CN106491834A - A kind of veterinary Chinese medicinal composition and its preparation method and application of granule - Google Patents

A kind of veterinary Chinese medicinal composition and its preparation method and application of granule Download PDF

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CN106491834A
CN106491834A CN201611126167.8A CN201611126167A CN106491834A CN 106491834 A CN106491834 A CN 106491834A CN 201611126167 A CN201611126167 A CN 201611126167A CN 106491834 A CN106491834 A CN 106491834A
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chinese medicine
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朱买勋
翟少钦
曹国文
郑华
张邑帆
闫志强
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Chongqing Academy of Animal Sciences
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/63Oleaceae (Olive family), e.g. jasmine, lilac or ash tree
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    • AHUMAN NECESSITIES
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    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/43Cuscutaceae (Dodder family), e.g. Cuscuta epithymum or greater dodder
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
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    • A61K36/48Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A61K36/481Astragalus (milkvetch)
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    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/81Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
    • A61K36/815Lycium (desert-thorn)
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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    • A61K2236/30Extraction of the material
    • A61K2236/39Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/51Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/53Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

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Abstract

This application discloses the preparation method and application of a kind of veterinary Chinese medicinal composition and its granule.Chinese medicine composition is by 40 parts of Fructus Ligustri Lucidi, 40 parts of the Radix Astragali, 10 parts of Fructus Lycii, 10 parts of Semen Cuscutae.Its preparation method includes:(1) research of extraction process by water, takes 40 parts of Fructus Ligustri Lucidi, and 40 parts of the Radix Astragali, 10 parts of Fructus Lycii, 10 parts of Semen Cuscutae carry out 3 decoctings and extract, and are concentrated into solid-liquid ratio 1:When 5, shell adding polysaccharide is clarified, and after filtration, is condensed into extractum.(2) selection of pharmaceutical drying condition, (1) gained extractum is dried, and is ground into medicated powder, crosses 60 mesh sieves.(3) screening of particle preparation condition, screens adjuvant, makes granule using single step.Chinese medicine composition of the present invention has functions that tonifying Qi and lifting yang, replenishing QI to invigorate the spleen, the medicine of enhancing chicken immuological function and treatment immune disease is can operate with, have the advantages that low production cost, reliable quality safety, colleges and universities' preventing and treating and immune disease and enhancing chicken immuological function.

Description

A kind of veterinary Chinese medicinal composition and its preparation method and application of granule
Technical field
The application belongs to medicine technology field, specifically, is related to the preparation of a kind of veterinary Chinese medicinal composition and its granule Methods and applications.
Background technology
The disease that fowl poultry immune disease is Organism immunoregulation disequilibrium and causes immune response disorderly, animal Immune system in body by immune organ (such as spleen, thymus, fabricius bursa etc.), immunocyte (as T lymphocytes, B lymphs thin Born of the same parents, macrophage, erythrocyte etc.), cytokine (such as IL, TNF, antibody etc.) composition, three is by blood and the mutual ditch of body fluid Logical, cooperate, play jointly the effect that safeguards body health and resist exopathogen invasion, as long as one part of any of which Damage or defect can all cause the not normal of body's immunity, so as to cause the reduction or forfeiture of immunologic function, body by Pathological changes are produced in protecting.If avian leukosis are that tumor change occur due to lymphocytes in blood, and then damage its immunity System, affects the g and D of chicken group.This para-immunity disease often make the impaired development of fowl poultry immune system or It is infected during late growing stage, easily causes the subinfection again such as antibacterial, virus, funguses, so as to forms multiple diseases simultaneously Infection is sent out, the development of animal farming industry is had a strong impact on.
Chinese herbal medicine is natural organic matter and mineral, with toxic and side effects little, be not likely to produce drug resistance and in poultry body The low feature of drug accumulation amount.Chinese medicine contains the effective ingredient such as substantial amounts of polysaccharide, flavone, alkaloid, not only can adjust poultry The effect of immunologic function, moreover it is possible to which the effects such as participating in body disease-resistant poison, antibacterial, is mainly to develop and apply to substitute antibiotics at present Key agents.In addition, Chinese medicine is again rich in abundant nutritional labeling, such as polysaccharide, protein, aminoacid, vitamin, digestion Enzyme, trace element, sterol and hormone etc., can be good at enriched animal immune organ, promote which to develop so as to possess good Opposing pathogen invasion.This is based on, Chinese herbal medicine is used for the immunologic function of enhancing body or some poultry Prevention of Infectious Diseases do not only have Good drug effect, and use to improve as feed additive and livestock and poultry production performance and improve meat, Egg Quality, with embodying medicine Effect globality and two big characteristics of medicine source natural sex.Surging with people's back to nature enthusiasm, Chinese herbal medicine is just with its uniqueness Effect and feature are widely paid attention to.
Content of the invention
In view of this, technical problems to be solved in this application are to provide a kind of Chinese medicine composition and preparation, preparation method And its application.
In order to solve above-mentioned technical problem, the application has opened a kind of veterinary Chinese medicinal composition, its include active component and/or Pharmaceutically acceptable carrier, described active component contain following crude drug:Fructus Ligustri Lucidi, the Radix Astragali, Fructus Lycii, Semen Cuscutae.
Further, veterinary Chinese medicinal composition as above, the active component are made up of the raw material of following mass ratioes: Fructus Ligustri Lucidi 10-80 parts, Radix Astragali 5-90 parts, Fructus Lycii 5-30 parts, Semen Cuscutae 5-30 parts.
Further, veterinary Chinese medicinal composition as above, the active component are made up of the raw material of following mass ratioes: 40 parts of Fructus Ligustri Lucidi, 40 parts of the Radix Astragali, 10 parts of Fructus Lycii, 10 parts of Semen Cuscutae.
The application also provides a kind of preparation method of veterinary Chinese medicine granule, comprises the following steps:
Step one:Veterinary Chinese medicinal composition is carried out water extraction;
Step 2:The extract that step one is obtained is dried;
Step 3:The preparation that dried for step 2 extract is carried out granule.
Further, method as above, the step one are comprised the following steps:
A, weigh Fructus Ligustri Lucidi, the Radix Astragali, Fructus Lycii, Semen Cuscutae in parts by mass, by solid-liquid ratio 1:10 add water, and soak 1h, decoct 1h is boiled, is filtered, is obtained medicinal liquid 1;
After b, step a, solid-liquid ratio 1 pressed by medicinal residues:8 add water, and decoct 1h, filter, obtain medicinal liquid 2;
After c, step b, solid-liquid ratio 1 pressed by medicinal residues:8 add water, and decoct 1h, filter, obtain medicinal liquid 3;
D, collection step a, b, c gained medicinal liquid carry out being concentrated into solid-liquid ratio 1:5,60 DEG C are cooled to, regulating liquid medicine pH is extremely 5.0, plus 0.01% chitosan solution carries out clarification 24h, is filtrated to get medicinal liquid;
E, step d gained medicinal liquid carry out being concentrated into solid-liquid ratio 1:1.
Further, method as above, the step 2 are comprised the following steps:By step e gained traditional Chinese drug inspissation thing Adding carries out infrared microwave drying inside drying baker, temperature 60 C, the 1st section of microwave drying 0min, the 2nd section of microwave drying 20min, 3rd section of microwave drying 0min, the 4th section of microwave drying 0min, the 5th section of microwave drying 120min, the 6th section of microwave drying 120min, 7th section of microwave drying 60min, period see drying the medicinal liquid drying regime in case, prevent medicinal liquid from splashing, and drying is finished, and is taken out Drying baker, is cooled to room temperature, takes out medicine, crushes, and crosses 60 mesh sieves, obtains Chinese medicine extract.
Further, method as above, the step 3 include:By the Chinese medicine extract for obtaining according to 1:2 add Enter adjuvant, adjuvant is soluble starch:Microcrystalline Cellulose is 7 in mass ratio:3 mix, and make starch slurry with soluble powder, 150mL/min is whitewashed, and 100 DEG C carry out boiling granulating and drying, is arranged granule and is crossed No. 1 sieve, but No. 5 are sieved.
Application of the veterinary Chinese medicinal composition as above in the medicine for the treatment of immunologic function disease.
Application of the veterinary Chinese medicinal composition as above in the medicine for strengthening chicken immuological function.
Beneficial effect:
Chinese medicine composition disclosed by the invention, its formula are unique, and raw material is easy to get, low cost, are good for tonifying Qi and lifting yang, QI invigorating Effect of spleen;The invention also discloses preparation prepared by Chinese medicine composition, its preparation method is simple, by Chinese medicine composition routinely Method is extracted, infrared microwave drying is mixed with adjuvant, single step spray granulation is sent out and makes granule, is in solid dispersible carrier High degree of dispersion state, increases the bioavailability of Chinese medicine composition;The traditional Chinese medicine particle of preparation can significantly improve chickling Immune Organs Index, the content of IgG, IgA in serum improve Immune Profile In Chicks newcastle and the antibody after infectious bursa of Fabricius vaccine Potency, and extend the lasting level of antibody.And the carbon clearance ability of hypoimmunity mice can be effectively improved, increase immune organ Index and serum hemolysin, improve serum in IL-2, IL-4, the content of IFN-γ and reach strengthen immunologic function effect.
Description of the drawings
Accompanying drawing described herein is used for providing further understanding of the present application, constitutes the part of the application, this Shen Schematic description and description please does not constitute the improper restriction to the application for explaining the application.In the accompanying drawings:
1 specnuezhenside standard substance canonical plotting of Fig. 1 embodiments;
1 specnuezhenside standard substance efficient liquid phase of Fig. 2 embodiments detection figure;
Fig. 3 embodiments 1 lack Fructus Ligustri Lucidi subsample (lacuna) efficient liquid phase detection figure;
Fig. 4 is 1 sample efficient liquid phase of embodiment detection figure.
Specific embodiment
Describe presently filed embodiment below in conjunction with drawings and Examples in detail, thereby how the application is applied Technological means come solve technical problem and reach technology effect realize that process can fully understand and implement according to this.
For the immunomodulating of poultry, theoretical according to Chinese veterinarian's compound compatibility, filter out with Fructus Ligustri Lucidi, the Radix Astragali, Fructus Lycii, Semen Cuscutae etc. with tonifying Qi and lifting yang, replenishing QI to invigorate the spleen Chinese medicine strengthening the immunologic function of chickling, carry out preventing and treating Immune Profile In Chicks and suppress Property the disease Chinese medicine and its technique of compound preparation, quality standard, pharmacology pharmacodynamic research.
Embodiment 1, Chinese medicine composition effective substance method determine
Method:Detected using HPLC, with principal agent Fructus Ligustri Lucidi effective ingredient specnuezhenside as detection object.Chromatostrip Part:C18 posts (46 × 150mm, 5 μm);Mobile phase:Methanol-water (40:60);Detection wavelength:224nm;Flow velocity::1.0mL/min; Column temperature:25℃;Standard solution is prepared:Precision weighs specnuezhenside in right amount, plus 50% methanol constant volume is configured to 10ml 0.25mg/ml concentration is obtained final product.Need testing solution is prepared:Accurate absorption test sample 2.0mL, 50% methanol solution are appropriate, at ultrasound Reason 20min, takes out, lets cool to room temperature, then be settled to 50mL, shake up, and filters (0.22um), takes subsequent filtrate molten as test sample Liquid.
Detection method:Accurate absorption reference substance solution and each 10 μ L of need testing solution, inject chromatograph of liquid, survey respectively Fixed, obtain final product.
As a result:Accurate 2,4,6,8,10 μ L of absorption specnuezhenside reference substance solution, determine peak area according to above-mentioned chromatographic condition, With peak area (Y) as vertical coordinate, the content (X, μ g) of special Fructus Ligustri Lucidi reference substance is abscissa, draws standard curve, and calculates recurrence Equation:Y=1186.41048X-1.36851, R2=1.0000, as a result show, in the range of 0.4865-2.4325 μ g, special Nuezhenoside integrating peak areas value y is linear good with content x, sees Fig. 1, Fig. 2, Fig. 3, Fig. 4.Precision is drawn the sample of 10 μ l and is lacked Item injection efficient liquid phase is detected that sample specnuezhenside peak occurs in 6.655min, and lacuna does not go out in corresponding time point Existing crest, the crest that remaining time point occurs are identical, illustrate that specnuezhenside content in the method detection sample has good special Attribute.
Embodiment 2, Chinese medicine composition water extraction technical study
Method:Fructus Ligustri Lucidi is the principal agent in Chinese medicine composition, and specnuezhenside is its main labeled composition, therefore with special female The content of loyal glycosides is inspection target, when decocting number of times, soak time, solid-liquid ratio, decoction with single factor test and orthogonal design screening Between etc. process conditions.According to literature survey and practical experience, solid-liquid ratio is usually the 1 of medical material weight:8-1:12 times, investigate 1:8, 1:10,1:12 3 levels;Immersion 0,0.5, tri- levels of 1.0h is investigated, decocting time is oversize then to waste the energy, therefore specifies 0.5h, 1h, 1.5h are three times to investigate level;Decoct, number of times take too much oversize, therefore regulation 1,2, Level is investigated for decocting number of times 3 times, respectively with single factor exploration each index, comprehensive the results of univariate logistic analysis, with soak time, Solid-liquid ratio, decocting time are factor, carry out orthogonal test analysis using L9 (34).
Table 1 extracts test quadrature factor level
As a result:
(1) different extraction time results:Take Fructus Ligustri Lucidi 100g, respectively according to solid-liquid ratio be 1:10、1:8、1:8 amount adds Water.Decocting and extract 3 times. each 1.0h, merging filtrate are concentrated into 100mL, determine the content for extracting specnuezhenside every time, knot respectively Fruit shows that the content of specnuezhenside in medicinal liquid is raised with increasing for extraction time.
The selection of 2 extraction time of table
(2) different soak time result of the tests:Chinese medicine composition 100g is taken, totally 3 parts, be 1 according to solid-liquid ratio:10 amount adds Hot water, soaks 0h, 0.5h, 1.0h respectively, filters, is concentrated into 100mL, determines respectively, as a result Show that soak time can affect the content of specnuezhenside in medicinal liquid.
The screening of 3 medicine soak time of table
(3) different extraction time result of the tests:Chinese medicine composition 100g is taken, totally 3 parts, be 1 according to solid-liquid ratio:10 amount adds Water, soaks 1h, decocts respectively and extracts 1.5h, 1.0h, 0.5h, filters, and is concentrated into 100mL, and measure is extracted specnuezhenside every time and contained Amount, as a result shows that decocting time can affect the content of specnuezhenside in medicinal liquid.
The screening of 4 extraction time of table
(4) different feed liquid compares result of the test:Take Chinese medicine composition 100g, totally 3 parts, respectively according to solid-liquid ratio be 1:12、1: 10、1:8、1:6 amount adds water, and soaks 1 hour, decocts respectively and extract 1.0 hours, filter, be concentrated into 100mL, determines special Fructus Ligustri Lucidi Glycosides content, as a result shows that solid-liquid ratio can affect the content of specnuezhenside in medicinal liquid.
Table 5 extracts the screening of amount of water
(5) orthogonal experiments:Test according to the arrangement of L9 (34) orthogonal table, water extraction is filtered, and is concentrated into 100mL, and measure is carried Specnuezhenside content in liquid is taken, result of the test is through range analysiss C (decocting time) > B (solid-liquid ratio) > A (soak time), and C2 > C1 > C3, B3 > B1 > B2, A3 > A1 > A2.The results of analysis of variance shows, difference not significantly (< between three factors 0.05) real cost of production and time is combined therefore, extraction process is determined for A3B3C2, i.e. immersion 1h, decoction three times, every time Solid-liquid ratio is respectively 1:10、1:8、1:8, decoct 1h every time.
6 orthogonal experiments of table
7 the results of analysis of variance of table
* Significant effect F 0.05 (2,2)=19.0
Embodiment 3, the screening of clarification factor level
Method:According to screening gained extraction process carry out extract 9 parts of Chinese medicine composition, every part of 100g, according to L9 (34) just Table arrangement is handed over, with clear and bright dose of 1% chitosan solution, the addition of shitosan, fluid temperature, medicinal liquid pH is investigated, designed Factor level table.
8 clearance quadrature factor level of table
As a result:Test according to the arrangement of L9 (34) orthogonal table, clarified using 1% shitosan, 5000r/min is centrifuged 10min, counts the weight of precipitate, and as a result range analysiss show B (fluid temperature) > C (medicinal liquid pH) > A (interpolations of shitosan Amount), and B3 > B2 > B1, C1 > C2 > C3, A2 > A3 > A1, the results of analysis of variance show that difference does not show between three factors Write (< 0.05), accordingly, it is determined that it is 60 DEG C that clarification condition is B3C1A2, i.e. fluid temperature, during pH5.0, add 0.10% shell Polysaccharide solution is clarified.
9 orthogonal experiments of table
10 the results of analysis of variance of table
* Significant effect F 0.05 (2,2)=19.0
The selection of embodiment 4, ratio of adjuvant
For the ease of storing and carrying, easy to use, the extract of Chinese medicine composition is made and various is controlled suitable for clinic The preparation for needing is treated, acceptable carrier on pharmaceuticss can be added in the formulation, the method for preparation is conventional method.Such as by medium-height grass Various Chinese crude drugs in drug composition are directly ground to fine powder, make powder, or add binding agent to make pill;Or by medium-height grass The extract of drug composition makes capsule, tablet, granule etc. with acceptable carrier on pharmaceuticss.These extracts and preparation There is identical therapeutical effect with herbal composite.So the Chinese medicine composition of test and Selection makes granule, granule is commonly used Adjuvant has starch, Icing Sugar, dextrin, Microcrystalline Cellulose and soluble starch etc..Due to Icing Sugar pelleting moisture absorption stronger, therefore not Use Icing Sugar.
Method:Added with 3 gained medicine of embodiment infrared microwave drying, temperature is carried out inside the drying baker of 40cm*50cm 60 DEG C, the 2nd section of microwave drying 20min, the 5th section of microwave drying 120min, the 6th section of microwave drying 120min, the 7th section of microwave drying 60min, period see drying the medicinal liquid drying regime in case, prevent medicinal liquid from splashing, and drying is finished, and is taken out drying baker, is cooled to Room temperature, takes out medicine, crushes, sieves, obtain Chinese medicine extract.Take extract in right amount, add the adjuvants of 2 times of amounts, mix respectively Uniformly, starch slurry is made with soluble powder, 150mL/min is whitewashed, carry out boiling granulating and drying according to 100 DEG C, arrange Granule crosses No. 1 sieve, but No. 5 are sieved, and the mouldability for investigating granule determines adjuvant used, selectes molding with mouldability as inspection target Adjuvant.
As a result:With soluble starch as adjuvant pelletize when, soft material stickiness is larger, is difficult to sieve.During with dextrin as adjuvant, auxiliary Material hygroscopicity is poor, is difficult to pelletize.Accordingly, it is considered to use soluble starch and microcrystalline cellulose fibres element mixing as adjuvant.And And the increase with Microcrystalline Cellulose materials, the mouldability of granule improves, but as soluble starch is compared with Microcrystalline Cellulose cost Low, it is thus determined that adjuvant is soluble starch:Microcrystalline Cellulose is 7:3.
The selection of 11 ratio of adjuvant of table
Embodiment 5, traditional Chinese medicine particle causes hypoimmunity mice immunomodulating to cyclophosphamide
According to 4 result of embodiment, it is dissolved as suspension shape and is carried out as follows enforcement.
Mice is randomly divided into 6 groups, respectively basic, normal, high dose of blank group, model group, positive drug group, Chinese medicine composition Amount group, 20 per group, does 1 repetition, compares according to EXPERIMENTAL DESIGN, blank group and model group gavage normal saline per group, in The basic, normal, high dosage group chickling of drug composition presses 0.5g/kg, and 1.0g/kg, 2.0g/kg gavage traditional Chinese medicine particle is used in conjunction 7 My god, once a day, positive controls are experimental control according to operation instruction feeding astragalus polysaccharidess, when being administered the 4th day, except blank Group is outer, and other every group of mice is according to 60mg/kg intraperitoneal injection of cyclophosphamide 1 time.
(1) mice phagocyte imdex determination test
Method:After each group mice last dose 24h, each repeats to randomly select 6, totally 12, with 0.1mL/10g body weight Dosage, the india ink after tail vein injection process, after injection 2,10min respectively with the suction pipe processed through heparin, from little Rathole vena orbitalis posterior takes 20 μ L of blood, is blown into the Na that 2mL mass fractions are 0.1% immediately2CO3Shake up in solution, take after blood is finished with Mass fraction is 0.1% Na2CO3Solution surveys absorbance (OD) compared with zero in 650nm, calculates phagocyte imdex (K) and correction Clean up index (α).
K=(lgA1- lgA2)/(t2- t1)
α=K1/3Body weight/(liver weight+spleen weight)
(A in formula1、A2For the absorbance that 2 institute's blood-sample withdrawals of priority are measured, t2- t1Time interval for 2 blood samplings.)
As a result:After mouse peritoneal injection cyclophosphamide, extremely significantly (P < 0.01) is less than blank group, gavage to phagocyte imdex Significantly raise (P < 0.01) compared with model group after medicine, and dosage is better than 0.5g/kg for the effect of 2.0g/kg and 1.0g/kg, its Between diversity significantly (0.01 < P < 0.05).
12 traditional Chinese medicine particle of table is affected on hypoimmunity mice phagocyte imdex
Note:The different lower case person significant difference (P of colleague's shoulder mark<0.05), different capitalization person differences extremely significantly (P< 0.01), without alphabetical person or the not notable (P of alphabetical identical person's difference>0.05).
(2) determination test of serum hemolysin
Method:After being administered the 7th day, each repeats to randomly select 6, totally 12, respectively lumbar injection 5%CRBC suspensions 0.2mL carries out immunity, immune 7 days, and after last dose 24h, eyeball is taken a blood sample, and separates serum.Serum normal saline is taken after centrifugation 100 times of dilution.Serum 1mL and 5%CRBC suspension 0.5mL, 10% complement 0.5mL after dilution is taken, is mixed, in 37 DEG C of calorstats 30min is after 0 DEG C of refrigerator (or in ice-water bath) terminating reaction for insulation.Supernatant is taken in ultraviolet spectrometry after 2000rpm centrifugation 10min Absorbance A is measured at photometer 540nm, and the value of hemolysin is with half hemolysis value HC50Represent.HC50=(sample absorbance value/ Absorbance during CRBC HD50s) × extension rate.
As a result:Model group mice serum hemolysin HC50Extremely significantly (P < 0.01) reduces;After gavage Chinese medicine composition, 1.0g/kg dosage groups effect is better than 2.0g/kg and 0.5g/kg dosage groups, and significant difference compared with 0.5g/kg dosage groups (0.01 < P < 0.05).
13 traditional Chinese medicine particle of table is affected on hypoimmunity mice serum hemolysin
Note:The different lower case person significant difference (P of colleague's shoulder mark<0.05), different capitalization person differences extremely significantly (P< 0.01), without alphabetical person or the not notable (P of alphabetical identical person's difference>0.05).
(3) in mice serum cytokine measure
Method:Each repeats to randomly select 6, and totally 12, after last dose 24h, eyeball takes blood, is prepared into serum, Carry out detecting corresponding index according to each kit specification respectively.
As a result:Model group mice IL-2, IL-4, IFN-γ content substantially reduce, after test, traditional Chinese medicine particle and In positive drug group serum, higher than model group (P < 0.01), IL-2 contents are raised not enough substantially, with mould for IL-4, IFN-γ content Type group compares difference not significantly (P > 0.05).
Impact of 14 traditional Chinese medicine particle of table to IL-2, IL-4, IFN-γ content in hypoimmunity mice serum
Note:The different lower case person significant difference (P of colleague's shoulder mark<0.05), different capitalization person differences extremely significantly (P< 0.01), without alphabetical person or the not notable (P of alphabetical identical person's difference>0.05).
(4) measure of mouse immune shoot formation
Method:To gather at the mice dislocation of blood, taken spleen and thymus, claim its quality, spleen index has been calculated by formula And thymus index.Index and spleen index=spleen/body weight;Thymus index=thymus/body weight.
As a result:Model group mouse spleen index and thymus index have downward trend, and compared with blank group, difference is extremely notable (P < 0.01);After gavage Chinese medicine, mouse spleen index is significantly raised, and in the range of finite concentration, has obvious dose-effect to close System;Thymus index also increases, and its drug concentration is that 1.0g/kg effects are preferable, significant difference compared with 0.5g/kg groups (0.01 < P < 0.05).
15 traditional Chinese medicine particle of table is affected on hypoimmunity mice Immune Organs Index
Note:The different lower case person significant difference (P of colleague's shoulder mark<0.05), different capitalization person differences extremely significantly (P< 0.01), without alphabetical person or the not notable (P of alphabetical identical person's difference>0.05).
Embodiment 6, traditional Chinese medicine particle causes hypoimmunity mice immunomodulating to cyclophosphamide
According to 4 result of embodiment, it is dissolved as suspension shape and is carried out as follows enforcement.
Test 3 age in days, 180 plumage SPF chickling is randomly divided into 6 groups, respectively blank group, positive controls, vaccine group, height, In, low dose group, per group of 30 plumages, adapt to 3 days after, according to EXPERIMENTAL DESIGN, basic, normal, high dosage group chickling presses 0.5g/kg, 1.0g/kg, 2.0g/kg gavage Chinese medicine compositions are used in conjunction 7 days, daily 2 times, and positive controls feed the Radix Astragali according to operation instruction Polysaccharide is experimental control, per 10 plumage of cage, equal free water and feeding, feeds identical basal diet.
(1) impact to growth performance
Method:At on-test, measurement each group chickling all the time, was weighed to chickling every 7 days respectively, test knot The indexs such as weightening, daily gain and the feed-weight ratio of each group chickling are calculated after beam, is observed per plumage chickling in units of cage during test Clinical change.
As a result:Before test, not significantly (P > 0.05), after test 35 days, high dose group is with for each group chickling weight differences The body weight of dosage group chickling is significantly raised, significant difference (P < 0.05) compared with blank group;As can be seen from Table 17, middle dose The weightening of amount group chicken feed is than substantially reduction, significant difference (P < 0.05) compared with blank group.
16 each test group chickling of table body weight change (unit not on the same day:Gram)
Body weight change (the unit not on the same day of table 17:Gram)
Note:The different capitalizations of same row shoulder mark represent, P < 0.01, different lowercase letter P < 0.05, do not mark or Person's table same letter represents P > 0.05.
(2) impact to hematological indices
Method:Every 7 days before immunity and after immunity, jugular vein gathers 200 μ L of blood to each group chickling, for detecting Hematological indices, erythrocyte, numeration of leukocyte adopt microscope count method;Differential blood count is using Rui Shi-Jim Sa dye Color method.
As a result:In test group chickling blood, erythrocyte number gradually rises, and assumes a certain amount effect relation, leukocyte count Amount, neutrophilia, acidophilia, basophilia, amount of mononuclear cells do not have significance change (P > 0.05), and high, middle dose group lymph is thin Born of the same parents are significantly higher than blank group (P < 0.05).
The change of 18 each test group chickling hemocyte of table
Note:The different capitalizations of same row shoulder mark represent, P < 0.01, different lowercase letter P < 0.05, do not mark or Person's table same letter represents P > 0.05.
(3) impact to Immune Organs Index
Method:After administration 2 weeks, per group of 10 plumage SPF chickens of random selection claim its body weight, after collection blood, take spleen, breast Gland and fabricius bursa, peel off fat, claim quality, calculate spleen, thymus and bursal index by formula, calculate organ index.Immune device Official's index=immune organ quality/body weight × 100%.
As a result:High, medium and low dosage group index and spleen index, thymus index and bursal index increase, middle dose group breast Gland index significant difference (P < 0.05) compared with blank group and low dose group with bursal index.
Change (the unit of 19 each test group Immune Organs Index of Chicks of table:mg/g)
Note:The different capitalizations of same row shoulder mark represent, P < 0.01, different lowercase letter P < 0.05, do not mark or Person's table same letter represents P > 0.05.
(4) impact to chicken Newcastle disease antibody horizontal
Method:Identical administration is carried out according to test requirements document, head on the 7th exempts from newcastle disease vaccine in addition to blank group, 30 Secondary immunity.Every 7 days before immunity and after immunity, jugular vein gathers blood 2mL to each group chickling, separates serum, adopts Blood clotting, hemagglutination inhibition test, detect ND antibody horizontals.
As a result show:The non-vaccination of chickling, antibody titres to newcastle disease virus have the trend of reduction, post-vaccine antibody potency It is obviously improved, on 14th, vaccinated chickling raised (P < 0.05) with blank group chickling antibody titer significant difference, respectively Medicine group antibody titer is also significantly greater than vaccine group (P < 0.05), and on 21st, each medicine group chickling also still kept higher Antibody titer, and vaccine group and blank group significant difference (P < 0.05), in second vaccination, the antibody titer of medicine group Pole is significantly higher than blank group (P < 0.01), 42, and high dose group and middle dose group chickling antibody titer are significantly higher than vaccine group With positive drug group (P < 0.05), and middle dosage effect is best.
The change (log2) of 20 each group chickling ND antibody titers of table
Note:The different capitalizations of same row shoulder mark represent, P < 0.01, different lowercase letter P < 0.05, do not mark or Person's table same letter represents P > 0.05.
(5) impact to infectious avian bursal disease virus antibody titer
Method:Identical administration is carried out according to test requirements document, head on the 14th exempts from IBD vaccines in addition to blank group, 32 days two Secondary immunity.Every 7 days before immunity and after immunity, jugular vein gathers blood 2mL to each group chickling, separates serum, agarose Diffusion method (AGPT) detects IBD antibody horizontals.
As a result:Before test, each group infectious avian bursal disease virus antibody positive rate is in the range of 34.8-40%, poor between group Different significantly (P > 0.05) during being administered, each group chickling antibody positive rate does not reduce, but not notable (the P > of group difference 0.05), 21 days when, medicine group antibody positive rate is significantly raised, and difference extremely significantly (P < 0.01), also shows compared with blank group It is higher than vaccine group (P < 0.05) to write;When 28 days, antibody has declined, but medicine group antibody positive rate is significantly higher than blank Group and vaccine group (P < 0.05);After two exempt from, medicine high dose group and middle dose group positive rate reach 100%, poor with vaccine group Different notable (P < 0.05), with blank group difference extremely significantly (P < 0.01);When 42 days, medicine group all reaches 100%, vaccine group 92.2% is only reached, with blank group difference extremely significantly (P < 0.01).
21 each group infectious avian bursal disease virus antibody positive rate of table
Note:Represent with the different capitalizations of a line shoulder mark, P < 0.01, different lowercase letter P < 0.05, do not mark or Person's table same letter represents P > 0.05.
As a result:When before test with 14 days, each group infectious avian bursal disease virus antibody titer group difference is not notable (P > 0.05), first immunity Seedling 7 days (21 age in days), antibody titer are significantly higher than blank group (P < 0.05), antibody effect when 28 days Valency has reduction trend, presents a certain amount effect relation, after 35 days, positive drug group and high, middle dose group antibody titer pole Blank group and vaccine group (P < 0.01) is significantly higher than, after 35 days, high, medium and low dosage group antibody titer pole is significantly higher than sky White group and vaccine group (P < 0.01),
The change of 22 each group infectious avian bursal disease virus antibody titer of table
Note:The different capitalizations of same row shoulder mark represent, P < 0.01, different lowercase letter P < 0.05, do not mark or Person's table same letter represents P > 0.05.
Conclusion
Modern pharmacological research shows, Chinese medicine prevent and treat immune disease and strengthen Immune Function In Animals application in, medicine Should be nontoxic as far as possible, damage of the medicine to histoorgan is reduced while disease is treated.Chinese medicine ingredients are complicated simultaneously Various, in order to be able to accurately study the effect of medicine, set up a perfect extraction and preparation technique and supporting examination criteria seems Particularly important.
From single factor test and orthogonal test, this test determines that Chinese medicine composition water extraction condition, orthogonal test determine clarification condition, Granule preparation condition, sets up a perfect extraction process and its supporting detection method.And by giving injected in mice ring phosphinylidyne Amine sets up immunodeficiency models, and gavage Chinese medicine composition carries out immunocompetence research;Add Chinese medicine group in chickling drinking-water simultaneously Polymer beads, detect growth performance of the Chinese medicine composition to chickling, the impact of newcastle and infectious bursa of Fabricius antibody titer.Knot Fruit shows that traditional Chinese medicine particle can effectively improve the carbon clearance ability of hypoimmunity mice, increase Immune Organs Index and Serum hemolysin, improve serum in IL-2, IL-4, the content of IFN-γ and reach strengthen immunologic function effect;And significantly The Immune Organs Index of chickling is improved, the content of IgG, IgA in serum improves Immune Profile In Chicks newcastle and infectious bursa of Fabricius epidemic disease The antibody titer of after seedling, and extend the lasting level of antibody.
Described above illustrates and describes some preferred embodiments of the present invention, but as previously mentioned, it should be understood that the present invention Be not limited to form disclosed herein, be not to be taken as the exclusion to other embodiment, and can be used for various other combinations, Modification and environment, and can be in invention contemplated scope described herein, by above-mentioned teaching or the technology or knowledge of association area It is modified.And change that those skilled in the art are carried out and change be without departing from the spirit and scope of the present invention, then all should be at this In the protection domain of bright claims.

Claims (9)

1. a kind of veterinary Chinese medicinal composition, which includes active component and/or pharmaceutically acceptable carrier, it is characterised in that institute The active component that states contains following crude drug:Fructus Ligustri Lucidi, the Radix Astragali, Fructus Lycii, Semen Cuscutae.
2. veterinary Chinese medicinal composition according to claim 1, it is characterised in that the active component is by following mass ratioes Raw material is constituted:Fructus Ligustri Lucidi 10-80 parts, Radix Astragali 5-90 parts, Fructus Lycii 5-30 parts, Semen Cuscutae 5-30 parts.
3. Chinese medicine composition according to claim 1, it is characterised in that raw material of the active component by following mass ratioes Composition:40 parts of Fructus Ligustri Lucidi, 40 parts of the Radix Astragali, 10 parts of Fructus Lycii, 10 parts of Semen Cuscutae.
4. a kind of preparation method of veterinary Chinese medicine granule, it is characterised in that comprise the following steps:
Step one:Veterinary Chinese medicinal composition is carried out water extraction;
Step 2:The extract that step one is obtained is dried;
Step 3:The preparation that dried for step 2 extract is carried out granule.
5. method according to claim 4, it is characterised in that the step one is comprised the following steps:
A, weigh Fructus Ligustri Lucidi, the Radix Astragali, Fructus Lycii, Semen Cuscutae in parts by mass, by solid-liquid ratio 1:10 add water, and soak 1h, decoct 1h, Filter, obtain medicinal liquid 1;
After b, step a, solid-liquid ratio 1 pressed by medicinal residues:8 add water, and decoct 1h, filter, obtain medicinal liquid 2;
After c, step b, solid-liquid ratio 1 pressed by medicinal residues:8 add water, and decoct 1h, filter, obtain medicinal liquid 3;
D, collection step a, b, c gained medicinal liquid carry out being concentrated into solid-liquid ratio 1:5, be cooled to 60 DEG C, regulating liquid medicine pH to 5.0, plus 0.01% chitosan solution carries out clarification 24h, is filtrated to get medicinal liquid;
E, step d gained medicinal liquid carry out being concentrated into solid-liquid ratio 1:1.
6. method according to claim 5, it is characterised in that the step 2 is comprised the following steps:By in step e gained Medicine concentrate is added and carries out infrared microwave drying inside drying baker, temperature 60 C, the 1st section of microwave drying 0min, and the 2nd section of microwave is done Dry 20min, the 3rd section of microwave drying 0min, the 4th section of microwave drying 0min, the 5th section of microwave drying 120min, the 6th section of microwave drying 120min, the 7th section of microwave drying 60min, period see drying the medicinal liquid drying regime in case, prevent medicinal liquid from splashing, have dried Finish, take out drying baker, be cooled to room temperature, take out medicine, crush, cross 60 mesh sieves, obtain Chinese medicine extract.
7. method according to claim 6, it is characterised in that the step 3 includes:By the Chinese medicine extract for obtaining According to 1:2 add adjuvant, and adjuvant is soluble starch:Microcrystalline Cellulose is 7 in mass ratio:3 mix, and make shallow lake with soluble powder Slurry, 150mL/min are whitewashed, and 100 DEG C carry out boiling granulating and drying, arrange granule and cross No. 1 sieve, but No. 5 are sieved.
8. application of the veterinary Chinese medicinal composition described in claim 1 in the medicine for the treatment of immunologic function disease.
9. application of the veterinary Chinese medicinal composition described in claim 1 in the medicine for strengthening chicken immuological function.
CN201611126167.8A 2016-12-09 2016-12-09 A kind of veterinary Chinese medicinal composition and its preparation method and application of granule Pending CN106491834A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107543893A (en) * 2017-08-24 2018-01-05 重庆市畜牧科学院 The discrimination method of Semen Cuscutae in a kind of female's Huang Zheng oral liquid
CN109893642A (en) * 2019-04-16 2019-06-18 重庆市畜牧科学院 The Chinese medicine composition for treating grice diarrhoea
CN111135251A (en) * 2020-01-09 2020-05-12 山西中医药大学 Veterinary drug for preventing and treating chicken bursal disease as well as preparation method and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101698027A (en) * 2009-10-30 2010-04-28 重庆市畜牧科学院 Traditional Chinese medicine composite for treating infectious bursal disease and preparation method thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101698027A (en) * 2009-10-30 2010-04-28 重庆市畜牧科学院 Traditional Chinese medicine composite for treating infectious bursal disease and preparation method thereof

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107543893A (en) * 2017-08-24 2018-01-05 重庆市畜牧科学院 The discrimination method of Semen Cuscutae in a kind of female's Huang Zheng oral liquid
CN109893642A (en) * 2019-04-16 2019-06-18 重庆市畜牧科学院 The Chinese medicine composition for treating grice diarrhoea
CN111135251A (en) * 2020-01-09 2020-05-12 山西中医药大学 Veterinary drug for preventing and treating chicken bursal disease as well as preparation method and application thereof
CN111135251B (en) * 2020-01-09 2021-11-26 太原市恩禾动物药业有限公司 Veterinary drug for preventing and treating chicken bursal disease as well as preparation method and application thereof

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Application publication date: 20170315