, adding 6, 6 and 6 times of water solution into the rhodiola rosea medicinal material, heating, refluxing and extracting for 3 times, each time for 2.0 hours, combining filter liquor, filtering, centrifuging, concentrating supernate to the relative density of about 1.02-1.05, adding 95% ethanol, stirring until the alcohol content is 70%, standing for 12 hours, filtering out precipitates, concentrating supernate, and drying under reduced pressure to obtain the rhodiola rosea extract, wherein the content of salidroside is 3%. 300g of hairyvein agrimony, adding 70% ethanol which is 4, 5 and 6 times the amount of the hairyvein agrimony, extracting for 3 times, 1.5 hours each time, concentrating the extracting solution until no ethanol exists, adding water, continuing to concentrate, standing, centrifuging to remove precipitates, taking supernate, concentrating and drying to obtain the hairyvein agrimony extract. Uniformly mixing the rhodiola rosea extract and the hairyvein agrimonia herb extract according to the ratio of 1.

Comparative example 3

Cutting 300g of rhizoma polygonati medicinal material into blocks, respectively carrying out reflux extraction for 2 times by using 6 and 8 times of water as raw materials, carrying out 1.5 hours each time, combining the two filtrates, concentrating until the relative density is 1.10, adding 70% ethanol for alcohol precipitation, carrying out magnetic stirring for 1 hour, standing for more than 24 hours, respectively taking the precipitate and carrying out spin drying to obtain the rhizoma polygonati extract, wherein the rhizoma polygonati polysaccharide content is 20%; uniformly mixing the rhodiola rosea extract and the polygonatum extract according to the ratio of 1. 300g of hairyvein agrimony medicinal material is added with 70 percent ethanol of 4, 5 and 6 times of the quantity of the hairyvein agrimony medicinal material for 3 times, each time lasts for 1.5 hours, the extracting solution is concentrated until no ethanol exists, water is added for continuous concentration, the mixture is kept stand, the precipitate is removed by centrifugation, and the supernatant is taken, concentrated and dried to obtain the hairyvein agrimony extract. Taking the rhizoma polygonati extract, namely the hairyvein agrimony extract, and uniformly mixing the extracts according to the ratio of 2.

Comparative example 4

The method comprises the steps of taking a red bull beverage (taurine: 125 mg, lysine: 50 mg, caffeine: 50 mg, inositol: 50 mg, vitamin PP:10 mg, vitamin B6:1 mg and vitamin B1:23 microgram. The red bull is prepared from the ingredients of water, white granulated sugar, citric acid, essence, taurine and lysine.) through heating at 60 ℃ and decompression concentration, and concentrating 500ml (to 180ml, containing 100mg of caffeine).

The beneficial effects of the traditional Chinese medicine composition on resisting fatigue and improving energy are further illustrated by test examples.

Experiment one:

1. materials and methods

1.1 Material

1.1.1 Instruments and consumables: swimming boxes (Shanghai Xin Soft information science and technology Co., ltd.), soda lime (Shanghai Wu Si chemical reagent Co., ltd., batch No. 20200713), white vaseline (Shandong Lin kang medical science and technology Co., ltd., batch No. 200701),

electronic balance (Sidoris, germany) and mouse gavage (Jinan Yiyan science and technology development Co., ltd.).

1.1.2 Medicine preparation: the test substance 1 (preparation in example 1), the test substance 2 (preparation in example 5), the test substance 3 (preparation in comparative example 1), the test substance 4 (preparation in comparative example 2), and the test substance 5 (preparation in comparative example 3).

1.1.3 Animals: SPF male KM mice, 20 + -2 g in body mass, 84 mice. Provided by the experimental animal breeding company Limited of Jinnanppon, license number: SCXK 2019-0003.

1.2 Method of producing a composite material

1.2.1 Feeding conditions and groups

5 mice in one cage, free diet drinking water raises in light and atmospheric control room: the temperature is 21 +/-2 ℃, the humidity is 50 +/-10%, and the light and shade period is 12h/12h (20.

Mice were acclimated for one week after arrival and entered the experiment. Each group is 14, and the groups are divided into 6 groups according to a random block design method: a normal control group; 1 group of test drugs; 2 groups of test drugs; 3 groups of test drugs; 4 groups of test drugs; 5 groups of test drugs.

1.2.2 Administration mode and course of treatment

The normal group is filled with purified water with the same volume, and the other groups are filled with the following pure water: the test agent 1 (0.2 g/kg, dissolved in purified water), the test agent 2 (0.2 g/kg, dissolved in purified water), the test agent 3 (0.2 g/kg, dissolved in purified water), the test agent 4 (0.2 g/kg, dissolved in purified water), and the test agent 5 (0.2 g/kg, dissolved in purified water). The mice were gavaged every morning at a fixed time (9: 00), with a gavage volume of 0.1 ml.10g ^-1 Administration continued for 28 days. And performing the behavioral detection 1h after the last intragastric administration.

1.2.3 Experimental method

1.2.3.1 Weight bearing swimming experiment

During the administration period, swimming adaptability training without load is performed once every 3 days and 5min every time. After each group of mice was subjected to intragastric administration for 1h on day 28, the mice were tied to the tail roots of the mice with a lead wire 7% of the body mass of the mice to cause the loading state of the mice, and immediately placed into a 50 cm × 40 cm × 40 cm water tank for swimming, the water depth was controlled to be not less than 30 cm, the water temperature was controlled to be 25 ± 1 ℃, and the time from the beginning of swimming to the time when the mice still could not float out of the water surface after sinking for 10 s was recorded by a time-second table as the mice exhaustion swimming time. The time to first sink and the time to exhaustion of the mice were recorded.

1.2.3.2 Experiment of oxygen deficiency resistance under normal pressure

After 1 day of rest after the last load swimming experiment, 1 hour after 29 days of intragastric administration, each group of mice is respectively placed into 250 ml ground bottles containing 5 g of soda lime, 1 mouse in each bottle is sealed by a bottle plug coated with vaseline in advance, so that the bottle is airtight, timing is carried out immediately, and the time of death of the mice due to oxygen deficiency is recorded by taking the respiratory arrest as an index. And hypoxia tolerance time calculation was performed according to the following criteria: t = (T1-T0)/(V0-W0/0.94) × 100 (where T1 is mouse death time, T0 is time to start sealing, V0 is effective vial volume, W0 is mouse body weight, and 0.94 is mouse density measured by drainage method).

1.2.4 Statistical method

Experimental data on

± SEStatistical analysis was performed using GraphPad Prism 8.02 Software (GraphPad Software, inc., san Diego, california, USA). All group data were tested for normality (Kolmogorov-Smirnov test) and for homogeneity of variance (Leven test) before parameter testing. All test parameters were compared pairwise between groups using unpaired T-test (single tail), with test levels set top < 0.05。

Results of the experiment

2.1 Weight bearing swimming experiment

TABLE 1 mouse weight bearing swimming experiment result recording TABLE: (

± SE）

Group of	Administration of drugs	Number of	Time of first subsidence	Exhaustion swimming time
					Normal control group	Purified water	11	15.49±2.62	92.63±17.23
1 group of test drugs	Preparation of example 1	12	23.76±3.09*	378.90±145.60*
					To-be-tested drugs 2 groups	Preparation of example 5	11	30.57±7.13*	423.20±164.20*
To-be-tested drugs 3 groups	Preparation of comparative example 1	12	17.56±2.65	193.40±35.69*
					4 groups of test drugs	Preparation of comparative example 2	13	17.81±2.82	154.07±91.942*
To-be-tested drugs 5 groups	Preparation of comparative example 3	13	16.44±2.08	102.38±48.19*

In a mouse weight swimming experiment, compared with a normal control group, the drugs to be tested 1 and 2 are administrated through gastric lavage, the first sinking time of the mouse weight swimming can be obviously prolonged, and the exhaustive swimming time of the mouse weight swimming can be obviously increased through the gastric lavage of the drugs to be tested.

2.2 Experiment of oxygen deficiency resistance under normal pressure

Table 2 mouse atmospheric hypoxia tolerance experiment result recording table (

± SE）

Group of	Administration of drugs	Number of	Time to hypoxia tolerance
				Normal control group	Purified water		10	509.90±15.14
To-be-tested drugs 1 group	Preparation of example 1	14	626.50±22.22***
				To-be-tested drugs 2 groups	Preparation of example 5	14	571.50±28.07*
To-be-tested drugs 3 groups	Preparation of comparative example 1	13	522.03±31.04
				4 groups of test drugs	Preparation of comparative example 2	13	509.32±36.65
5 groups of test drugs	Preparation of comparative example 3	14	517.40±22.74

In the mouse normal-pressure hypoxia tolerance experiment, compared with a normal control group, the gavage to-be-tested drugs 1 and 2 can obviously prolong the hypoxia tolerance time of the mouse.

Experiment summary

In the experiment of mouse weight swimming, the drugs to be tested (the drugs to be tested 1 and 5) in the embodiment can obviously increase the exhaustion swimming time of the mouse weight swimming, and also obviously prolong the first sinking time of the mouse weight swimming. The results show that the drugs 1 and 5 to be tested in the examples have obvious anti-fatigue drug effect, and the best effect is obtained by using the preparation method in the example 5. In the mouse normal-pressure hypoxia tolerance experiment, the preparations in examples 2 and 5 can obviously increase the hypoxia tolerance time of the mouse, and show that the medicines can obviously prolong the survival time of the mouse and improve the hypoxia tolerance capability of the mouse under the acute hypoxia condition. In the heavy-load swimming test and the hypoxia tolerance test, the effect of the preparation in the example group is more prominent than that of the preparation in the blank group and the comparison examples 1, 2 and 3.

Experiment two:

1. materials and methods

1.1 Material

1.1.1 Instruments and consumables: animal treadmill (mouse): chengduta alliance software, inc., model number FT-200; an electronic balance: mettler-toledo international trade (shanghai) ltd, model XSR205; mouse gavage device: jinan Yiyan science and technology development Limited.

1.1.2 Medicine preparation: the test drugs are administered to the group 1 (composition preparation in control example 4), the group 2 (composition preparation in example 1), the group 3 (composition preparation in example 2), the group 4 (composition preparation in example 3), the group 5 (composition preparation in example 4), and the group 6 (composition preparation in example 5).

1.1.3 Animals: SPF male Kunming mice, 6-8 weeks old, 18 + -3 g in body mass, 168. Provided by Beijing Weitonglihua laboratory animal technology Co., ltd, license number: SCXK (Jing) 2016-0006.

1.2 Method of producing a composite material

1.2.1 Feeding conditions and groups

After the mice are transported, the mice are numbered by using a tail mark instrument, and are raised in 6 cages to adapt to the environment for 7 days. After entering the laboratory, the light and shade cycle of 12h/12h (8 00 on, 20: the temperature is 21 +/-2 ℃, the humidity is 50 +/-10%, and all animal experiment operations are carried out according to the welfare of experimental animals issued by NIH and the use guiding principle.

After the adaptive feeding is finished, the feed is divided into 7 groups according to a random grouping design method (according to body mass), and each group comprises 24: normal control group, 1 group of medicines to be tested, 2 groups of medicines to be tested, 3 groups of medicines to be tested, 4 groups of medicines to be tested, 5 groups of medicines to be tested and 6 groups of medicines to be tested.

1.2.2 Administration mode and course of treatment

Administration: mice were gavaged at a fixed time (9 00) every morning for 28 days. Starting 3 days before the treadmill exhaustion test (days 20 to 22 of the gavage), daily treadmill adaptation training was performed, and a treadmill exhaustion test was performed for 6 days on days 23 to 28 of the gavage. Mouse body mass was recorded on days 1, 7, 14, 21, and 28, respectively.

The control group was gavaged with the same volume of saline. The other groups of gavage doses were as follows: 1 group of test drugs (30 g/kg), 2 groups of test drugs (0.2 g/kg, dissolved in purified water), 3 groups of test drugs (0.2 g/kg, dissolved in purified water), 4 groups of test drugs (0.2 g/kg, dissolved in purified water), 5 groups of test drugs (0.2 g/kg, dissolved in purified water), and 6 groups of test drugs (0.2 g/kg, dissolved in purified water).

1.2.3 Running test experiment method

All mice were first subjected to an adaptation training period of 15min each day for three days (days 20 to 22 of gavage) beginning 1h after gavage.

The treadmill exhaustion test experiment is carried out for 6 days on the 23 th day to 28 th day of the intragastric administration, the exhaustion test is carried out after the intragastric administration for 1h every day, the exhaustion speed is 18m/min (obtained by pre-experiment), and the speed parameter of 18m/min can enable the mouse to better complete the treadmill task and exhaust within 4 h. Recording the exhaustion time, wherein the exhaustion time standard is as follows: the mouse continuously falls off a power grid and is electrically shocked, is always positioned at a 1/3 section behind a runway, is rested to be in a lying position in the abdomen, breathes quickly, and cannot complete a running task.

1.2.4 tissue selection

And (3) anesthetizing the animals by using isoflurane 30 min after the test of the running platform of the mice of the second batch, and after the eyelid reflex detection animals enter a deep anesthesia state, taking blood from the abdominal aorta of all the animals. Standing the animal blood at room temperature for 30 min, centrifuging at 3500rpm/15min, collecting supernatant, and storing in a refrigerator at-80 deg.C for inspection. Then, the liver and the quadriceps femoris muscles on the left and right sides are accurately picked up and weighed, and the liver and the quadriceps femoris muscles are quickly placed into liquid nitrogen for freezing and storage for later use.

1.2.4.1 serum Urea Nitrogen and insulin assay (ELISA)

The content of urea nitrogen and insulin in serum are detected by a urea nitrogen detection kit (diacetyl monoxime method) and an insulin detection kit respectively. The frozen serum tissue is taken out and thawed (-20 ℃,4 ℃), centrifuged (13000 rpm/10 min) to take supernatant, and the detection is carried out according to the method of the relevant kit instruction. Within 5min after the reaction is terminated, the concentration of each standard substance on the test kit is inputted, and the Optical Density (OD) of each group is measured sequentially at the wavelength of 460nm by using a microplate reader. And after a standard curve is obtained, substituting the OD value of the sample into the obtained regression equation to calculate the concentration of each sample. If the test sample is diluted, the final sample concentration is multiplied by the dilution factor.

1.2.4.2 CRP, PG-E2, IL-6 and NPY content detection

The content of C-reactive protein (CRP), prostaglandin E2 (Prostaglandin E2, PG-E2), interleukin 6 (Interleukin-6, IL-6) and Neuropeptide Y (Neuropeptide Y, NPY) which is a pain marker in the serum of the mice is quantitatively detected by using an enzyme-linked immunosorbent assay (ELISA) kit. The frozen serum tissue is taken out and thawed (-20 ℃,4 ℃), centrifuged (13000 rpm/10 min) to take the supernatant, and the detection is carried out according to the method of the relevant kit instruction. And inputting the concentration value of each standard substance on the test kit within 5min after the reaction is terminated, and sequentially measuring the Optical Density (OD) of each group at the wavelength of 450nm by using a microplate reader. And (5) after a standard curve is obtained, substituting the OD value of the sample into the obtained regression equation to calculate the concentration of each sample. If the test sample is diluted, the final sample concentration is multiplied by the dilution factor.

1.2.4.3 detection of the content of Myolactone, myoglycogen, myoAMP

And (3) detecting the contents of mouse muscle glycogen, muscle lactic acid and AMP by using an ELISA kit, and calculating the ratio of AMP to ATP. The cryopreserved quadriceps tissue and liver of the mice were removed, thawed (-20 ℃,4 ℃), the tissue was rinsed with pre-cooled PBS (0.01M, pH = 7.4), residual blood was removed, and the tissue was minced. The minced tissue and the corresponding volume of PBS (weight to volume ratio of 1. Within 5min after the reaction is terminated, the concentration value of each standard substance on the test kit is input, and the Optical Density (OD) of each group is measured by a microplate reader at the wavelength of 450nm in sequence. And (5) after a standard curve is obtained, substituting the OD value of the sample into the obtained regression equation to calculate the concentration of each sample. If the test sample is diluted, the final sample concentration is multiplied by the dilution factor.

1.2.4.4 measurement of glycogen synthase and phosphofructokinase content in muscle tissue

Glycogen Synthase (GS) and Phosphofructokinase (PFK) contents in mouse muscle tissues were detected using an enzyme kit. Frozen mouse quadriceps femoris tissue samples were removed, thawed (-20 ℃,4 ℃), the tissue was rinsed with pre-cooled PBS (0.01M, pH = 7.4), residual blood was removed, and the tissue was minced. The minced tissue and the corresponding volume of PBS (weight to volume ratio of 1. Within 5min after the reaction is terminated, the concentration value of each standard substance on the test kit is input, and the Optical Density (OD) of each group is measured by a microplate reader at the wavelength of 450nm in sequence. And after a standard curve is obtained, substituting the OD value of the sample into the obtained regression equation to calculate the concentration of each sample. If the test sample is diluted, the final sample concentration is multiplied by the dilution factor.

1.2.4.5 measurement of the content of muscle tissue irisin, myoconjunctin, myostatin, and brain-derived neurotrophic factor

The ELISA kit is adopted to detect the content of mouse muscle tissue Irisin (Irisin), myonectin (Myonectin), myostatin (MSTN) and brain-derived neurotrophic factor (BDNF). The cryopreserved quadriceps tissue of the mouse was taken out, thawed (-20 ℃,4 ℃), the tissue was rinsed with pre-cooled PBS (0.01M, pH = 7.4), residual blood was removed, and the tissue was minced. The minced tissue and the corresponding volume of PBS (weight to volume ratio of 1. Within 5min after the reaction is terminated, the concentration value of each standard substance on the test kit is input, and the Optical Density (OD) of each group is measured by a microplate reader at the wavelength of 450nm in sequence. And after a standard curve is obtained, substituting the OD value of the sample into the obtained regression equation to calculate the concentration of each sample. If the test sample is diluted, the final sample concentration is multiplied by the dilution factor.

1.2.5 Statistical method

Data were analyzed using GraphPad Prism 8.02 Software (GraphPad Software, inc., san Diego, california, USA). All group tests data were tested for normality (Kolmogorov-Smirnov test) and homogeneity of variance (Leven test) before parameter testing. The single tail t-test was compared between groups, with the test level set at p < 0.05.

Results of the experiment

2.1 running-out time of mouse

TABLE 3 mouse running exhaustion test recording table: (

± SE）

Statistical results of running exhaustion time of mice ([ x ])p＜0.05；**p＜0.01；***p＜0.001；****p＜0.0001）

In the experiment of the running exhaustion test of the mouse, compared with a normal control group, the gastric lavage dry prognosis of each drug to be tested can effectively prolong the exhaustion time of the mouse in the running exhaustion test, and prompts that the drug has the effects of enhancing endurance and resisting fatigue. Compared with the efficacy result of the drug to be tested in the comparative example, the anti-fatigue effect of each compound in the example is better and more prominent.

2.2 Mouse serum urea nitrogen and insulin content

TABLE 4 mouse serum urea nitrogen and insulin content TABLE: (

± SE）

Group of	Administration of drugs	Number of	Serum urea nitrogen content	Serum insulin content
					Normal control group	Physiological saline	11	52.24±3.148	1.487±0.3158
1 group of test drugs	Comparative example 4 preparation of composition	12	47.02±5.809	2.487±0.4167*
					To-be-tested drugs 2 groups	EXAMPLE 1 preparation of the composition	11	56.34±7.148	2.175±0.6112
To-be-tested drugs 3 groups	Example 2 preparation of a composition	11	54.10±5.506	1.386±0.2758
					4 groups of test drugs	Example 3 preparation of a composition	11	44.03±3.559	2.410±0.2113*
To-be-tested drugs 5 groups	Example 4 preparation of a composition	11	33.58±7.080*	2.410±0.3770*
					6 groups of test drugs	EXAMPLE 5 preparation of the composition	11	47.85±5.001	2.578±0.5257*

In the test experiment of the urea nitrogen content in the mouse serum, compared with a normal control group, the urea nitrogen content in the mouse serum can be obviously reduced by the intragastric administration of the reagent 5 to be tested; in the insulin content test experiment, compared with a normal control group, the to-be-tested drugs 1, 4, 5 and 6 can obviously increase the serum insulin content of the mouse.

2.3 serum CRP, PG-E2, IL-6, NPY levels in mice

TABLE 5 mouse serum-related Biochemical index content List (

± SE）

Group of

Administration of drugs

Number of

CRP content

PG-E2 content

IL-6 content

NPY content

Normal control group

Physiological saline

11

12977±1816

2.207±0.3045

3.275±0.1779

3.174±0.9694

1 group of test drugs

Comparative example 4 preparation of composition

10

10410±1063

1.968±0.3294

3.357±0.5870

8.509±1.561*

To-be-tested drugs 2 groups

EXAMPLE 1 preparation of the composition

12

6733±611.6***

2.067±0.5241

3.048±0.3876

5.496±0.5958*

To-be-tested drugs 3 groups

EXAMPLE 2 preparation of the composition

9

7627±657.1**

1.930±0.4087

4.027±0.2372*

7.125±1.185*

4 groups of test drugs

Example 3 preparation of a composition

12

6152±629.8***

2.269±0.5087

3.093±0.2607

6.192±1.198*

To-be-tested drugs 5 groups

EXAMPLE 4 preparation of the composition

10

6072±911.9**

1.837±0.3958

4.654±0.1163***

10.11±1.741**

6 groups of test drugs

EXAMPLE 5 preparation of the composition

10

8780±809.5*

3.385±0.4981*

2.977±0.7641

6.840±1.251*

In the test experiment of the content of C-reactive protein, compared with a normal control group, the content of C-reactive protein in the serum of a mouse can be obviously reduced by intragastric administration of a to-be-tested drug 2, a to-be-tested drug 3, a to-be-tested drug 4, a to-be-tested drug 5 and a to-be-tested drug 6; in a prostaglandin content test experiment, compared with a normal control group, the content of prostaglandin in the serum of a mouse can be obviously increased by intragastric administration of a to-be-tested drug 6; in an interleukin 6 content test experiment, compared with a normal control group, the content of interleukin 6 in the serum of a mouse can be obviously increased by intragastric administration of a medicament 3 to be tested and a medicament 5 to be tested; in the experiment for testing the content of the neuropeptide Y, compared with a normal control group, the content of the neuropeptide Y in the serum of a mouse can be obviously increased by intragastric administration of a to-be-tested drug 1, a to-be-tested drug 2, a to-be-tested drug 3, a to-be-tested drug 4, a to-be-tested drug 5 and a to-be-tested drug 6.

2.4 Mouse serum MG, LA, AMP, ATP, LG content

TABLE 6 mouse Myoglycolic acid, myoglycogen, AMP content Table (A)

± SE）

Group of	Administration of drugs	Number of	Muscle glycogen content	Content of creatine in the body	AMP content
						Normal control group	Physiological saline	11	1.306±0.1689	2.140±0.1976	3783±75.59
1 group of test drugs	Medicament to be tested 1	11	1.017±0.03384	1.950±0.1456	3487±62.37**
						To-be-tested drugs 2 groups	Medicament to be tested 2	8	1.052±0.05257	1.611±0.1363*	3510±68.07**
To-be-tested drugs 3 groups	Medicament to be tested 3	11	1.048±0.05701	2.389±0.1706	3637±114.7
						4 groups of test drugs	Medicament to be tested 4	10	1.067±0.04406	2.099±0.2028	3466±59.20**
To-be-tested drugs 5 groups	Medicament to be tested 5	11	3.542±0.5861***	2.298±0.9514	3523±74.44*
						6 groups of test drugs	Medicament to be tested 6	10	4.308±0.2314****	1.802±0.1017	3524±80.33**

Statistical results of mouse contents of muscular lactic acid, muscular glycogen and AMP ([ sic ])p＜0.05；**p＜0.01；***p＜0.001；****p＜0.0001），

In the test experiment of the muscle glycogen content of the mouse muscle tissue, compared with a normal control group, the intramyoglycogen content of the mouse muscle tissue is obviously increased by intramyoglycogen content administration of a to-be-tested drug 5 and a to-be-tested drug 6 through intragastric administration; in a muscle lactic acid content test experiment, compared with a normal control group, the intramyolactic acid content of the mouse muscle tissue can be obviously reduced by intragastric administration of the test agent 2; in the adenine nucleotide content test experiment, compared with a normal control group, the AMP content in the muscle tissue of the mouse can be obviously reduced by intragastric administration of the to-be-tested drug 1, the to-be-tested drug 2, the to-be-tested drug 4, the to-be-tested drug 5 and the to-be-tested drug 6.

2.5 Serum GS and PFK content of mouse

TABLE 7 mouse muscle tissue glycogen synthase and phosphofructokinase content table: (

± SE）

Group of	Administration of drugs	Number of	Glycogen synthase	Phosphofructokinase
					Normal control group	Physiological saline	12	414.0±44.86	4.299±0.5540
To-be-tested drugs 1 group	Medicament to be tested 1	12	576.7±15.15***	3.951±0.6069
					To-be-tested drugs 2 groups	Medicament to be tested 2	11	514.3±19.62*	2.961±0.4193
To-be-tested drugs 3 groups	Medicament to be tested 3	11	545.2±36.99*	6.019±0.4930*
					4 groups of test drugs	Medicament to be tested 4	11	476.5±13.04	7.414±0.8391**
To-be-tested drugs 5 groups	Medicament to be tested 5	11	555.6±23.78**	3.402±0.5326
					6 groups of test drugs	Medicament to be tested 6	11	505.2±19.66*	3.917±0.7449

Statistical results of glycogen synthase and phosphofructokinase content in mouse muscle tissuep＜0.05；**p＜0.01；***p＜0.001），

In a glycogen synthase content test experiment, compared with a normal control group, the glycogen synthase content of the muscle tissue of a mouse can be obviously increased by intragastric administration of a to-be-tested drug 1, a to-be-tested drug 2, a to-be-tested drug 3, a to-be-tested drug 5 and a to-be-tested drug 6; in a phosphofructokinase content test experiment, compared with a normal control group, the content of phosphofructokinase in mouse muscle tissues can be obviously increased by intragastric administration of a to-be-tested drug 3 and a to-be-tested drug 4.

2.6 mouse serum Irisin, MSTN, BDNF content

TABLE 8 mouse muscle tissue irisin, myogenin, myostatin, and brain-derived neurotrophic factor content table (

± SE）

Group of

Administration of drugs

Number of

Irisin content

Content of myogenin

Content of myostatin

Brain-derived neurotrophic factor content

Normal control group

Physiological saline

12

127.7±8.723

22.66±0.9781

77.59±2.999

858.5±28.91

To-be-tested drugs 1 group

Medicament to be tested 1

12

244.6±6.301****

22.97±0.8865

83.27±2.201

985.0±43.21*

To-be-tested drugs 2 groups

Medicament to be tested 2

11

227.6±5.647****

18.27±0.2272***

71.54±1.505*

932.8±34.17

To-be-tested drugs 3 groups

Medicament to be tested 3

11

238.5±3.576****

24.32±0.5744

80.16±0.9674

960.3±27.42*

4 groups of test drugs

Medicament to be tested 4

11

213.6±3.837****

17.94±0.2688***

68.21±1.840**

836.0±28.11

To-be-tested drugs 5 groups

Medicament to be tested 5

11

240.8±5.221****

21.90±0.5899

80.46±1.511

1047±55.93**

6 groups of test drugs

Medicament to be tested 6

11

225.5±4.799****

20.72±1.250

65.75±1.410***

811.8±45.39

Statistical results of mouse muscle tissue irisin, myogenin, myostatin and brain-derived neurotrophic factor contentp＜0.05；**p＜0.01；***p＜0.001；****p＜0.0001）

In an irisin content test experiment, compared with a normal control group, the content of irisin in the muscle tissue of a mouse can be obviously increased by intragastric administration of a to-be-tested drug 1, a to-be-tested drug 2, a to-be-tested drug 3, a to-be-tested drug 4, a to-be-tested drug 5 and a to-be-tested drug 6; in the muscle union content test experiment, compared with a normal control group, the intragastric administration of the to-be-tested drug 2 and the to-be-tested drug 4 can obviously reduce the muscle union content of the mouse muscle tissue; in a myostatin content test experiment, compared with a normal control group, the content of myostatin in mouse musculature can be obviously reduced by intragastric administration of a to-be-tested drug 2, a to-be-tested drug 4 and a to-be-tested drug 6; in the experiment for testing the content of the brain-derived neurotrophic factor, compared with a normal control group, the contents of the mouse muscle tissue brain-derived neurotrophic factor can be obviously increased by intragastric administration of the to-be-tested drug 1, the to-be-tested drug 3 and the to-be-tested drug 5.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A preparation method of a traditional Chinese medicine compound composition with an anti-fatigue function comprises the following specific preparation steps:

taking the three extract powders, wherein the weight ratio of the rhodiola rosea extract, the sealwort extract, the hairyvein agrimonia herb and bud extract to the three formula compositions is 25 percent: 50%:25 percent, thus obtaining the product.

2. A preparation method of a traditional Chinese medicine compound composition with an anti-fatigue function is characterized by comprising the following specific preparation steps:

(5) Folium Ilicis extract is extracted with 4, 6 times of 70% ethanol for 2 times, each time for 2.0 hr, the extractive solution is concentrated to no ethanol, water is added for further concentration, and vacuum drying is carried out to obtain folium Ilicis extract containing chlorogenic acid 3%,

3. A preparation method of a traditional Chinese medicine compound composition with an anti-fatigue function is characterized by comprising the following specific preparation steps:

(2) Adding 6, 6 and 6 times of water solution into rhodiola rosea medicinal material, heating and refluxing for extraction for 3 times, wherein each time is 2.0 hours, combining filtrates, filtering, centrifuging, concentrating supernate to relative density of about 1.02-1.05, adding 95% ethanol, stirring until the alcohol content is 70%, standing for 12 hours, filtering out precipitate, concentrating supernate, and drying under reduced pressure to obtain rhodiola rosea extract with salidroside content of 3%;

(5) Extracting ramulus Cinnamomi with 4, 6 times of 70% ethanol for 2 times, each for 2.0 hr, concentrating the extractive solution until there is no ethanol, adding water, concentrating, drying under reduced pressure to obtain ramulus Cinnamomi extract containing cinnamic acid 2%,

taking the four extract powders, wherein the weight ratio of the four formula compositions of the rhodiola rosea extract, the rhizoma polygonati extract, the hairyvein agrimony extract and the cassia twig extract is 15 percent: 40%:31%: and (5) 14 percent, thus obtaining the product.

4. A preparation method of a traditional Chinese medicine compound composition with an anti-fatigue function is characterized by comprising the following specific preparation steps:

(5) Cutting rhizoma Zingiberis recens into pieces, adding 10-12 times of water, squeezing, filtering, drying under reduced pressure to obtain rhizoma Zingiberis recens extract,

5. Use of the Chinese medicinal compound composition with anti-fatigue effect prepared by the method of any one of claims 1 to 4 in preparing health food.

6. The use of claim 5, wherein the herbal composition is formulated as a capsule, tablet or granule for enhancing energy.