Background technique
With the increasingly quickening of social development, scientific and technological progress and rhythm of life, the pressure that people are born is being gradually increased,
The mental and manual labor of high-intensitive high load capacity make most of worker be chronically at fatigue or sub-health state, as colourless without
Power insomnia and dreamful sleep, is had a pain in the back, lather, memory loss, positive fistula premature ejaculation, it is prematurely senile phenomena such as.China or even full generation
Boundary has every year causes dead case because of long-time over fatigue.It is investigated according to WHO, the whole world has 35% or more people to be in tired
Labor state, if cannot eliminate in time after fatigue generation, gradually accumulation will appear overtraining syndrome (OTS) or chronic
The illnesss such as fatigue syndrome (CFS), while endocrine disturbance, immunity degradation occurs with body, or even occur organic
Lesion becomes an important factor for threatening human physical and mental health.Fatigue is damaged caused by muscle and bone organ being capable of direct shadow
The performance of skeletal muscle is rung, myocardial blood supply is blocked.Under work on hand environment and the unalterable situation of pressure, then fatigue is needed
Worker has the health food of anti-fatigue effect to alleviate and restore organism fatigue according to own situation timely and appropriate discovery supplement, mentions
High body anti-pressure ability.The mechanism of fatigue mainly includes (1) energy substance exhaustion hypothesis (2) metabolite Heaping doctrine
(3) interior environmental stability imbalance theory (4) protectiveness inhibits theory (5) endocrine system dissonance theory (6) catastrophe theory (7) certainly
It is influenced by base theoretical;Wherein, free radical theory increasingly obtains the approval of researcher, and a large amount of report display movements can induce a large amount of
The generation of free radical.Lead to blood lactase acid in Mice Body, blood urea nitrogen and hemoglobin as wang et al. discovery power exhausts swimming test
Content decline, it is believed that may be that function caused by the oxidative stress that movement generates declines;Nazeer et al. research discovery white flower fish
Protein enzymatic hydrolyzate has anti peroxidation of lipid ability and improves the vigor of antioxidase system, and it is anti-may to alleviate oxidative stress with it
Should be related to enhance tolerance of exercise, this also from certain angle reflect between antioxidant activity and antifatigue effect there is
Correlation.Therefore, how research is by preventing the generation of free radical from improving body movement endurance, to play tonifying kidney and strengthening yang
Effect is particularly important.
In recent years, on folk medicine and modern science experiment basis, many scholars are seeking antifatigue and Yang invigorating medicine
Aspect has done a large amount of work, and all there is many side effects and adverse reaction such as physical strength are excessively saturating for this kind of primary treatment drug
Branch, while drug cost is also high.Chinese traditional herbs become in recent years because of few side effects, without the excitant ingredient being harmful to the human body
The research hotspot for carrying out anti-fatigue medicament, due to " Diet cures more than the doctors ", the ideas such as " there's no such thing as a totally safe medicine " are rooted in the hearts of the people, and cause to resist tired
Labor functional health care food is more welcomed by the people compared with anti-fatigue medicament, therefore finds effective exploitation, safety and Small side effects
Functional food becomes a kind of very promising solution route.
Currently, the effect of Chinese medicine class anti-fatigue product is mainly relieved fatigue by following mechanism realization:
1, energy is supplemented, is reached by the trophic factors of supplement body consumption and is maintained body normal physiological function, relieves fatigue
Purpose such as increases glycogen storage level.
2, the intake of vitamin and microelement needed by human is supplemented.
3, the removing and discharge of acceleration bodies intracellular metabolite substance, reaches antifatigue purpose.
4, sex hormone level is adjusted, such as testosterone, progesterone, estradiol and cortisol equal size.
5, excessive free free radical is removed, antioxidant ability of organism is improved.
Existing anti-fatigue health-product containing in the market, kind is more, and function and effect are different, and quality is also irregular.According to
Known to applicant, general anti-fatigue medicament accelerates the clearance rate of metabolite, adjusts micro-loop by providing energy for body
Border, improves immunity or directly stimulation nervous system reaches antifatigue effect.Although this kind of drugs for nervous is very effective in cure,
But very big to the harm of cardiovascular and cerebrovascular and nervous system, there are certain risk, curative effect is up for further verifying.
HEIHUZHANG fungus scientific name jujube Sarcodon imbricatus (L. Ex Franch.) Karst (Sarcodon imbricatus(L.ex Fr.) Karst.), it is subordinate to
In Basidiomycetes Polyporus, it is the exclusive rare wild mushrooms in the area Yunnan Qian Zang, is rich in polysaccharide abundant, protein, amino
Acid, crude fibre, phenolic compound and several mineral materials microelement have very high tonic value and pharmacological activity.Motherland
Medicine thinks that HEIHUZHANG fungus is sweet natured, has the effect of relaxing tendons and activating collaterals, while can reduce cholesterol, QI invigorating tonifying Qi, Chinese medicine Zhuifengsan
It trembles with fear, controls wind blood-breaking, stomach function regulating defaecation, blood pressure lowering, the poison of solution food, controls the effects of headache and dizziness.However, at present to HEIHUZHANG fungus
Research primarily focuses on chemical composition analysis, Polyose extraction, the optimization of strain liquid fermentation culture conditions, In Vitro Bacteriostasis and resists in vivo
The research of tumor promotion, about HEIHUZHANG fungus it is antifatigue with it is anti-oxidant in terms of have not been reported, lack to its system comprehensively
Understanding.
Black pine be also known as ferfas (Tuber melanosporum) it is a kind of rare very your wild edible for being grown on underground
Fungi has high healthy nutritive value.Due to its growth conditions harshness, annual worldwide production is very rare, therefore price
Fairly expensive, comparable to gold, referred to as on dining table " Black Diamond ".It is domestic at present still few for the research of black pine, mainly
Be the Study on Physiological Activity of black pine polysaccharide is also only rested on anti-oxidant about kind classification and resource distribution etc., and its
He is rarely reported activity.Numerous studies prove that black pine contains 18 kinds of amino acid (required amino that cannot be synthesized including human body
Acid), protein abundant, vitamin, unsaturated fatty acid and mineral trace element, the mankind eat black pine existing more than 2000
Year history, in addition to playing an important role in delicious food other than, currently on the market one after another release with black pine for mainly at
The product divided, such as chocolate, bread, drink, pressed candy, facial mask etc..
So far, it is not yet found that closing with HEIHUZHANG fungus, black pine bacterium single component or by HEIHUZHANG fungus and black pine
The mutual compatibility of bacterium obtains the pertinent literature of ideal treatment with its synergistic effect for antifatigue, anti-oxidant and tonifying kidney and strengthening yang
Report.Therefore, it develops a kind of while there is antifatigue, anti-oxidant and tonifying kidney and strengthening yang effect functional health-care food to have well
Market prospects, composition provided by the invention have irreplaceable advantage and huge potentiality to be exploited.
Embodiment 7:
It is prepared as follows a kind of capsule with antifatigue and Yang strengthening function composition:
(1) it by after 55g HEIHUZHANG fungus and the drying of 35g black pine bacterium, is crushed through pulverizer, crosses 80 meshes, respectively obtain the black tiger palm
The composition of bacterium powder and black pine bacterium powder;
(2) the composition 80g for taking step (1) to obtain is added water to cook 2 times, and 2 hours every time, the quality that water is added every time was combination
10 times of amount of substance, obtain decoction liquor;
(3) decoction liquor is merged, is filtered, it is spray-dried at powder after reduced pressure;
(4) capsule is added in the powder obtained to step (3) and often uses auxiliary material, be uniformly mixed, be made by capsule common process
Capsule.
Comparative example 1
HEIHUZHANG fungus powder, crushes through pulverizer, crosses 80 meshes.
Comparative example 2
Black pine bacterium powder, crushes through pulverizer, crosses 80 meshes.
Test case 1:
1 experimental animal, reagent and instrument
1.1 experimental animals: SPF grades of healthy Kunming strain male mices, weight 18-22 g are purchased from Jilin University experimental animal
The heart (credit number: SCXK (Jilin) 2017-0003).Use 12/12 hour illumination/dark cycle (8 points at night 8 of morning
The illumination of point) mouse to be raised in controlled environment (23 ± 1 DEG C of temperature, humidity 50 ± 10%), diet is free.
1.2 main agents: urea nitrogen (BUN), lactic acid (LA), lactic dehydrogenase (LDH), superoxide dismutase (SOD),
Glutathione peroxidase (GSH-Px), active oxygen (ROS), muscle glycogen and hepatic glycogen assay kit build up life purchased from Nanjing
Object Science and Technology Ltd.;Progesterone (PROG), testosterone (T) and estradiol (E2) ELISA assay kit are purchased from Shanghai source leaf biology
Science and Technology Ltd.;0.9% physiological saline is purchased from Jilin Cologne Cornell pharmaceutical Co. Ltd;BCA determination of protein concentration kit
It is purchased from Merck Millipore, Billerica, MA.
1.3 key instruments: microplate reader, SYNERGY4, the U.S. is rich to rise;96 orifice plates, NEST;Eddy mixer, OL-901, river
Haimen City kylin medical apparatus factory of Soviet Union;Electric drying oven with forced convection, 101A-1E, Shanghai laboratory apparatus Co., Ltd., Factory;Water bath with thermostatic control
Pot, GSY-II, Beijing Medical Equipment Plant;Supercentrifuge, Mini Spin, German Eppendorf;Animal treadmill, FT-
200, Chengdu TME Technology Co., Ltd.;Autonomic activities recorder, ZZ-6, Chengdu TME Technology Co., Ltd.;Tired transfer rod instrument,
ZB-200, Chengdu TME Technology Co., Ltd..
2 experimental methods
2.1 groupings and administration
SPF grades of healthy male Kunming strain mices 70, weight 18-22g, the dry feed of sawtooth class animal adapts to according to national standards
Property feed 5 days after, mouse is divided into 7 groups, every group 10 at random: being Normal group, (ginseng soap is dry for positive controls
Powder, wherein content of ginsenoside is 80%, is purchased from Xi'an all places Biotechnology Co., Ltd) 0.05 g/kg, example 1 group, reality
2 groups of example, 3 groups of embodiment, comparative example 1 and comparative example 2 are applied, is prepared into 0.1 g/mL's with the preceding sodium carboxymethylcellulose with 0.5%
Suspension.Gastric infusion is carried out between the daily 9:00-10:00 period, stomach-filling volume is 0.2 mL/20g, Normal group
It fills with isometric solvent, continuous gavage 14 days.With reference to the Ministry of Public Health " health food is examined and assessment technique specification " (version in 2003)
The middle function of physical fatigue alleviation method of inspection carries out the measurement of every antifatigue index after experiment.
The test of 2.2 autonomic activities
Mouse after each group stomach-filling is placed in quiet shady place, 30 minutes is stood, is put into autonomic activities case, covers and be protected from light plate, make
After mouse adapts to 2 min, standing number and action number in record mouse 5 minutes, parallel 3 times.
2.3 tired transfer rod tests
After last gastric infusion 30 minutes, each group mouse is individually placed in transfer rod instrument (15 revs/min), its muscle is made to be in quiet
Power tense situation.Before each group mouse is tested, suitably train, after mouse falls 3 times on transfer rod, start recording is small
The time that mouse is fallen from transfer rod due to muscular fatigue, parallel 3 times.
2.4 endurance run test
After last gastric infusion 30 minutes, mouse is placed in FT-200 animal treadmill, after placing animal, mouse is carried out
Instrument revolving speed is set as 20 revs/min after so that its understanding is fled from charging zone by training, and electric current is turned up full blast, and record mouse is electric
The time for hitting 5 times exhausts the time in this, as power.
2.5 swimming with a load attached to the body test
According to dosage design continuous gavage is administered 14 days, and after last gastric infusion 30 minutes, mouse is placed in water swimming, the depth of water
30 cm, 25 ± 1.0 DEG C of water temperature, the lead button of 10% weight of rat-tail root load, record mouse exhausts since swimming to power, as
The swimming time of mouse.Being sunk with mouse head can not float more than 8 s after water, and is placed on the plane and cannot complete to right
Reflection exhausts judgment criteria as power.
At once BUN, LA and LDH assay in serum after the swimming of 2.6 mouse
After successive administration 14 days, last stomach-filling mouse tested material 30 minutes, do not born in the water that temperature is 25 ± 0.5 DEG C
It swims 30 minutes when resetting.It takes out mouse to dry, orbital venous plexus takes 0.8 mL of blood immediately, and centrifugation prepares serum and is placed on -80
DEG C refrigerator saves.It is detected according to the measuring method of respective kit specification.
At once SOD, GSH-Px and ROS assay in serum after the swimming of 2.7 mouse
After successive administration 14 days, last stomach-filling mouse tested material 30 minutes, do not born in the water that temperature is 25 ± 0.5 DEG C
It swims 30 minutes when resetting.It takes out mouse to dry, orbital venous plexus takes 0.8 mL of blood immediately, and centrifugation prepares serum and is placed on -80
DEG C refrigerator saves.It is detected according to the measuring method of respective kit specification.
Muscle glycogen, hepatic glycogen content measurement after the swimming of 2.8 mouse
Mouse liver and hindlimb muscle are won immediately, are rejected connective tissue and superabundant fats, are frozen -80 DEG C of refrigerators, it is spare.According to
The measuring method of respective kit specification is detected.
2.9 statistical methods: SPSS16.0 handles data, and using one-way analysis of variance, P < 0.05 is judged as
Difference has conspicuousness, and P < 0.01 is judged as that difference has extremely significant property.
3 experimental results
The experiment of 3.1 autonomic activities
As shown in table 3.1, compared with Normal group, embodiment 1,2,3, positive controls and comparative example 1 and 2 groups pairs of comparative example
The action number and standing number of mouse do not have conspicuousness influence, illustrate the composition or preparation to the nervous centralis of mouse
System does not influence, and not will cause damage.
Influence of the table 3.1 to mouse autonomic activities
Group |
Number of animals/only |
Action number |
Standing number |
P value |
Normal group |
10 |
10.6±3.7 |
57.8±17.9 |
|
Positive controls |
10 |
11.1±5.1 |
60.9±19.0 |
>0.05 |
Embodiment 1 |
10 |
10.0±3.3 |
59.2±14.8 |
>0.05 |
Embodiment 2 |
10 |
10.4±5.9 |
60.2±15.3 |
>0.05 |
Embodiment 3 |
10 |
10.2±3.3 |
56.1±18.8 |
>0.05 |
Comparative example 1 |
10 |
10.3±4.8 |
57.3±17.3 |
>0.05 |
Comparative example 2 |
10 |
11.6±5.3 |
56.9±15.8 |
>0.05 |
Note: P > 0.05, which is represented, does not have conspicuousness
3.2 tired turn-club tests
The raising of exercise tolerance is that anti-fatigue ability reinforces most strong macro manifestations, and transfer rod length of time can reflect animal fortune
Move the degree of fatigue.As shown in table 3.2, embodiment 1,2,3 and positive controls can significantly extend the transfer rod time of mouse,
It is respectively 1.2 times (P < 0.05) of Normal group mouse, 1.9 times (P < 0.01), 1.7 times (P < 0.05) and 1.1 times
(P < 0.05), comparative example 1 and 2 groups of comparative example it is not significant to the transfer rod time effects of mouse, do not show statistical significance.Phase
Compared with 1,2 and 3 group of embodiment, 2 groups of transfer rod times of comparative example 1 and comparative example be substantially less than composition of the invention (P <
0.05).
Influence of the table 3.2 to the mouse fatigue transfer rod time
Group |
Number of animals/only |
The transfer rod time/s |
P value |
Normal group |
10 |
284.6±18.1 |
|
Positive controls |
10 |
322.9±14.7 |
<0.05 |
Embodiment 1 |
10 |
348.7±20.9 |
<0.05 |
Embodiment 2 |
10 |
530.4±27.5 |
<0.01 |
Embodiment 3 |
10 |
474.7±17.2 |
<0.05 |
Comparative example 1 |
10 |
318.3±15.8 |
>0.05 |
Comparative example 2 |
10 |
290.3±22.1 |
>0.05 |
Note: P>0.05, which is represented, does not have conspicuousness, and P<0.05, which is represented, has conspicuousness, and P<0.01 represents extremely significant
3.3 endurance run experiment
If table 3.3 is shown, embodiment 1,2,3 and positive controls can significantly improve the running time of mouse, with normal control
Group is compared, and 14.8%(P < 0.05 has been respectively increased in the running time), 24.1%(P < 0.01), 30.0%(P < 0.01) and
35.4%(P < 0.01).Compared to 1 ~ 3 group of the embodiment of the present invention, the running time of 2 groups of extension mouse of comparative example 1 and comparative example
Significantly lower than composition (P < 0.05) of the invention, and 1 ~ 3 group of the embodiment of the present invention of endurance is run the time and is significantly higher than pair
The sum of ratio 1 and 2 group independent role.Wherein, 3 groups of effects of embodiment are best.
Table 3.3 runs the influence of time to Endurance
Group |
Number of animals/only |
Run time/min |
P value |
Normal group |
10 |
23.7±4.8 |
|
Positive controls |
10 |
32.1±6.4 |
<0.01 |
Embodiment 1 |
10 |
27.2±4.2 |
<0.05 |
Embodiment 2 |
10 |
29.4±4.9 |
<0.01 |
Embodiment 3 |
10 |
30.8±7.5 |
<0.01 |
Comparative example 1 |
10 |
25.3±7.1 |
>0.05 |
Comparative example 2 |
10 |
24.9±6.2 |
>0.05 |
Note: P>0.05, which is represented, does not have conspicuousness, and P<0.05, which is represented, has conspicuousness, and P<0.01 represents extremely significant
3.4 swimming with a load attached to the body experiment
Such as table 3.4,1 ~ 3 group of embodiment and positive controls can significantly extend the mice burden swimming time as the result is shown, compared with
Normal group is compared, and 17.2%(P < 0.05 has been respectively increased in swimming time), 20.7%(P < 0.05), 41.0%(P <
0.01) and 35.5%(P < 0.01), increased although the power of comparative example 1 and 2 group mouse exhausts the time, be significantly lower than this hair
1 ~ 3 group of bright embodiment, and 3 groups of influences of embodiment are extremely significant (P < 0.01), than 2 groups of independent roles of comparative example 1 and comparative example it
With improve 34.0%(P < 0.01).
Influence of the table 3.4 to mouse swimming time
Group |
Number of animals/only |
Power exhausts the time/s |
P value |
Normal group |
10 |
358.6±80.1 |
|
Positive controls |
10 |
485.8±63.9 |
<0.01 |
Embodiment 1 |
10 |
420.3±56.3 |
<0.05 |
Embodiment 2 |
10 |
432.7±54.1 |
<0.05 |
Embodiment 3 |
10 |
505.7±70.8 |
<0.01 |
Comparative example 1 |
10 |
382.1±66.3 |
>0.05 |
Comparative example 2 |
10 |
370.2±55.6 |
>0.05 |
Note: P>0.05, which is represented, does not have conspicuousness, and P<0.05, which is represented, has conspicuousness, and P<0.01 represents extremely significant
The influence of 3.5 compositions or preparation to BUN, LD and LDH in mice serum after swimming
Body BUN content increases with the increase of exercise load, and body is poorer to load performance, and the increase of BUN is brighter
It is aobvious.As shown in table 3.5, compared with Normal group, embodiment 1, embodiment 2 and comparative example 1 can be significantly reduced mouse trip
BUN content in serum, reduces 23.7%(P < 0.05 respectively after swimming), 28.8%(P < 0.01) and 18.7%(P < 0.05),
Embodiment 3 and positive controls effect are the most significant, reduce 30.9%(P < 0.001 than Normal group) and 33.8%(P <
0.001);LA removing accretion rate is faster, and aerobic capacity is higher, is conducive to the rapid elimination of fatigue.With Normal group
It compares, embodiment 1, embodiment 2 and embodiment 3 can significantly reduce after mouse swimming LA content in serum, reduce respectively
17.6% (P < 0.05), 14.7% (P < 0.05) and 18.4%(P < 0.01), comparative example 1 and 2 does not show statistics meaning
Justice;LDH can reduce accumulation of the LA in muscle after movement.The elimination of LA depends on the catalytic action of LDH after movement, and LDH is living
Property increase, excessive LA in muscle can be accelerated to be changed into pyruvic acid, thus the generation of delay fatigue.Compared with Normal group,
Embodiment 2, embodiment 3 and positive controls can significantly improve the content of LDH in serum after mouse is swum, be respectively increased
4.7%(P<0.05), 6.7%(P<0.05) and 7.0%(P<0.05), comparative example 1 and 2 influences not significant (P>0.05),
Wherein, the influence of LDH activity becomes apparent in embodiment 2 and 3 pair mice serum.
Influence of the table 3.5 to BUN, LA and LDH content in mice serum
Note: compared with Normal group, P < 0.001 * P < 0.05, * * P < 0.01, * * *
The influence of 3.6 compositions or preparation to SOD, GSH-Px and ROS content in mice serum after swimming
SOD is to the most important antioxidase of antioxidant radical in organism, is the important indicator of measurement antioxidant ability of organism,
Body damage as caused by oxidative stress can be prevented, plays vital work in the oxidative and anti-oxidative balance of body
With.GSH-Px can promote H2O2Decomposition, protect the structure and function of cell membrane not by the interference of peroxide and damage.Such as
Shown in table 3.6,1 ~ 3 group of embodiment and positive controls can significantly improve after swimming that SOD is horizontal in mice serum, and normal right
Compared according to group, 25.6%(P < 0.05 be respectively increased), 28.6%(P < 0.01), 32.1%(P < 0.01) and 20.0%(P <
0.05);Embodiment 2, embodiment 3 and positive controls significantly improve after swimming GSH-Px content in mice serum, and normal
Control group is compared, and 28.9%(P < 0.05 has been respectively increased), 33.7%(P < 0.01) and 35.9%(P < 0.01);Embodiment
1,2,3, positive controls and comparative example 1 can be significantly reduced after swimming that ROS is horizontal in mice serum.In summary experiment hair
Existing, the antioxidant activity that 1 ~ 3 group of embodiment is better than comparative example 1 and 2 groups of comparative example, and wherein 3 groups of embodiment of influence is more significant,
Effect is best, shows that the present composition or preparation can be significantly improved in the case where HEIHUZHANG fungus and black pine Cordycepps learn compatibility effect
The oxidation resistance of mouse after swimming, synergistic effect are greater than the sum of two kinds of raw material independent roles.
Influence of the table 3.6 to mice serum SOD, GSH-Px and ROS level
Note: compared with Normal group, P < 0.01 * P < 0.05, * *
The influence of 3.7 compositions or preparation to mouse muscle glycogen, hepatic glycogen level after swimming
The endurance when storage level and movement of muscle glycogen is positively correlated, and muscle glycogen deposit is more, and energy supply is more sufficient, and endurance is just
Better;It is tired to can be used as resisting for evaluation body for maintaining blood glucose level important in inhibiting when moving for the deposit of hepatic glycogen
Labor index.As shown in table 3.7, compared with Normal group, 1 ~ 3 group of embodiment can improve mouse muscle glycogen and glycogen after swimming
Raw water is flat, and effect, better than the sum of 2 groups of independent roles of comparative example 1 and comparative example, wherein 55.7%(P has been respectively increased in embodiment 3
< 0.01) and 24.3%(P < 0.05).
Influence of the table 3.7 to mouse muscle glycogen and hepatic glycogen level
Note: compared with Normal group, P < 0.01 * P < 0.05, * *
The influence of 3.8 compositions or preparation to PROG, T and E2 level in mice serum after swimming
Influence of the table 3.8 to three kinds of sex hormone levels in mice serum
Note: compared with Normal group, P < 0.01 * P < 0.05, * *
Testosterone can promote androgone continuously to break up, and stimulate the generation and maturation of sperm, and promote reproductive development, protein
Synthesis and the growth of bone.As shown in table 3.8, compared with Normal group mouse, embodiment 1 ~ 3 improves male mice serum
Middle PROG and T is horizontal, and wherein 3 effect of embodiment is most significant (P < 0.05), and 1 ~ 3 group of embodiment of promotion effect is obviously excellent
In comparative example 1 and 2 groups of comparative example, prompt the present composition or preparation can by stimulating the secretion and synthesis of male sex hormone,
Promote the growth and development of sexual organ indirectly.T content in serum can be reduced, the present composition with the gradually aging at age
Or this effect of preparation, especially embodiment 3 will be helpful to delay the aging of sexual organ, so that it is strong to play anti-aging and production of sperm
The effect of positive.
Experimental example 1
The composition or preparation that above-described embodiment 1 ~ 3 is obtained carry out toxicologic study.
1 acute toxicity test (uses MTD method)
1.1 experimental animal;SPF Kunming mice, 40, weight 18-22g, half male and half female is purchased from Jilin University experimental animal
Center (credit number: SCXK(is lucky) 2017-0001).Experimental animal use is carried out in accordance with experimental animal using regulations, and animal is raised
It supports and guarantees 12h/12h Dark-light cycle, temperature is 23 ± 1 DEG C, humidity 50 ± 10%, free diet, drinking-water.
1.2 drugs and reagent: composition or preparation prepared by Example 1,2,3, with 0.5% carboxymethyl fibre before experiment
It ties up plain sodium and is dissolved to 0.2 g/mL drug concentration.
1.3 methods and result: each stomach-filling amount is 0.6 ml/, gives tested material one at interval of 6 hours using in one day
Secondary, twice, 16 h of mouse fasting (can't help water), oral stomach-filling are given tested material 3 days, are observed continuously after administration before stomach-filling for stomach-filling altogether
14 days.Dead and abnormal phenomenon does not occur for conventinal breeding in 14 days after mouse administration.Prove the embodiment 1 ~ 3 that the present invention obtains
Composition or preparation have no toxic side effect, and can use safely.
2 long term toxicity tests
2.1 experimental animals: Wistar rat, 120, weight 180-220g, half male and half female, purchased from Jilin University experimental animal
The heart (credit number: SCXK(is lucky) 2016-0003).
2.2 drugs and reagent: preparation prepared by Example 3 is dissolved with 0.5% sodium carboxymethylcellulose when experiment.
2.3 groupings and medication: experimental animal is randomly divided into 4 groups, every group 30 by gender weight.1 group of stomach-filling is given
0.5% sodium carboxymethylcellulose, 0.2 ml/100g;2, test medicine is given in 3,4 groups of stomach-fillings, dosage be respectively as follows: 25 mg/100g,
50 mg/100g, 100 mg/100g, i.e., every 100 g weight give the crude drug amount of 25 mg, 50 mg or 100 mg, each group concentration
It is each configured to the given low of 0.2 ml/100g.The above each group is euthanized after continuously taking 6 months.
2.4 Indexs measure
2.4.1 general status detects: weight, diet, mood, movable energy.
2.4.2 biochemical index: red blood cell count(RBC), white blood cell count(WBC), platelet count, hemoglobin count, serum
Albumen, cholesterol and blood glucose etc..
2.4.3 pathological examination: the heart, liver, spleen, lung, kidney, brain, stomach, thymus gland, sexual gland etc..
2.5 the experimental results showed that, after rat continuously takes 6 months 3 preparations of embodiment, weight, diet, mood, movable energy
The general status such as amount are good, each main organs and Serum Indexes no abnormality seen, it was demonstrated that 3 preparation of embodiment is to body without obvious long-term
Toxic effect.
Experimental example 2
The present composition or preparation are subjected to clinical human's test
1. sample and grouping
1.1 samples: the powder of the composition of embodiment 1, comparative example 1, comparative example 2 and embodiment 2 ~ 3 is subjected to tabletting respectively, respectively
1200, tablet is made, separately takes placebo 1200 made of starch.By HEIHUZHANG fungus powder and black pine bacterium powder in prescription ratio
It is uniformly mixed, the magnesium stearate that 0.25% ratio is added, which is uniformly mixed, is pressed into 1200 to get tablet.
1.2 methods: all volunteers are randomly divided into 6 groups, every group of 12 people, wherein 3 groups take above-mentioned 3 embodiments respectively
Sample tablet, 2 groups take the tablet of 2 sample of comparative example 1 and comparative example respectively, 0.6 g/ piece, one at a time, 3 times a day.
Separately there is one group of 12 people, takes orally the placebo of equivalent.Continuously take surrounding.
2. test method
2.1 sports equipments: Spinning
2.2 exercise tests: subject carries out the measurement of Spinning breakdown test, when scrunching before taking sample, after taking four weeks
Between be set as 5min, scrunch load be 5 grades, measure stepping movement before and after blood lactase acid value.And blood is detected when taking end of a period
The case where uric acid.
3. experimental result
3.1 subject's distribution situations
1 subject age of table and gender analysis
Group |
Number |
Male |
Female |
Average age |
Embodiment 1 |
12 |
6 |
6 |
30.2±1.9 |
Embodiment 2 |
12 |
4 |
8 |
33.6±2.6 |
Embodiment 3 |
12 |
5 |
7 |
32.5±2.2 |
Comparative example 1 |
12 |
6 |
6 |
31.2±2.6 |
Comparative example 2 |
12 |
5 |
7 |
32.3±2.0 |
Placebo |
12 |
7 |
5 |
33.8±1.9 |
The result shows that between 3 embodiment groups, 2 comparative example groups and placebo group Sex distribution and men and women's age without obvious poor
It is different.
3.2 take blood uric acid value situation after four weeks
2 subject of table takes blood uric acid value (μm ol/L) after four weeks
Group |
Male |
Women |
Embodiment 1 |
364.0±79.5 |
220.8±54.2 |
Embodiment 2 |
373.8±82.3 |
256.1±58.7 |
Embodiment 3 |
377.2±68.6 |
237.5±66.0 |
Comparative example 1 |
380.6±59.6 |
251.5±45.2 |
Comparative example 2 |
379.5±62.8 |
232.8±39.7 |
Placebo |
385.8±90.0 |
248.4±56.9 |
The result shows that the lower 3 embodiment groups of Different Preparation, 2 comparative example groups are taken compared with placebo, blood after four weeks
Uric acid level no significant difference shows that the present composition or preparation will not cause internal uric acid metabolism to be lacked of proper care.
The difference of measurement blood lactase acid, the lift-off value as blood lactase acid after exercise load before and after 3.3 subject's stepping movements.
3 subject's blood lactase acid of table, which increases, changes situation (mmol/L)
Group |
Blood lactase acid lift-off value before testing |
Blood lactase acid lift-off value after experiment |
Blood lactase acid increases change value |
Embodiment 1 |
7.5±2.6 |
7.1±1.9 |
-0.4±1.1 |
Embodiment 2 |
7.9±1.8 |
7.6±2.4 |
-0.3±1.8 |
Embodiment 3 |
8.0±2.2 |
7.4±1.9 |
-0.6±1.2 |
Comparative example 1 |
7.7±1.6 |
7.6±1.8 |
-0.1±1.5 |
Comparative example 2 |
7.5±1.5 |
7.6±2.1 |
0.1±0.8 |
Placebo |
7.6±3.0 |
7.7±2.8 |
0.1±0.9 |
Experimental result is shown, compared with placebo, subject is can be significantly reduced in the present composition for taking different formulations
The raising of blood lactase acid value after movement, increases the anti-fatigue ability of body, wherein blood cream after 1 ~ 3 group of reduction subject motion of embodiment
The degree of acid value is apparently higher than comparative example 1 and 2 groups of comparative example (P < 0.05), and 3 effect of embodiment is best.
The resistance to experiment of machanics of animal and physiological and biochemical index are the results show that the present composition or preparation can significantly extend
The endurance of mouse runs time, transfer rod time and walking weight load, and SOD, GSH-Px and LDH contain in mice serum after raising swimming
Amount reduces BUN, LA and ROS level in serum, increases the storage level of hepatic glycogen and muscle glycogen, prompts the present composition or system
Agent, which has, adjusts human body energy metabolism and preferable oxidation resistance.According to the Ministry of Public Health promulgate China " health food examine with
Assessment technique specification " 2003 years versions regulation, if more than one is tested (swimming with a load attached to the body and pole-climbing) and two containing an endurance exercise
Or more biochemistry containing binomial to be designated as positive (blood lactase acid, serum urea nitrogen, muscle glycogen hepatic glycogen etc.) be with antifatigue effect
Criterion.Meanwhile clinical human's test result is shown, the present composition or preparation can significantly reduce human body blood after movement
Lactate level.Moreover it is possible to remarkably promote the secretion of T and PROG in male mice serum, improve male's sexual defect indirectly.
Disclosure of the invention will be the new selection of one kind that the infull patient of male's sexual finds safe and effective therapeutic agent.To sum up
It is described, the present composition or preparation realized after HEIHUZHANG fungus and black pine Cordycepps learn composition of prescription exercise tolerance promoted,
It relieves fatigue, is anti-oxidant, delaying senescence, the effect of strengthening yang and invigorating kidney, the synergistic effect shown, not only drug activity shows excellent,
It is also greatly reduced cost of material, is conducive to industrialized production, is particularly suitable for the male of fatigue crowd and kidney yang virtual loss and early ageing
It takes.