CN103251816A - Preparation for enhancing immunity and preparation method thereof - Google Patents
Preparation for enhancing immunity and preparation method thereof Download PDFInfo
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Abstract
The invention provides a preparation for enhancing immunity and a preparation method thereof. The preparation consists of the following components in parts by mass: 10-15 parts of dendrobium candidum powder, 4-10 parts of notoginseng powder, 1-1.5 parts of astragalus root extract, 0.2-0.5 part of fructus corni extract, 0.1-0.4 part of red sage root extract, and 98-105 parts of honey. The preparation method comprises the steps of uniformly mixing the dendrobium candidum powder, the notoginseng powder, the astragalus root extract, the fructus corni extract and the red sage root extract, then adding the honey, and uniformly stirring to obtain the preparation for enhancing the immunity. The preparation is suitable for persons with low immunity, can enhance the immunity of human bodies, and has the advantages of quick and obvious effect, high safety, low cost, simplicity and convenience in operation and the like.
Description
Technical field
The present invention relates to preparation of a kind of enhancing immunity and preparation method thereof, belong to the biological health-care product technical field.
Background technology
Immunity refers to the defense mechanism of human body self, be human body identification and any foreign body (virus, antibacterial etc.) of eliminating external intrusion, and handle self aging, damage, death, the cell of degeneration and the ability of identification and processing vivo mutations cell and virus infected cell.Modern immunology thinks that immunity is human body identification and the physiological reaction of getting rid of " dissident ".What carry out this function in the human body is immune system.For a long time, people live in the environment that not only had been fit to existence but also had been fraught with risks, and the mankind can survival, is because obtained outstanding immunity.Immunity is the product of biological evolution process thus.But along with the development of society, people's live and work rhythm is constantly accelerated, and pressure is also increasing, adds the pollution of living environment, causes physical function to descend, and hypoimmunity very easily causes infection such as antibacterial, virus, fungus.Therefore the most direct performance of hypoimmunity is exactly liable to illness; because of often ill; increased the weight of the consumption of body again; show as have a delicate constitution, malnutrition, lethargy, fatigue and weak, appetite depression, sleep disorder etc.; sick, have an injection and take medicine the normal potluck that just gets married; and each sickly all will could recover for a long time, and usually outbreak repeatedly.If things go on like this can cause health and intelligence development bad, also easily bring out major disease.
At present, have the health product of numerous enhancing immunity on the market, but great majority DeGrain or take effect slowlyer all even there is not health-care effect because of human body difference, does not satisfy numerous consumer demands.
Summary of the invention
The object of the present invention is to provide a kind of instant effect and outstanding effect can strengthen preparation of body immunity and preparation method thereof, to satisfy consumers in general's demand.
The present invention realizes by following technical proposal: a kind of preparation of enhancing immunity is characterized in that being made up of the component of following mass parts:
10～15 parts in Herba Dendrobii powder, 4～10 parts of Radix Notoginseng powder,
1～1.5 part of Radix Astragali extract, 0.2～0.5 part of Fructus Corni extract,
0.1～0.4 part of Radix Salviae Miltiorrhizae extract, 98～105 parts of Mel.
Described Herba Dendrobii powder is that Herba Dendrobii is crushed to fineness is 1200 purpose superfine powder.
Described Radix Notoginseng powder is that Radix Notoginseng powder is broken to fineness is 1200 purpose superfine powder.
Described Radix Astragali extract, Fructus Corni extract, Radix Salviae Miltiorrhizae extract are commercial products, perhaps extract through following conventional method:
A, respectively each crude drug of the described Radix Astragali, Fructus Corni, Radix Salviae Miltiorrhizae is pulverized after, decocting extracts 2～3 times, and each water consumption is 8～12 times of medical material gross mass, and each decocting extracts and keeps little boiling 0.5～1.5 hour, merge each time extracting solution, filter to isolate filtrate and filtering residue;
B, the filtrate of A step is concentrated into 1g medical material/ml, cools off, leaves standstill to room temperature, stir that slowly to add ethanol to ethanol final concentration down be 60wt%, leave standstill to precipitation fully, sucking filtration is isolated filtrate and filtering residue;
C, be 60% washing with alcohol with the filtering residue mass concentration of B step, colourless until cleaning mixture, cleaning mixture is incorporated in the filtrate of B step, reclaim ethanol, drying is pulverized, respectively extract---the dry extract of the Radix Astragali, Fructus Corni, Radix Salviae Miltiorrhizae.
Another object of the present invention is to provide a kind of preparation method of preparation of enhancing immunity, it is characterized in that through following each step:
(1) get the raw materials ready by following mass parts:
10～15 parts in Herba Dendrobii powder, 4～10 parts of Radix Notoginseng powder,
1～1.5 part of Radix Astragali extract, 0.2～0.5 part of Fructus Corni extract,
0.1～0.4 part of Radix Salviae Miltiorrhizae extract, 98～105 parts of Mel.
(2) fineness with step (1) is that 1200 purpose Herba Dendrobii superfine powder, fineness are 1200 purpose superfine notoginseng powders, with Radix Astragali extract, Fructus Corni extract and Radix Salviae Miltiorrhizae extract mix homogeneously, add Mel again and stir, the preparation of the immunity that namely is enhanced.
Above-mentioned preparation provided by the invention is prepared into the oral formulations such as capsule, granule, tablet, powder or pill with enhancing immunity effect by conventional moulding process.
Wherein, Herba Dendrobii: nature and flavor: sweet in the mouth, cold nature; Effect with the nourishing the stomach of promoting the production of body fluid, nourishing YIN and clearing away heat, lung moistening kidney tonifying, the strong waist that makes eye bright.Radix Notoginseng: nature and flavor: sweet in the mouth, little hardship, warm in nature; Return Liver Channel, stomach warp, heart channel, lung meridian, large intestine channel; Functions such as hemostasis, diffusing blood, analgesic therapy are arranged.The Radix Astragali: nature and flavor: sweet, slightly warm in nature; Return liver, spleen, lung, kidney channel; There are benefiting QI for strengthening the superficies, arresting sweating to take off, hold in the palm the effect of skin ulcer granulation promoting, inducing diuresis to remove edema admittedly.Fructus Corni: nature and flavor: sour, puckery, tepor; Return liver, kidney channel; Be used for liver and kidney tonifying, restrain astringent or styptic treatment for spontaneous sweating, controlling nocturnal emission with astringent drugs reducing urination leukorrhagia stopping only collapses, hidroschesis, the effect of promoting the production of body fluid to quench thirst in addition in addition.Radix Salviae Miltiorrhizae: nature and flavor: hardship is slightly cold; GUIXIN, Liver Channel; Be used for promoting blood flow to regulate menstruation, stasis-dispelling and pain-killing, removing heat from blood eliminating carbuncle, the relieving restlessness that clears away heart-fire, nourishing blood to tranquillize the mind.Mel: nature and flavor: sweet, property is put down; Return spleen, stomach, lung, large intestine channel; Split for harmonizing the spleen and stomach, relieving spasm to stop pain, nourishing the lung to arrest cough, loosening bowel to relieve constipation, skin moistening granulation promoting, detoxifcation, the empty pain of main gastral cavity abdomen, xeropulmonary cough, dryness of the intestine constipation, conjunctival congestion, aphtha, ulcer being unable to heal, rubella pruritus, burn due to hot liquid or fire, brothers.
The pharmacological action of medicine provided by the invention: Herba Dendrobii has the merit that strengthens immunologic function, facilitating digestion, hepatoprotective function of gallbladder promoting, rheumatism, lowering blood sugar and blood fat, antitumor, vision protection, nourishing skin, defying age.Radix Notoginseng has good hemostasia effect, significant hemopoietic function; Can tighten and improve the arteria coronaria microcirculation, the effect of blood vessel dilating; Stronger analgesic activity is arranged, the effect that have resisting fatigue, improves the learning and memory ability; Anti-inflammatory effect; Effect with immunomodulator can make too high or too low immunoreation return to normally, but the not normal immunoreation of interference body; Antitumor action; Defying age, antioxidation; Blood fat reducing and cholesterol effect.The Radix Astragali has the enhancing human body immunity function, protects the liver, diuresis, defying age, anti-stress, blood pressure lowering and antibacterial action more widely; Can eliminate experimental nephritic proteinuria, strengthen myocardial contraction, regulate blood sugar content; The Radix Astragali can not only coronary artery dilator, improves blood supply of cardiac muscle, improves immunologic function, and process that can delaying cell aging.Fructus Corni is used for deficiency of the liver and kindey, has a dizzy spell tinnitus, the soreness of waist; Can consolidating channel for hemostasis, availablely control diseases such as human female void, menorrhagia; Anti-diabetic, antibiotic effect are arranged, obvious to hemorrhagic shock, cardiac function and hemodynamic effect.Radix Salviae Miltiorrhizae has promoting blood circulation to remove blood stasis, detumescence hemostasis, anti-inflammatory analgetic, menstruction regulating and pain relieving, coronary artery dilator, improve the myocardial ischemia situation, with all worries set aside, blood sugar lowering and effect such as antibiotic bring high blood pressure down, calm the nerves, to menoxenia, the amenorrhea dysmenorrhea, lump in the abdomen, breast ventral spine pain, pyretic arthralgia pain, skin infection swells and ache, dysphoria and insomnia, hepatosplenomegaly, diseases such as angina pectoris have certain curative effect; Radix Salviae Miltiorrhizae also has anti-platelet aggregation, reduces the function of blood viscosity and the inside and outside blood coagulation system of adjusting.Mel has convergence, nutrition and promotion healing effect to wound surface, has lubricity to eliminate the phlegm and laxative effective.Each composition of the present invention is the national corporation of medical herbs commercially available prod.
The advantage that the present invention possesses and effect: said preparation is suitable for the immunocompromised person, has the effect of enhancing immunity.Can breed by the enhancing antibody cellulation, improve the serum hemolysin level, promote phagocytosis of macrophages, to body weight gain, thymus/body weight ratio, spleen/body weight ratio and spleen lymphocyte proliferation transformation, monokaryon-macrophage carbon is cleaned up with the NK cytoactive does not have obvious influence.Have instant effect, effect is obvious, safe, cost is low and advantage such as easy and simple to handle.
The specific embodiment
The present invention will be further described below in conjunction with embodiment.
Embodiment 1
(1) get the raw materials ready by the component of following mass parts:
Fineness is 12 parts of 1200 purpose Herba Dendrobii superfine powder
Fineness is 6.9 parts of 1200 purpose superfine notoginseng powders
1.356 parts of commercial Radix Astragali extracts
0.3 part of commercial Fructus Corni extract
0.228 part of commercial Radix Salviae Miltiorrhizae extract
101.76 parts of Mel;
(2) step (1) is equipped with Herba Dendrobii superfine powder, superfine notoginseng powder, Radix Astragali extract, Fructus Corni extract and Radix Salviae Miltiorrhizae extract mix homogeneously in the component, add Mel again and stir, moulding process is prepared into the powder with enhancing immunity effect routinely.
Embodiment 2
(1) get the raw materials ready by the component of following mass parts:
Fineness is 10 parts of 1200 purpose Herba Dendrobii superfine powder
Fineness is 4 parts of 1200 purpose superfine notoginseng powders
1.5 parts of commercial Radix Astragali extracts
0.5 part of commercial Fructus Corni extract
0.4 part of commercial Radix Salviae Miltiorrhizae extract
100 parts of Mel;
(2) step (1) is equipped with Herba Dendrobii superfine powder, superfine notoginseng powder, Radix Astragali extract, Fructus Corni extract and Radix Salviae Miltiorrhizae extract mix homogeneously in the component, add Mel again and stir, moulding process is prepared into the capsule with enhancing immunity effect routinely.
Embodiment 3
(1) get the raw materials ready by the component of following mass parts:
Fineness is 15 parts of 1200 purpose Herba Dendrobii superfine powder
Fineness is 10 parts of 1200 purpose superfine notoginseng powders
1.5 parts of commercial Radix Astragali extracts
0.5 part of commercial Fructus Corni extract
0.4 part of commercial Radix Salviae Miltiorrhizae extract
105 parts of Mel;
(2) step (1) is equipped with Herba Dendrobii superfine powder, superfine notoginseng powder, Radix Astragali extract, Fructus Corni extract and Radix Salviae Miltiorrhizae extract mix homogeneously in the component, add Mel again and stir, moulding process is prepared into the tablet with enhancing immunity effect routinely; Wherein:
Radix Astragali extract, Fructus Corni extract, Radix Salviae Miltiorrhizae extract extract through following conventional method:
A, respectively each crude drug of the described Radix Astragali, Fructus Corni, Radix Salviae Miltiorrhizae is pulverized after, decocting extracts 2 times, each water consumption is 12 times of medical material gross mass, each decocting extracts and keeps little boiling 1.5 hours, merges each time extracting solution, filters to isolate filtrate and filtering residue;
B, the filtrate of A step is concentrated into 1g medical material/ml, cools off, leaves standstill to room temperature, stir that slowly to add ethanol to ethanol final concentration down be 60wt%, leave standstill to precipitation fully, sucking filtration is isolated filtrate and filtering residue;
C, be 60% washing with alcohol with the filtering residue mass concentration of B step, colourless until cleaning mixture, cleaning mixture is incorporated in the filtrate of B step, reclaim ethanol, drying is pulverized, respectively extract---the dry extract of the Radix Astragali, Fructus Corni, Radix Salviae Miltiorrhizae.
Embodiment 4
(1) get the raw materials ready by the component of following mass parts:
Fineness is 13 parts of 1200 purpose Herba Dendrobii superfine powder
Fineness is 8 parts of 1200 purpose superfine notoginseng powders
1.0 parts of commercial Radix Astragali extracts
0.4 part of commercial Fructus Corni extract
0.3 part of commercial Radix Salviae Miltiorrhizae extract
101 parts of Mel;
(2) step (1) is equipped with Herba Dendrobii superfine powder, superfine notoginseng powder, Radix Astragali extract, Fructus Corni extract and Radix Salviae Miltiorrhizae extract mix homogeneously in the component, add Mel again and stir, moulding process is prepared into the pill with enhancing immunity effect routinely; Wherein:
Radix Astragali extract, Fructus Corni extract, Radix Salviae Miltiorrhizae extract extract through following conventional method:
A, respectively each crude drug of the described Radix Astragali, Fructus Corni, Radix Salviae Miltiorrhizae is pulverized after, decocting extracts 3 times, each water consumption is 8 times of medical material gross mass, each decocting extracts and keeps little boiling 0.5 hour, merges each time extracting solution, filters to isolate filtrate and filtering residue;
B, the filtrate of A step is concentrated into 1g medical material/ml, cools off, leaves standstill to room temperature, stir that slowly to add ethanol to ethanol final concentration down be 60wt%, leave standstill to precipitation fully, sucking filtration is isolated filtrate and filtering residue;
C, be 60% washing with alcohol with the filtering residue mass concentration of B step, colourless until cleaning mixture, cleaning mixture is incorporated in the filtrate of B step, reclaim ethanol, drying is pulverized, respectively extract---the dry extract of the Radix Astragali, Fructus Corni, Radix Salviae Miltiorrhizae.
In order to clearly demonstrate effect of the present invention, below that the brief introduction of enhancing immunity functional experiment is as follows:
1 material and method
The powder of the enhancing immunity of 1.1 sample: embodiment 1, specification: 120 grams/bottle, put shady and cool dry place and preserve.Human oral recommends consumption to be everyone (adult) every day 2 times, and each 3 grams become body weight for humans to press 60kg calculating, and amounting to dosage is 100 mg/kg BW.
1.2 laboratory animal and grouping: 240 of the healthy Male Kunming strain mice of SPF level, body weight is 18～22 grams, by the breeding of Guangxi Medical University Experimental Animal Center, laboratory animal production licence number: SCXK(osmanthus) 2009-0002, the laboratory animal certification of fitness number: 0006030.Per 40 mices are one group, totally 6 groups.Immunity I group is carried out the mouse spleen lymphocyte transformation experiment that ConA induces; Immunity II group is carried out the delayed allergy experiment; Immunity III group is carried out dirty/body ratio measurement, and serum hemolysin is measured and the antibody-producting cell number is measured; Immunity IV～VI group carries out respectively that carbon is cleaned up experiment, peritoneal macrophage is engulfed chicken red blood cell experiment and NK cytoactive mensuration.
1.3 experimental animal room environmental condition: Animal Lab. is barrier system, occupancy permit number: SYXK (osmanthus) 2011-0005.The zoopery room temperature: 22～25 ℃, relative humidity: 55～70%.
1.4 dosage is selected and is tried thing to give mode: recommend consumption according to human oral, be equivalent to human body respectively and recommend 5,10,20 times of consumption if the basic, normal, high dosage group of sample is respectively 500,1000,2000 mg/kg BW(), if a negative control group, every group of 10 animals.Take by weighing sample 2.5,5.0,10.0g respectively, respectively add pure water to 100mL, mixing, be made into 25,50, the solution of 100mg/mL, give the corresponding dosage treated animal by the volume of 0.2mL/10g BW and irritate stomach, negative control group gives isopyknic pure water, irritate stomach every day once, continuous irrigation stomach 30 days.
1.5 key instrument and reagent: animal balance, electronic analytical balance, centrifuge, clean bench, CO2 gas incubator, constant water bath box, microscope, semi-automatic biochemical analyzer, microplate reader etc.
Aseptic operation apparatus, microsyringe, cell counter, the flat Tissue Culture Plate in 24 holes and 96 holes, the U-shaped Tissue Culture Plate in 96 holes, glass dish, gauze, microscope slide, test tube, 200 eye mesh screens, timer, hematochrome suction pipe etc.
The RPMI1640 culture fluid, calf serum, 2-ME, penicillin, streptomycin, ConA, the HCL solution of 1mol/L, isopropyl alcohol, MTT, Hank ' s liquid (pH 7.2～7.4), PBS buffer (Ph7.2～7.4), platform phenol indigo plant, DNFB, acetone, barium sulfide, complement (guinea pig serum), the SA buffer, agarose, sheep red blood cell (SRBC) (SRBC), normal saline, india ink, 0.1% sodium carbonate, chicken red blood cell, methanol, the Gimsa dye liquor, the YAC-1 cell, EINECS 212-761-8, the nitro tetrazolium chloride, azophenlyene dimethyl ester sulfate, oxidized form of nicotinamide-adenine dinucleotide, 0.2mol/L the Tris-HCL buffer, 1% glacial acetic acid, 1% NP40 etc.
1.6 experimental technique
1.6.1 internal organs/weight ratio pH-value determination pH: mice is put to death in the back of weighing, and takes out thymus and spleen, weighs at electronic analytical balance, calculates dirty/body ratio.
1.6.2ConA the mouse spleen lymphocyte transformation experiment (mtt assay) of inducing
The aseptic spleen of getting places the plate that fills an amount of aseptic Hank ' s liquid, makes cell suspension, filters through 200 eye mesh screens.Use Hank ' s liquid to wash 2 times, each centrifugal 10min (1000 r/min).Then with cell suspension in the 1mL complete culture solution, the living cell counting number, it is dense to adjust cell with the RPMI1640 culture fluid
Degree is 3 * 106/mL.Again cell suspension is divided two holes to add in 24 well culture plates, every hole 1mL, a hole adds 75 μ L ConA liquid (being equivalent to 7.5 μ g/mL) therein, and 5 %CO2 are put in contrast in another hole, cultivate 72h in 37 ℃ of carbon dioxide incubators.
Cultivate and finish preceding 4h, supernatant 0.7mL is inhaled in every hole gently, adds the RPMI1640 culture fluid that 0.7mL does not contain calf serum, adds MTT (5 mg/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 are made in each hole, use microplate reader, measure optical density value with the 570nm wavelength.Lymphocytic multiplication capacity deducts the optical density value that does not add the ConA hole with the optical density value that adds the ConA hole and represents.
1.6.3 the mouse DTH that dinitrofluorobenzene is induced experiment (ear swelling method)
Experiment finish preceding 5 days with barium sulfide with mouse part skin depilation about 3cm * 3cm scope, evenly smear sensitization with 50 μ L DNFB solution, after 5 days 10 μ L DNFB evenly being applied in the mouse right ear two sides attacks, mice is put to death in the cervical vertebra dislocation after 24 hours, cut left and right sides auricular concha, with card punch take off the 8mm diameter auricle, weigh, represent the degree of DTH with the difference of left and right sides ear weight.
1.6.4 antibody-producting cell detects (Jerne improves slide method)
Get the Sanguis caprae seu ovis of defiber, with normal saline washing 3 times, each centrifugal 10min(2000r/min), with normal saline hematocrit SRBC is made into the cell suspension of 2% (v/v), every Mus lumbar injection 0.2mL.The mice that immunity is back 5 days is put to death, and gets spleen and puts in the plate that fills Hank ' s liquid, grinds spleen gently, make cell suspension, filter centrifugal 10min(2000 r/min through 200 eye mesh screens), use Hank ' s liquid washing 2 times, at last with cell suspension in 8mL Hank ' s liquid.After top layer culture medium (the 1g agarose adds distilled water to 100mL) heating for dissolving, put 45～50 ℃ of water bath heat preservations, with equivalent Ph7.2～7.4, Hank ' the s liquid of 2 times of concentration mixes, the packing small test tube, every pipe 0.5mL, in pipe, add 50 μ L10%SRBC (v/v again, with the preparation of SA buffer), 25 μ L splenocyte suspensions are poured on the slide of brushing the agarose thin layer behind the mixing rapidly, do parallel plate, after treating that agar solidifies, the flat button of slide is placed on the slide frame, puts into CO2 gas incubator and hatch 1.5h, in complement (1:8) the adding slide frame groove with the dilution of SA buffer, continue to hatch 1.5h, counting hemolysis plaque number.With plaque number/10
6Splenocyte is represented the antibody-producting cell number.
1.6.5 the mensuration of serum hemolysin (blood clotting method)
Get the Sanguis caprae seu ovis of defiber, with normal saline washing 3 times, each centrifugal 10min(2000r/min), with normal saline hematocrit SRBC is made into the cell suspension of 2% (v/v), every Mus lumbar injection 0.2mL.After the immunity 5 days, extract the eyeball of mice, get blood in centrifuge tube, place about 1h, the centrifugal 10min of 2000r/min separates, collects serum.With the serum doubling dilution, the dilution serum of difference is placed in the Microhemagglutination plate every hole 100 μ L respectively with normal saline, the SRBC suspension that adds 100 μ L0.5% (v/v) again, mixing is added a cover in the moistening square position of packing into, in 37 ℃ of incubation 3h, observe the hemagglutination degree.Be calculated as follows the antibody product:
Antibody product=(S
1+ 2S
2+ 3S
3NS
n)
In the formula 1,2,3 ... n is two-fold dilution's index, and S is the rank of coagulation degree.
1.6.6 mice carbon is cleaned up experiment
To the india ink of injected in mice with 4 times of normal saline dilutions, every 10g body weight is injected 0.1 mL through the tail vein, and timing immediately after prepared Chinese ink injects after injecting prepared Chinese ink the 2nd, 10min, is got blood 20 μ L from the angular vein clump respectively, joins 2 mL, 0.1% Na
2CO
3In the solution, shake up.With Na
2CO
3Solution is made blank, with 600 nm wavelength photometry density values (OD).Mice is put to death, get liver, spleen, weigh.Be calculated as follows phagocytic index a.
A=K
1/3* body weight/(liver weight+spleen is heavy)
K=(lgOD in the formula
1-lgOD
2)/(t
2-t
1).
1.6.7 Turnover of Mouse Peritoneal Macrophages is engulfed chicken red blood cell experiment (half intracorporal method)
To mouse peritoneal injection 20% (v/v, prepare with normal saline) the hematocrit chicken erythrocyte suspension, every Mus 1mL, at interval 30min, mice is put to death in the cervical vertebra dislocation, abdominal skin is cut off in the center, and the abdominal cavity injects the 2mL normal saline, rotates Mus plate 1min, sucking-off abdominal cavity washing liquid 1mL, mean droplet is put into the enamel box that is lined with wet gauze on 2 microscope slides, put 37 ℃ of incubator incubation 30min.Incubate completely, take out slide and dry after the rinsing in normal saline, use methanol: acetone (1:1) solution is fixed, and 4%(v/v) the Giemsa-phosphate buffer dyes, again with the pure water rinsing, dry.100 macrophages of every counting are calculated as follows phagocytic rate and phagocytic index under the oil mirror.
Phagocytic rate (%)=engulf macrophage number * 100% of the macrophage number/counting of chicken red blood cell
The macrophage number of chicken red blood cell sum/counting that phagocytic index=quilt is engulfed
1.6.8NK cytoactive is measured (LDH algoscopy)
The dislocation of mice cervical vertebra is put to death, and the aseptic spleen of getting is made splenocyte suspension, uses Hank ' s liquid to wash 2 times, each centrifugal 10min(1000r/min).Abandon supernatant, cytoplasm is upspring, added 0.5mL sterilization pure water 20 seconds, add 0.5mL 2 times of Hank ' s liquid and 8mL Hank ' s liquid after the splitting erythrocyte again, the centrifugal 10min of 1000r/min, 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, adjusting cell concentration with the RPMI1640 complete culture solution at last is 2 * 10
7Individual/mL, this is the effector lymphocyte.Get the well-grown YAC-1 cell of back 24h that goes down to posterity, adjusting cell concentration with the RPMI1640 complete culture solution is 4 * 10
5Individual/mL, this is target cell.Get each 100 μ L(of target cell and effector lymphocyte and imitate target than 50:1), add in U-shaped 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 3 parallel holes of respectively establishing are put 5%CO
2, cultivate 4h in 37 ℃ of carbon dioxide incubators.Then with 96 well culture plates with the centrifugal 5min of 1500r/min, in 96 well culture plates, add LDH substrate liquid 100 μ L simultaneously at the bottom of every hole absorption supernatant 100 μ L horizontalizations, reacted 8 minutes, every hole adds the HCL30 μ L of 1mol/L, measures optical density (OD) value at microplate reader 490nm place.Be calculated as follows the NK cytoactive:
NK cytoactive (%)=(reacting hole OD-nature release aperture OD)/(maximum release aperture OD-nature release aperture OD) * 100%
1.7 experimental data statistics: use the SPSS statistical software and carry out the variance analysis statistical disposition.
1.8 the result judges: any two aspects positive as a result aspect cellular immune function, humoral immune function, monokaryon-macrophage function, NK cytoactive four, can judge that this given the test agent has the enhancing immunity function.
2 results
2.1 sample is to the influence of mice body weight
The functional experiment mice body weight of table 1 Herba Dendrobii enhancing immunity agent (
± s)
Annotate: initial, mid-term of each dosage group mice and latter stage body weight and weightening finish and negative control group comparison, there are no significant for difference (P〉0.05).
The functional experiment mice body weight of table 2 Herba Dendrobii enhancing immunity agent (
± s)
Annotate: initial, mid-term of each dosage group mice of immune II～VI group and latter stage body weight and weightening finish and negative control group comparison, there are no significant for difference (P〉0.05).
By table 1, table 2 as seen, test between first, as to test mid-term, the mice body weight of testing each dosage group of sample in latter stage and experimental session mice body weight gain and negative control group and compare, there are no significant for difference (P〉0.05), show that this sample does not have obvious influence to the body weight gain of mice.
2.2 sample is to the influence of mouse immune organ internal organs/body weight ratio
Immune organ internal organs/body weight ratio of the functional experiment mice of table 3 Herba Dendrobii enhancing immunity agent (
± s)
By table 3 as seen, the cloud that per os gives the mice various dose is board Herba Dendrobii extractum 30 days still, thymus/body weight of mice and spleen/body weight ratio and negative control group compare, and difference that there are no significant (P〉0.05), show that this sample does not have obvious influence to the immune organ weight of mice.
2.3 sample is to the influence of the cellular immunization of mice
2.3.1 the influence of the mouse spleen lymphocyte conversion capability that sample is induced ConA
The mouse spleen lymphocyte transformation experiment result of table 4 Herba Dendrobii enhancing immunity agent (
± s)
As seen from Table 4, per os gives the Herba Dendrobii enhancing immunity agent 30 days of mice various dose, the splenocyte conversion capability of each dosage group all is higher than negative control group, but there are no significant for the difference of each dosage group and negative control group (P〉0.05), show that this sample does not have obvious facilitation to spleen lymphocyte proliferation, the conversion capability of mice.
2.3.2 sample is to the influence of mice delayed allergy (DTH)
Mice delayed allergy (DTH) experimental result of table 5 Herba Dendrobii enhancing immunity agent (
± s)
As seen from Table 5, per os gives the Herba Dendrobii enhancing immunity agent 30 days of mice various dose, the left and right sides auricle weight difference of each dosage group all is higher than negative control group, and the difference of height, middle dosage group and negative control group has significance (P＜0.01 and P＜0.05 respectively), shows that this sample has the effect of the delayed allergy that promotes mice.
2.4 sample is to the influence of the humoral immunization of mice
2.4.1 sample is to the influence of the antibody-producting cell number of mice
As seen from Table 6, per os gives the Herba Dendrobii enhancing immunity agent 30 days of mice various dose, the antibody-producting cell number average of each dosage group is higher than negative control group, and the difference of high dose group and negative control group has significance (P＜0.05), shows that this sample has the effect of the antibody-producting cell propagation that promotes mice.
The antibody-producting cell test experience result of table 6 Herba Dendrobii enhancing immunity agent (
± s)
2.4.2 product are to the influence of mice serum hemolysin
As seen from Table 7, per os gives the Herba Dendrobii enhancing immunity agent 30 days of mice various dose, the antibody product of each dosage group all is higher than negative control group, and the difference of high dose group and negative control group has significance (P＜0.05), shows that this sample has the effect of the serum hemolysin level that improves mice.
The mice hemolysin measurement result of table 7 Herba Dendrobii enhancing immunity agent (
± s)
2.5 sample is to the influence of monokaryon-macrophage phagocytic function of mice
2.5.1 sample is to the influence that monokaryon-macrophage carbon is cleaned up of mice
Mouse monokaryon-macrophage the carbon of table 8 Herba Dendrobii enhancing immunity agent clean up experimental result (
± s)
As seen from Table 8, per os gives the Herba Dendrobii enhancing immunity agent 30 days of mice various dose, the phagocytic index of each dosage group all is higher than negative control group, but there are no significant for the difference of each dosage group and negative control group (P〉0.05), show this sample to mice monokaryon-macrophage carbon is cleaned up function does not have obvious facilitation.
2.5.2 sample is engulfed the influence of chicken red blood cell ability to the peritoneal macrophage of mice
The Turnover of Mouse Peritoneal Macrophages of table 9 Herba Dendrobii enhancing immunity agent engulf the chicken red blood cell experimental result (
± s)
As seen from Table 9, per os gives the Herba Dendrobii enhancing immunity agent 30 days of mice various dose, the peritoneal macrophage of each dosage group all is higher than negative control group to phagocytic rate and the phagocytic index of chicken red blood cell, and the phagocytic rate of height, middle dosage group and the difference of negative control group have significance (P＜0.01 and P＜0.05 respectively), the phagocytic index of each dosage group and the difference of negative control group all have significance (P＜0.01 or P＜0.05), show that this sample has the effect of the peritoneal macrophage phagocytic activity that promotes mice.
2.6 sample is to the influence of the NK cytoactive of mice
The NK cells in mice determination of activity result of table 10 Herba Dendrobii enhancing immunity agent (
± s)
As seen from Table 10, per os gives the Herba Dendrobii enhancing immunity agent 30 days of mice various dose, the NK cytoactive of each dosage group and negative control group approach, there are no significant for the difference of each dosage group and negative control group (P〉0.05), show that this sample does not have obvious facilitation to the NK cytoactive of mice.
3 brief summaries
Per os gives mice 500,1000,2000 mg/kg BW (are equivalent to human body and recommend consumption 5,10,20 times) the Herba Dendrobii enhancing immunity agent of dosage 30 days, can promote the delayed allergy of mice, promote the antibody-producting cell propagation of mice, improve the serum hemolysin level of mice, promote the phagocytosis of macrophages of mice, body weight gain to mice, thymus/body weight ratio, spleen/body weight ratio and spleen lymphocyte proliferation transformation, monokaryon-macrophage carbon is cleaned up with the NK cytoactive does not have obvious influence
Point out this sample to have the enhancing immunity function.
Claims (5)
1. the preparation of an enhancing immunity is characterized in that being made up of the component of following mass parts:
10～15 parts in Herba Dendrobii powder, 4～10 parts of Radix Notoginseng powder,
1～1.5 part of Radix Astragali extract, 0.2～0.5 part of Fructus Corni extract,
0.1～0.4 part of Radix Salviae Miltiorrhizae extract, 98～105 parts of Mel.
2. the preparation of enhancing immunity according to claim 1 is characterized in that: described Herba Dendrobii powder is that Herba Dendrobii is crushed to fineness is 1200 purpose superfine powder.
3. the preparation of enhancing immunity according to claim 1 is characterized in that: described Radix Notoginseng powder is that Radix Notoginseng powder is broken to fineness is 1200 purpose superfine powder.
4. the preparation method of the preparation of an enhancing immunity is characterized in that through following each step:
(1) get the raw materials ready by following mass parts:
10～15 parts in Herba Dendrobii powder, 4～10 parts of Radix Notoginseng powder,
1～1.5 part of Radix Astragali extract, 0.2～0.5 part of Fructus Corni extract,
0.1～0.4 part of Radix Salviae Miltiorrhizae extract, 98～105 parts of Mel;
(2) fineness with step (1) is that 1200 purpose Herba Dendrobii superfine powder, fineness are 1200 purpose superfine notoginseng powders, mixes with Radix Astragali extract, Fructus Corni extract and Radix Salviae Miltiorrhizae extract, adds Mel again and stirs, the preparation of the immunity that namely is enhanced.
5. the preparation method of the preparation of enhancing immunity according to claim 4 is characterized in that: with step (2) gained preparation routinely moulding process be prepared into capsule, granule, tablet, powder or the pill preparation with enhancing immunity effect.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103705566A (en) * | 2013-12-30 | 2014-04-09 | 陈静 | Healthy food prepared from plateau bee products and Chinese herbal medicines |
| CN103768406A (en) * | 2014-01-22 | 2014-05-07 | 福建永耕农业开发有限公司 | Blooming dendrobium candidum pill |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101322774A (en) * | 2008-03-25 | 2008-12-17 | 深圳力瑞医药科技有限公司 | Medicament composition with fatigue-resisting function and preparation method and application thereof |
| CN101850066A (en) * | 2009-11-06 | 2010-10-06 | 中山市中智药业集团有限公司 | Chinese medicinal composition granules and preparation method thereof |
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2013
- 2013-05-25 CN CN2013101977974A patent/CN103251816A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101322774A (en) * | 2008-03-25 | 2008-12-17 | 深圳力瑞医药科技有限公司 | Medicament composition with fatigue-resisting function and preparation method and application thereof |
| CN101850066A (en) * | 2009-11-06 | 2010-10-06 | 中山市中智药业集团有限公司 | Chinese medicinal composition granules and preparation method thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103705566A (en) * | 2013-12-30 | 2014-04-09 | 陈静 | Healthy food prepared from plateau bee products and Chinese herbal medicines |
| CN103705566B (en) * | 2013-12-30 | 2016-04-13 | 陈静 | The health food of a kind of plateau bee product and prepared from traditional Chinese medicines |
| CN103768406A (en) * | 2014-01-22 | 2014-05-07 | 福建永耕农业开发有限公司 | Blooming dendrobium candidum pill |
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Application publication date: 20130821 |