CN103405590B - A kind of Chinese medicine preparation and preparation method with immunoregulation effect - Google Patents

Abstract

The invention belongs to oral preparation of Chinese traditional medicinal technical field, in order to avoid body's immunity imbalance, make cancer patient's life span extension, hepatitis hepatic injury alleviates, extend survival rate, improve and strengthen the immunoreation etc. of various diseases patient, provide a kind of Chinese medicine preparation and the preparation method with immunoregulation effect.Be prepared from according to a certain percentage by the Radix Astragali, Radix Ginseng, Cordyceps, Cornu Cervi Degelatinatum, Poria, Rhizoma Alismatis, Fructus Lycii, Ganoderma, Fructus Corni.Chinese medicine preparation safety non-toxic of the present invention, there is effect of QI invigorating primordial QI reinforcing, for malignant tumor and tumor Radiotherapy chemotherapy, the body's immunity imbalances such as viral hepatitis, make cancer patient's life span extension, hepatitis hepatic injury alleviates, particularly the adjustment for the treatment of of the various diseases such as immunologic hypofunction, extends survival rate.

Description

A kind of Chinese medicine preparation and preparation method with immunoregulation effect

Technical field

The invention belongs to oral preparation of Chinese traditional medicinal technical field, be specifically related to a kind of Chinese medicine preparation and the preparation method with immunoregulation effect.

Background technology

Modern medicine study thinks that immunity is the anti-pre-identification foreign body of body self and the process that reacts with it, it growth, heredity, old and feeble, infect, important role in the generation of tumor and autoimmune disease.Along with developing rapidly of modern medicine, the disease relevant with immune dysfunction increases relatively increasingly.When being caused kidney and gastroenteropathy, exfoliative dermatitis, congenital heart disease by dysfunction, immunoglobulin content is significantly lower than normal level: digestive tract malabsorption, respiratory repeated infection, rheumatoid arthritis, systemic lupus erythematosus, thyroiditis, pernicious anemia often accompany the defect in humoral immunization; Late tumor, Hokdkin disease, chronic lymphocytic leukemia people, cellular immunization often shows defectiveness; When acute, chronic infection, severe malnutrition, also often occur that secondary immunodeficiency disease poison infects immunne response capable of inhibiting cell, tuberculin as measles patient can be converted into feminine gender, the immunoreactivity of tuberculosis, leprosy, chronic hepatitis B patients is normal lower, when disease recovery, immunologic function is also just recovered; Life-time service immunosuppressant, during as cyclophosphamide, ammonia methylpurine, vincristine or adrenocortical hormone, the immunologic function of body is in low state, and the frequency that infection or malignant tumor occur is often higher.In addition, the various antibiotic of life-time service also has the possibility causing immunodeficiency.

Polysaccharide, with small-molecular peptides, neuroendocrine agent there is immunization.Polysaccharide generic nonspecific immunity promoter, be also the factor of determination of many microorganism immunological characteristics, its action intensity is different with its chemical characteristic.The larger effect of molecular weight is stronger, water miscible by force more non-water-soluble, and various polysaccharide has again its respective immunological characteristic, promotes the immunoregulation effect such as nonspecific immunity and cellular immunization as astragalus polysaccharides and Hedysamn polysaccharide have; Lentinan is T cell specific immunity adjuvant, from activating T cell, is remake for B cell by t helper cell.Small-molecular peptides section has the immunoreactive effect of adjustment body, and if muramyldipeptide and a series of derivant thereof are class synthesis glycopeptides, the immunoreation of energy irritation cell, enhancing antibody generate, have nonspecific anti-infectious function; Thymus Toplink makes lymphoid stem cell maturation become the lymphocyte with immunologic function, and can improve immunity, mainly strengthens cellular immune function.Neuroendocrine agent such as choline, epinephrine, dopamine and opiate receptor etc. are all present in lymphocyte.Epinephrine is a kind of immunosuppressant; Dopamine has excitation to beta-receptor; H2 antagonist as cimetidine and metiamide can improve lymphocytic propagation, Ig generation, eliminate T suppression cell (Ts) and enhancing macrophage function.

Alkylating agent, corticoid, antibacterials etc. have immunosuppressive action.Alkylating agent is a para-immunity inhibitor, all has inhibitory action, as cyclophosphamide to primary immune, secondary immunoresponse and cellular immunization, humoral immunization.Corticosteroids is a class major effect sugar, protein, lipometabolic medicine, be widely used in allergic disease, as cortisone, prednisone, omcilon, dexamethasone etc. can suppress the pathological change caused by anaphylaxis, suppress the effect of macrophage phagocytic and " process " antigen, the destruction of immunocompetent lymphocytes can be accelerated, make lymphopenia, thus T suppression cell immunity and humoral immunization.Antibacterials are as the metabolite polyactin, Tremella fuciformis spores polysaccharide, krestin, fragrant glucosides class etc. of penicillins, cephalosporins, Tetracyclines, Macrolide, antitumor antibiotics class, aminoglycosides and microorganism, most suppress lymphoblastic transformation effect, and to engulf and the effect of macrophage has show as strengthen have for weakening.

Interference element has immunoregulation effect, regulate multiple lymphocyte and macrophage function, that a class can suppress virus at the protein of cellular proliferative, the propagation of multiple virus can be suppressed, but there is certain selectivity to acted on cell, namely the zooblast of interferon is produced, only to of the same race or have antiviral effect with generic zooblast, interferon does not directly suppress virus, but inducing cell synthesis another kind ofly has inhibiting protein to virus multiplication, the method having adopted genetic engineering to recombinate at present is produced in a large number, and learn that plant cell also can be induced to produce interferon.Immunopharmacology result of study confirms the medicine not yet obtained so far several the pointed effects of immunologic function, with regard to all kinds of immune drugs used at present, their chemical property and pharmacology have sizable difference, therefore, the mechanism of various kinds of drug and the problem of regularity also treat continuous research and exploitation.

Chinese medicine is promoting and research in conditioner body immunity function constantly makes further progress, and all conducts in-depth research chemical composition, model of action and site of action contained by them and inquires into.Situation about activating by it is different, can be divided into promotion reticuloendothelial system, adjustment humoral immunization is more answered and cellular immunization more should wait three class medicines.The medicine of reticuloendothelial system effect is inquired in recent years and more has Qi-tonifying drug, drugs for nourishing yin, drug for invigorating blood circulation and eliminating stasis and heat and toxic materials clearing away medicine etc. as Radix Ginseng, the Radix Astragali, Radix Codonopsis, Cordyceps, the Rhizoma Atractylodis Macrocephalae, Radix Glycyrrhizae, Ganoderma, Rhizoma Dioscoreae, Rhizoma Polygonati etc., all there is the effect significantly strengthening reticuloendothelial system phagocytic function and spleen sterilizing ability, visible righting QI invigorating class Chinese medicine is from immunology viewpoint, and its mechanism is the nonspecific immunity reaction of enhancing body; Drugs for nourishing yin Carapax Et Plastrum Testudinis, Carapax Trionycis, Concha Ostreae etc. also have the effect significantly stimulating reticuloendothelial system phagocytic activity; Drug for invigorating blood circulation and eliminating stasis Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Semen Persicae, Rhizoma Sparganii, Rhizoma Curcumae etc. can excited reticuloendothelial systems and discharge a large amount of macrophage, improve the opsonic vigor of serum, strengthen the effect of leukocytes phagocytic and bacterial digestion and sheep red blood cell (SRBC); Directly antibacterial or antivirus action was all thought mainly in the effect of heat and toxic materials clearing away medicine in the past, recently find that its anti-infectious function plays, as Flos Lonicerae, Herba Andrographis, Herba Houttuyniae, Radix Sophorae Tonkinensis, Flos Chrysanthemi Indici, Rhizoma Coptidis, Radix Scutellariae etc. have the function promoting macrophage phagocytic effect due to the immunologic function of enhancing body by immunological investigation.Humoral immunization is played to the Chinese medicine of activation, mainly contain qi-benefiting and spleen-strengthening drug and reinforcing the kidney and supporting YANG medicine, what research report was more is the Radix Astragali, it can not only promote humoral immunization, improve the content of immunoglobulin, increases the titre of blood plasma agglutination element, and can improve cytotrophy, promote protein synthesis and energy metabolism, expansion peripheral blood vessel, promotes blood circulation, is all conducive to resistance against diseases.The said healthy energy of the traditional Chinese medical science is deficient, go down with the said body's immunity of modern medicine, especially cellular immune level lowly has substantial connection, test report factually, Radix Codonopsis, the Radix Astragali, Radix Ginseng, Rhizoma Polygonati, the Rhizoma Atractylodis Macrocephalae, Cornu Cervi Pantotrichum, Ganoderma, Rhizoma Dioscoreae, Radix Rehmanniae Preparata, Herba Ecliptae, Fructus Schisandrae Chinensis, Semen Cuscutae etc. can increase T cell ratio; Herba Taxilli, Herba Ecliptae, Herba Violae, Herba Ephedrae, Cornu Bubali, Rhizoma Coptidis, Radix Scutellariae, Fructus Schisandrae Chinensis, the Radix Paeoniae Alba etc. can promote lymphocytic conversion.The research of drug for invigorating blood circulation and eliminating stasis thinks all there is certain inhibitory action to humoral immunization and cellular immunization, as Herba Leonuri produces obvious inhibitory action to mouse immune antibody capable, the antibody generated can not only be reduced, and suppress antibody-producting cell: Caulis Spatholobi, Flos Carthami, Radix Salviae Miltiorrhizae etc. have penetration enhancement and Digestion to the antigen antibody complex deposited; And for example removing heat from blood drugs for nourishing yin Radix Rehmanniae, Cortex Moutan, Fructus Ligustri Lucidi, Radix Ophiopogonis, Radix Scrophulariae, Radix Asparagi etc. can Immunosuppression hyperfunctioning: the generation that can suppress neutralizing antibody had, the rising that can reduce serum immune globulin had and promote albuminous synthesis, some can T suppression cell immunity.Radix Glycyrrhizae is also immunosuppressant very good simply.Compound preparation BUZHONG YIQI TANG, Huangqi Jianzhong Tang, Radix Angelicae Sinensis decoction for tonifying blood, LIUWEI DIHUANG TANG, spleen kidney soup, warming YANG soup, body resistance-strengthening Zhenqi soup, YUPINGFENG SAN, GUHAN YANGSHENG JING etc., confirm through experiment the effect all having promotion or immunity moderation.

Along with the development of science and technology, medical research shows that many theories of Chinese medicine and immunologic argument have identical part, as traditional Chinese medical science vigour and Immunology Today immunity of organism have very close relationship.The concept of Chinese medicine " gas " comprises modern medicine immunity of organism basic conception.Thus the various diseases such as immune state and malignant tumor, viral hepatitis, nephritis of body is closely related.Therefore, improve and strengthen the effect that the immunoreation of patient improves operation, radiotherapy or chemotherapy to a great extent.Immune dysfunction, particularly immunologic hypofunction, Chinese medicine thinks the deficiency of vital energy or insufficiency of primordial QI in belong to more, first weakness of QI then disease by giving birth to.Combine with modern medicine according to theory of Chinese medical science, the invigorating the spleen and benefiting QI of application Chinese medicine, QI invigorating, with the rule for the treatment of of primordial QI reinforcing, is recovered body vigour, is reached the object of strengthening vital QI to eliminate pathogenic factors, and with modern medicine technique study QI invigorating primordial QI reinforcing and immunopharmacology relation, and develop new drug QI invigorating primordial QI reinforcing compound Chinese medicinal preparation, and for malignant tumor and tumor Radiotherapy chemotherapy, the body's immunity imbalances such as viral hepatitis, make cancer patient's life span extension, hepatitis hepatic injury alleviates.Have research data to show: reinforcement has not only played certain effect on the comprehensive therapy of tumor, and can not undergo surgery to some, the end-stage patients of radiotherapy or chemotherapy, also obtain good therapeutic effect, stablize the development of tumor, extend survival rate.

Summary of the invention

The present invention is in order to avoid body's immunity imbalance, make cancer patient's life span extension, hepatitis hepatic injury alleviates, and extends survival rate, improve and strengthen the immunoreation etc. of various diseases patient, provide a kind of Chinese medicine preparation and the preparation method with immunoregulation effect.

The present invention is realized by following technical scheme: a kind of Chinese medicine preparation with immunity moderation effect, is made up of the raw material of following weight proportion: Radix Astragali 9-30 part, Radix Ginseng 10-20 part, Cordyceps 2-5 part, Cornu Cervi Degelatinatum 10-20 part, Poria 10-20 part, Rhizoma Alismatis 9-20 part, Fructus Lycii 9-20 part, Ganoderma 15-30 part, Fructus Corni 9-20 part.

The optimum ratio scheme of each component of Chinese medicine preparation of the present invention is: the Radix Astragali 10 parts, Radix Ginseng 15 parts, Cordyceps 3 parts, Cornu Cervi Degelatinatum 15 parts, 15 parts, Poria, Rhizoma Alismatis 10 parts, Fructus Lycii 10 parts, Ganoderma 20 parts, Fructus Corni 10 parts.

The preparation method of Chinese medicine preparation of the present invention is: the powder of Fructus Corni of Cordyceps, Poria, 1/3 weight portion is broken into 80-100 order fine powder;

Radix Ginseng adds raw material weight 25-35 soak by water 2-4h doubly, and filter, filtrate is for subsequent use; Medicine residues of Radix Ginseng after decoction and all the other medicines add raw material weight 20-25 soak by water doubly 1-3 time, each 1-3h, and collecting decoction also filters, and filtrate and Radix Ginseng decoct filtrate merging, and 80 DEG C are concentrated into the clear paste that relative density is 1.34-1.38;

Add above-mentioned Cordyceps, Poria, 1/3 weight portion Fructus Corni pulverize the fine powder made; Drying under reduced pressure, is ground into 80-100 order fine powder, and mixing, is prepared into peroral dosage form, Co ₆₀sterilizing.

Described conventional peroral dosage form is pill, powder, tablet, capsule, oral liquid etc.It is emphasized that for a person skilled in the art, it still can be modified to technical solutions according to the invention, or carries out equivalent replacement to wherein portion of techniques feature.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.

In Chinese medicine preparation of the present invention, Cordyceps, QI invigorating kidney tonifying, helps unit, is equipped with Cornu Cervi Degelatinatum and jointly plays fonifying life-gate fire, help kidney yang, effect of tonifying YANG gas." kind YANG invigorating person, must in treating YANG within YIN " be so except with Cordyceps in side, outside the YANG invigorating gas medicines such as Cornu Cervi Degelatinatum, being aided with Fructus Lycii, Fructus Corni negative and positive with mending, making the meaning of the party's tool YANG invigorating gas and " its source is inexhaustible "; By the gas of Radix Ginseng, the Radix Astragali, Ganoderma lucidum seu Japonicum tonifying the lung kidney, invigorate vital energy and yang, the unboiled water by gold, mends soil and helps life fire, and Poria, Rhizoma Alismatis diuretic and to rush down kidney turbid, yang-energy is also mended, negative and positive syngenesis, jointly reaches the effect helping Yuanyang to order fire.Safety non-toxic, has effect of QI invigorating primordial QI reinforcing, for malignant tumor and tumor Radiotherapy chemotherapy, the body's immunity imbalances such as viral hepatitis, make cancer patient's life span extension, hepatitis hepatic injury alleviates, the particularly adjustment for the treatment of of the various diseases such as immunologic hypofunction, extends survival rate.

Accompanying drawing explanation

The capsule of Fig. 1 prepared by the embodiment of the present invention 2 is to In vitro culture HL ₆₀tumor cell growth inhibitory action;

The capsule of Fig. 2 prepared by the embodiment of the present invention 2 is to In vitro culture K ₅₆₂tumor cell growth inhibitory action;

Fig. 3 is the liver Electronic Speculum figure of normal control mice;

Fig. 4 is model comparison mouse liver Electronic Speculum figure;

Fig. 5 is capsule low dosage process mouse liver micro-organization chart of the present invention;

Fig. 6 is capsule in high dose process mouse liver micro-organization chart of the present invention;

Fig. 7 is Lentinula edodes mycelium polysaccharide sheet process mouse liver micro-organization chart.

Detailed description of the invention

embodiment 1

There is a Chinese medicine preparation for immunity moderation effect, be made up of the raw material of following weight proportion: Radix Astragali 9g, Radix Ginseng 10g, Cordyceps 2g, Cornu Cervi Degelatinatum 10g, Poria 10g, Rhizoma Alismatis 9g, Fructus Lycii 9g, Ganoderma 15g, Fructus Corni 9g.The powder of Fructus Corni of Cordyceps, Poria, 3g is broken into 80 order fine powders; People participates in the soak by water 2h of raw material weight 25 times, and filter, filtrate is for subsequent use; Medicine residues of Radix Ginseng after decoction and all the other medicines add the soak by water 1 time of raw material weight 20 times, each 1h, collecting decoction also filters, and filtrate and Radix Ginseng decoct filtrate merging, 80 DEG C are concentrated into the clear paste that relative density is 1.34, add the fine powder of Cordyceps, Poria, Fructus Corni pulverizing; Drying under reduced pressure, is ground into 90 object fine powders, and mixing, is prepared into powder, Co in conventional manner ₆₀sterilizing.

embodiment 2

There is a Chinese medicine preparation for immunity moderation effect, be made up of the raw material of following weight proportion: Radix Astragali 10g, Radix Ginseng 15g, Cordyceps 3g, Cornu Cervi Degelatinatum 15g, Poria 15g, Rhizoma Alismatis 10g, Fructus Lycii 10g, Ganoderma 20g, Fructus Corni 10g.The powder of Fructus Corni of Cordyceps, Poria, 3.33g is broken into 100 order fine powders; People participates in the soak by water 3h of raw material weight 30 times, and filter, filtrate is for subsequent use; Medicine residues of Radix Ginseng after decoction and all the other medicines add the soak by water 2 times of raw material weight 22 times, each 2h, collecting decoction also filters, and filtrate and Radix Ginseng decoct filtrate merging, 80 DEG C are concentrated into the clear paste that relative density is 1.34-1.38, add the fine powder of Cordyceps, Poria, Fructus Corni pulverizing; Drying under reduced pressure, is ground into 80 order fine powders, and mixing, is prepared into capsule, Co in conventional manner ₆₀sterilizing.

embodiment 3

There is a Chinese medicine preparation for immunity moderation effect, be made up of the raw material of following weight proportion: Radix Astragali 30g, Radix Ginseng 20g, Cordyceps 5g, Cornu Cervi Degelatinatum 20g, Poria 20g, Rhizoma Alismatis 20g, Fructus Lycii 20g, Ganoderma 30g, Fructus Corni 20g.The powder of Fructus Corni of Cordyceps, Poria, 6.67g is broken into 90 order fine powders; People participates in the soak by water 4h of raw material weight 35 times, and filter, filtrate is for subsequent use; Medicine residues of Radix Ginseng after decoction and all the other medicines add the soak by water 3 times of raw material weight 25 times, each 3h, collecting decoction also filters, and filtrate and Radix Ginseng decoct filtrate merging, 80 DEG C are concentrated into the clear paste that relative density is 1.34-1.38, add the fine powder of Cordyceps, Poria, Fructus Corni pulverizing; Drying under reduced pressure, is ground into 100 order fine powders, and mixing, is prepared into tablet, Co in conventional manner ₆₀sterilizing.

embodiment 4

There is a Chinese medicine preparation for immunity moderation effect, be made up of the raw material of following weight proportion: Radix Astragali 15g, Radix Ginseng 10g, Cordyceps 4g, Cornu Cervi Degelatinatum 20g, Poria 10g, Rhizoma Alismatis 15g, Fructus Lycii 20g, Ganoderma 25g, Fructus Corni 15g.The powder of Fructus Corni of Cordyceps, Poria, 5g is broken into 95 order fine powders; People participates in the soak by water 2h of raw material weight 28 times, and filter, filtrate is for subsequent use; Medicine residues of Radix Ginseng after decoction and all the other medicines add the soak by water 2 times of raw material weight 20 times, each 3h, collecting decoction also filters, and filtrate and Radix Ginseng decoct filtrate merging, 80 DEG C are concentrated into the clear paste that relative density is 1.34-1.38, add the fine powder of Cordyceps, Poria, Fructus Corni pulverizing; Drying under reduced pressure, is ground into 85 order fine powders, and mixing, is prepared into pill, Co in conventional manner ₆₀sterilizing.

Below by way of zoopery, effect of the present invention is described, in experiment, Chinese medicine preparation used is Chinese medicine preparation described in embodiment 2.

experimental example 1: immunoregulation effect

One, on the impact of mice reticuloendothelial system phagocytic function

Tested material: the capsule prepared by the embodiment of the present invention 2; Reagent: india ink, chemical plant in West Beijing, faces in order to normal saline 20% india ink normal saline solution; Instrument: torsion balance, 7210 type spectrophotometers, Shanghai analytical tool factory; Animal subject: Kunming mouse, laboratory animal room of Shanxi Institute of Traditional Chinese Medicine provides.

Test method and result:

Get mice 42, body weight 22 ± 2g, male and female half and half, be divided into 4 groups at random: normal control, capsule in high dose of the present invention, low dosage, Lentinula edodes mycelium polysaccharide sheet.Gastric infusion, Normal group to equivalent distilled water, 1 time on the one, continuous 10 days.11st day, after mice is weighed, inject 20% india ink normal saline solution 0.1ml/10g by tail vein, after injection, 2min and 22min to take a blood sample 20 μ l from vena orbitalis posterior clump respectively, is placed in spectrophotometer at 570nm place its absorbance of mensuration.After getting blood, mice dislocation is put to death, and win liver, spleen torsion balance is weighed, calculate phagocytic index, computing formula is: .Experimental result is in table 1.Result shows capsule in high dose administration of the present invention and Lentinula edodes mycelium polysaccharide sheet all can significantly improve mouse monokaryon phagocytic function.

Table 1 capsule of the present invention is on the impact (X ± SD) of mouse monokaryon phagocytic function

Compare with Normal group: * P<0.05, * * P<0.01.

Two, on the impact of caused by cyclophosphamide mouse humoral immune hypofunction

Tested material and Lentinula edodes mycelium polysaccharide the same; Reagent: cyclophosphamide, Shanghai No.12 Pharmaceutical Factory, lot number 920205, faces used time normal saline; A1serer, Dou Shi liquid agents useful for same is analytical pure; Instrument: 7210 type spectrophotometers, Shanghai analytical tool factory; Animal subject: Kun Ming mice, laboratory animal room of Shanxi Institute of Traditional Chinese Medicine provides.

Test method and result:

Get mice 60, male, body weight 23 ± 3g, is divided into 5 groups: normal control, model comparison, capsule in high dose of the present invention, low dosage, Lentinula edodes mycelium polysaccharide sheet.Gastric infusion, Normal group and model control group to equivalent distilled water, 1 time on the one, continuous 10 days.In 7,8,9 three days except Normal group, equal subcutaneous injection cyclophosphamide 20mg/Kg.In the 7th day equal lumbar injection sheep red blood cell (SRBC) of each treated animal: normal saline is the solution of 3:5,0.2ml/ only.11st day, pluck eyeball and get blood, separation of serum, measure absorbance in 540nm place, and record SRBC HD50 absorbance 0.55 simultaneously, calculate half hemolysis value (HC ₅₀), computing formula is: HC ₅₀absorbance × extension rate during=sample absorbance value/sheep red blood cell HD50.Experimental result is in table 2.Result shows capsule in high dose administration of the present invention and Lentinula edodes mycelium polysaccharide sheet all can significantly improve sensitized mice humoral immune function.

Table 2 capsule of the present invention is on the impact (X ± SD) of Acu-point plaster therapy antibody tormation

Compare with model control group: * P<0.05.

Three, right ⁶⁰co irradiates the impact of immune function of mice

(i), right ⁶⁰co irradiates the impact of mouse lymphocyte conversion ratio, IL-2, NK cytoactive

Tested material and Lentinula edodes mycelium polysaccharide the same; Reagent: RPMll640t nutritional solution completely, F1owLabScoland; Concanavalin A, Con A (ConA), Sigma; Phytohaemagglutinin (PHA), Guangzhou institute of Pharmaceutical Industry; ³h-TdR, Shanghai Institute for Atomic Research; Cell strain YaK-1, Shanxi Tumors Inst. provides; Instrument: torsion balance, Shanghai analytical tool factory; Cell sample collector with multiple heads (DYQ-2 type), Shaoxing Po Kuang medical apparatus and instruments factory; LS5000CE liquid scintillation instrument, U.S. Beckman; Animal: Kunming mouse, laboratory animal room of Chinese Radiation Protection Research Inst provides.

Test method and result:

Get mice 34, male, body weight 24 ± 2g, is divided into 5 groups at random: normal control, model comparison, capsule in high dose of the present invention, low dosage, Lentinula edodes mycelium polysaccharide sheet.Gastric infusion, normal control and model comparison are to equivalent distilled water, and 1 time on the one, continuous 21 days, within the 8th day, except Normal group, respectively group was equal for all the other ⁶⁰co irradiates once, close rate 1.05GY/min, accumulated dose 3GY.22nd day, pluck eyeball blood-letting, asepticly get spleen, spleen is put into plate, filter through grinding, be made into cell suspension with complete RPMll640 culture fluid, in order to lymhocyte transformation rate, IL-2, NK cytoactive detection.

(1), lymhocyte transformation rate measures

Get 1 × 10 ⁵/ 0.1ml spleen cell cultures liquid add respectively 0.1ml concentration be the concanavalin A, Con A (ConA) of 5 μ g/ml in 96 well culture plates, in CO ₂cultivate 72 hours in incubator, gather in the crops and within first 12 hours, add 0.2 Μ ci/ hole ³h-TdR, collecting cell, survey cpm, experimental result is in table 3.Result display capsule low dosage high, of the present invention administration and Lentinula edodes mycelium polysaccharide sheet all significantly raise ⁶⁰co irradiates mouse lymphocyte conversion ratio.

Table 3 capsule pair of the present invention ⁶⁰co irradiates the impact (X ± SD) of mouse lymphocyte conversion ratio

Compare with model control group, * P<0.05, * * P<0.01.

(2), IL-2 measures

IL-2 supernatant is induced: by 1 × 10 ⁷it is that the ConA of 5 μ g/m1 is sub-packed in 25ml culture bottle that/m1 splenocyte suspension adds 0.1ml concentration, is placed in the CO containing 5% ₂, moisture-saturated incubator in, 37 DEG C of incubations 48 hours; Harvested by centrifugation supernatant, bottle subpackage-20 DEG C preservation.

The preparation of the mouse spleen lymphocyte of ConA activation: make 5 × 10 ⁶/ ml splenocyte suspension, add the ConA that 0.1ml concentration is 10 μ g/ml, be placed in 25ml culture bottle incubation 96h, Hank ' the s liquid of cell l0mg/ml methyl mannose glycoside α-mm washes secondary, and wash once containing 10% people AB pooled serum RPMll640, regulate cell concentration to be 1 × 10 ⁶/ ml is as the reacting cells measuring IL-2.

IL-2 determination of activity: in 96 hole microflat bottom Tissue Culture Plates, every hole adds reacting cells 1 × 10 ⁵the IL-2 supernatant that/ml and equivalent 1/8 are diluted ,-20 DEG C are preserved, adds 0.1ml culture fluid in control wells, experimental group and matched group all establish 2 multiple holes, and every hole adds the α-mm20 μ l of l00mg/ml, is placed in the CO containing 5% ₂incubator in, 37 DEG C of incubation 48h; In the little 0.5 μ ci/ hole of adding constantly of cultivation 32 ³h-TdR, is harvested from fibrous paper with cell sample collector with multiple heads, and survey cpm with Beckman liquid scintillation meter enumerator, experimental result is in table 4.Result shows capsule high and low dose of the present invention administration and Lentinula edodes mycelium polysaccharide sheet all significantly raises ⁶⁰co irradiates mice IL-2 effect.

Table 4 capsule pair of the present invention ⁶⁰co irradiates the impact (X ± SD) of mice IL-2

Compare with model control group, * P<0.05.

(3), NK cytoactive detection: with ³h-TdR labelling, YaK-1 is as target cell, and splenocyte is effector lymphocyte.Effect target ratio E/T=20:1, Dual culture 16 hours, collecting cell, surveys cpm.Calculate NK active, computing formula is: NK activity (%)=(contrast of 1-processed group (splenocyte+oncocyte)/oncocyte) × 100%, experimental result is in table 5.Result display model control group mice warp ⁶⁰after Co irradiates, NK cytoactive has no significant effect, and capsule high and low dose of the present invention administration and Lentinula edodes mycelium polysaccharide sheet are significantly increased NK cytoactive.

Table 5 capsule pair of the present invention ⁶⁰co irradiates the impact (X ± SD) of NK cells in mice activity

* P<0.05 is compared, * * P<0.01 with Normal group

(ii), right ⁶⁰co irradiates the heavy impact of murine interleukin number, thymus and spleen

Tested material and Lentinula edodes mycelium polysaccharide the same; Animal subject: Kunming mouse, laboratory animal room of Chinese Radiation Protection Research Inst provides; Instrument: Cai Shi microscope, Germany; Torsion balance, Shanghai analytical tool factory.

Test method and result:

Get mice 34, body weight 24 ± 2g, male, be divided into 5 groups at random: normal control, model comparison, capsule in high dose of the present invention, low dosage, Lentinula edodes mycelium polysaccharide sheet.Gastric infusion, normal control and model comparison are to equivalent distilled water, and 1 time on the one, continuous 21 days, within the 8th day, except Normal group, respectively group was equal for all the other ⁶⁰co irradiates once, close rate 1.05GY/min, accumulated dose 3GY.Irradiate latter one week, model control group mice occurs that hair color is gloomy, Capsules group hair color of the present invention is glossy close to Normal group, Herba Glossogynes tenuifoliae granulose sheet administration group, between capsule administration group of the present invention and model group, the 22nd day, is got blood 0.01ml from eyeball rear vein beard and is added 1.5%HAC0.2m1, leukocyte (WBC) counting under light microscopic, experimental result is in table 6, and cut open the belly and win thymus, spleen torsion balance is weighed and is converted into ponderal index, experimental result is in table 7.Result shows, and capsule height dosed administration of the present invention and Lentinula edodes mycelium polysaccharide sheet are to being subject to ⁶⁰co irradiates mice WBC rising trend, and to being subject to ⁶⁰co irradiates mouse thymus weight rising trend, and capsule in high dose administration of the present invention significantly raises mice spleen weight.

Table 6 capsule pair of the present invention ⁶⁰co irradiates the impact (X ± SD) of murine interleukin number

Compare with model control group; * * P<0.001.

Table 7 capsule pair of the present invention ⁶⁰co irradiates the impact (X ± SD) of mouse thymus, spleen weight

Compare with model control group, * P<0.05.

experimental example 2: antitumor action

One, to the tumor-inhibiting action of Ehrlich ascites carcinoma

Tested medicine: capsule of the present invention is the same; Tegafur (FNFNMD), Jinan pharmaceutical factory, lot number 910315; Animal subject: Ehrlich ascites carcinoma, Kunming mouse, provides by laboratory animal room of Shanxi Tumors Inst..

Test method and result:

Get Kunming mouse 48, male and female half and half, body weight 23 ± 2g, aseptically, take out Ehrlich ascites carcinoma ascites, do 1:2 dilution with normal saline, every mouse peritoneal injection 0.2ml, is divided into 4 groups at random by the mice processed: ehrlich ascites carcinoma matched group, capsule in high dose of the present invention, low dosage administration group, Tegafur group.Gastric infusion, ehrlich ascites carcinoma matched group is to equivalent distilled water, 1 time on the one, drug withdrawal after continuous 7 days, records Mouse Weight day by day, after 11 days, mice weightening finish and the heavy situation of tumor are in table 8, and observe death condition, observe 31 days time limits, calculate increase in life span, tumor control rate, computing formula is: increase in life span=(treatment group Average Survival natural law-matched group Average Survival natural law)/matched group Average Survival natural law × 100%; Tumor control rate=(matched group tumor weight-administration group tumor weight)/matched group tumor heavy × 100%; Experimental result is in table 9.Experimental result shows: capsule in high dose administration of the present invention and Tegafur all delay ehrlich ascites carcinoma mice tumors grew speed, and all can extend ehrlich ascites carcinoma mouse survival sweet food, improves tumor control rate.

The impact (X ± SD) that table 8 capsule of the present invention is heavy on Ehrlich ascites carcinoma tumor

Compare with ehrlich ascites carcinoma matched group: * P<0.05, * * P<0.01.

Table 9 is on the impact of Ehrlich ascites carcinoma increase in life span, tumor control rate

Two, to the In-vitro Inhibitory Effect of tumor strain

Culture fluid: 10% calf serum (CS), Hangzhou Ilex purpurea Hassk.[I.chinensis Sims engineering material institute; RPMll640J culture fluid: FlowLabScoland; Tumor cell line K ₅₆₂, HL ₆₀, Shanxi Tumors Inst., with 10%CSl640 culture fluid adjustment cell concentration to 5 × 10 ⁶/ ml; Experimental drug: capsule of the present invention described in embodiment 2, the dry liquid of capsule of 20% is mixed with 10%CSl640 culture fluid, adjust PH to 7.2 with 1NNaOH, then be diluted to variable concentrations with 10%CSl640 culture fluid: 10 μ g/ml, 100 μ g/ml, 1000 μ g/ml, hereinafter referred to as the first culture fluid of Ji; ³h-thymidine ( ³h-TdR), Chinese Atomic Energy Research Institute, faces with being mixed with 20% μ ci/ml's ³h-TdR.

Test method and result:

Tumor cell suspension 100 μ l, adds in 96 well culture plates, preculture 30 minutes, adds 10 μ g/ml, 100 μ g/ml, the Ji unit culture fluid 100 μ l of 1000 μ g/ml concentration makes final concentration be 5 μ g/ml, 50 μ g/ml, 500 μ g/ml, at 5%CO ₂, saturated humidity 37 DEG C cultivation.

1, cell row dye test:

Respectively at cultivation rear 4, 24, the tumor cell suspension getting 10 μ 1 cultivations for 48 hours adds equivalent 0.4% Trypan Blue liquid, mixing, instillation blood counting chamber, stablize after 2 minutes and observe, count Normal group respectively, high, in, low dosage Capsules group staining cell of the present invention and dead cell, with cell and the living cells of being unstained, try to achieve cell survival rate, computing formula is: cell survival rate=cell number of being unstained/(staining cell number+cell number of being unstained) × 100%, non-transfect cell number (living cells) R value in statistics four large lattice, try to achieve every ml cells number, computing formula is: every ml cells number=R × 1/2 × 104, try to achieve inhibitory rate of cell growth, computing formula is: inhibitory rate of cell growth=(matched group viable count-experimental group viable count)/matched group viable count × 100%.Capsule of the present invention is to the HL of In vitro culture ₆₀the impact of viability, the results are shown in Table 10, to the K of In vitro culture ₅₆₂the impact of viability the results are shown in Table 11.Result shows high, medium and low three the administration groups of capsule of the present invention all obviously can suppress HL at 4,24,48 hours ₆₀and K ₅₆₂viability, and along with dosage increasing, extended durations of action, presents certain timeliness, dose-effect relationship.

Table 10 capsule of the present invention is to the HL of In vitro culture ₆₀impact (%) (X ± SD) of viability

Compare with matched group: * * * P<0.001.

Table 11 capsule of the present invention is to the K of In vitro culture ₅₆₂impact (%) (X ± SD) of viability

Compare with matched group: * P<0.05, * * * P<0.001.

Capsule of the present invention is to HL ₆₀oncocyte, K ₅₆₂the effect of tumor cell growth suppression ratio is in table 12 and Fig. 1,2, and result shows high, medium and low three dosage of capsule of the present invention to HL ₆₀, K ₅₆₂tumor cell growth all has obvious inhibitory action.

Table 12 capsule of the present invention is to In vitro culture 48 hours HL ₆₀, K ₅₆₂the impact of tumor cell growth suppression ratio

2, ³h-TdR mixes the mensuration of DNA:

4 dosage group process tumor cells of variable concentrations pressed by medicine of the present invention, and concentration for the treatment of, in table 13, is cultivated 48 hours, and when processing 44h, every hole adds ³h-TdR1 μ ci, cultivate the ice trichloroacetic acid adding equivalent 20% after terminating, precipitate collecting cell after 10 minutes, add 5% trichloroacetic acid washing, absolute ethanol washing 1 time, surveys cpm, and compares with normal control, what calculate thymus pyrimidine mixes suitable rate and suppression ratio, and computing formula is: mix suitable rate=experimental group cpm/ matched group cpm × 100%; Suppression ratio=(matched group cpm-experimental group cpm)/matched group cpm × 100%.Experimental agents concentration with the results are shown in Table 13.Result shows capsule of the present invention 4 dosage groups pair ³h-TdR mixes HL ₆₀oncocyte DNA all has obvious inhibitory action.

Table 13 capsule of the present invention is to external ³h-TdR mixes HL ₆₀the impact (X ± SD) of oncocyte DNA

Compare with matched group: * * * P<0.001.

experimental example 3: lower cyclophosphamide toxic and side effects

Tested medicine: capsule of the present invention, Lentinula edodes mycelium polysaccharide sheet are the same; Cyclophosphamide, Hualian Pharmaceutical Co., Shanghai, lot number: 950907; Reagent: DAM.α-one glutaraldehyde etc. is analytical pure; Instrument: 7210 type spectrophotometers, Shanghai analytical tool factory; Animal subject: Kun Ming mice, laboratory animal room of Chinese Radiation Protection Research Inst provides.

Test method and result:

Get mice 44, male and female half and half, body weight 25 ± 4g, be divided into 5 groups at random: normal control, model comparison, capsule in high dose of the present invention, low dosage, Lentinula edodes mycelium polysaccharide sheet.Gastric infusion, Normal group and model control group to equivalent distilled water, 1 time on the one, continuous 11 days.8th, within 9 two days, except Normal group, all the other respectively organize mice all by 100mg/kg intraperitoneal injection of cyclophosphamide.Killed inspection in the 12nd day, pluck eyeball and get blood, separation of serum, carry out Liver and kidney function mensuration, the results are shown in Table 14; Cut open the belly and win liver, kidney is placed in 10% formalin fixative and fixes, routine pathology is cut into slices, and HE dyes, and mirror undertissue morphological observation, the results are shown in Figure 3,4,5,6,7.Result shows, and 100mg/Kg cyclophosphamide gives mouse peritoneal and injects twice on renal function without impact, and has certain damage to liver function.Capsule height low dosage of the present invention and Lentinula edodes mycelium polysaccharide sheet all can have protective effect to the hepar damnification of caused by cyclophosphamide.

Table 14 capsule of the present invention is on the impact (X ± SD) of injection Acu-point plaster therapy Liver and kidney function

Compare with model control group: * P<0.05, * * P<0.01.

From Fig. 3,4,5,6,7, Normal group Mouse Liver leaflet structure is clear, has no necrosis region, and portal area is without cell infiltration; Model control group some animals lobules of liver is still clear, the downright bad and companion's connective tissue proliferation of visible lamellar, and some animals lobules of liver is still clear, visible spotty necrosis, and companion's connective tissue proliferation and portal area visible inflammatory cell infiltrate; Capsule low dose group Mouse Liver lobule part-structure of the present invention is still clear, is dispersed in a lamellar necrosis, and infiltrates with connective tissue proliferation and portal area visible inflammatory cell, individual mice lobules of liver clear in structure, has no downright bad and cell infiltration; Capsule in high dose group of the present invention, most lobules of liver clear in structure, has no necrosis region, individual mice lobules of liver clear in structure, is dispersed in a lamellar necrosis, a little cell infiltration in portal area; Herba Glossogynes tenuifoliae granulose sheet group, most Mouse Liver leaflet structure is clear, has no necrosis region, and portal area has no cell infiltration and oozes out, and the still clear visible point lamellar necrosis region of individual mice lobules of liver structure, there is cell infiltration portal area.

experimental example 4: to external hbs antigen (HBsAg) activity inhibition

Tested medicine: capsule prepared by the embodiment of the present invention 2, centrifugal 15 minutes of 3000rpm before use; Purifying antigen, is provided by Ministry of Public Health Clinical Laboratory center; Instrument: 0D3022 microplate reader, state-run East China Electronics Co., Ltd pipe factory.

Test method and result:

By HBsAg purifying antigen, 40ng, 20ng, 10ng tri-concentration are diluted to 10% calf serum PBS, capsule PH7.2 buffer solution of the present invention is diluted to 508 μ g (high dose), 254 μ g (middle dosage), 127 μ g (low dosage), three concentration, join in the HBsAg diluted, act on 12 and 24 hours.With ELISA method operation, microplate reader 450hm place measures its absorbance, and adopt Shanghai to found readable method, Cutoff=sample OD value/negative control mean OD value >0.1, the results are shown in Table 15.

Table 15 capsule of the present invention is to hbs antigen activity 12,24 hours inhibitory action

Note: "+", for invalid, "-" is effective.

Result shows capsule of the present invention and HBsAg acts on 12 hours, the HBsAg activity of capsule in high dose of the present invention to 10ng and 20ng has inhibitory action, measure agent in capsule of the present invention and only have inhibitory action to the HBsAg activity of 10ng, and capsule of the present invention is low dose of to the active unrestraint effect of HBsAg; Capsule of the present invention and HBsAg act on 24 hours, and the HBsAg activity of high dose to 10ng, 20ng and 40ng has inhibitory action, and middle dosage only has inhibitory action to the HBsAg activity of 10ng, and low dose is to the active unrestraint effect of HBsAg.

Claims (3)

1. there is a Chinese medicine preparation for immunity moderation effect, it is characterized in that: be made up of the raw material of following weight proportion: Radix Astragali 9-30 part, Radix Ginseng 10-20 part, Cordyceps 2-5 part, Cornu Cervi Degelatinatum 10-20 part, Poria 10-20 part, Rhizoma Alismatis 9-20 part, Fructus Lycii 9-20 part, Ganoderma 15-30 part, Fructus Corni 9-20 part.

2. a kind of Chinese medicine preparation with immunity moderation effect according to claim 1, is characterized in that: be made up of the raw material of following weight proportion: the Radix Astragali 10 parts, Radix Ginseng 15 parts, Cordyceps 3 parts, Cornu Cervi Degelatinatum 15 parts, 15 parts, Poria, Rhizoma Alismatis 10 parts, Fructus Lycii 10 parts, Ganoderma 20 parts, Fructus Corni 10 parts.

3. there is a preparation method for the Chinese medicine preparation of immunity moderation effect as claimed in claim 1 or 2, it is characterized in that: concrete steps are: the powder of Fructus Corni of Cordyceps, Poria, 1/3 weight portion is broken into 80-100 order fine powder;

Radix Ginseng adds raw material weight 25-35 soak by water 2-4h doubly, and filter, filtrate is for subsequent use; Medicine residues of Radix Ginseng after decoction and all the other medicines add raw material weight 20-25 soak by water doubly 1-3 time, each 1-3h, and collecting decoction also filters, and filtrate and Radix Ginseng decoct filtrate merging, and 80 DEG C are concentrated into the clear paste that relative density is 1.34-1.38;

Add above-mentioned Cordyceps, Poria, 1/3 weight portion Fructus Corni pulverize the fine powder made; Drying under reduced pressure, is ground into 80-100 order fine powder, and mixing, is prepared into peroral dosage form, ⁶⁰co sterilizing.