CN103239628B - Wine preparation for warming and tonifying kidney yang, enhancing immunity and warding off diseases and strengthening body - Google Patents
Wine preparation for warming and tonifying kidney yang, enhancing immunity and warding off diseases and strengthening body Download PDFInfo
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Abstract
The invention discloses a wine preparation for warming and tonifying kidney yang, enhancing immunity and warding off diseases and strengthening body. The wine preparation is prepared from 150-210 parts of American ginseng, 80-110 parts of mulberry, 80-110 parts of radix polygonati officinalis, 45-75 parts of fructus lycii, 15-35 parts of chrysanthemum, 15-35 parts of poria cocos, 15-35 parts of radix paeoniae alba, 15-35 parts of oyster, 15-35 parts of kelp, 8-16 parts of Chinese angelica, 8-16 parts of gecko, 3-9 parts of tangerine peel, 6,000-7,000 parts of 50-degree base liquor and 3,000-4,000 parts of water by grinding, soaking extraction, filtration, blending and filling. The wine preparation disclosed by the invention has the beneficial effects that a unique efficient molecular group acts on a human body and has the functions of warming and tonifying kidney yang, improving immunity of the human body, improving blood circulation, repairing nervous tissue, supplementing new energy of the human body, strengthening the kidney and consolidating root and beautifying and lengthening life, and is suitable for the symptoms such as deficiency of kidney yang, aching lumbus and limp legs, weak limbs, lassitude, low immunity and the like.
Description
Technical field
The present invention relates to the liquor preparation that a kind of Chinese prescription is made, for warming and recuperating the kidney-YANG, strengthen immunity, strong body gets rid of illness.
Background technology
Kidney yang is each dirty positive the dominator of QI, and each internal organs, histoorgan are played a part to promotion, warm, and kidney yang is sufficient, function of human body activity is vigorous, and kidney-yang deficiency internal organs loses temperature, and functional activity is low, can bring many inconvenience to people's life, affect quality of life.Society, nervous work, busy dinner party, brings unavoidable pressure to people, and therefore there is the symptoms such as soreness of the waist and knees, ache all over the body, limbs fatigue, lassitude in many people, and these are all the performances of kidney-yang deficiency.Because kidney yang has the effect of warm whole body internal organs, therefore if syndrome of deficiency of kidney-YANG mistake is controlled, with the passing of time wrong treatment is delayed, can there are many deteriorated cases, as immunocompromised, soreness of the waist and knees, ache all over the body etc.This invention preparation is developed according to syndrome of deficiency of kidney-YANG, and the efficient micel that it contains can improve body immunity, improves blood circulation, repair nervous tissue, supplements human body new forms of energy, kidney tonifying consolidates, face nursing and life prolonging.
Summary of the invention
The present invention overcomes the deficiency in prior art, and a kind of liquor preparation for warming and recuperating the kidney-YANG, enhancing immunity, the strong body that gets rid of illness is provided, and not only therapeutic effect is good, and cheap, taking convenience.
The technical solution adopted for the present invention to solve the technical problems is: a kind of for warming and recuperating the kidney-YANG, strengthen immunity, the liquor preparation of strong body gets rid of illness, it adopts Radix Panacis Quinquefolii, Fructus Mori, Rhizoma Polygonati Odorati, Fructus Lycii, Flos Chrysanthemi, Poria, the Radix Paeoniae Alba, Concha Ostreae, Thallus Laminariae (Thallus Eckloniae), Radix Angelicae Sinensis, Gecko, Pericarpium Citri Reticulatae, 50 ° of base wine are raw material, by following quality proportioning: 150～210 parts of Radix Panacis Quinquefoliis, 80～110 parts, Fructus Mori, 80～110 parts of Rhizoma Polygonati Odorati, 45～75 parts of Fructus Lycii, 15～35 parts of Flos Chrysanthemis, 15～35 parts, Poria, 15～35 parts of the Radix Paeoniae Albas, 15～35 parts of Concha Ostreaes, 15～35 parts of Thallus Laminariae (Thallus Eckloniae)s, 8～16 parts of Radix Angelicae Sinensis, 8～16 parts of Geckos, 3～9 parts of Pericarpium Citri Reticulataes, 6000～7000 parts of 50 ° of base wine, 3000～4000 parts, water is through pulverizing, dipping extracts, filter, blend, liquor preparation is made in fill.Its concrete preparation technology is: Radix Panacis Quinquefolii, Fructus Mori, Rhizoma Polygonati Odorati, Fructus Lycii, Flos Chrysanthemi, Poria, the Radix Paeoniae Alba, Concha Ostreae, Thallus Laminariae (Thallus Eckloniae), Radix Angelicae Sinensis, Gecko, Pericarpium Citri Reticulatae are ground into coarse powder, put into extraction pot, add 50 ° of base wine sealing dippings 30 days, stir once every day, after one week, stir once weekly, filter, filtrate water is blent into 28 °～35 °, sealing and standing 7 days, filter fill and get final product.
Of the present invention part is mass parts, is the mass units such as the conventional microgram in this area, milligram, gram, kilogram.
It is 50ml that preparation suggestion day of the present invention takes effective dose, at twice, and one after each meal.
The invention has the beneficial effects as follows: unique efficient micel acts on human body have warming and recuperating the kidney-YANG, improve body immunity, improve blood circulation, repair nervous tissue, supplement human body new forms of energy, kidney tonifying consolidates, the function of face nursing and life prolonging.Be applicable to the diseases such as kidney-yang deficiency, waist soreness, limbs fatigue, lassitude, hypoimmunity.
Compound basis:
Radix Panacis Quinquefolii, sweet in the mouth, micro-hardship, cool in nature.GUIXIN, lung, kidney channel.There is boosting qi and nourishing yin, the effect of clearing away heat and promoting production of body fluid.For the deficiency of vital energy is cloudy, lose, interior-heat, cough with asthma expectorant blood, deficiency-heat is tired tired, quenches one's thirst, the diseases such as dryness of the mouth and throat.Main chemical compositions has Chikusetsusaponin V, Rb1, Rb2, Rc, Rd, Re, Rg1 and Panax pseudoginseng Wall. Saponin F1; 18 seed amino acids such as arginine, aspartic acid; Again containing volatile oil, resin etc.Archaism cloud: " Radix Panacis Quinquefolii is cool in nature and mend, all wishs with Radix Ginseng, be not subject to the warm person of Radix Ginseng all available it ".Not dry therefore mend is the special feature of Radix Panacis Quinquefolii.Modern medicine proves: Radix Panacis Quinquefolii has the ability that improves muscle power and mental work, reduces fatigue strength and regulates the pharmacological actions such as central nervous system; The heart diseases such as, coronary heart diseases and angina pectoris bad to hypertension, myocardial nutrition all have good curative effect, be particularly useful for improving agitation that heart disease causes, sultry, thirsty, can alleviate the untoward reaction that cancer patient's radiotherapy and chemotherapy cause, as dry pharynx, feel sick, become thin, leukopenia, have no appetite, atrophy of salivary gland, and can change physical stress state, alleviate the effects such as thymus, lymph gland Telatrophy.
Fructus Mori, sweet in the mouth, acid, cold nature.GUIXIN, liver, kidney channel.The YIN nourishing of enriching blood, promoting the production of body fluid to quench thirst, the effect that intestine moistening is dry.Sugary in Fructus Mori, tannic acid, malic acid and vitamin B1, B2, C and carotene etc., have the skin of improvement (comprising scalp) blood supply, nutrition skin, makes the effects such as the delicate and black hair of skin, and can slow down aging.The traditional Chinese medical science thinks, Fructus Mori is sweet in flavor and cold in property, there is the liver and the kidney tonifying, the intestine moistening of promoting the production of body fluid, black hair tomorrow etc. effect, be healthy and strong U.S. face, antidotal good fruit and good medicine of middle-aged and elderly people.
Rhizoma Polygonati Odorati, sweet in the mouth is flat.Attach to the lung and stomach meridians.Main chemical compositions is Rhizoma Polygonati Odorati mucopolysaccharide and 4 kinds of Rhizoma Polygonati Odorati levan, azetidine-2-carboxylic acid etc.There is yin nourishing, moisturize, relieving restlessness, the function of quenching the thirst.For the cloudy wound of calentura, cough excessive thirst, deficient fever, rapid digestion of food is easily hungry, the diseases such as frequent micturition.This product has the laboratory animal of promotion antibody and generates, improve phagocytic percentage and the phagocytic index of macrophage, promote interferon synthetic, suppress growth of bacillus tubercle, blood sugar lowering, blood fat reducing, alleviates atherosclerotic plaque and forms, and makes peripheral blood vessel and coronary dilation, extend hypoxia endurance time, heart tonifying, antioxidation, the anti-ageing effect of waiting for a long time.
Fructus Lycii, sweet in the mouth is flat.Return liver, kidney channel.There is nourishing the liver and kidney, the effect of replenishing vital essence to improve eyesight.Contain the nutritional labelings such as betaine, polysaccharide, crude fat, crude protein, carotene, multivitamin and calcium, phosphorus, ferrum, zinc, manganese, linoleic acid.Fructus Lycii has rising peripheral leukocytes, strengthens reticuloendothelial system phagocytic activity, strengthens again the effect of cell and humoral immunization; Hemopoietic function is had to the effect of promotion; Can also defying age, mutation, antitumor, protect the liver and blood sugar lowering etc.For asthenia damage of essence, soreness of waist and knee joint, vertigo and tinnitus, interior-heat is quenched one's thirst, blood deficiency and yellow complexion, blurred vision is not clear.
Flos Chrysanthemi, acrid in the mouth, sweet, hardship; Cold nature.Return lung, Liver Channel.There is loose wind heat clearing away, the merit of suppressing the hyperactive liver improving eyesight.Main chemical compositions diosmetin.Can be used for anemopyretic cold, have a headache dizzy, conjunctival congestion and swelling pain, eyes is dim-sighted.Flos Chrysanthemi has some inhibitory action to gram-positive bacterium (staphylococcus aureus and beta hemolytic streptococcus), Bacillus tuberculosis in vitro; Also can strengthen capillary resistance.
Poria, sweet in the mouth, light is flat.Return liver, stomach warp.There is eliminating dampness and diuresis, strengthening the spleen stomach function regulating, the effect of mind tranquilizing and the heart calming.Sclerotium accounts for dry weight 93% and triterpenoid compound Pachymic acid, pachymic acid, 3 beta-hydroxy Pilus Caprae seu Ovis steroid trienic acids containing β-pachyman.In addition still gummy, chitin, protein, fat, sterol, lecithin, glucose, adenine, histidine, choline, β-pachyman catabolic enzyme, lipase, protease etc..For dysuria, edema distension, phlegm retention cough with dyspnea, the sound of vomiting of vomitting, has loose bowels, seminal emission, stranguria with turbid discharge, palpitation with fear, forgetful disease such as grade.
The Radix Paeoniae Alba, sweet in the mouth is flat; Return spleen, stomach, large intestine channel.There is nourishing blood to suppress the hyperactive liver, slow middle pain relieving, yin fluid astringing is received the effect of antiperspirant.Mainly contain BAIYAO alcohol, oleanolic acid, longispinogenin and the normal ruscogenin of Mexico celestial being.For the breast abdomen pain over the hypochondriac region, dysentery stomachache, spontaneous sweating, fever due to yin deficiency, menoxenia, metrorrhagia, is with inferior disease.
Radix Angelicae Sinensis, sweet, pungent, warm in nature.Return liver, the heart, spleen channel.Enrich blood and invigorate blood circulation, menstruction regulating and pain relieving, loosening bowel to relieve constipation.Containing Rhizoma Ligustici lactone, n-butene acyl lactone, ferulic acid, nicotinic acid, sucrose and several amino acids, and sesquiterpenoids etc.For blood deficiency and yellow complexion, dizzy cardiopalmus, menoxenia, amenorrhea dysmenorrhea, asthenia cold abdominalgia, dryness of the intestine constipation, rheumatic arthralgia, injury from falling down, ulcer sores.Radix Angelicae Sinensis (processed with wine) promoting blood circulation to restore menstrual flow.For amenorrhea dysmenorrhea, rheumatic arthralgia, injury from falling down.Due to Radix Angelicae Sinensis to women through, band, tire, product various diseases, have therapeutic effect, so the traditional Chinese medical science claims when being classified as " panacea of female section ".
Pericarpium Citri Reticulatae, bitter in the mouth, pungent, warm in nature.Return lung, spleen channel.Regulating qi-flowing for strengthening spleen, drying dampness to eliminate phlegm.Containing compositions such as volatile oil, hesperidin, vitamin B, C.For fullness in the epigastrium and chest, lack of appetite and vomiting, cough with copious phlegm.
Concha Ostreae, salty in the mouth, puckery, cold in nature.Return liver, kidney channel.There is suppressing the hyperactive liver and subsiding YANG, tranquillization with heavy prescription, hard masses softening and resolving, restrains astringent or styptic treatment for spontaneous sweating effect.Containing glycogen, taurine, 10 kinds of essential propylhomoserins, inorganic salt, iodine, lipid etc.For vertigo and tinnitus, palpitation with fear insomnia, scrofula goiter, lump in the abdomen mass in the abdomen, spontaneous sweating, seminal emission, metrorrhagia, is with inferior disease.
Thallus Laminariae (Thallus Eckloniae), salty in the mouth, cold in nature; Enter liver, stomach, kidney channel.Eliminating phlegm and softening indurated mass, inducing diuresis to remove edema.Containing algin element, mannitol, galactan, laminine, Thallus Laminariae (Thallus Eckloniae) polysaccharide, glutamic acid, aspartic acid, proline, vitamin B1, C, P and sulphur, potassium etc.For goiter scrofula, the diseases such as edema.
Gecko, salty in the mouth, property is flat.Enter lung, kidney channel.There is invigorating the lung and the kidney, improving inspiration by invigorating kidney-QI Dingchuan, the effect of supporing yang benefit essence.Containing compositions such as carnosine, choline, carnitine, guanine, protein, fat.For dyspnea due to deficiency tachypnea, chronic cough hemoptysis, impotence and seminal emission.
In sum, preparation of the present invention has following effect:
1, blood circulation promoting and blood stasis dispelling, improves circulation, and regulative mechanism promotes immunity
Phagocytic percentage and phagocytic index that macrophage can improve in the Chinese traditional medicine molecule group that liquor preparation of the present invention contains, improve immunity, and peculiar efficient micel can be enriched blood and be invigorated blood circulation in addition, blood circulation promoting, and two-ways regulation kidney equilibrium between yin and yang, regulates internal organs yang blood and qi balance.
2, the strong body that gets rid of illness, consolidates and keeps unit
Preparation of the present invention has easypro through blood stasis dispelling, the active wind that goes, analgesia itching-relieving action, the discomfort that rheumatic arthritis, rheumatoid arthritis, lumbar vertebra pain, cervical vertebra pain, sciatica, hyperosteogeny, scapulohumeral periarthritis, numb hand and foot, gout traumatic injury etc. are caused has the effect of the health-care recovery of promotion.
3, warming and recuperating the kidney-YANG, benefiting vital QI for tranquillizing
The efficient micel that liquor preparation of the present invention contains can effectively regulate the diseases such as waist soreness, limbs fatigue, hands and feet being not warm, lassitude, sexual impotence erectile problem, the scrotum of kidney-yang deficiency, essence and blood consumption wound, deficient qi and blood appearance is clammy, seminal emission premature ejaculation, soreness of waist cold, women's clear and thin leucorrhea.
The specific embodiment
Below in conjunction with embodiment, the invention will be further described:
Embodiment 1: a kind of for warming and recuperating the kidney-YANG, strengthen the liquor preparation of immunity, the strong body that gets rid of illness, by following quality proportioning: Radix Panacis Quinquefolii 180g, Fructus Mori 96g, Rhizoma Polygonati Odorati 96g, Fructus Lycii 60g, Flos Chrysanthemi 24g, Poria 24g, Radix Paeoniae Alba 24g, Concha Ostreae 24g, Thallus Laminariae (Thallus Eckloniae) 24g, Radix Angelicae Sinensis 12g, Gecko 12g, Pericarpium Citri Reticulatae 6g, 50 ° of base wine 6400ml, water 3600ml through pulverizing, dipping extracts, filter, blend, liquor preparation is made in fill.Take 50ml every day, divide and take for 2 times.
Embodiment 2: a kind of for warming and recuperating the kidney-YANG, strengthen the liquor preparation of immunity, the strong body that gets rid of illness, by following quality proportioning: Radix Panacis Quinquefolii 210g, Fructus Mori 80g, Rhizoma Polygonati Odorati 110g, Fructus Lycii 70g, Flos Chrysanthemi 20g, Poria 30g, Radix Paeoniae Alba 20g, Concha Ostreae 35g, Thallus Laminariae (Thallus Eckloniae) 15g, Radix Angelicae Sinensis 10g, Gecko 10g, Pericarpium Citri Reticulatae 9g, 50 ° of base wine 7000ml, water 4000ml through pulverizing, dipping extracts, filter, blend, liquor preparation is made in fill.Take 50ml every day, divide and take for 2 times.
Embodiment 3: a kind of for warming and recuperating the kidney-YANG, strengthen the liquor preparation of immunity, the strong body that gets rid of illness, by following quality proportioning: Radix Panacis Quinquefolii 150g, Fructus Mori 110g, Rhizoma Polygonati Odorati 80g, Fructus Lycii 50g, Flos Chrysanthemi 35g, Poria 15g, Radix Paeoniae Alba 30g, Concha Ostreae 20g, Thallus Laminariae (Thallus Eckloniae) 35g, Radix Angelicae Sinensis 15g, Gecko 12g, Pericarpium Citri Reticulatae 5g, 50 ° of base wine 6000ml, water 3000ml through pulverizing, dipping extracts, filter, blend, liquor preparation is made in fill.Take 50ml every day, divide and take for 2 times.
Embodiment 4: use the preparation of the present invention of making in embodiment 1, the present invention is strengthened to immune function and verify.
1. material
1.1 samples:
Brownish red clear liquor preparation, human body recommended amounts 50ml/60kg BW, every day 50ml, room temperature preservation, is for experiment.
1.2 laboratory animals:
Select 60 of the female ICR mices of the healthy secondary of 17.0～19.7g, be divided into 4 groups, every group 15, as immune I group, carry out organ weight ratio pH-value determination pH, delayed allergy experiment, Turnover of Mouse Peritoneal Macrophages and engulf chicken red blood cell experiment, half hemolysis value (HC
50) mensuration and the mensuration of antibody-producting cell number; 60 of the female ICR mices of the healthy secondary of 16.0～17.7g, are divided into 4 groups, 15 every group, as immune II group, carry out carbon and clean up experiment; 60 of the female ICR mices of the healthy secondary of 16.0～16.9g, are divided into 4 groups, 15 every group, as immune III group, carry out mouse lymphocyte transformation experiment and the NK cytoactive detection of ConA induction.
1.3 dosage group selections and tested material give mode:
Everyone (pressing 60kg weighing machine) 50ml every day of recommended dose preparation of the present invention, is equivalent to 0.83ml/kg BW.It is 8.33ml/kg BW every day that 10 times of human body recommended amounts are established in experiment, upper and lowerly respectively establishes a dosage group: 15.00ml/kg BW and 0.83ml/kg BW.0.00ml/kg BW dosage group replaces tested material with water.Experiment mice is with outbreeding system mice PP Pipe Compound feed, and per os once a day gavage gives mice tested material, gives continuously to survey indices after 30d.
1.4 key instruments and reagent:
Animal balance, analytical balance, clean bench, CO2 gas incubator, centrifuge, spectrophotometer, water bath with thermostatic control, microplate reader, microscope etc.
Aseptic operation apparatus, slide gauge (precision 0.01mm), microsyringe (25 μ L), cell counter, 24 holes and 96 level land, hole Tissue Culture Plates, the 96 U-shaped Tissue Culture Plates in hole, glass dish, gauze, test tube, slide frame, 200 order mesh screens, timer, hemoglobin pipet, microscope slide etc.
Mianyang erythrocyte (SRBC), normal saline, Hank ' liquid (PH7.2～7.4), RPMI1640 culture fluid, calf serum, mycillin, concanavalin A, Con A (ConA), 1% glacial acetic acid, the HCl solution of 1moL/L, acid isopropyl alcohol (96mL isopropyl alcohol adds the HCl solution 4mL of 1moL/L), MTT, PBS buffer (PH7.2～7.4), complement (guinea pig serum), SA buffer, agarose, Dou Shi reagent (sodium bicarbonate 1.0g, high-potassium ferricyanide 0.2g, potassium cyanide 0.05g, adding distil water is to 1000mL), YAC-1 cell, sodium lactate, nitro tetrazolium chloride, PMS, oxidized form of nicotinamide-adenine dinucleotide, the Tris-HCl buffer (PH8.2) of 0.2mol/L, 2.5%Triton, india ink, 0.1%NaCO
3, chicken red blood cell, methanol, Giemsa dye liquor etc.
1.5 experimental techniques:
1.5.1 the mensuration of organ weight ratio value
Mice dislocates and puts to death after weighing, and gets spleen and thymus, removes most fascia, with filter paper, blots organ surface blood stains, weighs, and calculates spleen body weight ratio and thymus body weight ratio.
1.5.2 delayed allergy (the sufficient sole of the foot thickens method) experiment (DTH)
Get Sanguis caprae seu ovis, normal saline washing 3 times, every Mus is through lumbar injection 2% (v/v prepares with normal saline) hematocrit SRBC (2000rpm, 10min) 0.2mL, to quick rear 4d, measures left back sufficient sole of the foot portion thickness, and same position is measured 3 times, averages.Then at measuring point subcutaneous injection 20% (v/v prepares with normal saline) hematocrit SRBC20 μ L, after injection, 24h measures left back sufficient sole of the foot portion thickness, represents the degree of DTH with the difference (swelling degree of the paw) of sufficient sole of the foot thickness before and after attacking.
The data obtained is measurement data, if tested material group is attacked the difference of the sufficient sole of the foot thickness in front and back apparently higher than 0.000g/kg BW group, and difference has significance (P < 0.05), can judge that this tested material is improved the effect of the delayed allergy ability of mice.
The mouse lymphocyte transformation experiment (mtt assay) of 1.5.3ConA inducing
The aseptic spleen of getting, is placed in the little plate that fills appropriate aseptic Hank ' s liquid, makes cell suspension.With Hank ' s liquid, wash 3 times each centrifugal 10min (1000rpm).Then by cell suspension in the complete culture solution of 2mL, living cell counting number, adjusts cell concentration 2 * 10
6individual/mL.Again cell suspension is divided two holes to add in 24 well culture plates, every hole 1mL, a hole adds 50 μ L ConA liquid (being equivalent to 5 μ g/mL) therein, and 5%CO in contrast, is put in another hole
2, cultivate 72h for 37 ℃.Cultivation finishes front 4h, and every hole sucks supernatant 0.7mL gently, adds 0.7mL not containing the RPMI1640 culture fluid of calf serum, adds MTT (5mg/mL) 50 μ L/ holes simultaneously, continues to cultivate 4h.After cultivation finishes, every hole adds 1mL acid isopropyl alcohol, and piping and druming mixes, and purple crystal is dissolved completely.Then this liquid is moved on 96 well culture plates, 3 parallel holes are made in every hole, measure wavelength 570nm by microplate reader.Lymphocytic multiplication capacity deducts by the optical density value that adds ConA hole the optical density value that does not add ConA hole and represents.
The data obtained is measurement data, if the lymphocytic multiplication capacity of tested material group is apparently higher than 0.000g/kg BW group, and difference has significance (P < 0.05), can judge that this tested material is improved the effect of the mouse lymphocyte conversion capability of ConA induction.
1.5.4 the mensuration of antibody-producting cell number (Jerne improves slide method)
Get Sanguis caprae seu ovis, normal saline washing 3 times, every Mus is through lumbar injection 2% (v/v prepares with normal saline) hematocrit SRBC0.2mL.Mice after SRBC immunity 5d is put to death, get spleen, make cell suspension.By after agarose heating for dissolving, mix with the double Hank ' s of equivalent liquid, subpackage small test tube, every pipe 0.5mL, in pipe, add 10% (v/v again, with the preparation of SA liquid) hematocrit SRBC50 μ L, splenocyte suspension 20 μ L, after mixing rapidly, be poured on the slide of brushing agarose thin layer, after agar solidifies, slide level buckled and is placed on horse, put into CO2 gas incubator incubation 1.5h, then with the complement (1: 10) of SA buffer dilution, join in slide frame groove, continue after incubation 1.5h, counting hemolysis plaque number.
The data obtained is measurement data, if the hemolysis plaque number of tested material group apparently higher than 0.000g/kg BW group, and difference has significance (P < 0.05), can judge that this tested material has the effect of the antibody-producting cell number that increases mice.
1.5.5 half hemolysis value (HC
50) mensuration
Get Sanguis caprae seu ovis, normal saline washing 3 times, every Mus carries out immunity through lumbar injection 2% (v/v prepares with normal saline) hematocrit SRBC0.2mL.After 5d, 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 2000rpm, collects serum.With SA buffer, by serum dilution, be 400 times, get 1mL and put in vitro, add successively 10% (v/v, with the preparation of SA buffer) hematocrit SRBC0.5mL, complement 1mL (pressing dilution in 1: 10 with SA buffer).Separately establish the not control tube of increase serum (replacing with SA buffer).Put in 37 ℃ of constant temperature waters and be incubated after 30min, ice bath cessation reaction.The centrifugal 10min of 2000rpm, gets supernatant 1mL, adds Dou Shi reagent 3mL.(v/v, with the preparation of SA buffer) the hematocrit SRBC0.25mL that simultaneously gets 10%, add Dou Shi reagent to 4mL in another test tube, fully mix, place after 10min, in 540nm, sentence control tube and do blankly, measure and respectively manage optical density value respectively.The amount of hemolysin is with half hemolysis value (HC
50) represent, be calculated as follows.
Optical density value * extension rate during sample half hemolysis value=sample optical density value/SRBC HD50
The data obtained is measurement data, if the half hemolysis value of tested material group apparently higher than 0.000g/kg BW group, and difference has significance (P < 0.05), can judge that this tested material is improved the effect of the half hemolysis value of mice.
1.5.6 mice carbon is cleaned up experiment
The india ink of mouse tail vein injection dilution in 1: 3.5, treats that prepared Chinese ink injects, timing immediately.Inject after prepared Chinese ink 2,10min, from angular vein clump, get blood 20 μ L respectively, and be added to 2mLNa
2cO
3in solution.With 721 spectrophotometers 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 is weighed.Be calculated as follows a:
K=(lgOD
1-lgOD
2)/(t
2-t
1) a=body weight ÷ (liver weight+spleen weight) * k
1/3
The data obtained is measurement data, if the phagocytic index of tested material group is apparently higher than 0.000g/kg BW group, and difference has significance (P < 0.05), can judge that this tested material is improved the effect that the carbon of mouse monokaryon-macrophage is cleaned up ability.
1.5.7 Turnover of Mouse Peritoneal Macrophages is engulfed chicken red blood cell experiment (dripping sheet method)
With cervical vertebra dislocation method, put to death mice, lumbar injection adds Hank ' s liquid 4mL/ of calf serum, rubs gently abdominal part 20 times, fully to wash out peritoneal macrophage, then stomach wall is cut off to an osculum, with glue head straw, drawn abdominal cavity washing liquid 2mL in vitro (or with syringe).With 1mL sample injector absorption abdominal cavity washing liquid 0.5mL, add and fill 0.5mL1% Sanguis Gallus domesticus red cell suspension in vitro, mix.With syringe (filling large syringe needle), draw 0.5mL mixed liquor, add in the agar circle of slide.Placing interior 37 ℃ of incubator hatches 15 minutes.Hatch after end rapidly with normal saline not attached cell wash out, in methanol solution, fix 1 minute, Giemsa liquid dyeing 15 minutes.Clean with distilled water flushing, dry, with 40 * microscopic counting phagocytic rate and phagocytic index.
Phagocytic rate %=engulfs macrophage number * 100 of the macrophage number/counting of chicken red blood cell
The macrophage number of the chicken red blood cell sum/counting of phagocytic index=engulfed
The data obtained is measurement data, if the phagocytic rate of tested material group or phagocytic index are apparently higher than 0.000g/kg BW group, and difference has significance (P < 0.05), can judge that this tested material is improved the effect that mouse macrophage is engulfed chicken red blood cell ability.
1.5.8NK the mensuration of cytoactive (lactate dehydrogenase L DH algoscopy)
Before experiment, 24h, by target cell YAC-1 cultivations of going down to posterity, washes 3 times with Hank ' s liquid before application, with the RPMI1640 complete culture solution adjustment cell concentration that contains 10% calf serum, is 4 * 10
5individual/mL.Tested mice draws neck to put to death, the aseptic spleen of getting, make splenocyte suspension, with Hank ' s liquid, wash 3 times, the centrifugal 10min of 1500rpm, resuspended containing the RPMI1640 complete culture solution of 10% calf serum with 2mL again, with the blue dyeing counting of platform phenol (viable count should more than 95%), adjusting cell concentration be 2 * 10
7individual/mL, making to imitate target ratio is 50: 1.Get each 100 μ L of target cell and effector lymphocyte, add in U-shaped 96 well culture plates; Target cell Spontaneous release hole 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, 37 ℃, 5%CO of all establishing
2in incubator, cultivate 4h, by 96 orifice plates with the centrifugal 5min of 1500rpm, every hole is drawn supernatant 100 μ L and is put in ELISA Plate, add LDH substrate liquid 100 μ L, reaction 10min, then every hole adds the HCl solution 30 μ L cessation reactions of 1mol, surveys OD value at microplate reader 490nm place, and NK cytoactive is calculated as follows:
NK%=(reacting hole OD-Spontaneous release hole OD)/(maximum release aperture OD-Spontaneous release hole OD) * 100
The preparation of LDH substrate liquid: sodium lactate 5 * 10
-2mol/L
Nitro tetrazolium chloride 6.6 * 10
-4mol/mL
PMS 2.8 * 10
-4mol/mL
Oxidized form of nicotinamide-adenine dinucleotide 1.3 * 10
-3mol/mL
Mentioned reagent is dissolved in the Tris-HCl buffer of 0.2mol/L (pH8.2)
The data obtained is measurement data, if the NK cytoactive of tested material group apparently higher than 0.000g/kg BW group, and difference has significance (P < 0.05), can judge that this tested material is improved the ability of NK cells in mice activity.
1.6 experimental data statistics:
Experimental data is carried out homogeneity test of variance with Stata software to each experiment initial data, and the data information that meets " variance is neat " requirement carries out statistical disposition with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group; The data information of nonnormal distribution or heterogeneity of variance is carried out to suitable variable conversion, after meeting " normal state variance is neat " requirement, with the data meter of conversion gained, process.
1.7 results are judged:
Enhancing immunity function is judged: aspect four of cellular immune functions, humoral immune function, monocytes/macrophages function, NK cytoactive, any two aspect results are positive, can judge that this given the test agent has enhancing immunity function.
Wherein two experimental results in cellular immune function assay project are all positive, or an above dosage group result of arbitrary experiment is positive, can judge that cellular immune function assay result is positive.Two experimental results in humoral immune function mensuration project are all positive, or the more than one dosage group result of arbitrary experiment is positive, can judge that humoral immune function measurement result is positive.Above dosage group result of monocytes/macrophages functional examination experiment is positive, can judge that monocytes/macrophages functional examination result is positive.An above dosage group result of NK cytoactive detection experiment is positive, can judge that NK cytoactive result is positive.
2 results
2.1 impacts of preparation of the present invention on Mouse Weight
The impact of table 1 preparation of the present invention on Mouse Weight
I group
II group
III group
Initial data carries out after homogeneity test of variance, meets the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carries out statistical disposition.From table 1, between each dosage group and 0.00ml/kg BW group relatively, there are no significant for difference (P > 0.05) for initial body weight, and the initial body weight of mice is comparatively balanced between each group.Per os gives the preparation 30d of the present invention of mice various dose, end-body focuses between its corresponding 0.00ml/kg BW of each dosage group group and compares, there are no significant for difference (P > 0.05), preparation of the present invention on weight of mice without impact.
2.2 impacts of preparation of the present invention on mice organs body weight ratio
The impact of table 2 preparation of the present invention on mouse spleen body weight ratio
Per os gives the preparation 30d of the present invention of mice various dose, and initial data carries out after homogeneity test of variance, meets the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carries out statistical disposition.From table 2, between each dosage group and 0.00ml/kg BW group, relatively, there are no significant for difference (P > 0.05), preparation of the present invention on mouse spleen body weight ratio without impact.
The impact of table 3 preparation of the present invention on mouse thymus body weight ratio
Per os gives the preparation 30d of the present invention of mice various dose, and initial data carries out after homogeneity test of variance, meets the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carries out statistical disposition.From table 3, between each dosage group and 0.00ml/kg BW group, relatively, there are no significant for difference (P > 0.05), preparation of the present invention on mouse thymus body weight ratio without impact.
The impact of table 4 preparation of the present invention on mice delayed allergy
**P＜0.01
Per os gives the preparation 30d of the present invention of mice various dose, and initial data carries out after homogeneity test of variance, meets the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carries out statistical disposition.From table 4, between each dosage group and 0.00ml/kg BW group, compare, difference all has utmost point significance (P < 0.01), and preparation of the present invention all can improve the delayed allergy ability of mice in 0.83ml/kg BW group, 8.33ml/kg BW group, 15.00ml/kg BW group.
The impact of table 5 preparation of the present invention on the mouse lymphocyte transformation experiment of ConA induction
Per os gives the preparation 30d of the present invention of mice various dose, and initial data carries out after homogeneity test of variance, meets the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carries out statistical disposition.From table 5, between each dosage group and 0.00ml/kg BW group, relatively, there are no significant for difference (P > 0.05), and the mouse lymphocyte conversion capability that preparation of the present invention is induced ConA is without impact.
From table 4,5, preparation immune function of mice measurement result of the present invention is positive.
2.4 impacts of preparation of the present invention on mouse humoral immune
The impact of table 6 preparation of the present invention on mouse antibodies cellulation number
*P＜0.01
Per os gives the preparation 30d of the present invention of mice various dose, and initial data carries out after homogeneity test of variance, meets the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carries out statistical disposition.From table 6, between 15.00ml/kg BW group and 0.00ml/kg BW group, compare, difference all has significance (P < 0.05), and between all the other each dosage groups and 0.00ml/kg BW group, relatively, there are no significant for difference (P > 0.05).Be that preparation of the present invention can improve mouse antibodies cellulation number in 15.00ml/kg BW group.
The impact of table 7 preparation of the present invention on mice half hemolysis value
*P＜0.05
Per os gives the preparation 30d of the present invention of mice various dose, and initial data carries out after homogeneity test of variance, meets the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carries out statistical disposition.From table 7, between 8.33ml/kg BW group and 0.00ml/kg BW group, compare, difference has significance (P < 0.05), and between all the other each dosage groups and 0.00ml/kg BW group, relatively, there are no significant for difference (P > 0.05).Be that preparation of the present invention can increase the half hemolysis value of mice in 8.33ml/kg BW group.
From table 6,7, preparation mouse humoral immune functional examination result of the present invention is positive.
2.5 impacts of preparation of the present invention on mouse monokaryon-macrophage phagocytic function
The impact that table 8 preparation of the present invention is cleaned up ability to mouse monokaryon-macrophage carbon
Per os gives the preparation 30d of the present invention of mice various dose, and initial data carries out after homogeneity test of variance, meets the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carries out statistical disposition.From table 8, between each dosage group and 0.00ml/kg BW group, relatively, there are no significant for difference (P > 0.05).Be that preparation of the present invention is cleaned up ability without impact to mouse monokaryon-macrophage carbon.
The impact of table 9 preparation of the present invention on mouse macrophage chicken red blood cell phagocytic rate
*P＜0.05 **P＜0.01
Per os gives the preparation 30d of the present invention of mice various dose, initial data carries out after the conversion of square root arcsine, through homogeneity test of variance, meet the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carry out statistical disposition.From table 9, between 8.33ml/kg BW group and 0.00ml/kg BW group, compare, difference has significance (P < 0.05), between 15.00ml/kg BW group and 0.00ml/kg BW group, compare, difference has utmost point significance (P < 0.01), between 0.83ml/kg BW group and 0.00ml/kg BW group, compare no significant difference (P > 0.05).Be that preparation of the present invention can improve mouse macrophage chicken red blood cell phagocytic rate in 8.33ml/kg BW group, 15.00ml/kg BW group.
The impact of table 10 preparation of the present invention on mouse macrophage chicken red blood cell phagocytic index
*P＜0.05 **P＜0.01
Per os gives the preparation 30d of the present invention of mice various dose, and initial data carries out after homogeneity test of variance, meets the neat requirement of variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carries out statistical disposition.From table 10, between 8.33ml/kg BW group and 0.00ml/kg BW group, compare, difference has significance (P < 0.05), between 15.00ml/kg BW group and 0.00ml/kg BW group, compare, difference has utmost point significance (P < 0.01), between 0.83ml/kgBW group and 0.00ml/kg BW group, compare no significant difference (P > 0.05).Be that preparation of the present invention can improve the phagocytic index of mouse macrophage chicken red blood cell in 8.33ml/kg BW group, 15.00ml/kg BW group.
From table 8,9,10, preparation mouse monokaryon-macrophage function measurement result of the present invention is positive.
2.6 impacts of preparation of the present invention on NK cells in mice activity
The impact of table 11 preparation of the present invention on NK cells in mice activity
Per os gives the preparation 30d of the present invention of mice various dose, initial data nonnormal distribution, carry out, after the conversion of square root arcsine, meeting the neat requirement of normal state variance, with the comparative approach between two of mean between one factor analysis of variance method and a plurality of experimental group and a matched group, carry out statistical disposition.From table 11, between each dosage group and 0.00ml/kg BW group, relatively, there are no significant for difference (P > 0.05).Be that preparation of the present invention is active in impact on NK cells in mice, i.e. NK cells in mice determination of activity result is negative.
3 brief summaries
Per os gives the preparation 30d of the present invention of mice various dose, on mouse thymus body weight ratio, spleen body weight ratio without impact.Body weight to mice has no adverse effects.The monocytes/macrophages function result of the cellular immune function of mice, humoral immune function, mice is all positive, and NK cytoactive detection result is negative.As can be seen here, preparation of the present invention has the function of enhancing immunity.
Claims (1)
1. for a liquor preparation for warming and recuperating the kidney-YANG, enhancing immunity, the strong body that gets rid of illness, its formula adopting is: 150～210 parts of Radix Panacis Quinquefoliis, 80～110 parts, Fructus Mori, 80～110 parts of Rhizoma Polygonati Odorati, 45～75 parts of Fructus Lycii, 15～35 parts of Flos Chrysanthemis, 15～35 parts, Poria, 15～35 parts of the Radix Paeoniae Albas, 15～35 parts of Concha Ostreaes, 15～35 parts of Thallus Laminariae (Thallus Eckloniae)s, 8～16 parts of Radix Angelicae Sinensis, 8～16 parts of Geckos, 6000～7000 parts of 3～9 parts, 50 ° base wine of Pericarpium Citri Reticulatae, 3000～4000 parts, water; Concrete preparation technology is: Radix Panacis Quinquefolii, Fructus Mori, Rhizoma Polygonati Odorati, Fructus Lycii, Flos Chrysanthemi, Poria, the Radix Paeoniae Alba, Concha Ostreae, Thallus Laminariae (Thallus Eckloniae), Radix Angelicae Sinensis, Gecko, Pericarpium Citri Reticulatae are ground into coarse powder, put into extraction pot, add 50 ° of base wine sealing dippings 30 days, stir once every day, after one week, stir once weekly, filter, filtrate water is blent into 28 °～35 °, sealing and standing 7 days, filter fill and get final product.
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