CN107279497A - Application of the Aspartame in young age ruminant intestinal development, the small intestinal barrier function of enhancing is promoted - Google Patents
Application of the Aspartame in young age ruminant intestinal development, the small intestinal barrier function of enhancing is promoted Download PDFInfo
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- 229960003438 aspartame Drugs 0.000 title claims abstract description 42
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Birds (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses application of the Aspartame in animal intestinal development, the small intestinal barrier function of enhancing is promoted；Aspartame can be used as additive in young age ruminant opened guideway, pass through a series of signal pathway stimulation such as sweet receptor GLP 2 secretion, after being combined afterwards with GLP 2R acceptors, activate ERK1/2 signal paths, so as to promote the expression of tight junction protein, strengthen the barrier function of small intestine epithelium, expression by adjusting young age ruminant cyclin and relative dependencies kinases promotes the development of cell cycle progression promotion small intestine epithelium, strengthen small intestinal barrier function, prevent the generation of the young age ruminants such as lamb diarrhoea, improve the survival rate of the young age ruminants such as lamb, be conducive to young age ruminant wean early stage intestinal health, weaken ablactation stress.
Description
Technical field
The invention belongs to feed additive field, and in particular to Aspartame promotes dynamic in young age ruminant opened guideway
The application of thing intestinal development, the small intestinal barrier function of enhancing.
Background technology
After birth to wean before be the young age ruminant digestion tract disease stage occurred frequently.By taking lamb as an example, correlation study shows
Show, the diarrhea rate of China nursing period lamb is about 50%, and the death rate of these diarrhoea lambs is up to 20%, causes to surpass every year
Cross 1,000,000,000 yuan of economic loss.The pathogenic factor of lamb diarrhoea is extremely complex, and ewe is malnutritive, lamb supplementary feeding feeding quality
Difference, stress and cause pathogeny imcrobe infection etc. be all probably the factor for inducing lamb diarrhoea.All being at present can by application antibiotic
It is still, residual with the disabling and antibiotic of antibiotic to treat or slow down the diarrhoea situation during the wean of young age ruminant
Problem is stayed, it is that society is required to find a kind of safer, effective and economic mode.Meanwhile, in recent years many studies have shown that, children
It is also the key factor for causing animal susceptible or even suffering from diarrhoea that age animal, which not yet develops perfect gut barrier function,.But,
Up to the present, research is there is no from the angle of protection gut epithelium barrier to prevent the generation that young age ruminant is suffered from diarrhoea.Therefore,
Corresponding nutrition regulation means are found to promote the development of young age intestine of ruminants barrier function and improve to ruminate reduction young age
Animal diarrhea risk is significant.
Aspartame molecular formula is：C14H18N2O5, structural formula is as follows：
A kind of natural function oligosaccharide, will not carious tooth, sweet taste be pure, hygroscopicity is low, without stickiness.It will not draw
The significantly raised of blood glucose is played, is adapted to patients with diabetes mellitus.Aspartame can be used for cake, biscuit, bread, assembled alcoholic drinks, ice cream,
Ice lolly, beverage, candy, consumption are needed by normal production.In terms of feed, the use on Aspartame at present is all compound
Application in terms of sweetener, not on the report to young age intestine of ruminants epithelial protective.
Goal of the invention
The content of the invention：The problem of existing for prior art, the present invention provide Aspartame promote animal intestinal development,
Strengthen the application of small intestinal barrier function.The present invention proposes a kind of New function of Aspartame, can be opened in young age ruminant
As additive in foodstuff, the generation that the young age such as lamb ruminant is suffered from diarrhoea is prevented from the angle of protection gut epithelium barrier,
Promote animal intestinal development, strengthen small intestinal barrier function.
Technical scheme：To achieve these goals, Aspartame as described in the present invention is promoting young age ruminant small
Intestines develop, strengthen the application in small intestinal barrier function.
Further, the Aspartame in animal feed as additive promote young age ruminant intestinal development,
Strengthen the application in small intestinal barrier function.
Wherein, described feed is opened guideway.
Further, the addition of Aspartame is the 0.08-0.16% of opened guideway quality in the opened guideway.It is preferred that
Addition be opened guideway quality 0.08%.
Preferably, the young age ruminant is lamb or calf.It is lamb generally from young age ruminant.
Wherein, the growth phase of described animal is to be arrived after being born before wean.
The present invention is used raw material and instrument is all commercially available.
Aspartame is purchased from Niu Te companies of the U.S.
Portable pH meter (HI 9024C；HANNA Instruments, the U.S.)
Plasma glucose kit is purchased from ShangHai RongSheng Biology Pharmacy Co., Ltd
GLP-2 fluorescence Enzymoimmune reagent kit is purchased from Germany Phoenix Europe GmbH
Reverse transcription reagent box is purchased from Takara companies
Q5Real-time PCR instruments (Applied Biosystems, the U.S.).
Beneficial effect：Compared with prior art, the invention has the advantages that：
Aspartame used in the present invention is a kind of cost-effective sweetener, and the present invention proposes a kind of Aspartame
New function, lamb etc. can be prevented from protection gut epithelium barrier in young age ruminant opened guideway as additive
The generation of young age ruminant diarrhoea, improves the survival rate of the young age ruminants such as lamb；Aspartame used in the present invention
Can be by a series of signal pathway stimulation such as sweet receptor GLP-2 secretion, after being combined afterwards with GLP-2R acceptors, activation
ERK1/2 signal paths, so as to promote the expression of tight junction protein, strengthen the barrier function of small intestine epithelium, by adjusting young age
The development of the expression promotion cell cycle progression promotion small intestine epithelium of ruminant cyclin and relative dependencies kinases,
Strengthen small intestinal barrier function, be conducive to young age ruminant wean early stage intestinal health, weaken ablactation stress.
Brief description of the drawings
Fig. 1 is that Aspartame adds the influence graph of a relation changed to lamb weight；
Fig. 2 is influence relation of the Aspartame addition to lamb intestinal epithelial cell cyclin mRNA relative expression quantities
Figure；
Fig. 3 is influence relation of the Aspartame addition to lamb small intestine epithelium tight junction protein mRNA relative expression quantities
Figure；
Fig. 4 is that Aspartame is added to lamb small intestine epithelium GCG, GLP-2R, IGF-1 and IGF-1RmRNA relative expression quantity
Influence graph of a relation.
Embodiment
Below in conjunction with drawings and examples, the present invention will be further described.
Embodiment 1
Experiment 12 body conditions of selection are good, and parity is consistent, and the close newborn lamb of body weight (sheep) is randomly divided into control group
With sweetener (Aspartame) treatment group, every group 6.During 10 age in days, lamb starts supplementary feeding opened guideway, and 8 points of every morning is with
5 points of noon respectively feeds once.During 14 age in days, start to add the A Siba of opened guideway quality 0.08% in test group lamb opened guideway
Sweet tea, during experiment, lamb follows ewe Free nursing and drinking-water, free choice feeding clover and oat hay, and lamb body is monitored weekly
Change again.Before on-test, the body weight of two groups of lambs is not significantly different (6.18 ± 0.47vs.5.80 ± 0.26kg, P=
0.504).Aspartame is purchased from Niu Te companies of the U.S..Opened guideway is according to sheep opened guideway nutrition requirements (NY/Y816-
2004；The Ministry of Agriculture, 2004) design opened guideway formula, constitute and be shown in Table 1.
The experiment opened guideway composition of table 1 and trophic level
Embodiment 2
Embodiment 2 is using being that identical opened guideway coordinates and test method, and difference is that subjects are body weight phase
The Aspartame of addition opened guideway quality 0.16% near newborn calf, treatment group opened guideway.
Embodiment 3
Control group and the change of test group lamb weight of embodiment 1 are monitored during experiment weekly, as a result as shown in Figure 1.
Two groups of lamb weights are without significant difference during whole experiment.
Embodiment 4
During 42 age in days of lamb of Example 1, morning feeding is butchered after two hours, butchers preceding jugular vein blood collection, blood
Liquid is stored in -20 DEG C in 4 DEG C of centrifugation 15min of 1000g is used for blood routine detection.Cud and small intestine are immediately disconnected after butchering and is claimed
Weight, collects representational rumen content and determines pH value, has determined four layers of filtered through gauze of the rumen content after pH value and has collected
Rumen fluid, -20 DEG C preserve Vfa Concentration to be measured.Collection representative duodenum, jejunum and ileal tissue
After being cleaned 3 times in the phosphate buffer solution of ice, the fine grained chippings loading cryopreservation tube for being cut into 0.5cm × 0.5cm is placed in Liquid nitrogen storage,
Extraction for follow-up RNA.
Embodiment 5
Detect that sweetener Aspartame adds the influence to each organ weight of lamb：
Sampling is butchered in 42 age in days of lamb, lamb is immediately disconnected each organ and weighed after butchering, as a result as shown in table 2.
The sweetener of table 2. adds the influence to each organ weight of lamb
Note：Data represent in average value ± standard error form,n=5
As shown in Table 2, sweetener Aspartame addition significantly improves lamb jejunum weight and caecum weight, to cud weight in wet base,
Cud dry weight, omasum weight in wet base, omasum dry weight, duodenum weight, ileum weight, colon weight and liver weight are without conspicuousness influence.It is above-mentioned
As a result illustrate：Sweetener Aspartame improves the weight of jejunum, has facilitation to intestinal development.
Embodiment 6
Blood routine detection is carried out to the blood of embodiment 4 and rumen content determines pH value, sweetener A Siba is investigated
Influence of the sweet tea to lamb cud and blood parameters, as a result as shown in table 3.
The plasma glucose kit of routine analysis of blood is purchased from ShangHai RongSheng Biology Pharmacy Co., Ltd；GLP-2 luciferases are exempted from
Epidemic disease kit is purchased from Germany Phoenix Europe GmbH.The measurement range of blood glucose and GLP-2 kits is 3.89 respectively~
6.11mmol/L and 0-10000pg/ml.
Rumen physiology parametric measurement, with portable pH meter (HI 9024C；HANNA Instruments, the U.S.) determine knurl
Stomach pH value.Gas Chromatographic Determination (GC-14B, Shimadzu, Japan；Capillary column:30m × 0.32 mm × 0.25mm film thicknesses) VFA
Concentration, is beautiful jade (the Improvement Agricultural University Of Nanjing of application gas Chromatographic Determination cud volatile fatty acid method with reference to the Qin
Journal, 1982 (4):Method (column temperature=110 DEG C, temperature of vaporization chamber=180 DEG C, detector=180 DEG C) 110-116).
The sweetener of table 3 adds the influence to lamb cud parameter and blood parameters
As shown in table 3, sweetener addition is to lamb cud butyric acid density, other Vfa Concentrations and blood plasma GLP-
2 concentration have elevated trend；Knurl pH, general volatile fatty acid concentration, acetic acid concentration, propionate concentration, second third are compared and blood plasma Portugal
Grape sugar concentration influences without conspicuousness.The above results explanation：Sweetener Aspartame promotes lamb itself GLP-2 secretion.
Embodiment 7
Sweetener Aspartame adds the influence to lamb small intestine epithelium mRN relative expression quantities.
RNA extraction and cDNA synthesis：Extract embodiment 4 and obtain small intestine epithelium total serum IgE, use NanoDrop light splitting light
Degree meter detection RNA concentration and purity.The assimilation ratio (260/280nm) of all samples is all between 1.8~2.0, it was demonstrated that RNA
Purity is higher.RNA integrality is detected with 1.4% agarose gel.All RNA concentration is adjusted to 1 μ g/ μ L, -80 are placed in
DEG C refrigerator is saved backup.CDNA synthesis is carried out using the reverse transcription reagent box purchased from Takara companies enzyme containing geneome RNA.
The synthesis of primer and real-time quantitative PCR：Utilize Q5Real-time PCR instruments (Applied Biosystems, U.S.
State) target gene and reference gene GAPDH, 18S rRNA are quantified, GAPDH primer sequences reference Wang etc. (Wang A,
Gu Z,Heid B,et al.Identification and characterization of the bovine G
protein-coupled receptor GPR41and GPR43genes 1[J].Journal of Dairy Science,
2009,92(6):2696-2705.), Real-time PCR Analysis the primer uses the software designed, designeds of Primer 5.0, by upper
Extra large JaRa bioengineering Co., Ltd synthesis, primer SQE ID NO.1-36, as shown in table 4.SYBR is purchased from Takara companies,
Reaction condition is：95 DEG C of 30s pre-degenerations, 95 DEG C of 5s, 60 DEG C of 30s repeat 40 circulations, 95 DEG C, 15s；60 DEG C, 1min；95,
15s.The μ L of reaction volume 20：10.4ul SYBR, 10umol/L upstream and downstream primers each 0.4ul, 2ul sample cDNA and 6.8ul without
Bacterium water.All samples set 3 repetitions.The relative expression quantity of target gene is corrected using house-keeping gene GAPDH as internal reference, number
2 are used according to analysis-△△CTMethod.
The amplimer sequence of table 4
As a result it is indicated with average value ± standard error (means ± SE).Data use the independent sample in SPSS 20.0
T, which is examined, carries out significance analysis.Using GraphPad Prism 6.01 (www.graphpad.com) software analysis target gene
Mrna expression amount.P<0.05 represents significant difference.
Sweetener Aspartame adds the influence to lamb intestinal epithelial cell cyclin mRNA relative expression quantities：Cell
The main regulation and control by cyclin (cyclins) and its kinases (CDKs) of correlation of the propulsion in cycle, thus cyclins and
CDKs expression can influence cell cycle progression.As shown in Fig. 2 the addition of sweetener Aspartame significantly improves lamb ten
Two duodenum 12 Cyclin D1, CDK1 and CDK6 mrna expression amount；Jejunum CyclinA, Cyclin B1, CDK1 and CDK6 mRNA
Expression quantity；Ileum Cyclin A, Cyclin D1 and CDK4 mrna expression amount.Result above shows：Aspartame addition promotes
The GLP-2 of lamb itself synthesis promotes the mRNA expression of intestinal epithelial cell cyclin and relative dependencies kinases, necessarily
Cell cycle progression has been promoted in degree, the propagation of small intestine epithelium is promoted.
Sweetener Aspartame adds the influence to lamb small intestine epithelium tight junction protein mRNA relative expression quantities：Epithelium
The complete talin for mainly passing through anchor frame albumen between cell and intercellular connection albumen and cell and matrix of barrier
Complex is maintained.As a result as shown in figure 3, Aspartame addition, which significantly improves lamb 12, refers to Claudin-4's and ZO-1
Mrna expression amount；Jejunum Claudin-1, Occludin and ZO-1 mrna expression amount；Ileum Claudin-4's and Occludin
Mrna expression amount.Claudin family proteins are topmost transmembrane proteins, between its major function closing cell and cell
Gap.Occludin is a kind of transmembrane protein directly acted on claudins and actin.Zo-1 is a kind of peripheral protein, for
Very crucial effect is played in close-connected distribution and maintenance.The above results show：Aspartame addition promotes lamb itself
The GLP-2 of secretion promotes the mrna expression amount of small intestine tight junction protein, is conducive to the development of the small intestinal barrier function of lamb, dimension
Hold intestinal health.
Sweetener Aspartame is added to lamb small intestine epithelium GCG, GLP-2R, IGF-1 and IGF-1RmRNA relative expression
The influence of amount：As shown in figure 4, compared with control group, Aspartame addition significantly increases duodenum GCG and IGF-1；Jejunum
GCG, IGF-1, IGF-1R and GLP-2R；Ileum GCG and IGF-1 mrna expression amount.The studies above result shows：Aspartame
Addition promotes the expression of lamb small intestine epithelium GLP-2 acceptors, while promoting the expression of IGF-1 and IGF-1 acceptors, implies
GLP-2 is, by IGF-1 signal path cell cycle regulation processes, to promote the propagation of small intestine epithelium.Embodiment 2 is used simultaneously
Calf is experimental subjects, as a result similar with above-described embodiment result.
SEQUENCE LISTING
<110>Agricultural University Of Nanjing
<120>Aspartame is promoting the application of young age ruminant intestinal development, the small intestinal barrier function of enhancing
<130> 2017
<160> 36
<170> PatentIn version 3.3
<210> 1
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<212> DNA
<213>It is artificial synthesized
<400> 1
cggaagaagt caacatcgt 19
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<400> 2
taaagtctcg ggtagcaag 19
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<400> 3
ggcctacaga tactgcctgt ct 22
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<400> 5
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<400> 24
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<400> 25
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<400> 27
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<400> 28
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<400> 29
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<400> 32
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<210> 33
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<213>It is artificial synthesized
<400> 33
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<400> 34
aatcggtagt agcgacgggc ggtg 24
<210> 35
<211> 20
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<213>It is artificial synthesized
<400> 35
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<400> 36
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Claims (6)
1. application of the Aspartame in young age ruminant intestinal development, the small intestinal barrier function of enhancing is promoted.
2. application according to claim 1, it is characterised in that the Aspartame promotees in animal feed as additive
The application entered in young age ruminant intestinal development, the small intestinal barrier function of enhancing.
3. application according to claim 2, it is characterised in that described feed is opened guideway.
4. application according to claim 3, it is characterised in that the addition of Aspartame is opened guideway in the opened guideway
The 0.08-0.16% of quality.
5. application according to claim 1, it is characterised in that the young age ruminant is lamb or calf.
6. application according to claim 1, it is characterised in that the growth phase of described animal weans to be arrived after birth
Before.
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| Application Number | Priority Date | Filing Date | Title |
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| CN110178991A (en) * | 2019-05-09 | 2019-08-30 | 中国农业大学 | A kind of nutrition regulation method promoting lamb intestinal growth |
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| CN106858081A (en) * | 2017-01-24 | 2017-06-20 | 南宁学院 | A kind of beef cattle special feed and preparation method |
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2017
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| CN106858081A (en) * | 2017-01-24 | 2017-06-20 | 南宁学院 | A kind of beef cattle special feed and preparation method |
Non-Patent Citations (1)
| Title |
|---|
| 李享等: "肠内分泌细胞表面甜味与苦味受体的研究进展", 《上海交通大学学报(医学版)》 * |
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