CN104473872B - Plant polyose oil emu and its production and use - Google Patents
Plant polyose oil emu and its production and use Download PDFInfo
- Publication number
- CN104473872B CN104473872B CN201410759414.2A CN201410759414A CN104473872B CN 104473872 B CN104473872 B CN 104473872B CN 201410759414 A CN201410759414 A CN 201410759414A CN 104473872 B CN104473872 B CN 104473872B
- Authority
- CN
- China
- Prior art keywords
- oil
- emu
- plant polyose
- polyose
- polysaccharide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of preparation method of plant polyose oil emu, Bao includes Ru Xia Bu Sudden：1), by sorbester p17 and edible vegetable oil with volume ratio 10:90~16:84 mix, and obtain oil phase；2), plant polyose is dissolved in the Tween 80 aqueous solution that volumetric concentration is 5~7%, acid-base value is then adjusted to pH2~7, obtains every milliliter of plant polyose aqueous solution containing 8~64 milligrams of plant polyoses, be used as aqueous phase；3), by aqueous phase and oil phase with 1:1~2 volume ratio is sufficiently mixed, emulsification, obtains plant polyose oil emu.Plant polyose is, for example, astragalus polyose, soluble polysaccharide, Codonopsis pilosula polysaccharide or licorice polysaccharide.The plant polyose oil emu can serve as immunostimulant, or make the medicine of mastitis for milk cows.
Description
Technical field
The present invention relates to a kind of plant polyose oil emu and its production and use.
Background technology
Polysaccharide is the polymer that a kind of monosaccharide groups by more than 10 are linked together by glycosidic bond, is widely present in
In the cell membrane of the internal and microorganism of animal and plant.The polysaccharide obtained is extracted from Chinese herbal medicine has various biological function.Radix glycyrrhizae
Polysaccharide can play immunological regulation and make by reducing the Treg cell proportions of tumor-bearing mice and improving spleen lymphocyte proliferation ability
With [1].LBP-X can raise CD4+T cell proportions in cancer of the esophagus rat peripheral blood, under CD4+CD25+Tr cell proportions
Drop, IL-10 secretions are reduced and IL-2 secretion increases, and improve experimental cancer of the esophagus rat cell immunologic function [2].Six drugs containing rehmanniae
Polyoses oral administration can antagonism endoxan to mouse boosting cell antibody tormation inhibitory action, external application can promote mice spleen thin
Born of the same parents' breeder reaction, rise splenocyte produces the level of antibody, increases the number [3] of spleen antibody-producting cell.Porphyra haitanensis polysaccharide
And its catabolite can remarkably promote T cell propagation, suppress B cell proliferation [4].The ganoderma lucidum of various concentrations is added in NK cells
Polysaccharide can improve NK cytoactives, promote TNF ɑ, IL-2, interferon gamma release [5].LBP-X can promote production
Raw a variety of lymphocyte factor such as macrophage shifter factors, lymphotoxin transfer factor etc..These factors can promote in vivo
The division growth of immunocompetent cell, improves the phagocytosis of macrophage and the killing ability [6] of T cell.Saline cistanche is more
Sugar can substantially increase RAW264.7 cells TNF-α and IL-1 release [7].Astragalus polyose (APS) [8] can activate mouse abdominal cavity macrophage
Cell induction NO is produced, and the transcription of NO synthase (iNOS) can be induced by activating transcription factor NF- κ B, is played Immune-enhancing effect and is made
With.Wheat-based diet can be by improving macrophage phagocytosis and secreting function, and activated macrophage strengthens immunologic function and played
Antitumor action [9].Due to the excellent biological function of polysaccharide, polysaccharides of traditional Chinese medicine is widely applied in field of medicaments, for preventing
Control human diseases.Polysaccharides of traditional Chinese medicine is also widely used in animal farming industry, prevents and treats Animal diseases, reduces making for chemicals
With the residual of reduction medicine in animal body.
As medical industry and animal farming industry are to the growing day by day of Chinese medicine demand, natural resources of Chinese medicinal materials faces huge pressure
Power.According to investigations, resources of medicinal plant radix glycyrrhizae, glycyrrhiza glabra, saline cistanche, radix stellariae dichotomae, Asian puccoon, Radix Astragali etc. 100 several kinds of Chinese medicinal materials
Stock number is generally reduced, and has influence on the pharmaceutical drugs of more than 60 medicinal material kind.Dysosma versipellis, the bark of eucommia, upas, wild mountain ginseng,
More than the 30 kinds of medicinal materials such as ribbed hedyotis herb, small hook catechu, Magnolia bilola are rare because of wild resource amount, in endangered edge, and nothing
Method provides commodity or can only provide a small amount of commodity.By taking wild licorice as an example, the fifties in last century China wild resource reserves
For 2,000,000, and at present less than 350,000；The coverage of many local wild licorices drops to fragmentary distribution from more than 90%
[10].Therefore, the method for studying saving Chinese medicaments material, while medical industry and animal farming industry is ensured to Chinese medicine demand,
The utilization rate for improving Chinese medicine is extremely urgent.
Bibliography
[1] Li Xiaobing, He little Juan, Liu Biao etc.；Immunoregulation effect of the licorice polysaccharide to H22 tumor-bearing mices；Traditional Chinese and western medicine knot
Close journal, 2010,8 (4):363-367；
[2] Dan Tieying, Qiao Junhong, Yang Shuliang etc.；LBP-X is exempted to experimental cancer of the esophagus rat peripheral blood cell immunity
The influence of epidemic disease function；When precious traditional Chinese medical science traditional Chinese medicines, 2010,21 (4):1016-1017；
[3] Qi Chunhui, Fu Yanrong, Zhang Yongxiang etc.；Effect China of the Liuwei Dihuang polysaccharides CA4-3 to mouse B cell function
Pharmacological Bulletin, 2001,17 (4):469-472.；
[4] Tingting ZHAO, Zhang Quanbin, Li Zhien etc.；The influence of Porphyra haitanensis polysaccharide and its degradation products to immune cell propagation；
New Chinese medicine and clinical pharmacology, 2009,20 (1):30-32；
[5] Liu Yuan, Geng Yue；The antineoplastic cell mechanism of GL-B and effect path research；Food and medicine, 2008,
10(7):48-51；
[6] Li Haibo, Mei Zhinan, Zhu Fan；The discussion of immune mechanism of anti-tumor by lycium barbarum polysaccharides；Chinese Hospitals pharmacy
Magazine, 2005,25 (2):115-7；
[7] Wang Xiangyan, Qi Yun, Cai Runlan etc.；The macrophage activation effect of wheat-based diet；Chinese Pharmacological Bulletin,
2009,25(6):787-790；
[8]Paulsen B S.Plant polysaccharides with immunostimulatory
activities.Curr Org Chem,2001,5(9):939-950；
[9] Wang Xiangyan, Qi Yun, Cai Runlan etc.；The macrophage activation effect Chinese Pharmacological Bulletins of wheat-based diet,
2009,25(6):787-790；
[10] plum gets the better of, Zhang Wensheng, Wang Yongyan；The wild natural resources of Chinese medicinal materials protection of China is in urgent need of strengthening；Chinese traditional Chinese medicine information
Magazine, 2008,15 (10)：4-6.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of plant polyose oil emu and its production and use.
In order to solve the above-mentioned technical problem, the present invention provides a kind of preparation method of plant polyose oil emu, including as follows
Bu Sudden：
1), the preparation of oil phase：
By Arlacel-80 and edible vegetable oil with volume ratio 10:90~16:84 mix；Obtain oil phase；
That is, volume content of the Arlacel-80 in oil phase is 10~16% (preferably 10~14%)；
2), the preparation of aqueous phase：
Plant polyose is dissolved in the Tween-80 aqueous solution that volumetric concentration is 5~7%, then adjusts acid-base value to pH2
~7 (preferably pH6~7), obtain every milliliter of plant polyose aqueous solution containing 8~64 milligrams of plant polyoses, are used as aqueous phase；
3), by aqueous phase and oil phase with 1:1~2 volume ratio is sufficiently mixed, emulsification, obtains plant polyose oil emu.The plant
Polysaccharide oil emu is the oil emu that outward appearance is creamy white.
The present invention is also simultaneously there is provided the plant polyose oil emu being prepared using the above method.
A kind of purposes of the present invention also simultaneously there is provided the plant polyose oil emu being prepared using the above method：It is used as
Immunostimulant.
Another purposes of the present invention also simultaneously there is provided the plant polyose oil emu being prepared using the above method：With
Make the medicine of mastitis for milk cows.I.e. there is provided the plant polyose oil emu in the medicine for preparing treatment mastitis for milk cows
Application.
The plant polyose oil emu of the present invention is a kind of outward appearance containing plant polyose, edible vegetable oil and surfactant
The oil emu being creamy white.
In the present invention：
Plant polyose includes astragalus polyose, soluble polysaccharide, Codonopsis pilosula polysaccharide, licorice polysaccharide.
Vegetable oil include all edible plants oil, such as rapeseed oil, corn oil, soybean oil, tea oil, sesame oil, peanut oil,
Olive oil, palm oil, sunflower oil etc..
The regulation of pH value belongs to routine techniques, for example, can use the hydrochloric acid or hydrogen-oxygen that concentration is added dropwise for 1mol/L~5mol/L
Change sodium solution to be adjusted.
Emulsification refers to：Two kinds of mutual exclusive liquid are mutually mixed, common method includes strong stirring, or addition
Emulsifying agent changes the surface tension of one of them liquid, promotes it to may be uniformly distributed in another liquid.This falls within routine
Technology.
The present invention has following technical advantage：
It is eighth with only aqua effective dose the invention provides a kind of preparation method of plant polyose preparation
Oil emu injection animal is made in plant polyose, and the stimulation to immune system is not reduced.Said preparation can be used as milk cow's milk
The scorching medicine in room, each case shot.Injection system and consumption are：At milk cow Ipsilateral lymphonodi mammarici position
The oil emu of the plant polyose containing 8mg~32mg is subcutaneously injected；Or at the bilateral lymphonodi mammarici position of recessive mastitis milk cow
It is subcutaneously injected, per the oil emu of side injection plant polyose containing 16mg.
Brief description of the drawings
The embodiment to the present invention is described in further detail below in conjunction with the accompanying drawings.
Fig. 1 is the influence comparison diagram of astragalus polyose aqua and oil emu hypodermic injection to mouse spleen index；
Fig. 2 Codonopsis pilosula polysaccharides, licorice polysaccharide, astragalus polyose and soluble polysaccharide oil emu and aqua injection are to Mouse interferon gamma
Rise action diagram；
Fig. 3 is the comparison diagram that astragalus polyose aqua and oil emu injection produce interferon gamma effect to mouse.
Remarks explanation：In figure, the different numerical tabular of Superscript letters is shown with significant difference.
Embodiment
The preparation of embodiment 1, astragalus polyose oil emu
1. the preparation of oil phase：1 milliliter of Arlacel-80 and 9 milliliters of edible rapeseed oils are uniformly mixed, oil phase is obtained；
2. the preparation of aqueous phase：Astragalus polyose is dissolved in 6% (volume %) the Tween-80 aqueous solution, acid-base value is adjusted
To pH7, every milliliter of aqueous solution containing 8 milligrams of astragalus polyoses is made, aqueous phase is used as；
3. by both aqueous phase and oil phase according to volume ratio 1:1 is sufficiently mixed, emulsification, i.e. equal using high cut disperse emulsification
Matter machine (b25 types, spy's electromechanical equipment Science and Technology Ltd. of Shanghai Bell product) is stirred 30 seconds under rotating speed 22000rpm, is made outer
The oil emu being creamy white is seen, astragalus polyose oil emu is produced.
The preparation of embodiment 2, licorice polysaccharide oil emu
1. the preparation of oil phase：1 milliliter of Arlacel-80 and 9 milliliters of edible rapeseed oils are uniformly mixed, oil phase is obtained；
2. the preparation of aqueous phase：Licorice polysaccharide is dissolved in the 5% Tween-80 aqueous solution, regulation acid-base value to pH6, system
Into every milliliter of aqueous solution containing 16 milligrams of licorice polysaccharides, aqueous phase is used as；
3. by both aqueous phase and oil phase according to volume ratio 1:1 is sufficiently mixed, emulsification, and the oil breast that outward appearance is creamy white is made
Agent, produces licorice polysaccharide oil emu.
Embodiment 3：The preparation of Codonopsis pilosula polysaccharide oil emu
1. the preparation of oil phase：1 milliliter of Arlacel-80 and 9 milliliters of edible rapeseed oils are uniformly mixed；Obtain oil phase；
2. the preparation of aqueous phase：Codonopsis pilosula polysaccharide is dissolved in the 7% Tween-80 aqueous solution, regulation acid-base value to pH6, system
Into every milliliter of aqueous solution containing 32 milligrams of Codonopsis pilosula polysaccharides, aqueous phase is used as；
3. by both aqueous phase and oil phase according to volume ratio 1:1 is sufficiently mixed, emulsification, and the oil breast that outward appearance is creamy white is made
Agent, produces Codonopsis pilosula polysaccharide oil emu.
Embodiment 4：The preparation of soluble polysaccharide oil emu
1. the preparation of oil phase：1 milliliter of Arlacel-80 and 9 milliliters of edible rapeseed oils are uniformly mixed；Obtain oil phase；
2. the preparation of aqueous phase：Soluble polysaccharide is dissolved in the 5% Tween-80 aqueous solution, regulation acid-base value to pH6, system
Into every milliliter of aqueous solution containing 64 milligrams of soluble polysaccharides, aqueous phase is used as；
3. by both aqueous phase and oil phase according to volume ratio 1:1 is sufficiently mixed, emulsification, and the oil breast that outward appearance is creamy white is made
Agent, produces soluble polysaccharide oil emu.
Embodiment 5：Immunostimulation of the various dose astragalus polyose oil emu to mouse
Material and method
1. the preparation of astragalus polyose oil emu
(1) preparation of oil phase：1 milliliter of Arlacel-80 and 9 milliliters of edible rapeseed oils are uniformly mixed；Obtain oil phase；
(2) preparation of aqueous phase：Astragalus polyose is dissolved in 6% (volume %) the Tween-80 aqueous solution, acid-base value is adjusted
To pH 7, every milliliter of two kinds of aqueous solution containing 5 milligrams and 10 milligrams of astragalus polyose are made, aqueous phase is used as；
(3) by both aqueous phase and oil phase according to volume ratio 1:1 is sufficiently mixed, emulsification, is made every milliliter and contains astragalus polyose 2.5
Milligram and 5 milligrams of two kinds of astragalus polyose oil emu.
2. medication and sampling
By ICR male mices 40,5 groups, every group 8 are randomly divided into.5 groups of mouse distinguish subcutaneous shot 0.5ml lifes
Manage salt solution, astragalus polyose aqua (containing astragalus polyose 10mg), (contain astragalus polyose without medicine oil emu, astragalus polyose oil emu 1
2.5mg), astragalus polyose oil emu 2 (1.25mg).The 8th day after injection, blood sampling prepared serum, detects serum interferon γ water
It is flat；Mouse is put to death, weighs, takes spleen to weigh, spleen index is calculated according to formula：Spleen index=spleen weight (mg)/body weight (g).
Remarks explanation：
Refer to the use for cancelling astragalus polyose in above-mentioned steps (2) without medicine oil emu；Remaining is with above-mentioned preparation method
Gains.
As a result and analysis
As shown in Figure 1, astragalus polyose aqua (10mg) and without medicine oil emu be once subcutaneously injected mouse generation spleen refer to
Number is respectively 4.14 ± 0.61 and 5.51 ± 0.30；When astragalus polyose and vegetable oil are mixed, astragalus polyose oil emu 1 is prepared into
(contain astragalus polyose 2.5mg) and astragalus polyose oil emu 2 (1.25mg) inject mouse afterwards, and the spleen index of generation is respectively 6.45 ±
1.07 and 5.97 ± 0.68.Although the astragalus polyose content in astragalus polyose oil emu 1 (containing astragalus polyose 2.5mg) is only aqua
The a quarter of (containing astragalus polyose 10mg), but caused spleen index is significantly higher than astragalus polyose or oil emu is used alone and caused
Spleen index (P<0.05)；Astragalus polyose content in astragalus polyose oil emu 2 (containing astragalus polyose 1.25mg) is the eight of aqua
/ mono-, but caused spleen index is significantly higher than astragalus polyose aqua (P<0.05), and numerically it is higher than without medicine oil emu
Spleen index (P caused by being used alone>0.05).The result shows astragalus polyose, which is prepared into after oil emu, produces immunostimulation
Effect is significantly higher than aqua.
From the figure 3, it may be seen that astragalus polyose aqua (10mg) and without medicine oil emu be once subcutaneously injected mouse generation serum
Interferon gamma level and saline control group are not more raised；When astragalus polyose and vegetable oil are mixed, the Radix Astragali is prepared into many
Sugar and oil emulsion 1 (containing astragalus polyose 2.5mg) and astragalus polyose oil emu 2 (1.25mg) inject mouse, the serum interference of generation afterwards
Plain γ levels are significantly higher than astragalus polyose aqua and without medicine oil emu group (P<0.05).The result shows astragalus polyose system
It is standby to be significantly higher than aqua into the interferon gamma level induced after oil emu.
Embodiment 6：The influence of astragalus polyose, licorice polysaccharide, Codonopsis pilosula polysaccharide, soluble polysaccharide emulsion to mice serum interferon
Material and method
56 ICR mouse are randomly divided into 8 groups, every group 7.Every mouse difference subcutaneous abdomen shot 0.2ml is white
Art polysaccharide (1.6mg), astragalus polyose (1.6mg), the aqua or oil emu of licorice polysaccharide (1.6mg) and Codonopsis pilosula polysaccharide (1.6mg).
Every mouse blood about 0.35ml is extracted after injecting 2 days, with ELISA method detection part serum interferon γ contents.
Remarks explanation：
First, the preparation method of soluble polysaccharide oil emu is：
1. the preparation of oil phase：1 milliliter of Arlacel-80 and 9 milliliters of edible rapeseed oils are uniformly mixed；Obtain oil phase；
2. the preparation of aqueous phase：Soluble polysaccharide is dissolved in the 5% Tween-80 aqueous solution, regulation acid-base value to pH7, system
Into every milliliter of aqueous solution containing 16 milligrams of soluble polysaccharides, aqueous phase is used as；
3. by both aqueous phase and oil phase according to volume ratio 1:1 is sufficiently mixed, emulsification, and the oil breast that outward appearance is creamy white is made
Agent, produces soluble polysaccharide oil emu.
2nd, the preparation method of astragalus polyose oil emu is：
1. the preparation of oil phase：1 milliliter of Arlacel-80 and 9 milliliters of edible rapeseed oils are uniformly mixed；Obtain oil phase；
2. the preparation of aqueous phase：Astragalus polyose is dissolved in 6% (volume %) the Tween-80 aqueous solution, acid-base value is adjusted
To pH7, every milliliter of aqueous solution containing 16 milligrams of astragalus polyoses is made, aqueous phase is used as；
3. by both aqueous phase and oil phase according to volume ratio 1:1 is sufficiently mixed, emulsification, and the oil breast that outward appearance is creamy white is made
Agent, produces astragalus polyose oil emu.
3rd, the preparation method be the same as Example 2 of licorice polysaccharide oil emu.
4th, the preparation method of Codonopsis pilosula polysaccharide oil emu is：
1. the preparation of oil phase：1 milliliter of Arlacel-80 and 9 milliliters of edible rapeseed oils are uniformly mixed；Obtain oil phase；
2. the preparation of aqueous phase：Codonopsis pilosula polysaccharide is dissolved in the 7% Tween-80 aqueous solution, regulation acid-base value to pH6, system
Into every milliliter of aqueous solution containing 16 milligrams of Codonopsis pilosula polysaccharides, aqueous phase is used as；
3. by both aqueous phase and oil phase according to volume ratio 1:1 is sufficiently mixed, emulsification, and the oil breast that outward appearance is creamy white is made
Agent, produces Codonopsis pilosula polysaccharide oil emu.
As a result and it is analyzed as follows：
From Figure 2 it can be seen that mouse shot soluble polysaccharide, astragalus polyose, the aqua of licorice polysaccharide and Codonopsis pilosula polysaccharide are to small
The interferon gamma level rise of mouse serum is little, but the serum interferon γ levels of the oil emu generation of four kinds of polysaccharide of injection are notable
Higher than aqua, illustrate that polysaccharide oil emu produces immunostimulation and causes the effect of interferon gamma to be significantly higher than aqua.
Embodiment 7：Various dose soluble polysaccharide oil emu Ipsilateral lymphonodi mammarici position is subcutaneously injected to milk cow recessive breast
The scorching therapeutic action in room
Purpose
Various dose soluble polysaccharide oil emu is examined to be subcutaneously injected at Ipsilateral lymphonodi mammarici position to milk cow recessive breast
Scorching therapeutic action.
Material and method
1. soluble polysaccharide oil emu：(1) preparation of oil phase：1.4 milliliters of Arlacel-80s and 8.4 milliliters of edible rapeseed oils are equal
Even mixing；Obtain oil phase；(2) preparation of aqueous phase：Soluble polysaccharide is dissolved in the 6% Tween-80 aqueous solution, regulation acid-base value to
PH 6, is made every milliliter of aqueous solution containing 24 milligrams of soluble polysaccharides, is used as aqueous phase；(3) by both aqueous phase and oil phase according to volume
Than 1:2 are sufficiently mixed, emulsification, and the oil emu that outward appearance is creamy white is made, every milliliter of 8mg containing soluble polysaccharide soluble polysaccharide is produced
Oil emu.
2. recessive mastitis infected cattle is selected：To being examined in the black-and-white flower cow in milk of Hangzhou suburbs cattle farm with Hangzhou mammitis
Test agent (HMT) filters out the newborn area of positive (++) and strong positive (+++), then the milk sample in the newborn area of aseptic collection, and low temperature is transported
Milk somatic cell counting (SCC) and bacteria distribution identification are carried out to laboratory, one weekly check of interval twice, filters out at least one
Ge Ru areas Somatic Cell Count SCC twice>500000/ml, and the positive milk cow 24 of bacteriology checking.
3rd, the treatment of recessive mastitis milk cow：24 cow heads are randomly divided into 4 groups, every group 6 by more than.Group 1~3 exists respectively
The subcutaneous shot soluble polysaccharide oil emu 1ml (8mg) of ill breast district side lymphonodi mammarici, 2ml (16mg) or 4ml
(32mg)；Control group milk cow is not treated.
4. treat post-sampling：1 after medication, 2,3 weeks, it is thin that the newborn area's milk sample of aseptic collection experiment carries out Somatic Cell Count, milk sample
Mycology is checked and NAGase determinations of activity.
5. recessive mastitis detects (HMT methods)：Scrubbed each newborn area of ox to be checked with hot towel, discard first three
Milk, squeezes 4 small interiors in diagnosis disk by the breast in 4 Ge Ru areas respectively, and each cell about 2ml containing milk is separately added into 2ml HMT
Diagnostic reagent, diagnosis disk, result of determination are shaken in concentric circles (criterion is shown in Table 1).
The HMT diagnostic reagent criterion of table 1
6. milk somatic cell (SCC) is counted：Carried out according to instrument specification.Instrument is corrected with standard items first,
Then by milk sample heating water bath to 40-42 DEG C, detected after shaking up, record data.
7. milk bacteriology checking：Carried out according to NMC (National Mastitis Council) recommended program.Specifically
Operating procedure is as follows：
(1) take 10 μ l milk samples to be inoculated in 6% Sheep Blood plate, put 37 DEG C of incubator cultures 24~48 hours.
(2) colony morphology characteristic is observed, picking colonies typical is inoculated in ordinary broth and carries out Zengjing Granule, 37 DEG C of vibration trainings
Support 24 hours.Smear, Gram's staining, microscopy carry out preliminary judgement.
(3) biochemical identification is carried out.Gram-positive cocci is carried out first catalase test determine the bacterium for staphylococcus or
Streptococcus.Staphylococcus is divided into staphylococcus aureus and coagulase-negative staphylococci with coagulase test of blood plasma
(CNS)；Streptococcus is divided into agalasisa with experiments such as CAMP test, aesculin hydrolysis, sodium hippurate hydrolysis, the reduction of methylene blue cow's milk
Streptococcus, streptococcus dysgalactiae or streptococcus uberis.Be not belonging to above-mentioned five kinds of bacteriums is classified as other classes.
8. milk NAGase Enzyme assays：Operate according to the following steps：
(1) centrifuge tube of the milk containing 5ml is put into multigelation 3 times in refrigerator, fully discharges the NAGase of intracellular.
(2) milk sample melted is centrifuged into 20min in 4000rpm, removes upper strata butterfat.Then dripped in the milk of degreasing
Plus 10% glacial acetic acid, pH to 4.6 is adjusted, in being stored at room temperature；Then 4000rpm, centrifuges 20min, separates out whey.
(3) the NAGase activity in whey is detected according to kit specification.Concrete operations are：Add and steam in blank tube
Distilled water (0.1ml)；Paranitrophenol titer (0.6mmol/L, 0.1ml) is added in standard pipe；It is each in control tube and measure pipe
Add 0.1ml wheys to be measured.It is each in blank tube and standard pipe to add 0.5ml reagents one；0.5ml substrates are added in pipe is determined
Buffer solution.Each pipe is put after 37 DEG C of water-bath 15min, adds 2ml reagents three；0.5ml substrate buffer solutions are added in control tube.Finally exist
It is each in each test tube to add 0.05ml reagents four, mix, OD values are determined at 400nm.NAGase activity is calculated according to formula：Breast
NAGase vigor (U/L)=(determining pipe OD values-control tube OD values)/(standard pipe OD values-blank tube OD values) × standard items in clear
Sample extension rate before concentration (0.6mmol/L) × 1/15 × 1000 × test
9. statistical analysis：Data are represented with Mean ± S.E..Examined with t compare between same period group, the difference before and after medication in group
The opposite sex, before statistical analysis, SCC, NAGase numerical value is carried out logarithmic transformed.
As a result with analysis
1. the treatment of soluble polysaccharide oil emu reduces milk somatic cell content
Milk somatic cell number before treatment between each group is without significant difference, and after being treated through soluble polysaccharide oil emu, three are controlled
Somatic number in treatment group milk has declined, and the milk somatic cell number of wherein treatment group 2 (soluble polysaccharide breast 16mg) is being controlled
2 after treatment, it is substantially less than concurrent control group within 3 weeks, also there is significant difference compared with pre-treatment；And control group ox somatic cell count in milk exists
It is basically unchanged (table 2) before and after treatment.
Table 2 treats front and rear each group milk sample somatic number (average × 104±S.E./ml)
Represent before treatment；A represents that treatment group compares significant difference (P with concurrent control group<0.05)；* table is distinguished with * *
Show compared with pre-treatment compared with significant difference (P<0.05) with extremely significantly (P<0.01).
2. soluble polysaccharide oil emu treats the activity for reducing milk NAGase enzymes
The NAGase enzymatic activitys of each group milk are without significant difference, after being treated through soluble polysaccharide oil emu, three before treatment
NAGase activity in treatment group's milk has declined, wherein (16mg) milk for the treatment of group 2 NAGase activity after the treatment 2,3
It is substantially less than in week before treating, after the treatment 1,2,3 weeks substantially less than control group；(32mg) the milk NAGase for the treatment of group 3 activity exists
It is substantially less than control group within 2 weeks after treatment.
The NAGase that table 3. treats front and rear each group milk sample is active (average ± S.E.U/L)
Represent before treatment；A represents that treatment group compares significant difference (P with concurrent control group<0.05)(P<0.01)；*
Represented respectively compared with pre-treatment compared with significant difference (P with * *<0.05) with extremely significantly (P<0.01)
This result of study shows the bighead atractylodes rhizome is once subcutaneously injected at the lymphonodi mammarici position of recessive mastitis milk cow Ipsilateral many
Sugar and oil emulsion, can significantly reduce the activity of the Somatic Cells Content and NAGase enzymes in the newborn area's milk of recessive mastitis.Milk cow suffers from breast
The activity rise of Somatic Cells Content and NAGase enzymes when room is scorching in milk.Injection soluble polysaccharide emulsion can make Bovine Mastitis Milk area ox
Somatic Cells Content and NAGase activity reductions in milk, illustrate that soluble polysaccharide oil emu has treatment to make to cow subclinical mastitis
With.And when injecting the aqua of the soluble polysaccharide containing 64mg as control (experiment method is ibid), the control group is to milk cow recessive breast
The scorching therapeutic effect in room is nothing like " treatment group 1 (8mg) " of present invention therapeutic effect.
Embodiment 8：Soluble polysaccharide oil emu bilateral lymphonodi mammarici position is subcutaneously injected and cow subclinical mastitis is controlled
Treatment is acted on
Material and method
1. prepared by soluble polysaccharide oil emu, recessive mastitis infected cattle is selected, recessive mastitis detects (HMT methods), milk body
Cell (SCC) counting, milk bacteriology checking, milk NAGase Enzyme assays be the same as Example 7.
2. the treatment of recessive mastitis milk cow：The recessive mastitis milk cow that will be singled out is randomly divided into two groups：Treatment group, 11
Cow head (has 14 to suffer from recessive mastitis) in 41 Ge Ru areas, and at the lymphonodi mammarici position of milk cow, the subcutaneous shot bighead atractylodes rhizome is more
Sugar and oil emulsion, per side 2ml (16mg)；Control group, 11 cow heads (have 12 to suffer from recessive mastitis) in 44 Ge Ru areas, do not treat.
3. treat post-sampling：1 after medication, 2,3 weeks, it is thin that the newborn area's milk sample of aseptic collection experiment carries out Somatic Cell Count, milk sample
Mycology is checked and NAGase determinations of activity.
4. statistical analysis：Data are represented with Mean ± S.E..Examined with t compare between same period group, the difference before and after medication in group
The opposite sex, before statistical analysis, SCC, NAGase numerical value is carried out logarithmic transformed.
As a result with analysis
1st, the body cell for reducing ill breast district milk is subcutaneously injected in soluble polysaccharide oil emu bilateral lymphonodi mammarici position
Content：
The milk somatic cell number of first two groups for the treatment of is without significant difference, after being treated through soluble polysaccharide oil emu, treatment group's milk
Somatic number after the treatment 2, be substantially less than concurrent control group within 3 weeks, be significantly reduced compared with pre-treatment (table 4).
Change (average × 10 of ill breast district milk sample somatic number before and after the treatment of the soluble polysaccharide oil emu of table 44±
S.E./ml)
Check that milk somatic cell number is all higher than 500,000/ml and the positive newborn area of bacteriology checking before treatment twice.
#Before treatment.
aP<0.05, and concurrent control group compares.
*P<0.05 and group in treat before compare.
2nd, the NAGase for reducing ill breast district milk is subcutaneously injected in soluble polysaccharide oil emu bilateral lymphonodi mammarici position
Enzymatic activity：
The NAGase enzymatic activitys of the preceding two groups of milk for the treatment of are without significant difference, after being treated through soluble polysaccharide oil emu, treatment
2,3 weeks were substantially less than concurrent control group to group milk NAGase activity after the treatment, 1,2,3 weeks substantially less than treatment after the treatment
Before.
Change (average × 10 of ill breast district milk sample NAGase enzymatic activitys before and after the treatment of the soluble polysaccharide oil emu of table 54
±S.E./ml)
Check that milk somatic cell number is all higher than 500,000/ml and the positive newborn area of bacteriology checking before treatment twice.
#Before treatment.
aP<0.05, and concurrent control group compares.
*P<0.05, P<Compare before being treated in 0.01, and group.
3. the body cell for reducing newborn area's mixing milk is subcutaneously injected in soluble polysaccharide oil emu bilateral lymphonodi mammarici position
Number
The somatic number of the newborn area's mixing milk of the preceding two groups of milk cows four for the treatment of is treated without significant difference through soluble polysaccharide oil emu
Afterwards, the SCC for the treatment of group milk cow is substantially less than the somatic number (table 6) before being treated in concurrent control group and group.
The soluble polysaccharide oil emu of table 6 treats the somatic number (average ± S.E.10 of preceding rear milk area mixing milk4/ml)
| Group | Milk cow number | Before treatment | After treatment |
| Soluble polysaccharide oil emu | 11 | 62.55±25.38 | 29.21±6.85a* |
| Control group | 11 | 56.66±32.34 | 54.89±11.88 |
aP<0.05, and concurrent control group compares.
*P<Compare before being treated in 0.05, and group.
4. the quantity for reducing the newborn area of bacterium infection is subcutaneously injected in soluble polysaccharide oil emu bilateral lymphonodi mammarici position
Treatment group You21Ge Ru areas bacterium infection before treatment, control group You23Ge Ru areas bacterium infection, without aobvious between two groups
Write difference.After being treated through soluble polysaccharide oil emu, the newborn area's number of infection for the treatment of group is reduced to 9 (P<0.05), concurrent control
The newborn area of the infection of group is not reduced (table 7) still more than 20.
Number (tested breast area number in bacterioscopy positive udder region before and after table 7, the treatment of soluble polysaccharide oil emu：Treatment group=41；It is right
According to group=44)
#Before treatment.
*P<Compare before being treated in 0.05, and group.
This result of study shows subcutaneously once to be subcutaneously injected in vain at bilateral lymphonodi mammarici position in recessive mastitis milk cow
Art polysaccharide oil emu, can significantly reduce the Somatic Cells Content of milk, the activity of NAGase enzymes and the bacterial infections levels of mammary gland,
Illustrate that soluble polysaccharide oil emu has therapeutic action to cow subclinical mastitis.
Finally, in addition it is also necessary to it is noted that listed above is only several specific embodiments of the invention.Obviously, this hair
It is bright to be not limited to above example, there can also be many deformations.One of ordinary skill in the art can be from present disclosure
All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Claims (4)
1. the preparation method of the plant polyose oil emu with immunostimulation performance or mastitis for milk cows curative properties, its feature exists
In comprising the following steps：
1) preparation of oil phase：
By Arlacel-80 and edible vegetable oil with volume ratio 10：90~14：86 mix；Obtain oil phase；
The edible vegetable oil is rapeseed oil, corn oil, soybean oil, tea oil, sesame oil, olive oil, palm oil or sunflower oil；
2) preparation of aqueous phase：
Plant polyose is dissolved in the Tween-80 aqueous solution that volumetric concentration is 5-7%, then adjusts acid-base value to pH6-7, obtain
The plant polyose aqueous solution of every milliliter of plant polyose containing 8-64mg, is used as aqueous phase；
The plant polyose is astragalus polyose, soluble polysaccharide, Codonopsis pilosula polysaccharide or licorice polysaccharide；
Regulation acid-base value uses dropwise addition concentration to be adjusted for 1mol/L~5mol/L hydrochloric acid or sodium hydroxide solution；
3) by aqueous phase and oil phase with 1：2 volume ratio mixing, emulsification obtains the plant polyose oil emu that outward appearance is creamy white, described
Emulsification is stirred 30 seconds using high cut disperse emulsification homogenizer under rotating speed 2200rpm.
2. the plant polyose oil emu that method as claimed in claim 1 is prepared.
3. application of the plant polyose oil emu that method as claimed in claim 1 is prepared in immunostimulant is prepared.
4. the plant polyose oil emu that method as claimed in claim 1 is prepared is in the medicine of mastitis for milk cows is prepared
Application.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410759414.2A CN104473872B (en) | 2014-12-11 | 2014-12-11 | Plant polyose oil emu and its production and use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410759414.2A CN104473872B (en) | 2014-12-11 | 2014-12-11 | Plant polyose oil emu and its production and use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104473872A CN104473872A (en) | 2015-04-01 |
| CN104473872B true CN104473872B (en) | 2017-09-26 |
Family
ID=52748578
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410759414.2A Active CN104473872B (en) | 2014-12-11 | 2014-12-11 | Plant polyose oil emu and its production and use |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104473872B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000245378A (en) * | 1999-03-02 | 2000-09-12 | Tsukishima Shokuhin Kogyo Kk | Water in oil type emulsion composition |
| CN101108195A (en) * | 2006-07-19 | 2008-01-23 | 天津天士力制药股份有限公司 | Medicament for curing tumour, improving immunity of organism and method of manufacturing the same |
| CN101428145A (en) * | 2007-11-05 | 2009-05-13 | 北京生泰尔生物科技有限公司 | Novel vaccine adjuvant |
| CN102058530A (en) * | 2010-12-14 | 2011-05-18 | 于荣敏 | Ganoderma lucidum polysaccharide oral nanoemulsion and preparation method thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080242739A1 (en) * | 2007-03-30 | 2008-10-02 | Gijsbert Kroon | Mixed hydrophobe polysaccharide as polymeric emulsifier and stabilizer |
-
2014
- 2014-12-11 CN CN201410759414.2A patent/CN104473872B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000245378A (en) * | 1999-03-02 | 2000-09-12 | Tsukishima Shokuhin Kogyo Kk | Water in oil type emulsion composition |
| CN101108195A (en) * | 2006-07-19 | 2008-01-23 | 天津天士力制药股份有限公司 | Medicament for curing tumour, improving immunity of organism and method of manufacturing the same |
| CN101428145A (en) * | 2007-11-05 | 2009-05-13 | 北京生泰尔生物科技有限公司 | Novel vaccine adjuvant |
| CN102058530A (en) * | 2010-12-14 | 2011-05-18 | 于荣敏 | Ganoderma lucidum polysaccharide oral nanoemulsion and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 黄芪多糖油乳佐剂新城疫灭活苗对鸡免疫功能的影响;王学斌，等;《河南农业科学》;20071231(第4期);第103-105页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104473872A (en) | 2015-04-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106942724B (en) | Composition with function of regulating intestinal flora structure and preparation method and application thereof | |
| CN108244432A (en) | A kind of fermentation Cordyceps militaris probiotic beverage and preparation method thereof | |
| CN104739919B (en) | A kind of fermentation Radix Astragali compound formulation and the preparation method and application thereof | |
| KR20130077802A (en) | The preparing method of immune improving agents | |
| CN100420448C (en) | Health-care food with function for improving immunity and its prepn. method | |
| CN101125151A (en) | Bacteroides signal molecule microecological preparation and its preparation method | |
| CN105661515A (en) | Enzyme lozenges for three-hypers and preparation method thereof | |
| CN101224279B (en) | Medicine compound for treating urinary infection and preparing method thereof | |
| CN103083510A (en) | Chinese medicinal oral liquid for improving poultry immune function and its preparation method | |
| CN104352521B (en) | Prepare the method for calf spleen extract and the application in antitumor and immunological regulation | |
| CN109527099A (en) | A kind of fructus lycii compound probiotic goat milk piece and preparation method thereof | |
| CN102688280A (en) | Preparation method for radiation-resistant Chinese medicine fermentation liquor | |
| CN104473872B (en) | Plant polyose oil emu and its production and use | |
| CN107254501A (en) | It is a kind of have significantly increase Cordyceps militaris protein peptides of immunity and preparation method thereof | |
| CN111228316A (en) | Composite probiotics for improving diabetes | |
| CN106389477B (en) | A kind of preparation method and application of the full cellular plant oil extract of Gordonia terrae | |
| CN113186118B (en) | Composite probiotic preparation for improving sow productivity and preparation method thereof | |
| CN102451221A (en) | Astragalus polysaccharides microecological regulator | |
| CN109329207A (en) | A kind of experimental animal model for anticancer agents and its construction method | |
| CN107893035A (en) | A kind of culture medium of Hericium erinaceus, sealwort bioconversion mycelium, the mycelial extract of sealwort bioconversion and application thereof | |
| CN1443542A (en) | Nocardia rubra actinomycete cell wall skeleton preparation | |
| CN111195298B (en) | Composition for regulating intestinal flora and preventing and treating food allergy and preparation method thereof | |
| CN106668077A (en) | Marine bioactive composition and pharmaceutical preparation | |
| CN113967243A (en) | Fermented traditional Chinese medicine preparation for preventing and treating poultry diarrhea and preparation method thereof | |
| CN100363012C (en) | Cordyceps sinensis polysaccharide emulsion and preparing method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20160706 Address after: 310000 2, West 1101, Zhenhua Road West Harbor Development Center, Hangzhou, Zhejiang, Xihu District Applicant after: Hangzhou vision Wang Biological Technology Co. Ltd. Address before: 310058 Xihu District, Zhejiang, Yuhang Tong Road, No. 866, No. Applicant before: Zhejiang University |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |