CN105943820A - Traditional Chinese medicine compound for resisting swine influenza and porcine reproductive and respiratory syndrome and extraction method thereof - Google Patents
Traditional Chinese medicine compound for resisting swine influenza and porcine reproductive and respiratory syndrome and extraction method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/35—Caprifoliaceae (Honeysuckle family)
- A61K36/355—Lonicera (honeysuckle)
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/11—Pteridophyta or Filicophyta (ferns)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/233—Bupleurum
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/484—Glycyrrhiza (licorice)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/78—Saururaceae (Lizard's-tail family)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8964—Anemarrhena
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation 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
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation 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
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
Abstract
The invention provides a traditional Chinese medicine compound for resisting H1N1 and H3N2 subtype swine influenza and porcine reproductive and respiratory syndrome. The compound is prepared from the following medicinal materials in parts by weight: 20-30 parts of lonicera japonica, 10-20 parts of rhizoma anemarrhenae, 10-15 parts of radix paeoniae rubra, 10-15 parts of cyrtomium fortunei, 10-15 parts of radix bupleuri, 10-15 parts of cordate houttuynia and 5-15 parts of licorice roots. The traditional Chinese medicine compound can be used for resisting swine influenza virus and porcine reproductive and respiratory syndrome virus at the same time, especially H1N1 and H3N2 subtype swine influenza, has the advantages of relatively low or no toxin remaining, small side effect, hardly drug resistance generation and the like, and can be used for regulating body immunity, inhibiting virus replication and improving clinical symptoms.
Description
Technical field
The invention belongs to veterinary Chinese medicine field, relate to a kind of anti-swine flu and the Chinese medicine compound of reproductive and respiratory syndrome and extraction side
Method.
Background technology
Swine flue (swine influenza, SI), also known as swine influenza, is by orthomyxovirus section influenza A
The one that the swine influenza virus (swine influenza virus, SIV) belonged to causes is acute, hot, high degree in contact respiratory tract
Infectious disease, clinical with burst, high heat, cough, dyspnea, recurrent exerbation, sickness rate height, mortality rate is low is characterized.This disease exists
Having generation all over the world, harm is serious, and economic loss is huge, and constitutes a threat to the health of the mankind.Its prominent popular spy
Point is often to there is and often cause other respiratory tract antibacterials and the secondary of virus or mixed infection with subclinical form, as pig breeding with
Breathing syndrome, Porcine circovirus desease and porcine contagious pleuropneumonia etc., make epidemic situation become complicated, and mortality rate increases.
Porcine reproductive and respiratory syndrome (Porcine Reproductive and Respiration Syndrome
Virus, PRRS) also known as " reproductive and respiratory syndrome ", be the pig that caused by porcine reproductive and respiratory syndrome virus (PRRSV) with breeding difficulty and
Respiratory symptom is a kind of hyperinfection disease of principal character.This condition of disease multipath is propagated, and can not only cause in-pig
Generate heat, miscarry, produce mummy tire and weak son, also can induce the respiratory system disease of piglet and bred pigs.PRRSV may result in disease pig
Inappetence, weight loss, respiratory system disease also produce interstitial pneumonia.Serious threat and huge is caused to National Show industry
Big economic loss.
In the preventing and treating of clinical practice swine flue, it is widely used owing to antibiotic has the feature of control complication,
Coordinate the medicines such as substantial amounts of antipyretic-antalgic, glucocorticoids simultaneously.These processing modes are same suppression body inflammatory reaction
Time also increased the weight of immunosuppressant, cause the state of an illness more to deteriorate.And Porcine reproductive and respiratory syndrome relies primarily on vaccine and prevents
Controlling, but inactivated vaccine needs in a large number, repeatedly immunity, produces immunization time long, there is again virulence and returns strong danger in attenuated vaccine.
Chinese herbal medicine, as natural product, has medicine source and enriches, and residual hazard is relatively low or without residual hazard, side effect is little, is not likely to produce drug resistance etc. excellent
Point；Body's immunity can be regulated simultaneously, suppress virus replication, improve clinical symptoms, should on treatment disease of viral infection
With extensively.Therefore, application herbal control swine flue and Porcine reproductive and respiratory syndrome, prospect is the most wide.
Make a general survey of the experimentation of present Chinese medicine compound antiviral, comprehensive with breathing to anti-swine flu virus respectively and pig breeding
Levying viral folk prescription research more, the most not working out a compound recipe anti-swine flu virus and pig can breed and exhale simultaneously
Inhale syndrome virus.And mostly it is respectively adopted the method independent studies of in vivo test or in vitro tests, it is impossible in thoroughly evaluating
Recurrence due to taking drug side's anti-swine flu virus and the effect of porcine reproductive and respiratory syndrome virus.
For solving the problems referred to above, in the urgent need to filtering out the Chinese medicine effectively preventing and treating swine flue and Porcine reproductive and respiratory syndrome
Compound recipe.And using in vitro tests and in vivo test, from the mechanism of action and clinical practice, comprehensive study Chinese medicine compound resists simultaneously
H1N1 and H3N2 hypotype swine influenza virus and the effect of reproductive and respiratory syndrome virus, provide theoretical and actual foundation for clinical prevention.
Summary of the invention
Instant invention overcomes shortcoming of the prior art, it is provided that a kind of anti-swine flu and the Chinese medicine compound of reproductive and respiratory syndrome and
Extracting method, can prevent and treat swine influenza virus and porcine reproductive and respiratory syndrome virus simultaneously, and it is relatively low or without residual that it has residual hazard
Poison, side effect is little, is not likely to produce the advantages such as drug resistance, can regulate body's immunity simultaneously, suppresses virus replication, improves clinic
Symptom.
In order to solve above-mentioned technical problem, the present invention is achieved by the following technical solutions:
A kind of anti-swine flu and the Chinese medicine compound of reproductive and respiratory syndrome, comprise following medicine: Flos Lonicerae 20～30 weight portion, the Rhizoma Anemarrhenae
10～20 weight portions, Radix Paeoniae Rubra 10～15 weight portion, Rhizoma Osmundae 10～15 weight portion, Radix Bupleuri 10～15 weight portion, Herba Houttuyniae 10～15
Weight portion, Radix Glycyrrhizae 5～15 weight portion, described weight portion refers to that in compound recipe, various medicines account for the part by weight of whole compound recipe.
Further, following medicine is comprised: Flos Lonicerae 25 weight portion, the Rhizoma Anemarrhenae 15 weight portion, Radix Paeoniae Rubra 12.5 weight portion, Rhizoma Osmundae
12.5 weight portions, Radix Bupleuri 12.5 weight portion, Herba Houttuyniae 12.5 weight portion, Radix Glycyrrhizae 10 weight portion.
The extracting method of the Chinese medicine compound of a kind of anti-swine flu and reproductive and respiratory syndrome, comprises the following steps:
(1) each medicine is weighed by said ratio；
(2) the recurrence due to taking drug side after weighing is crushed to 20 mesh, adds the water of doubling dose, soaks 1h, decocting 2 times, boils every time
30min, merges 2 decocting liquids；
(3) by decocting liquid in normal-pressure vacuum sucking filtration and concentration, being settled to 100mL decocting liquid, every milliliter is equivalent to 1g crude drug (i.e.
1000mg/mL)；
(4) decocting liquid is carried out autoclaving, save backup in 4 DEG C of refrigerators
Compared with prior art, the invention has the beneficial effects as follows:
A kind of anti-swine flu of the present invention and the Chinese medicine compound of reproductive and respiratory syndrome and extracting method, comprise following medicine: gold
Flos Lonicerae 20～30 weight portion, the Rhizoma Anemarrhenae 10～20 weight portion, Radix Paeoniae Rubra 10～15 weight portion, Rhizoma Osmundae 10～15 weight portion, Radix Bupleuri 10～
15 weight portions, Herba Houttuyniae 10～15 weight portion, Radix Glycyrrhizae 5～15 weight portion, anti-swine flu virus and pig can breed and breathe simultaneously
Syndrome virus, especially H1N1 and H3N2 hypotype swine influenza virus, and have that residual hazard is relatively low or without residual hazard, side effect is little,
It is not likely to produce the advantages such as drug resistance, body's immunity can be regulated simultaneously, suppress virus replication, improve clinical symptoms.
Accompanying drawing explanation
Accompanying drawing is used for providing a further understanding of the present invention, is used for together with embodiments of the present invention explaining the present invention,
It is not intended that limitation of the present invention, in the accompanying drawings:
Fig. 1 is the change of H1N1 hypotype swine influenza virus infection Mouse Weight；
Fig. 2 is the change of H3N2 hypotype swine influenza virus infection Mouse Weight；
Fig. 3 A represents the lung pathologies section of the H1N1 hypotype swine influenza virus infection mice of Chinese medicine compound group；
Fig. 3 B represents the lung pathologies section of the H1N1 hypotype swine influenza virus infection mice of MAXINGSHIGANGSAN compound recipe group；
Fig. 3 C represents the lung pathologies section of the H1N1 hypotype swine influenza virus infection mice of ribavirin group；
Fig. 3 D represents the lung pathologies section of the H1N1 hypotype swine influenza virus infection mice of model control group；
The lung pathologies section of the H1N1 hypotype swine influenza virus infection mice of Fig. 3 E Normal group；
Fig. 4 A represents the lung pathologies section of the H3N2 hypotype swine influenza virus infection mice of Chinese medicine compound group,
Fig. 4 B represents the lung pathologies section of the H3N2 hypotype swine influenza virus infection mice of MAXINGSHIGANGSAN group,
Fig. 4 C represents the lung pathologies section of the H3N2 hypotype swine influenza virus infection mice of ribavirin group,
Fig. 4 D represents the lung pathologies section of the H3N2 hypotype swine influenza virus infection mice of model control group,
Fig. 4 E represents the lung pathologies section of the H3N2 hypotype swine influenza virus infection mice of Normal group.
Detailed description of the invention
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are illustrated, it will be appreciated that preferred reality described herein
Execute example be merely to illustrate and explain the present invention, be not intended to limit the present invention.
A kind of anti-swine flu of the present invention and the Chinese medicine compound of reproductive and respiratory syndrome, comprise following medicine: Flos Lonicerae 25 weight
Part, the Rhizoma Anemarrhenae 15 weight portion, Radix Paeoniae Rubra 12.5 weight portion, Rhizoma Osmundae 12.5 weight portion, Radix Bupleuri 12.5 weight portion, Herba Houttuyniae 12.5 weight
Part, Radix Glycyrrhizae 10 weight portion, weight portion refers to that in compound recipe, various medicines account for the part by weight of whole compound recipe.
Flos Lonicerae: sweet, cold；Return lung, the heart, stomach warp；Primary efficacy master is heat-clearing and toxic substances removing and dispelling wind and heat pathogens.Containing chlorogenic acid,
The pharmacological components such as luteolin glycosides, to the various pathogens such as Hemolytic streptococcus, staphylococcus aureus and upper respiratory tract sense
Dye Causative virus etc. has stronger restraint, is monarch drug in compound recipe.The Rhizoma Anemarrhenae: bitter, cold；Return lung, stomach, kidney channel；Primary efficacy is
Clearing away heat-fire, nourshing Yin and drynsessmoistening prescription, it is ministerial drug in compound recipe.Radix Bupleuri: nature and flavor are bitter, be slightly cold, returns liver, gallbladder meridian；There are reconciling superficies and interior, soothing the liver liter
Effect of sun.Herba Houttuyniae: acrid in the mouth, cold, return lung meridian；Energy energy heat-clearing and toxic substances removing, inducing diuresis to remove edema, Radix Bupleuri and Herba Houttuyniae are all assistant
Medicine.Radix Glycyrrhizae: feeble QI, taste is sweet and special, and primary efficacy is invigorating the spleen and replenishing QI, heat-clearing and toxic substances removing, expelling phlegm for arresting cough, and relieving spasm to stop pain, at compound recipe
In for making medicine.
In following embodiment, experiment material used is as follows:
(1) experimental animal
9-11 age in days SPF Embryo Gallus domesticus is purchased from Guangdong great Hua Nong fowl egg company limited；SPF level 15g ± 2g BALB/c mouse is purchased from
Guangdong Medical Lab Animal Center.
(2) solution or preparation of reagents
The preparation of 1.PBS: weigh NaCl 8.0g, Na2HPO4 3.63g、KCl 0.2g KH2PO40.24g is dissolved in right amount
Ultra-pure water, adjusts pH value to 7.4, is settled to 1000mL；100ml/ bottle subpackage, 15 pounds of autoclaving 30min, 4 DEG C save backup.
The preparation of 2.1% chicken erythrocyte suspension: (its consumption is first to draw 3.8% liquor sodii citratis with sterilizing syringe
The 1/5 of required blood volume), from chicken wing vein haemospasia to required blood volume, put in sterile centrifugation tube, adding sterilizing PBS is anticoagulant 2
Times, it being centrifuged 10min with 2000r/min, abandon supernatant, then add normal saline suspension blood cell, ibid method centrifugation, so by red
Cell washs three times, finally according to institute's expense, is made into 1% suspension with sterilizing PBS, now with the current.
3. the extraction of Chinese medicine: use soak by water method slightly to carry, comprise the concrete steps that:
(1) by Flos Lonicerae 25 weight portion, the Rhizoma Anemarrhenae 15 weight portion, Radix Paeoniae Rubra 12.5 weight portion, Rhizoma Osmundae 12.5 weight portion, Radix Bupleuri
12.5 weight portions, Herba Houttuyniae 12.5 weight portion, Radix Glycyrrhizae 10 weight ratio, weigh each Chinese medicine, form 100g Chinese medicine compound；
(2) 100g Chinese medicine compound is crushed to 20 mesh, adds 2 times of water, soak 1h, decocting 2 times, boil 30min every time, close
And 2 decocting liquids；
(3) by decocting liquid in normal-pressure vacuum sucking filtration and concentration, being settled to 100mL decocting liquid, every milliliter is equivalent to 0.51g crude drug
(i.e. 1000mg/mL)；
(4) decocting liquid is carried out autoclaving, preserve in being 4 DEG C of refrigerators to temperature standby.
In Vitro Anti porcine reproductive and respiratory syndrome virus (PRRSV) effect of compound Chinese medicinal preparation
The median lethal dose(LD 50) (TCID50) of MARC 145 cell is measured by 1.PRRSV
By virus to be measured by 10 Concentraton gradient of 10 multiple proportions serial dilution, covering with 96 orifice plates of Marc-145 cell, pressing
The dosage in 100 μ l/ holes adds virus liquid, and each gradient does 8 repetitions, and last 2 are classified as cell controls group, after 37 DEG C of absorption 1.5h
Discard virus liquid, and add maintenance liquid 100 μ l, place CO2 cell culture incubator and cultivate.
Every day, observation of cell pathological changes situation, measured each hole OD value with mtt assay after pathological changes reaches 75%.Virus is to cell
Growth inhibition ratio=(cell controls group OD value virus control group OD value)/cell controls group OD value.The half toxicity of virus is dense
Degree (TCID50) uses Reed-Muench Liang Shi method to calculate.
2. the compound Chinese medicinal preparation mensuration to the maximum safe concentration of MARC-145 cell
96 well culture plates are cultivated monolayer Marc-145 cell, discards stock solution, utilize PBS to wash 1 time.1st to the 12nd
Hole is separately added into 100 μ l DMEM and maintains liquid, and the stock solution then taking 100 μ l Chinese medicine ingredients carries out 2 from the 1st hole successively to the 10th hole
Times doubling dilution, the 10th hole discards 100 μ l, the 11st, 12 holes be cell controls, each dilution factor repeats 4 holes.It is placed in 37 DEG C, dioxy
Change and carbon incubator cultivated 72h, every day observation of cell growing state and metamorphosis.Discard culture fluid after 72h, use mtt assay
OD value is measured under 490nm wavelength.According to the following formula calculating Chinese medicine ingredients safe concentration to Marc-145 cell:
Variable concentrations Chinese medicine ingredients growth inhibition ratio=(cell controls group OD value medicine group OD value)/cell to cell
Matched group OD value, being 0 to determine the safe concentration of Chinese medicine ingredients to the growth inhibition ratio of cell.
3. the mensuration of compound Chinese medicinal preparation In Vitro Anti PRRSV effect
Cell is passed to 96 well culture plates, uses three kinds of dosing methods:
Mode one, first add Chinese medicine and add virus (Chinese medicine blocking effect to PRRSV) again: on 96 well culture plates, cultivate growth
To the Marc-145 cell of monolayer, add with safe concentration for the Chinese medicine extract begun, every hole 100 μ L, 37 DEG C, carbon dioxide
Incubator discards medicinal liquid after cultivating 4h, and every hole adds the 100TCID50PRRSV liquid of 100 μ l, 37 DEG C, CO2 gas incubator training
Discarding virus liquid after supporting 2h, last every hole adds the DMEM culture fluid of 100 μ l.
Mode two, first add virus and add Chinese medicine (Chinese medicine inhibitory action to PRRSV) again:
After on 96 well culture plates, cultivation grows to the Marc-145 cell of monolayer, every hole adds 100 μ l's
100TCID50PRRSV liquid, 37 DEG C, CO2 gas incubator cultivate after 2h and discard virus liquid, add initial with safe concentration
Chinese medicine extract, every hole 100 μ l, 37 DEG C, CO2 gas incubator cultivates 4h after discard medicinal liquid, last every hole adds 100 μ l
DMEM culture fluid.
Add together (the Chinese medicine direct killing action to PRRSV) after the mixing of mode three, Chinese medicine virus:
Cultivating the Marc-145 cell growing to monolayer on 96 well culture plates, preparation rises with Banlangen Polysaccharide safe concentration
Begin, doubling dilution and the culture fluid of the variable concentrations Chinese medicine extract containing 100TCID50 virus liquid, it is sequentially added into 96 well culture plates
In, every hole 100 μ l, 37 DEG C, CO2 gas incubator cultivates 4h after abandon medicinal liquid, last every hole adds DMEM and cultivates 100 μ l.
Set up virus control and negative control simultaneously.
Carrying out statistical analysis with SPSS19.0 software, first calculate the mean OD value of each group of 490nm, it is right to calculate further according to formula
The protective rate of cell, computing formula is: protective rate=(the medicine group OD value-disease of the Chinese medicine Marc-145 cell to infecting virus
Poison matched group OD value)/(cell controls group OD value-virus control group OD value).Each Chinese medicine is evaluated finally according to cytoprotective rate
The antiviral effect of extract, cytoprotective rate is the biggest, shows that medicine anti-PRRSV effect is the strongest.
4. data process
Data withRepresent, analyze by SPSS19.0 software statistics.
Table 1: the Chinese medicine compound blocking effect to PRRSV
Table 2: the Chinese medicine compound inhibitory action to PRRSV
Table 3: the Chinese medicine compound direct killing action to PRRSV
Note: A represents P 0.01 compared with virus control, and difference is extremely notable, and a represents P 0.05, significant difference；B represent with
Blank compares P 0.01, and difference is extremely notable, and b represents P 0.01, significant difference.
Table 1 shows that Chinese medicine compound of the present invention is obvious to the direct killing action of PRRSV on MARC-145 cell
It is better than ribavirin and MAXINGSHIGANGSAN, although the strongest blocking-up and inhibitory action are slightly weaker than ribavirin and MAXINGSHIGANGSAN,
But the effect of the anti-PRRSV of Chinese medicine compound is highly stable, the most drastically decline with its antivirus action of reduction of concentration, and
Each concentration in each model of action has extremely significantly difference compared with virus group, illustrates that it has obvious anti-PRRSV
Effect.
Chinese medicine compound is to mouse infection H1N1 and the prevention effect of H3N2 hypotype swine influenza virus
1.H1N1 and H3N2 hypotype swine influenza virus is to Embryo Gallus domesticus median infective dose (EID50) mensuration
Virus liquid being made 10 times of doubling dilutions successively, is inoculated in 10 age in days chick embryo allantois intracavity, each dilution factor is inoculated
0.2ml/ piece, make 4 repetitions, set the comparison of normal Embryo Gallus domesticus simultaneously.Put hatching 72h in 37 DEG C of calorstats, dead embryo in discarding 24h,
After results 24h, dead embryo and the allantoic fluid of 72h embryo alive, carry out the mensuration of HA titer.Calculate by Reed-Muench Liang Shi method
EID50。
2. the Chinese medicine compound impact on H1N1 and H3N2 hypotype swine influenza virus infection mice pneumonia
(1) laboratory animal packet
BALB/c mouse is randomly divided into Chinese medicine compound group, ribavirin group, MAXINGSHIGANGSAN group, model control group, just
Often matched group, totally 5 groups, often group 10, and be marked.
(2) laboratory animal is administered
Respectively group mice is all with gastric infusion, 2 times/d, 0.2ml/ time.Each group of Chinese medicine 1d before infecting starts to be administered, continuously
7d.Meanwhile, Normal group, model control group give equivalent distilled water.
(3) laboratory animal counteracting toxic substances
After mice is anesthetized with ether, in addition to Normal group, each group collunarium inoculation 106EID50 influenza virus 50 μ l, just
Often matched group inoculates 50 μ l sterilizing PBS.
(4) the 8th days (more than fasting 8h) weighs, and plucks eyeball sacrificed by exsanguination mice, aseptic wins lungs, rinses also with PBS
Lung weight is weighed respectively after sucking excess surface liquid with aseptic filter paper.
Lung Exponent is as follows with Lung Exponent inhibition percentage computing formula:
Lung Exponent=lung weight (g)/mice weights (g) × 100
Lung Exponent inhibition percentage=(model control group Lung Exponent mean-experimental group Lung Exponent mean)/model control group
Number × 100%
(5) statistical analysis: analyze by SPSS17.0 software statistics, refer to table 3:
Table 4: the compound Chinese medicinal preparation impact on H1N1 hypotype swine influenza virus infection mice pneumonia
Table 5: the compound Chinese medicinal preparation impact on H3N2 hypotype swine influenza virus infection mice pneumonia
Note: data above is compared with model control group, and A represents P 0.01, difference is extremely notable, and a represents P 0.05, and difference shows
Write.
From table 4, it can be seen that the not only Lung Exponent inhibition percentage of Chinese medicine compound group has 8.73%, and Lung Exponent is the lowest
In MAXINGSHIGANGSAN classics compound recipe group, illustrate that it is wanted for the protective effect of the mice through H1N1 hypotype swine influenza virus infection
It is better than classical compound recipe group.
3. the Chinese medicine compound impact on swine influenza virus infection Mouse Weight
(1) laboratory animal packet, administration, modeling are with 2 (1) (2) (3)
(2) mensuration of body weight
From being administered from the beginning of the 1st day, before being administered every morning, weigh each Mouse Weight, totally seven days, to each group of mice
Body weight change be analyzed.
(3) statistical analysis: Mouse Weight changes as illustrated in fig. 1 and 2, the column in figure represents experiment the most respectively
1st to the 7th day.
It will be seen from figure 1 that Chinese medicine compound group is relatively strong, only at the 4th day to the resistance of H1N1 hypotype swine influenza virus
Time body weight occur alleviating, but recover fast, body weight increasing degree is big, close with Normal group to the 7th day average weight.And it is numb
Fructus Pruni stone sweet scattered group and ribavirin group are substantially affected very big by H1N1 hypotype swine influenza virus, and weight loss amplitude is big, and
Time is long, recovers slow, and MAXINGSHIGANGSAN group is until the 7th talent occurs that body weight is gone up.As can be seen from Figure 2, H3N2 hypotype swine flue is sick
The virulence relatively H1N1 hypotype swine influenza virus of poison is strong, and significantly change occurs in the Mice Body weight average of all groups, but from figure
It can be seen that Chinese medicine compound group in four infected group still resistance recover more by force very fast, although just occur to the 6th day body weight
Rise, but still much stronger than MAXINGSHIGANGSAN group.Therefore, the experimental result that comprehensive two-strain infects understands, Chinese medicine compound group
There is obvious anti-swine flu effect.
4. the lung pathologies after mouse infection swine influenza virus is changed by Chinese medicine compound
(1) laboratory animal packet, administration, modeling are with 1 (1) (2) (3)
(2) the 8th days (more than fasting 8h) weighs, and plucks eyeball sacrificed by exsanguination mice, aseptic lungs of winning, and makes paraffin and cut
Sheet.
(3) section interpretation of result:
The main generation interstitial pneumonia of the metainfective mice of swine flue also has the pathological change such as hyperemia, edema.From Fig. 3 A
To Fig. 4 E it can be seen that compared with Normal group, model control group alveolar septum is broadening, and alveolar wall is imperfect, and with
Bleeding.Although Chinese medicine compound group also occurs in that hemorrhage, but manifest symptom is weak compared with MAXINGSHIGANGSAN group and ribavirin group,
And alveolar wall is complete.Thus, it can be known that Chinese medicine compound group has prevention effect to the swine flue of two kinds of hypotypes of H1N1 and H3N2, and
It is better than MAXINGSHIGANGSAN group and ribavirin group.
Finally it is noted that these are only the preferred embodiments of the present invention, it is not limited to the present invention, although
Being described in detail the present invention with reference to embodiment, for a person skilled in the art, it still can be to aforementioned
Technical scheme described in each embodiment is modified, or wherein portion of techniques feature carries out equivalent, but all
Within the spirit and principles in the present invention, any modification, equivalent substitution and improvement etc. made, should be included in the protection of the present invention
Within the scope of.
Claims (3)
1. an anti-swine flu and the Chinese medicine compound of reproductive and respiratory syndrome, it is characterised in that:
Comprise following medicine: Flos Lonicerae 20～30 weight portion, the Rhizoma Anemarrhenae 10～20 weight portion, Radix Paeoniae Rubra 10～15 weight portion, Rhizoma Osmundae 10
～15 weight portions, Radix Bupleuri 10～15 weight portion, Herba Houttuyniae 10～15 weight portion, Radix Glycyrrhizae 5～15 weight portion.
2. there are a kind of anti-swine flu described in 1 and the Chinese medicine compound of reproductive and respiratory syndrome according to right, it is characterised in that: comprise such as prescribe medicine
Thing: Flos Lonicerae 25 weight portion, the Rhizoma Anemarrhenae 15 weight portion, Radix Paeoniae Rubra 12.5 weight portion, Rhizoma Osmundae 12.5 weight portion, Radix Bupleuri 12.5 weight portion,
Herba Houttuyniae 12.5 weight portion, Radix Glycyrrhizae 10 weight portion.
3. having the extracting method of the Chinese medicine compound of a kind of anti-swine flu and reproductive and respiratory syndrome described in 1 or 2 according to right, its feature exists
In, comprise the following steps:
(1) each medicine is weighed by said ratio；
(2) the recurrence due to taking drug side after weighing is crushed to 20 mesh, adds the water of doubling dose, soaks 1h, decocting 2 times, boils every time
30min, merges 2 decocting liquids；
(3) by decocting liquid in normal-pressure vacuum sucking filtration and concentration, being settled to 100mL decocting liquid, every milliliter is equivalent to 1g crude drug；
(4) decocting liquid is carried out autoclaving, save backup in 4 DEG C of refrigerators.
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| CN115350238A (en) * | 2022-09-26 | 2022-11-18 | 宁波科瑞特动物药业有限公司 | Traditional Chinese veterinary medicine granules and preparation method thereof |
| CN115887572A (en) * | 2022-12-05 | 2023-04-04 | 广东海大集团股份有限公司 | Preparation method and application of compound traditional Chinese medicine soluble powder for preventing and treating porcine reproductive and respiratory syndrome |
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| CN115350238A (en) * | 2022-09-26 | 2022-11-18 | 宁波科瑞特动物药业有限公司 | Traditional Chinese veterinary medicine granules and preparation method thereof |
| CN115350238B (en) * | 2022-09-26 | 2023-09-22 | 宁波科瑞特动物药业有限公司 | Chinese veterinary medicine granule and preparation method thereof |
| CN115887572A (en) * | 2022-12-05 | 2023-04-04 | 广东海大集团股份有限公司 | Preparation method and application of compound traditional Chinese medicine soluble powder for preventing and treating porcine reproductive and respiratory syndrome |
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