CN1857350A - Immunological function strengthening medicine and health product and their preparing method - Google Patents
Immunological function strengthening medicine and health product and their preparing method Download PDFInfo
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Abstract
The present invention relates to medicine and health product technology, and is especially one kind of medicine or health product capable of strengthening immunologic function. The medicine or health product capable of strengthening immunologic function consists of earthworm extract 10~80 wt% and manyprickle acanthopanax root extract 10~70 wt%, as well as starch 5~30 wt%, mannitol 4~20 wt% and magnesium stearate 1~10 wt%.
Description
Technical field
The present invention relates to pharmaceutical products or field of health care products, particularly a kind of medicine of energy raise immunity or health product and preparation method thereof.
Background technology
Immunity is a kind of physiological function of human body; Human body relies on the composition of this function identification " oneself " and " non-own ", thereby destruction and repulsion enter intravital antigenic substance, as pathogen etc., or damaging cells that human body itself produced and tumor cell etc., keep balance of inside of human body environment and stable with this.This shows that the major function of immunity has: defence infection, homeostasis and immune surveillance.Prevention infection makes human body can resist the invasion and attack of pathogen etc., the generation that wards off disease, thus safeguard the health of human body; Homeostasis makes human body can in time remove intravital aging, death and damaging cells; Immune surveillance makes human body can discern and remove the abnormal cell (as tumor cell) that produces in the body at any time.Yet, raising along with people's living standard, the progressively increase of the adjustment of variation, the growth at age of life structure, the deficiency of sleep, dietary habit and the pressure of spirit constantly increases the load of our health, and self epidemic prevention ability but is to descend gradually; Simultaneously, along with the deterioration of environment, various viruses, antibacterial are really more and more various and have an attack; In order to keep the physique of a health, we are except needs are cultivated the immunity of self, and we also are necessary to carry out necessary replenishing by the medicine of enhancing immunity, particularly to the crowd of hypoimmunity.
Summary of the invention
The object of the present invention is to provide a kind of medicine or health product of energy raise immunity.
Another object of the present invention is to provide a kind of method for preparing said medicine or health product.
The objective of the invention is to be achieved through the following technical solutions:
A kind of medicine of raise immunity or health product, its active component comprises Pheretima extract and Radix Et Caulis Acanthopanacis Senticosi extract.
Described medicine or health product also comprise supplementary product starch, mannitol, magnesium stearate.
The weight percentage of described medicine or each component of health product is:
Pheretima extract 10～80%
Radix Et Caulis Acanthopanacis Senticosi extract 10～70%
Starch 5～30%
Mannitol 4～20%
Magnesium stearate 1～10%
Pheretima extract effective ingredient described in described medicine or the health product is the Pheretima polypeptide; The Radix Et Caulis Acanthopanacis Senticosi extract effective ingredient is a total saponins.
A kind of method for preparing said medicine or health product, it comprises following process,
With Pheretima extract and Radix Et Caulis Acanthopanacis Senticosi extract pulverize separately, sieve;
Mannitol and magnesium stearate are mixed;
Mixture and starch mixing granulation with Pheretima extract, Radix Et Caulis Acanthopanacis Senticosi extract, mannitol and magnesium stearate; Make required dosage form then.
Pheretima (earthworm) is commonly called as " Lumbricus ", and " bent meals ", another name " Pheretima " is an annelid.It is one of animal that has at first in the biological evolution process immunological memory and identification.Its salty in the mouth is cold in nature, returns liver, spleen, urinary bladder channel.Have the heat clearing away arresting convulsion, collateral dredging is relievingd asthma, the effect of diuresis.Extracted the peptide class of multiple biologically active in the Pheretima body, promptly the Pheretima peptide (lumbricus peptide, LP), it is the general name of micromolecule polypeptide in the Pheretima body, a large amount of documents shows that Pheretima extract has the effect of raise immunity.
Prove the effect that the Pheretima peptide has raise immunity by following experimentation:
1. the Pheretima Toplink of various dose strengthens mice M φ cytotoxic activity, and the cytotoxic activity of 3 dosage groups of LP M φ all strengthens to some extent, compare with the normal control group, 0.1,1 and the cytotoxic activity of 10mg/l LP group M φ obviously strengthen (P＜0.01)
2. the Pheretima peptide is to mice M φ, the influence of splenocyte TNF secretion-α, compare with the normal control group, 0.1,0.5mg/l LP group M φ level of TNF secretion-α after LPS induces obviously increases (P＜0.01), compares with the cyclophosphamide group, cyclophosphamide+LP group M φ level spontaneous and TNF secretion-α after LPS induces all obviously increases (P＜0.01).
3. the Pheretima peptide is compared with the normal control group to the influence of mouse boosting cell secretion IL-2, and 0.1g/l LP group splenocyte is spontaneous and all obviously increase (P＜0.01) through the amount that ConA induces secretion IL-2; The spontaneous secretion of 1g/l and 0.5g/l LP group IL-2 also increases (being respectively P＜0.01 and P＜0.05).After LPS induces, though the level of secretion IL-2 increases the no significant difference (P＞0.05) of comparing with the normal control group to some extent.Cyclophosphamide+LP group is compared with the cyclophosphamide group, all obviously risings (P＜0.01) of splenocyte level spontaneous and secretion IL-2 after ConA induces
4. each component of Pheretima peptide is to the influence of NK cytoactive, the Pheretima peptide is 0.1,0.5 and during 1mg/l in concentration, all can significantly improve NK cell activity (P＜0.001), and the big more effect of concentration is obvious more, the Pheretima peptide is the inhibitory action of antagonism cyclophosphamide (P＜0.05) obviously.
5. the Pheretima peptide is to the influence of normal M φ of mice and normal splenocyte secretion NO, and NO is generated by inducible nitric oxide synthase (iNOS) catalysis L arginine and oxygen molecule reaction in the M φ.NO plays an important role in the effect of activatory M φ cell killing tumor cell, and NO is a kind of newfound antibumor molecules, and 0.1g/l LP can make the level of M φ and splenocyte secretion NO obviously improve (p＜0.01).
6. the Pheretima Toplink significantly strengthens obviously the raise immunologic function of (P＜0.05) illustrative experiment group white mice of experimental group white mice spleen index and thymus index.The result show clearance in mice index and phagocytic index obviously raise (P＜0.01) illustrate that LP strengthens immune function of mice and obviously is better than Radix Astragali electuary.The plain measurement result of mice HD50 obviously raises, and illustrates that the humoral immune function of mice also has remarkable enhancing (P＜0.05).
In addition, abundant fatty acid, nucleic acid and derivant free amino acid thereof are arranged in the Lumbricus, also have trace element phosphorus, calcium, ferrum, potassium, zinc, copper and multivitamin, can be used as the ideal source of nutrition of human body.Particularly its protein content is very abundant, far above egg and general meat, 8 seed amino acids of needed by human is arranged wherein.In the U.S., Japan, Oceania and Africa the custom that Lumbricus is eaten as food is arranged all.Japan is the maximum country of consumption of present Lumbricus, also is maximum Lumbricus importer.Japanese food industrial applicability modern technologies are developed out the Lumbricus powder health food.This after measured Lumbricus health product contain protein 58.5%, fat 6.3%, 110 milligrams of linoleic acids, VB11 .3 milligram, 3.3 milligrams of vitamin Bies, multiple nutritional components such as ash 5.9%.Result of the test shows, this health food has the physical fatigue of adjusting situation, reduces the too high cholesterol of blood, brings high blood pressure down and prevents neuralgia and effect such as just secrete.Lumbricus also can be extracted aminoacid, as the raw material of light industry, produces skin Caring cosmetics and various food and health food.Is that the health food that primary raw material exploitation has an immunoregulation effect is feasible according to the modern medicine and pharmacology theory with the Pheretima peptide material.
Radix Et Caulis Acanthopanacis Senticosi extract is the extract of the root and rhizome of Araliaceae Radix Et Caulis Acanthopanacis Senticosi.Nature and flavor with return through acrid in the mouth, little hardship, warm in nature, return spleen, kidney, heart channel.Function with cure mainly: replenishing kidney, strengthening waist, benefiting vital QI for tranquillizing, promoting blood circulation to remove obstruction in the collateral.Cure mainly kidney deficiency and bodily weakness, soreness of the waist and knees, children's's walking retardation in children, insufficiency of the spleen weak, deficiency of vital energy edema, inappetence, insomnia and dreamful sleep, forgetful, thoracic obstruction pain, anemofrigid-damp arthralgia, treating swelling and pain by traumatic injury.
Chemical constituent: root contains eleutheroside A, B, B
1, C, D, E.Rhizome contains alkali solubility Radix Et Caulis Acanthopanacis Senticosi polysaccharide AS-II, AS-III and water solublity Radix Et Caulis Acanthopanacis Senticosi polysaccharide PES-A, the PES-B of tool immunologic enhancement.
The pharmacological action of Radix Et Caulis Acanthopanacis Senticosi extract:
1, to the effect of nonspecific stimulation
(1) antifatigue effect (2) resisting oxygen lack (3) anti-high and low-temp and anti-radiation function (4) anti-stress effect (5) Detoxication
2, to Immune Effects
(1) irritates stomach Radix Et Caulis Acanthopanacis Senticosi alcohol extraction aqueous solution 50g (crude drug)/kg1 time every day for mice or Cavia porcellus to the influence of monokaryon-macrophage system phagocytic function, continuous 14 days, 15, obviously increase monokaryon, phagocyte and peritoneal macrophage phagocytic activity.
(2) irritate stomach Radix Et Caulis Acanthopanacis Senticosi extract 0.02ml for every of mice every day to the influence of lymphocyte function, continuous 9 days, obviously stop because of T, bone-marrow-derived lymphocyte, killer cell, non-specific immunity due to the fatigue of swimming to descend.
Radix Et Caulis Acanthopanacis Senticosi polysaccharide obviously increases mouse spleen cell number and mesenteric lymph node cell number, and the cell number minimizing of caused by cyclophosphamide is had remarkable antagonistic effect.Radix Et Caulis Acanthopanacis Senticosi polysaccharide can make mouse spleen white pulp cumulative volume and cortex of lymph node cumulative volume obviously increase, and can significantly resist the effect that cyclophosphamide reduces spleen white pulp cumulative volume and cortex of lymph node cumulative volume.As immunomodulator, Radix Et Caulis Acanthopanacis Senticosi polysaccharide can promote lymphopoiesis, and it is active to make it metabolism, and increased functionality based on this morphological change, has promoted body's immunological function.Effect such as the slow down aging of Radix Et Caulis Acanthopanacis Senticosi polysaccharide, antitumor may be inseparable to immune effect with it.
Radix Et Caulis Acanthopanacis Senticosi polysaccharide has an effect that the inducing mouse splenocyte produces IL-2 external, and the mechanism of prompting polysaccharide enhancing human body immunity may be with to induce body to produce IL-2 relevant.IL-2 is a kind of very important lymphokine, has the Th of inducing cell and Tc cell proliferation, stimulates the B cell to produce antibody, and activated macrophage strengthens NK cell and the active effect of LAK.
Macrophage is the strongest class cell of effect in the mononuclear phagocyte system, also is the antineoplastic effector lymphocyte.Radix Et Caulis Acanthopanacis Senticosi extractum and ethanol extract have the phagocytosis of obvious enhancing macrophage, and Turnover of Mouse Peritoneal Macrophages phagocytic function under the stress state is studied, and the result shows that the main component that works is a Radix Et Caulis Acanthopanacis Senticosi glucoside.From stress learning angle, because the adaptation former state effect of Radix Et Caulis Acanthopanacis Senticosi makes the secretion of stress state mice adrenocortical hormone down be controlled at appropriate level, and then the phagocytic function of promotion mononuclear phagocyte system.Thereby the raising immunologic function is improved the body health situation.
(3) influence that forms of antagonist to LACA system and C57BL/6 mouse inbred lines lumbar injection every day Radix Et Caulis Acanthopanacis Senticosi polysaccharide injection 12.5mg/kg, 25mg/kg, 50mg/kg, 100mg/kg1 time, after the administration 9 days, 25-100mg/kg adds the slender acanthopanax polysaccharide obviously increases the antibody secreting cell of mice IgG secretion and IgM (PFC), and prompting strengthens the specific humoral immunity function.
(4) to the influence of the interferon culture medium with the Radix Et Caulis Acanthopanacis Senticosi polysaccharide that contains 10 μ g/ml, will bring out the cell pretreatment after 24-48 hour, and start 2 hours with the 150U/ml interferon, the result proves that Radix Et Caulis Acanthopanacis Senticosi polysaccharide significantly improves the ability that cell produces interferon.
(5) effect of leukocyte increasing is arranged.
The above results prompting Radix Et Caulis Acanthopanacis Senticosi extract is many-sided to Immune Effects.
Further specify effect of the present invention below by concrete pharmacological evaluation.
The report of dragon peptide immunity capsule enhancing immunity function animal experiment
1 laboratory animal
SPF level Kunming kind female mice, 18-22g, laboratory animal production licence number: SCXK (Hubei Province) 2003-0005; Laboratory animal occupancy permit number: SYXK (Hubei Province) 2003-0014.The animal feeding room temperature is 22-26 ℃, and humidity is 47-67%.
2 sample surveys and foundation
A kind of by the capsular hard capsule of imperial peptide immunity of being named as of supplementary materials such as Pheretima extract, Radix Et Caulis Acanthopanacis Senticosi extract, starch, mannitol, magnesium stearate making, press the relevant regulations of " health food check and assessment technique standard " the middle health food evaluation test project of version (function assessment assessment process), test principle and result's judgement in 2003 and test.
The grouping of 3 dosage
Recommend daily intaking amount 0.05g/kg BW to enlarge 10,20,30 times by the crowd and set up basic, normal, high three dosage groups, 1.5g/kg BW promptly 0.5,1.0,, be divided into four experimental grouies, every experimental group comprises blank group, basic, normal, high dosage group, and each dosage group laboratory animal number is 10.One group of immunization experiment carries out delayed allergy and Turnover of Mouse Peritoneal Macrophages is engulfed the chicken red blood cell experiment; Immunization experiment carries out inductive mouse lymphocyte transformation experiment of ConA and NK cytoactive mensuration for two groups; Three groups of immunization experiments carry out lymphatic organ/weight ratio pH-value determination pH and carbon is cleaned up experiment; Four groups of mensuration and antibody-producting cells that carry out serum hemolysin of immunization experiment detect.Mice continuous irrigation stomach begins test after 30 days.Each experimental group is all irritated stomach by 0.20ml/10g BW capacity per os and is given.
4 test methods and result
4.1 method
4.1.1 the inductive mouse spleen lymphocyte conversion test of ConA
Method: the aseptic spleen of getting, place to fill an amount of aseptic Hank ' s liquid plate, gently spleen is ground with tweezers, make the individual cells suspension.Filter through 200 eye mesh screens, use Hank ' s liquid to wash 2 times, each centrifugal 10min (1000r/min).Then with cell suspension in the complete culture solution of 1mL, with the blue dyeing counting viable count of platform phenol (should more than 95%), adjusting cell concentration be 3 * 10
6Individual/mL.Divide two holes to add in 24 well culture plates each part splenocyte suspension, every hole 1mL, a hole adds 75 μ LConA liquid (being equivalent to 7.5 μ g/mL), and 5%CO is put in contrast in another hole
2, cultivate 72h in 37 ℃ of incubators.Cultivate and finish preceding 4h, supernatant 0.7mL is inhaled in every hole gently, adds the RPMl1640 culture fluid that 0.7mL does not contain calf serum, adds MTT (5mg/mL) 50 μ L/ holes simultaneously, continues to cultivate 4h.After cultivating end, every hole adds 1mL acid isopropyl alcohol, and the piping and druming mixing dissolves purple crystal fully.Divide then to install in 96 well culture plates, 3 parallel holes (100 μ L/ hole) are made in each hole, measure optical density value with microplate reader with the 570nm wavelength.
4.1.2 dinitrofluorobenzene (DNFB) is induced delayed allergy (DTH)
Method: ear swelling method.Behind 1%DNFB (with 1: 1 acetone Oleum Sesami solution preparation) sensitized mice, reuse DNFB attacked auris dextra in the 5th day, put to death animal behind the 24h and cut left and right sides auricular concha and take off directly through the auricle of 8mm, weigh, represent the degree of DTH with the difference of the weight of left and right sides ear with card punch.
4.1.3 the mensuration of serum hemolysin
Method: blood clotting method.Get Sanguis caprae seu ovis, with normal saline washing 3 times, centrifugal at every turn (2000r/min) 10min.Hematocrit SRBC is made into the cell suspension of 2% (v/v) with normal saline, and every Mus lumbar injection 0.2mL carries out immunity.After 4～5 days, extract eyeball and get blood in centrifuge tube, place about 1h, solidification blood and tube wall are peeled off, serum is fully separated out, the centrifugal 10min of 2000r/min collects serum.
Agglutination: with normal saline with the serum doubling dilution, the dilution serum of difference is placed respectively in the Microhemagglutination brassboard, every hole 100 μ L, add 100 μ L 0.5% (v/v) SRBC suspensions again, mixing, put into moistening square position and add a cover,, observe the hemagglutination degree in 37 ℃ of incubation 3h.Level calculation according to serum cohesion degree goes out the antibody product.
4.1.4 peritoneal macrophage is engulfed the chicken red blood cell test
Method: half intracorporal method.The chicken erythrocyte suspension of preparation 20%; Every this suspension of Mus lumbar injection 1mL is put to death animal behind the 30min, it is faced upward the position be fixed on the Mus plate, opens abdomen, injects normal saline 2mL through the abdominal cavity, rotates Mus plate 1min, and then, sucking-off abdominal cavity washing liquid 1mL, average mark drip on 2 microscope slides, 37 ℃ of incubation 30min; Educate to finish and use the normal saline rinsing, dry, fix with 1: 1 acetone methanol solution, 4%Giemsa-phosphate buffer dyeing 3min, the rinsing of reuse distilled water is dried.The oil mirror is 100 macrophages of counting down, are calculated as follows phagocytic rate and phagocytic index:
4.1.5 mice carbon clearance test
Method: inject the india ink (10mL/kg) of dilution by body weight from mouse tail vein, treat that prepared Chinese ink injects, timing immediately injects behind the prepared Chinese ink 2,10min, gets blood 20 μ L from the angular vein clump respectively, exists side by side to be about to it and to be added to 2mL0.1% Na
2CO
3In the solution.Each inhales 0.1mL in 96 hole ELISA Plate, uses microplate reader at 600nm wavelength place's photometry density value (OD), with Na
2CO
3Solution is made blank.
Mice is put to death, get liver and spleen, blot the organ surface blood stains, weigh respectively with filter paper.
Represent the ability that mice carbon is cleaned up with phagocytic index.Be calculated as follows phagocytic index a.The phagocytic index of given the test agent group is significantly higher than matched group, this experimental result positive of decidable.
4.1.6 antibody-producting cell detects
Get the Sanguis caprae seu ovis of defiber, with normal saline washing 3 times, centrifugal at every turn (2000r/min) 10min, every Mus is through lumbar injection 2% (v/v) SRBC 0.2mL.The mice cervical vertebra dislocation of SRBC immunity after 4～5 days put to death, take out spleen, be placed in the little plate that is loaded with Hank ' s liquid, grind spleen gently, make cell suspension, filter through 200 eye mesh screens, centrifugal (1000/min) 10min uses Hank ' s liquid to wash 2 times, at last with cell suspension in 5mL RPMI1640 culture fluid, counting cells, and cell concentration is adjusted into 5 * 10
6Individual/mL.
The mensuration of plaque: after top layer culture medium (the 1g agarose adds distilled water to 100mL) heating for dissolving, put into 45～50 ℃ of water bath heat preservations, mix with Hank ' the s liquid of equivalent pH7.2～7.4,2 times concentration, the packing small test tube, every pipe 0.5mL, in pipe, add 50 μ L10%SRBC (v/v is with the preparation of SA buffer) again, 20 μ L splenocyte suspensions (5 * 10
6Individual/mL), rapid mixing, be poured on the slide of oneself brush agarose thin layer, do parallel plate, treat that agar solidifies after, the slide level buckled be placed on the horse, put into CO2 gas incubator and hatch 1.5h, complement (1: 8) with the dilution of SA buffer adds in the slide frame groove then, behind the continuation incubation 1.5h, and counting hemolysis plaque number.
4.1.7 the NK cytoactive is measured
Method: lactic acid dehydrogenase (LDH) algoscopy.
Go down to posterity (the YAC-1 cell) of target cell:
24h is with the target cell cultivation of going down to posterity before the experiment.Wash 3 times with Hank ' s liquid with preceding, cultivating the adjustment cell concentration fully with RPMI1640 is 4 * 10
5Individual/mL.
The preparation of splenocyte suspension (effector lymphocyte):
The aseptic spleen of getting places the little plate that fills an amount of aseptic Hank ' s liquid, gently spleen is ground with tweezers, makes single cell suspension.Filter through 200 eye mesh screens, use Hank ' s liquid to wash 2 times, each centrifugal 10min (1000r/min).Abandoning supernatant upsprings cell, added the 0.5mL aquesterilisa 20 seconds, add 0.5mL2 times of Hank ' s liquid and 8mLHanks liquid after the splitting erythrocyte again, 1000r/min, 10min is centrifugal, and the RPMI1640 complete culture solution that contains 10% calf serum with 1mL is resuspended, with 1% glacial acetic acid dilution back counting (viable count should more than 95%), with the blue dyeing counting viable count of platform phenol (should more than 95%), be 2 * 10 with RPMI 1640 complete culture solutions adjustment cell concentration at last
7Individual/mL.
The NK cytoactive detects:
Get each 100 μ L of target cell and effector lymphocyte (imitating target than 50: 1), add in U type 96 well culture plates: target cell nature release aperture adds target cell and each 100 μ L of culture fluid, and the maximum release aperture of target cell adds target cell and each 100 μ L of 1%NP40; Above-mentioned every three parallel holes of all establishing are in 37 ℃, 5%CO
2Cultivate 4h in the incubator, then with 96 well culture plates with the centrifugal 5min of 1500r/min, draw at the bottom of the supernatant 100 μ L horizontalizations in 96 well culture plates in every hole, add LDH substrate liquid 100 μ L simultaneously, according to room temperature differential responses 3～10min, every hole adds the HCl30 μ L of 1mol/L, measures optical density value (OD) at microplate reader 490nm place.
Be calculated as follows the NK cytoactive, the NK cytoactive of given the test agent group is significantly higher than the NK cytoactive of matched group, i.e. this experimental result positive of decidable.
4.2 result
4.2.1 the weight of animals result before and after the test: see Table 1, body weight no significant difference between table 2, table 3, each treated animal of table 4.
The weight of animals record before and after the table 1 experiment battery of tests (x ± SD)
| Group | Dosage (g/kg BW) | Number of animals (only) | Body weight (gram) | Weightening finish (gram) | |
| Before the test | After the test | ||||
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 20.0±1.0 19.5±0.9 19.9±1.0 19.7±1.0 | 32.2±0.8 32.0±1.0 32.3±0.7 32.4±0.8 | 12.2±0.8 12.5±0.7 12.4±0.9 12.7±1.0 |
The weight of animals record before and after two groups of tests of table 2 experiment (x ± SD)
| Group | Dosage (g/kg BW) | Number of animals (only) | Body weight (gram) | Weightening finish (gram) | |
| Before the test | After the test | ||||
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 19.7±1.2 20.2±1.0 19.8±0.9 19.7±1.0 | 32.2±0.9 32.5±0.8 32.3±0.7 32.4±0.7 | 12.5±1.2 12.3±1.1 12.5±0.7 12.7±1.0 |
The weight of animals record before and after three groups of tests of table 3 experiment (x ± SD)
| Group | Dosage (g/kg BW) | Number of animals (only) | Body weight (gram) | Weightening finish (gram) | |
| Before the test | After the test | ||||
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 19.9±1.0 20.1±1.1 19.8±1.0 20.1±0.9 | 32.4±0.8 32.6±0.7 32.7±0.6 32.8±0.8 | 12.4±0.8 12.5±0.8 12.9±1.2 12.7±1.1 |
The weight of animals record before and after four groups of tests of table 4 experiment (x ± SD)
| Group | Dosage (g/kg BW) | Number of animals (only) | Body weight (gram) | Weightening finish (gram) | |
| Before the test | After the test | ||||
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 20.3±1.0 19.6±0.8 19.5±1.0 19.8±0.6 | 32.5±0.9 32.0±0.7 32.3±0.9 32.3±0.7 | 12.2±0.9 12.4±1.1 12.8±1.1 12.5±1.0 |
4.2.2 the influence to animal lymph organ/body weight ratio: see Table 5.Thymus between each treated animal/body weight ratio and spleen/body weight ratio no significant difference (P＞0.05, P＞0.05).
The imperial peptide of table 5 immunity capsule is to the influence of animal lymph organ/body weight ratio (x ± SD)
| Group | Dosage (g/kg BW) | Number of animals (only) | Thymus/body weight ratio (* 10 3) | The P value | Spleen/body weight ratio (* 10 3) | The P value |
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 2.90±0.26 2.88±0.27 2.87±0.21 2.86±0.23 | - 0.840 0.797 0.713 | 3.70±0.21 3.74±0.21 3.73±0.19 3.72±0.25 | - 0.727 0.760 0.913 |
4.2.3 splenocyte transformation experiment result: see Table 6.As seen from Table 6, compare with the blank group, imperial peptide immunity capsule in high dose group significance strengthens the inductive spleen lymphocyte proliferation ability of ConA P＜0.05).
4.2.4 delayed allergy experimental result: see Table 6.As seen from Table 6: compare with the blank group, imperial peptide immunity capsule in high dose group significance strengthens the DTH reaction (p＜0.05) that mice is brought out DNFB.
The imperial peptide immunity of table 6 capsule is to the mouse cell Immune Function
| Group | Dosage (g/kg BW) | Number of animals (only) | ConA induces spleen lymphocyte proliferation OD difference | The P value | DNFB induces DTH left and right sides ear weight difference (mg) | The P value |
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 0.054±0.019 0.069±0.020 0.072±0.023 0.080±0.017 | - 0.243 0.116 0.016 | 12.63±2.74 15.10±2.41 15.43±3.09 16.24±3.13 | - 0.149 0.089 0.021 |
4.2.5 serum hemolysin determination experiment result: see Table 7.As seen from Table 7, compare with the blank group, but imperial peptide immunity capsule in high dose group significance rising serum hemolysin content (P＜0.05).
4.2.6 peritoneal macrophage is engulfed the chicken red blood cell experimental result: see Table 8.As seen from Table 8, compare with the blank group, imperial peptide immunity capsule in high dose group obviously improves phagocytic rate, phagocytic index (P＜0.05, P＜0.05).
The imperial peptide immunity of table 7 capsule is to the influence of mouse humoral immune function
| Group | Dosage (g/kg BW) | Number of animals (only) | Serum hemolysin (antibody product) | The P value |
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 l0 | 81.2±11.9 93.8±18.9 96.4±13.7 103.2±18.3 | - 0.204 0.101 0.011 |
The imperial peptide immunity of table 8 capsule is to the influence of Turnover of Mouse Peritoneal Macrophages phagocytic function
| Group | Dosage (g/kg BW) | Number of animals (only) | Phagocytic percentage (%) | The P value | Phagocytic index | The P value |
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 25.500±2.173 27.200±1.874 27.600±2.066 28.800±2.658 | - 0.209 0.127 0.011 | 0.345±0.025 0.363±0.029 0.370±0.022 0.378±0.028 | - 0.300 0.099 0.021 |
4.2.7 carbon is cleaned up experimental result: see Table 9.As seen from Table 9, compare, clean up phagocytic index (P＜0.05) but imperial peptide immunity capsule in high dose group significance improves mice carbon with the blank group.
The imperial peptide immunity of table 9 capsule is cleaned up function to mice carbon influence
| Group | Dosage (g/kg BW) | Number of animals (only) | Phagocytic index a | The P value |
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 4.63±0.99 5.38±0.89 5.51±0.92 5.75±0.61 | - 0.155 0.076 0.018 |
4.2.8 antibody-producting cell test experience result: see Table 10.See from table 10, compare that imperial peptide immunity capsule in high dose group can obviously strengthen antibody-producting cell ability (P＜0.05) with the blank group.
The influence of the imperial peptide immunity of table 10 capsule antagonist cellulation function
| Group | Dosage (g/kg BW) | Number of animals (only) | Hemolysis plaque number (/ 10 6Splenocyte) | The P value |
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 261.4±20.64 276.5±26.82 279.6±21.60 291.9±17.12 | - 0.296 0.170 0.010 |
4.2.9 NK cytoactive experimental result: see Table 11.As seen from Table 11, compare with the blank group, imperial each dosage group of peptide immunity capsule all can not strengthen NK cells in mice activity (P＞0.05).
The imperial peptide immunity of table 11 capsule is to the active influence of NK cells in mice
| Group | Dosage (g/kg BW) | Number of animals (only) | Cytoactive (%) | The P value |
| Dosage group high dose group in the blank group low dose group | 0.0 0.5 1.0 1.5 | 10 10 10 10 | 14.56±2.78 16.24±3.19 16.49±2.47 16.90±2.02 | - 0.371 0.247 0.125 |
5 conclusions
The crowd recommends daily intaking amount 0.05g/kg BW to enlarge 10,20,30 times and sets up basic, normal, high three dosage groups, and promptly 0.5,1.0,1.5/kg BW, other establishes the blank group.Adopt SPF level Kunming mouse, per os filling stomach gives continuously, begins experiment after 30 days.Experimental result is judged as significant difference with P≤0.05, and the result shows: 1) each dosage group and blank group comparison, and the mice body weight, lymphatic organ/there are no significant for the body weight ratio difference; 2) high dose group can obviously strengthen the inductive mice spleen lymphocytes proliferation ability of ConA; 3) high dose group can obviously strengthen the inductive mice delayed allergy of dinitrofluorobenzene; 4) the high dose group mice serum hemolysin content that can obviously raise; 5) high dose group can obviously strengthen Turnover of Mouse Peritoneal Macrophages and engulfs the chicken red blood cell function; 6) high dose group can obviously strengthen mice carbon and cleans up ability; 7) high dose group can obviously strengthen the antibody-producting cell ability; 8) each dosage group all can not strengthen the NK cells in mice activity.According to enhancing immunity function assessment process regulation, this is tried thing and is had the enhancing immunity function.
The specific embodiment:
Embodiment one
Get 10 parts of Pheretima extracts (its effective ingredient is the Pheretima polypeptide) and Radix Et Caulis Acanthopanacis Senticosi extract 70 (effective ingredient is a total saponins) pulverize separately, sieve;
Get 5 parts in mannitol and 10 parts of mixing of magnesium stearate;
With 5 parts of the mixture of Pheretima extract, Radix Et Caulis Acanthopanacis Senticosi extract, mannitol and the magnesium stearate of above-mentioned deal and starch mixing granulation at normal temperatures and pressures.Carry out capsule then and fill, make capsule.
Embodiment two
Get 80 parts of Pheretima extracts and Radix Et Caulis Acanthopanacis Senticosi extract 10 pulverize separately, sieve;
Get 4 parts in mannitol and 1 part of mixing of magnesium stearate;
With 5 parts of the mixture of Pheretima extract, Radix Et Caulis Acanthopanacis Senticosi extract, mannitol and the magnesium stearate of above-mentioned deal and starch mixing granulation at normal temperatures and pressures.Tabletting gets tablet then.
Embodiment three
Get 25 parts of Pheretima extracts and Radix Et Caulis Acanthopanacis Senticosi extract 25 pulverize separately, sieve;
Get 20 parts in mannitol and 10 parts of mixing of magnesium stearate;
With 30 parts of the mixture of Pheretima extract, Radix Et Caulis Acanthopanacis Senticosi extract, mannitol and the magnesium stearate of above-mentioned deal and starch mixing granulation at normal temperatures and pressures.Carry out capsule then and fill, make capsule.
Embodiment four
Get 40 parts of Pheretima extracts and Radix Et Caulis Acanthopanacis Senticosi extract 10 pulverize separately, sieve;
Get 20 parts in mannitol and 10 parts of mixing of magnesium stearate;
With 30 parts of the mixture of Pheretima extract, Radix Et Caulis Acanthopanacis Senticosi extract, mannitol and the magnesium stearate of above-mentioned deal and starch mixing granulation at normal temperatures and pressures.Carry out capsule then and fill, make capsule.
Embodiment five
Get 40 parts of Pheretima extracts and Radix Et Caulis Acanthopanacis Senticosi extract 40 pulverize separately, sieve;
Get 4 parts in mannitol and 10 parts of mixing of magnesium stearate;
With 6 parts of the mixture of Pheretima extract, Radix Et Caulis Acanthopanacis Senticosi extract, mannitol and the magnesium stearate of above-mentioned deal and starch mixing granulation at normal temperatures and pressures.
Claims (10)
1, a kind of medicine of raise immunity or health product is characterized in that, its active component comprises Pheretima extract and Radix Et Caulis Acanthopanacis Senticosi extract.
2, the medicine of a kind of raise immunity according to claim 1 or health product is characterized in that, the weight percentage of described each component is: Pheretima extract 10～80%,
Radix Et Caulis Acanthopanacis Senticosi extract 10～70%.
3, the medicine of a kind of raise immunity according to claim 1 and 2 or health product is characterized in that, described medicine or health product also comprise supplementary product starch, mannitol, magnesium stearate.
4, the medicine of a kind of raise immunity according to claim 3 or health product is characterized in that, the weight percentage of described each component of adjuvant is:
Starch 5～30%
Mannitol 4～20%
Magnesium stearate 1～10%
5, the medicine of a kind of raise immunity according to claim 1 and 2 or health product is characterized in that, described Pheretima extract effective ingredient is the Pheretima polypeptide.
6, the medicine of a kind of raise immunity according to claim 1 and 2 or health product is characterized in that, described Radix Et Caulis Acanthopanacis Senticosi extract effective ingredient is a total saponins.
7, the medicine of a kind of raise immunity according to claim 1 and 2 or health product is characterized in that, described medicine or health product can be made acceptable any solid preparation on the materia medica.
8, the medicine of a kind of raise immunity according to claim 7 or health product is characterized in that, described solid preparation is a capsule.
9, a kind of method for preparing the medicine or the health product of any one described raise immunity among the claim 1-8, it comprises following process: with Pheretima extract and Radix Et Caulis Acanthopanacis Senticosi extract pulverize separately, sieve; Mannitol and magnesium stearate are mixed; Mixture and starch mixing granulation with Pheretima extract, Radix Et Caulis Acanthopanacis Senticosi extract, mannitol and magnesium stearate; Make required dosage form then.
10, preparation method according to claim 9 is characterized in that, made dosage form is tablet or granule or capsule.
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| CNB2006100345784A CN100544739C (en) | 2006-03-27 | 2006-03-27 | A kind of medicine of raise immunity or health product and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2006100345784A CN100544739C (en) | 2006-03-27 | 2006-03-27 | A kind of medicine of raise immunity or health product and preparation method thereof |
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| CN1857350A true CN1857350A (en) | 2006-11-08 |
| CN100544739C CN100544739C (en) | 2009-09-30 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CZ302051B6 (en) * | 2010-02-08 | 2010-09-15 | Enzymix S. R. O. | Immunomodulating biological composition exhibiting regenerative and vitalization activity and process for preparing thereof |
| CN103169943A (en) * | 2013-03-01 | 2013-06-26 | 珠海博康药业有限公司 | Anti-fatigue compound earthworm protein preparation |
| CN105535026A (en) * | 2016-01-08 | 2016-05-04 | 中国人民解放军第二军医大学 | Application of lumbricus in preparing anti-radiation product |
| CN107007558A (en) * | 2017-04-20 | 2017-08-04 | 成都农业科技职业学院 | A kind of sustained release preparation for animals containing Pheretima extract and preparation method thereof |
-
2006
- 2006-03-27 CN CNB2006100345784A patent/CN100544739C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CZ302051B6 (en) * | 2010-02-08 | 2010-09-15 | Enzymix S. R. O. | Immunomodulating biological composition exhibiting regenerative and vitalization activity and process for preparing thereof |
| CN103169943A (en) * | 2013-03-01 | 2013-06-26 | 珠海博康药业有限公司 | Anti-fatigue compound earthworm protein preparation |
| CN103169943B (en) * | 2013-03-01 | 2014-04-16 | 珠海博康药业有限公司 | Anti-fatigue compound earthworm protein preparation |
| CN105535026A (en) * | 2016-01-08 | 2016-05-04 | 中国人民解放军第二军医大学 | Application of lumbricus in preparing anti-radiation product |
| CN107007558A (en) * | 2017-04-20 | 2017-08-04 | 成都农业科技职业学院 | A kind of sustained release preparation for animals containing Pheretima extract and preparation method thereof |
| CN107007558B (en) * | 2017-04-20 | 2020-07-03 | 成都农业科技职业学院 | A veterinary sustained release preparation containing Lumbricus extract, and its preparation method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100544739C (en) | 2009-09-30 |
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