CN101322711B - Health care medicament for resisting fatigue and enhancing immunity and preparation thereof - Google Patents
Health care medicament for resisting fatigue and enhancing immunity and preparation thereof Download PDFInfo
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- CN101322711B CN101322711B CN200810013722A CN200810013722A CN101322711B CN 101322711 B CN101322711 B CN 101322711B CN 200810013722 A CN200810013722 A CN 200810013722A CN 200810013722 A CN200810013722 A CN 200810013722A CN 101322711 B CN101322711 B CN 101322711B
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- ubiquinone
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Abstract
The invention discloses a health care medicine for resisting fatigue and strengthening human immunity and a preparation method thereof. The health care medicine is essentially made from the pharmaceutical raw materials according to the following parts by weight: 5-60 parts of coenzyme Q10, 12.5-200 parts of vitamin C, 1.5-50 parts of vitamin E, and 0.04-0.6 part of folic acid. The health care medicine has simple components, reliable curative effect and the health care functions of relieving fatigue and strengthening immunity.
Description
(1) technical field
The present invention relates to a kind of health care medicine, the health care medicine of particularly a kind of resisting fatigue and enhancing human immune.
(2) background technology
Along with the quickening of modern life rhythm, the aggravation of social competition, tired, hypoimmunity becomes the problem that people often run into.Fatigue is a kind of physiological phenomenon, is a kind of mechanism of protectiveness to the people.This is that health sends the signal that have a rest to us.If no matter ignore, health will suffer damage, and falls sick from overwork at last to it.It is tired that the immunocompromised person then feels easily, but can not check organic disease; Often catch a cold; Wound infects easily, and healing is slow; Gastrointestinal function poor (mainly be meant here and eat improperly a little) with regard to vomiting and dirarrhea.
Learn that from State Food and Drug Administration's basic data library inquiry the health product of declaring health care and be alleviating physical fatigue, enhancing immunity have 23, wherein 20 is the Chinese medicine health care product of Chinese crude drug prescription.Mostly Chinese medicine health care product is that medical material is prepared from through suitable technology extraction; Extractum after the medicinal material extract is not refining through further mostly, thereby its dose is bigger, moreover because the source of medical material is different; Thereby the product function curative effect may be variant; Its quality can not reach control fully, because also there is the indefinite shortcoming of some functional component in the Chinese crude drug complicated component.
(3) summary of the invention
The objective of the invention is to overcome deficiency of the prior art, provide a kind of curative effect reliable, the health care medicine of the simple alleviating physical fatigue of composition, enhancing immunity.
The present invention realizes through following measure:
The health care medicine of resisting fatigue of the present invention and enhancing immunity is to be processed by the medicinal raw material of following parts by weight:
Ubiquinone
1015-60 vitamin C 12.5-200
Vitamin E 1.5-50 folic acid 0.04-0.56
The health care medicine of resisting fatigue of the present invention and enhancing immunity, described medicinal raw material weight proportion can be preferably:
Ubiquinone
1020 vitamin Cs 50
Vitamin E 7.5 folic acid 0.15
The health care medicine of resisting fatigue of the present invention and enhancing immunity, said medicine can be made different dosage forms such as powder, pill, tablet capsule.
Resisting fatigue of the present invention and enhancing immunity health care medicine, can process with raw material by the auxilliary of following parts by weight:
Soybean oil 90-1500 Cera Flava 5-150
Resisting fatigue of the present invention and enhancing immunity health care medicine, can process by the raw material of following parts by weight:
Ubiquinone
1020 vitamin Cs 50
Vitamin E 7.5 folic acid 0.15
Soybean oil 307 Cera Flavas 16
Aforesaid health care medicine preparation of soft capsule method of the present invention may further comprise the steps:
A) take by weighing ubiquinone
10, vitamin C, vitamin E, folic acid, soybean oil, Cera Flava, subsequent use;
B) after the soybean oil heating with said weight proportion, add the Cera Flava of said weight proportion;
C) with the ubiquinone of said weight proportion
10, vitamin E, join in the above-mentioned fluid, stir and to make dissolving, cool to appropriateness after, add folic acid, the vitamin C of said weight proportion, mix, grind evenly with colloid mill, fill promptly gets.
Functional component is a ubiquinone in this prescription
10, vitamin E, vitamin C, folic acid, wherein ubiquinone
10, vitamin C, vitamin E, be antioxidant, can remove free radical; Improve immunity, folic acid can be removed free radical, the protection cell membrane; Various compositions cooperatively interact, through the control free radical, and the protection human immune system; Just can ease people psychology and physiological pressure, thereby reach alleviating physical fatigue, the health care of enhancing immunity.In this product prescription, with ubiquinone
10Be main, be equipped with other three kinds of compositions, they do not have incompatibility each other, and can synergism.
Described ubiquinone
10, it is natural antioxidant and the cellular metabolism activator that cell self produces, has protection and recovers biomembrane integrity, the effect of stabilising membrane current potential, and be the nonspecific immunity strengthening agent of body, therefore demonstrate fabulous antifatigue effect, ubiquinone
10Make the state of cell maintenance good health, thereby body is dynamic, full of vitality, mental abundant, in addition, it can also promote that heat is converted into energy, transforms the human heat in (for energy) process to play a significant role.
Described vitamin E is a kind of natural fatsoluble vitamin; Its basic physiological effect is an anti-oxidation function, strengthens the invasion and attack that cell membrane is avoided free radical, mainly act as the protection cell; Avoid the infringement of its oxidated effect; Thereby reach the antifatigue effect, in addition can also human body immunity improving power, humoral immunization and cellular immunization all there are obvious facilitation.
Described vitamin C is as natural free radical scavenger; Can directly or indirectly eliminate free radical, thereby can protect the biomembrane integrity, improve the body ability to work; Postpone or the generation that lessens fatigue; Accelerate fatigue recovery, improve the utilization of oxygen in the body, have certain function improving aerobic sport ability.In addition, vitamin C can also promote the formation of immunoglobulin (antibody) and strengthen humoral immunization and cellular immune function, thus human body immunity improving power.
Described folic acid is the important vitamin of needed by human body, participates in the generation of nucleic acid, protein and phospholipid and penetrates, and is closely related with differentiation, the propagation of cell.Folic acid can improve anti-oxidative defense system in the cell, reduces the generation of homocysteine; Strengthen BH
4Combine with eNOS; And direct anti-radical action arranged.Folic acid is protected cell function at free radical resisting, and aspects such as raising immunity play an important role.
The invention has the beneficial effects as follows: composition is simple, and curative effect is reliable, has alleviating physical fatigue, the health care of enhancing immunity.
(4) description of drawings
Fig. 1 is SOD vigor and a MDA content sketch map in the mice serum
Fig. 2 is a ubiquinone
10High-efficient liquid phase chromatogram
Fig. 3 is a ubiquinone in mice plasma and the tissue
10The content sketch map
(5) specific embodiment
Embodiment 1:
Take by weighing the coenzyme ubiquinone
1020g, vitamin C 50g, vitamin E 7.5g, folic acid 0.15g, subsequent use, take by weighing 307g soybean oil heating proper temperature, the Cera Flava that takes by weighing 16g adds in the oil, and making slowly, dissolving is transparence; Ubiquinone with said weight
10, vitamin E, join in the above-mentioned fluid, stir and to make whole dissolvings, after temperature is fallen, add folic acid, the vitamin C of said weight, stir and make mix homogeneously, grind evenly with colloid mill, in the capsule that is filled to, promptly get soft capsule.
Embodiment 2:
Take by weighing ubiquinone
105g, vitamin C 15g, vitamin E2 g, folic acid 0.5g, appropriate amount of auxiliary materials mix homogeneously in addition, compacting promptly gets tablet in flakes.
Embodiment 3:
Take by weighing ubiquinone
1055g, vitamin C 180g, vitamin E 50g, folic acid 0.04g, in addition adjuvant fully stirs, granulate, drying, granulate is packaged into bag and gets final product, and promptly gets a kind of granule.
For proving effect of the present invention, make the sample experiment Analysis with soft capsule of the present invention:
Test one: health care medicine of the present invention is to the antifatigue effect of mice
This test has been studied the antifatigue effect of health care medicine of the present invention through mouse test, has studied ubiquinone simultaneously
10Distribution in mice plasma, liver and heart tissue, being intended to provides scientific basis for developing health care medicine of the present invention.
1 materials and methods
1.1 reagent and instrument
Ubiquinone
10(98.0% ~ 101.0%): day clear pharmacy;
Ubiquinone
10Standard substance: Sgima company;
Normal saline: state-run Zhangjiagang pharmaceutical factory;
Heparin: biochemical reagents, China Medicine (Group) Shanghai Chemical Reagent Co.;
VCXSO type ultrasonic processor: U.S. SONICS & MATERIALS company;
Nano-ZS90 type particle size analyzer: Britain Mavlern company;
7170A automatic clinical chemistry analyzer: HIT;
Superoxide dismutase (superoxdiedsimutase, SOD), malonaldehyde (mlaondaildehyde, MDA), liver glycogen, blood Lactic acid Kit: build up bio-engineering research institute available from Nanjing;
Highly effective liquid phase chromatographic system (being furnished with 1525 binary pump, 2996 PDADs and Empower chromatographic work station): U.S. Waters company.
1.2 experimental animal and feedstuff
Male mice in kunming: body weight (20 ± 2) g, 80: available from Shanghai Slac Experimental Animal Co., Ltd., credit number SCXK (Shanghai) 2003-0003;
The common size Mus produces breeding feed: Chinese Academy of Sciences's Shanghai Experimental Animal Center development, Shanghai Slac Experimental Animal Co., Ltd. produces.
1.3 test preparation method for preparing
The preparation of health care medicine of the present invention: with the soybean oil heating, add an amount of Cera Flava, make dissolving be transparence, then with the recipe quantity ubiquinone
10, vitamin E, join in the above-mentioned fluid, stir and to make whole dissolvings, after temperature reduces, add folic acid, vitamin C, stir and make mix homogeneously, with grinding evenly, fill promptly gets.
Positive control: Liquid CoQ
10(ubiquinone
10Content is 10mg/ml): Nurittional Specaliites company produces.
1.4 test method
1.4.1 ubiquinone in blood plasma, heart and the liver
10Determination on content
(1) chromatographic condition:
Chromatograph: Waters2690; Detector: Waters996 UV-detector; Chromatographic column: SunFrie
TMC
18ODS (150 * 4.6mm, 5 μ m); Flow velocity: 1ml/min; Detect wavelength: UV275nm; Column temperature: 35 ℃; Sample size: 20 μ l.
Mobile phase: methanol: ethanol=30: 70 (V: V) (blood plasma, heart), methanol: ethanol=50: 50 (V: V) (liver),
(2) drafting of standard curve
Get ubiquinone
10Ethanol standard solution (108.9ug/ml) 0,0.02,0.05,0.1,0.3; 0.5,1.0,2.0,3.0,5.0ml places the 10ml volumetric flask; Be settled to scale with ethanol and get standard series, sample introduction 20 μ l are abscissa with concentration (C) respectively, and peak area (A) is a vertical coordinate, get regression equation:
A=18959C-5776.2 (R=0.9990) mobile phase: methanol: ethanol=30: 70 (V: V);
A=16987C+422.45 (R=0.9997) mobile phase: methanol: ethanol=50: 50 (V: V).
(3) sample pretreatment
The list of references method is also improved: taking heparin anticoagulant plasma sample 500ul adds dehydrated alcohol 1ml, vortex vibration 1min; Add normal hexane 3ml; Add normal hexane 0.5ml again behind the vortex vibration 3min, leave standstill 5min, high speed centrifugation 15min (2500r/min); Shift the supernatant in centrifuge tube, N is led in (45 ± 0.5) ℃ water-bath
2Dry up, residue together with centrifuge tube in-20 ℃ of freezing preservations of refrigerator lucifuge, to be measured.With 100 μ l anhydrous alcohol solutions, sample introduction 20 μ l make HPLC and analyze before measuring.The record ubiquinone
10Peak area, adopt external standard method to carry out quantitative analysis.Because of ubiquinone
10Contain isopentene group in the structure, see that light is prone to degraded, under the lucifuge condition so The pretreatment is all carried out.
Core dirty and liver tissue homogenate 2ml, processing and assay method are with above-mentioned plasma sample.
1.4.2 tried mice group and irritated the stomach dosetest
The mice adaptability is divided into 5 groups after feeding 3d at random, i.e. negative control group, positive controls, low dosage test group, middle dosetest group, high dose test group, 16 every group.Duration of test is respectively organized the mice normal feedstuff and is fed, free drinking public water supply.Adopt the quantitative stomach mode of irritating directly sample to be poured into, irritating the stomach amount is 0.1ml/10g, regularly irritates stomach every day 1 time.Negative control group is irritated stomach with normal saline, and it is 0.1ml/10g/d that positive controls and basic, normal, high dose groups mice bland are irritated the stomach total amount, sees table 1.
Table 1 mouse stomach dose form
+
| Group | Normal saline/ml | LiquidCo Q 10/mL | Health care medicine/mL of the present invention | Sport type beverage/mL | Ubiquinone 10Dosage/(mgkg -1· d -1) |
| Negative control group | 0.10 | - | - | - | 0 |
| Positive controls | - | 0.02 | - | 0.08 | 20 |
| Low dose group | - | - | 0.02(LC＝10 ％) | 0.08 | 4.3 |
| Middle dose groups | - | - | 0.02(LC＝20 ％) | 0.08 | 8.6 |
| High dose group | - | - | 0.02(LC＝30 ％) | 0.08 | 12.9 |
+ mouse stomach dosage (0.1mL/10g*d)
Consider that phosphide also has antifatigue effect, therefore basic, normal, high 3 dosage are that the health care medicine of the present invention with 3 carrying capacity levels makes an experiment under the identical prerequisite of adjuvant concentration such as assurance phosphide, further study ubiquinone
10Antifatigue effect.The continuous irrigation stomach in the time of the 30th day every component be that 3 inferior groups are carried out the exercise tolerance test and measured corresponding index.
1.4.3 exercise tolerance test
Set up tired model with the mode of swimming with a load attached to the body, after last is irritated stomach 30min, at the bear a heavy burden galvanized wire of 5% (g/g) body weight of mouse tail root, put into water temperature and be the swimming case that 25 ± 1 ℃, the depth of water are not less than 30cm, every case is once put into 4 mices.Clock with stopwatch, swimming beginning to the dead time is mice swimming with a load attached to the body time (s).
1.4.4 ubiquinone in SOD vigor, MDA content and the tissue in hepatic glycogen, serum urea nitrogen content, the serum
10Determination on content
After last is irritated stomach 30min, place water temperature to be 30 ℃ not swimming with a load attached to the body of swimming case 90min the mice after, pluck the eyeball blood sampling, separation of serum adopts the 7170A automatic clinical chemistry analyzer to measure serum urea nitrogen; SOD vigor, MDA assay are pressed the operation of test kit description.
Take off simultaneously cervical vertebra execution immediately and get liver and heart,, blot with filter paper with normal saline flushing.Accurately take by weighing liver 50mg, press test kit description time-and-motion study hepatic glycogen content; To remain liver and heart tissue and claim to add an amount of cold saline respectively after the gross weight, and in ice bath, prepare certain density tissue homogenate with glass homogenizer, ubiquinone in liver and the heart is measured and calculated to sample pretreating method among the employing 1.4.1
10Content.
1.4.5 ubiquinone in variation of blood lactic acid content and the blood plasma
10Determination on content
After last is irritated stomach 30min; Mouse orbit is got blood; Measure basic lactic acid content, again mice is put the people galvanized wire swimming 10min of 4% (g/g) body weight of bearing a heavy burden in (depth of water 30cm, 30 ℃ of water temperatures) in the case that swims; Pluck eyeball behind blood and the rest 20min and get blood respectively at immediately mouse orbit being got after the swimming, press test kit mensuration full blood lactic content is described.Calculate blood lactic acid TG-AUC, in order to judge motion bleeding from anus lactic acid situation of change:
Blood lactic acid TG-AUC=5 * (20min of blood lactic acid value+2 * swimming rest afterwards at once blood lactic acid value after blood lactic acid value before the swimming+3 * swimming)
Blood lactic acid content situation of change before and after the comparing motion.Adopt among the 1.4.1 sample pretreating method to measure ubiquinone in the blood plasma simultaneously
10Content.
1.4.7 test data statistical analysis
Data are represented with x ± S.D., carry out date processing with SPSS10.0 software, adopt one factor analysis of variance (ANOVA), and P＜0.05 has significance for difference.
2 results and discussion
2.1 health care medicine of the present invention is to the influence of mice body weight
Can know by table 2, respectively organize the mice body weight behind the filling stomach raising 30d and all do not have significant difference that this shows that test preparation can not produce obviously influence to the normal growth of mice.
Table 2 is respectively organized variation (n=5, the x ± S.D.) of mice body weight
| Group | Before the test/g | Behind the 30d/g |
| Negative control group | 21.02±1.40 | 36.25±2.43 |
| Positive controls | 20.84±1.65 | 36.12±2.30 |
| Low dose group | 21.28±1.31 | 36.58±1.92 |
| Middle dose groups | 20.92±1.54 | 37.38±2.23 |
| High dose group | 20.72±1.20 | 37.75±2.72 |
2.2 health care medicine of the present invention is to the influence of mouse movement endurance and biochemical indicator
The raising of exercise tolerance is the direct embodiment of anti-fatigue ability.Mice swimming with a load attached to the body result of the test shows (seeing table 3): test group is compared with negative control group and all can be prolonged the mice swimming with a load attached to the body time, improves exercise tolerance, and has significant difference (P＜0.05) between middle and high dose groups and the negative control group.
It is relevant that minimizings such as energy i (in vivo) material such as glycogen behind tired generation and high strength or the prolonged exercise, metabolite such as serum urea nitrogen, lactic acid such as accumulate at factor.Hepatic glycogen be keep the important stock of glucose normal level in the blood, the source of energy when also being muscle fibers contract.Can find out from table 3; 3 dose groups hepatic glycogen contents all are higher than negative control group; Wherein there is significant difference (P＜0.05) between positive controls and middle and high dose groups and the negative control group, explains that health care medicine of the present invention helps the minimizing of hepatic glycogen consumption after the mice swimming with a load attached to the body.Motion back middle and high dose groups serum urea nitrogen (BUN) level is starkly lower than negative control group (P＜0.05), and middle and high dose groups blood lactic acid TG-AUC is compared obvious reduction (P＜0.05) with negative control group.Wang Qirong etc. also find through human trial, contain ubiquinone
10Composite antioxidant can reduce serum BUN level, have to promote the athlete that sports load is adapted to and improve the effect that physical function recovers.Researchs such as YanJ are thought, ubiquinone
10Biological activity mainly come from the physicochemical property of redox characteristic He its side chain of its quinone ring; Its quinone ring works to transmit electronics and proton in oxidation-respiration chain; It is requisite that all life forms are not only in this effect, but also be the key that forms ATP.And ATP is the main storage form of human body energy, also is the important foundation that all cells function is rely and normally brought into play.
Table 3 mouse anti-reflecting fatigue index of correlation (n=5, x ± S.D)
| Group | Swimming with a load attached to the body time/s | Hepatic glycogen/(mg*g -1Liver) | Serum urea nitrogen/(mmolL -1) | Blood lactic acid TG-AUC/(mmol*L -1*min -1) |
| Negative control group | 457.75±91.77 | 13.59±12.25 | 9.00±1.03 | 334.62±37.60 |
| Positive controls | 638.00±76.68 | 17.96±1.78* | 8.80±0.77 | 302.06±31.32 |
| Low dose group | 545.25±64.75 | 16.75±2.63 | 8.64±0.89 | 308.57±29.08 |
| Middle dose groups | 713.00±157.11. | 16.75±2.63 | 7.60±10.38* | 257.97±14.75** |
| High dose group | 1130.75±195.23* | 19.56±1.74* | 6.82±0.92** | 218.36±36.46** |
2.3 health care medicine of the present invention is to the influence of SOD vigor and MDA content
SOD is the enzyme that unique direct specificity is removed free radical in the body, and the ability that the high more representative body of its activity is removed free radical is strong more, and this enzymatic activity descends, and causes body free radical surplus and oxidative damage.Unsaturated fatty acid in the too much free radical attack cells film starts lipid peroxidation and forms MDA, and MDA content can be represented level of lipid peroxidation in the body.The remarkable rising of a large amount of generations of free radical and blood plasma lipide peroxide in the motion is the major reason that causes sports fatigue to produce.Fig. 2 result shows that there are significant difference apparently higher than negative control group (P＜0.05) in positive controls and basic, normal, high dose groups mice serum active sod (SOD) vigor but have only between high dose group and the positive controls.The content of malonaldehyde (MDA) also significantly is lower than negative control group (P＜0.05) in positive controls and the basic, normal, high dose groups serum, but basic, normal, high dose groups and positive controls it. a no significant difference.This shows, supplemented with exogenous property ubiquinone
10The trend that reduces tissue oxidizing damage is arranged, avoid lipid peroxidation, thus the generation of delay fatigue or the tired elimination in back that helps to move.
There are some researches show: contain ubiquinone
10Composite antioxidant can reduce serum MDA level behind the training athlete, improve serum activity of SOD, this possibly be that the recovery and the antioxidation of motion back physical function is closely related.Research is thought, ubiquinone
10Can get into as oxygen free radical scavenger and non-ly in the cell to combine with each position of cell specifically, reduce the generation of oxygen-derived free radicals, suppress lipid peroxidation, protection SOD active center and structure thereof are avoided free-radical oxidation damage, SOD activity improving.
2.5 health care medicine of the present invention is to ubiquinone in blood plasma and the tissue
10Influence
The sample analysis method specificity is measured the result and is seen Fig. 2.The result shows, under the chromatographic condition that test is confirmed, analyzes, when the mobile phase condition is a methanol: ethanol=30: 70 (V: in the time of V), health care medicine ubiquinone that newly can be former
10Retention time be about 7.7min; When the mobile phase condition is a methanol: ethanol=50: 50 (V: in the time of V), ubiquinone
10Retention time be about 12.8min, the free from admixture peak disturbs.
Compare ubiquinone in positive controls, 3 dose groups blood plasma and the heart with negative control group
10Content all have and raise but do not have significant difference (seeing Fig. 3 a and Fig. 3 b).Ubiquinone in positive controls, 3 the dose groups livers
10Content compare with negative control group all significantly and raise (P＜0.05), and ubiquinone in middle and high dose groups and the positive controls liver
10Content compare remarkable rising (P＜0.05) (seeing Fig. 3 c), explain with health care medicine be that carrier can improve ubiquinone
10Content in blood plasma, liver and heart.
There are some researches show: different with the mankind, give birth to ubiquinone in the mice
10Content be lower than the homologue ubiquinone of another kind of quinone
9, but through long-term supplemented with exogenous property ubiquinone
10Can significantly improve its content in mice mitochondrion and tissue, most of ubiquinone
10Absorbed and accumulate by liver after getting into blood circulation, also can be distributed in positions such as heart, muscle.Exogenous ubiquinone
10Can be converted into the reduction form by the oxidoreductase on the cell membrane and diaphorase and prevent kinetic cellular oxidation damage, intravital oxidative phosphorylation reaction possibly make ubiquinone
10Consumption increase, cause that content reduces ubiquinone in heart, the liver organization in the blood plasma
10With the radical reaction that motion produces, therefore, it might be ubiquinone in blood plasma, the heart that its content reduces to a certain extent
10Concentration does not have one of reason of significant difference.It should be noted that ubiquinone in the liver
10Content is significantly raise by very big its content of the influence of intake.
3 conclusions
Health care medicine of the present invention can prolong the mice swimming with a load attached to the body time, keeps swimming Mouse Liver glycogen levels, and CKIs matter is decomposed, and reduces blood lactic acid content in the serum, has certain antifatigue effect.Ubiquinone
10As biological anti-oxidant, can improve SOD vigor in the serum through eliminating the peroxy radical that the body endogenous movement produces, reduce MDA content in the serum, thus the generation of delay fatigue, the tired elimination in back helps to move.Compare with positive controls (external emulsion product), health care medicine of the present invention can effectively improve the accumulation of coenzyme Q10 in mouse liver.
Test two: health care medicine of the present invention is to the enhancing immunity effect of mice
The enhancing immunity effect of health care medicine of the present invention has been studied in this test through mouse test.
1. laboratory animal: ICR kind white mice, credit number: SCXK (Shanghai) 2002-0010, body weight 18～22 grams, female, by the Chinese Academy of Sciences's Shanghai Experimental Animal Center cleaning level animal is provided.The temperature 20--25 of laboratory animal breeding room ℃; Relative humidity 40--70%, laboratory animal credit number SYXK (Shanghai) 2004-0011, animal feeding material; Two lion laboratory animal feed technology Services Co., Ltd provide registration card number by Suzhou: the E of Soviet Union raises new word (2002) 006.
2. dosage design: human body RD 0.8g/60kg every day of the present invention, basic, normal, high three dose groups are established in this test, and other establishes distilled water blank group.
3. give the appearance approach: irritate stomach.
4. experimental technique and result:
4.1 body weight: animal is weighed, and presses the body weight random packet, is divided into 20 groups, 10 every group, and per four groups of unique experimental grouies, design was according to dosage fed 30 days continuously, claimed once weekly, and experiment with computing is first, neutralization experiment opisthosoma is heavy.Immunity is carried out the NK cell for one group and is changeed test with drenching, and immune two groups are carried out DTH, spleen index, thymus index test, and immune three groups are carried out hemolysis plaque and hemolysin test, and immune four groups are carried out the carbon clearance test, and immune five groups are carried out phagocytosis test.
The present invention's (immune one group) is to the influence of the weight of animals (gram)
Visible by last table, each dose groups of sample is compared with matched group, does not all have significant difference.
The present invention's (immune two groups) is to the influence of the weight of animals (gram)
Visible by last table, each dose groups of sample is compared with matched group, does not all have significant difference.
The present invention's (immune three groups) is to the influence of the weight of animals (gram)
Visible by last table, each dose groups of sample is compared with matched group, does not all have significant difference.
The present invention's (immune four groups) is to the influence of the weight of animals (gram)
Visible by last table, each dose groups of sample is compared with matched group, does not all have significant difference.
The present invention's (immune five groups) is to the influence of the weight of animals (gram)
Visible by last table, each dose groups of sample is compared with matched group, does not all have significant difference.
4.2 serum hemolysin test:
Animal is given appearance 30 days continuously, and after five days, eye socket is taken a blood sample with 2% (v/v) SRBC immune animal, and separation of serum is put to death in the cervical vertebra dislocation.On 96 hole Microhemagglutination plates, add 25ul serum, each row opposes and doubly dilutes later on, and every hole adds 1%SRBC100ul, shakes back 37 ℃ and places 3 hours, observed result after sinking appears in the blood cell contrast.
The serum hemolysin test
*(through variance analysis) compared in p＜0.05 with matched group
Statistical analysis: dosage, high dose group are compared with matched group and are all had significant difference among the present invention.
4.3 generating, antibody detects test:
Animal is given appearance 30 days continuously, and after five days, the dislocation of animal cervical vertebra is put to death, and removes spleen system splenocyte suspension with lower limb fiber sheep cellular immunization, adjustment cell concentration 5 * 10
6Individual/ml, with the surface medium heating for dissolving, put 45 ℃ of water insulations, mix with the Hanks liquid of equivalent pH7.2-7.4,2 times of concentration; Be divided into the every test tube of 0.5ml, add 50ul100%SRBC again, the 20ul splenocyte suspension; The speed mixing is inclined on the 6cm plate of brushing thin agar layer, puts into CO
2Incubator incubation 1.5h, the complement (1: 10) of reuse buffering dilution is added to plate, incubation 1.5h, counting hemolysis plaque number, the result adds up with variance analysis.
Antibody generates and detects test
Statistical analysis: high dose group of the present invention is compared with matched group all has significant difference.
4.4 the thymus index spleen index is measured:
Animal is given appearance 30 days continuously, weighs, and the cervical vertebra dislocation is put to death, and gets thymus, spleen is weighed, and calculates the thymus index spleen index.
Thymus index, spleen index
Statistical analysis: difference that each dose groups of the present invention compares with matched group that there are no significant.
4.5 mice carbon clearance test:
Animal is given appearance 30 days continuously, for the india ink of intravenous injection dilution in 1: 3, gets 20ul from vena ophthalmica after 2 minutes and adds Na
2CO
3In the solution, survey the OD value.Calculate phagocytic index, the result adds up with variance analysis.
The carbon clearance test
*(through variance analysis) compared in p＜0.05 with matched group
Statistical analysis: dosage, high dose group are compared with matched group and are all had significant difference among the present invention.
4.6ConA inductive mouse lymphocyte conversion test:
Animal is given appearance 30 days continuously, and the cervical vertebra dislocation is put to death, and gets spleen system splenocyte suspension, adjustment cell concentration 2 * 10
6Individual/ml, divide two empty 24 well culture plates that add with cell suspension, every hole 1ml, a hole adds 50ulConA, and another hole contrast is in 37 ℃, 5%CO
2Cultivate 72h, cultivate and finish preceding 4h, supernatant 0.7ml is drawn in every hole; Add the PRMI1640 culture fluid that does not contain calf serum, add MTT (5mg/ml) 50ul/ hole simultaneously, after continuing to cultivate 4h; Every hole adds 1ml acid isopropyl alcohol, and the piping and druming mixing is treated the purple crystal dissolving; With 570nm wavelength colorimetric, the result adds up with variance analysis.
The inductive mouse lymphocyte conversion test of ConA
*(through variance analysis) compared in p＜0.05 with matched group
Statistical analysis: dosage, high dose group are compared with matched group and are all had significant difference among the present invention.
4.6DTH measure:
Animal is given appearance 30 days continuously, and QUMAO is smeared sensitization with 1%DNFT50ul.Smear in mouse right ear with 1%DNFT10ul after five days.Dislocation is put to death after 24 hours, gets two ears about 8mm, weighs, and calculates the swelling degree.
DTH measures
Statistical analysis: difference that each dose groups of the present invention compares with matched group that there are no significant.
4.7NK cytoactive is measured:
Animal is given appearance 30 days continuously, and the cervical vertebra dislocation is put to death, and gets spleen, system splenocyte suspension (effector lymphocyte), and the back 24hYAC-1 cell that goes to go down to posterity adds 1640 complete culture solutions, and making cell concentration is 1 * 10
5Individual/ml (target cell) got target cell, each 100ul of effector lymphocyte, adds U type 96 well culture plates; The automatic release aperture of target cell adds target cell and each 100ul of culture fluid; Maximum release aperture adds target cell and each 100ul of 1%NP40, and above-mentioned each item is all established three multiple holes, in 37 ℃, 5%CO
2In the incubator four hours, every hole draw supernatant 100ul to the level land 96 well culture plates, add LDH substrate liquid 100ul simultaneously, reacted 3 minutes, every hole adds 1mol/L's
HCL 30ul stops, with ELIASA at 490mm place photometry density value (OD).
The NK cytoactive is measured
*(through variance analysis) compared in p＜0.05 with matched group
Statistical analysis: dosage, high dose group are compared with matched group and are all had significant difference among the present invention.
4.7 Turnover of Mouse Peritoneal Macrophages is engulfed the chicken red blood cell test:
Animal is given appearance 30 days continuously, every Mus lumbar injection 20% chicken erythrocyte suspension 1, and at a distance from 30 minutes, the cervical vertebra dislocation was put to death, and is fixed in the Mus plate, cuts off abdominal skin, and injecting normal saline 2 rotated the Mus plate 1 minute, and sucking-off abdominal cavity washing liquid 1 is divided and is dripped in 2 pieces of slides, and 37 ℃ of incubators are wet
Incubated 30 minutes, and used the normal saline rinsing, dry, fix with 1: 1 butanone methanol solution, 4%--phosphate buffer dyeing three minutes is dried with the distilled water rinsing, and oily mirror microscopy calculates and engulfs % and phagocytic index.
The macrophage phagocytic test
*(through variance analysis) compared in p＜0.05 with matched group
Statistical analysis: dosage, high dose group are compared with matched group and are all had significant difference among the present invention.
5. conclusion: the animal per os gives the effect that the present invention has enhancing immunity.
Claims (6)
1. the health care medicine of resisting fatigue and enhancing immunity is characterized in that it being that crude drug by following parts by weight is processed:
Coenzyme Q10 5-60 vitamin C 12.5-200
Vitamin E 1.5-50 folic acid 0.04-0.6.
2. the health care medicine of resisting fatigue according to claim 1 and enhancing immunity is characterized in that the weight proportion of said crude drug is:
Coenzyme Q10 20 vitamin Cs 50
Vitamin E 7.5 folic acid 0.15.
3. the health care medicine of resisting fatigue according to claim 1 and enhancing immunity is characterized in that: the dosage form of said medicine is soft capsule, tablet, granule or capsule.
4. resisting fatigue according to claim 1 and enhancing immunity health care medicine is characterized in that also comprising the auxilliary raw material of using of following parts by weight:
Soybean oil 90-1500 Cera Flava 5-150.
5. resisting fatigue according to claim 4 and enhancing immunity health care medicine is characterized in that it being to be processed with raw material with auxilliary by the crude drug of following parts by weight:
Coenzyme Q10 20 vitamin Cs 50
Vitamin E 7.5 folic acid 0.15
Soybean oil 307 Cera Flavas 16.
6. method for preparing like claim 4 or 5 described health care medicines may further comprise the steps:
A) take by weighing coenzyme Q10, vitamin C, vitamin E, folic acid, soybean oil, Cera Flava, subsequent use;
B) after the soybean oil heating with said weight proportion, add the Cera Flava of said weight proportion;
C) with coenzyme Q10, the vitamin E of said weight proportion, join in the above-mentioned fluid, stir and make dissolving; After cooling to appropriateness, add folic acid, the vitamin C of said weight proportion, mix; Grind evenly with colloid mill, be filled in the capsule, promptly get soft capsule.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810013722A CN101322711B (en) | 2008-01-09 | 2008-01-09 | Health care medicament for resisting fatigue and enhancing immunity and preparation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN102949411A (en) * | 2012-11-23 | 2013-03-06 | 西安泰科迈医药科技有限公司 | Antioxidative health care product and preparation method thereof |
| CN103494202B (en) * | 2013-10-18 | 2015-08-19 | 济南老来寿生物股份有限公司 | A kind of health food with anti-oxidation function |
| CN104857180B (en) * | 2015-05-29 | 2020-02-14 | 山东金城生物药业有限公司 | Composition for resisting fatigue and improving immunity and preparation method and application thereof |
| CN115554029A (en) * | 2022-11-07 | 2023-01-03 | 佛山市顺德区州福慈鑫无纺布有限公司 | Production process of coenzyme Q10 of sanitary towel |
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| CN1686547A (en) * | 2005-03-30 | 2005-10-26 | 淮北市辉克药业有限公司 | Long time use compound preparation for treating diabetes |
| HU0600428D0 (en) * | 2006-05-23 | 2006-07-28 | Szuecs Miklos Dr | Vitamine product |
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| CN1686547A (en) * | 2005-03-30 | 2005-10-26 | 淮北市辉克药业有限公司 | Long time use compound preparation for treating diabetes |
| HU0600428D0 (en) * | 2006-05-23 | 2006-07-28 | Szuecs Miklos Dr | Vitamine product |
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