CN109833314A - Application of the glutamic acid binding peptide in the preparation that the intestinal tract injury of animal vomitoxin induction is alleviated in preparation - Google Patents
Application of the glutamic acid binding peptide in the preparation that the intestinal tract injury of animal vomitoxin induction is alleviated in preparation Download PDFInfo
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- CN109833314A CN109833314A CN201910173215.6A CN201910173215A CN109833314A CN 109833314 A CN109833314 A CN 109833314A CN 201910173215 A CN201910173215 A CN 201910173215A CN 109833314 A CN109833314 A CN 109833314A
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Abstract
The invention belongs to Animal nutritions and intestinal health technical field, specifically disclose application of the glutamic acid binding peptide in the preparation that the intestinal tract injury of animal vomitoxin induction is alleviated in preparation.The main component of the glutamic acid binding peptide is glutamic acid and glutamine, it can be when enteron aisle be damaged by vomitoxin, it is horizontal to raise Wnt/ β-catenin signaling pathway protein, improve intestinal stem cell proliferation and differentiation capability, reinforce gut barrier function, to promote enteric epithelium to regenerate, gut integrity is safeguarded, alleviate and damage caused by vomitoxin.The glutamic acid binding peptide has apparent remission effect the damage of the mouse intestinal as caused by vomitoxin by nutrition regulation means, and adds relatively low-dose glutamic acid binding peptide and can be obtained better effects, reduces the toxic reaction to mouse intestinal.Therefore, the glutamic acid binding peptide is applicable not only to the property diarrhea prevention and control of animal feed source, also provides new approach for the exploitation of mankind's intestinal health adjusting control agent.
Description
Technical field
The present invention relates to Animal nutritions and intestinal health technical field, and in particular, to glutamic acid binding peptide is slow in preparation
Solve the application in the preparation of the intestinal tract injury of animal vomitoxin induction.
Background technique
Vomitoxin is the major pollutants of cereal preparation and feed.No matter in developing country or in developed country,
Vomitoxin pollution problem has received widespread attention.After the feed of feed intake vomitoxin pollution, it frequently can lead to grow
Performance decline, serious person vomit, or even dead.Enteron aisle is the primary target of vomitoxin attack, and vomitoxin can destroy
Enteron aisle structure reduces intestinal stem cell activity, inhibits epithelial cell proliferation, induce cell apoptosis, damage barrier function, reduces intestines
Road eventually leads to animal growth and is obstructed to the utilization rate of nutrient.In addition, the vomitoxin in food can also cause diarrhea
And vomiting, seriously endanger human health.
Although traditional physics, chemistry and biological detoxication method can reduce vomitoxin content in feed to a certain extent,
But a series of problem is also brought along simultaneously.Such as: feed palatability and nutritive value do not reduce, there are bio-safety hidden danger and not
It is produced on a large scale.Therefore, more effectively developing or find at present one kind can be effectively relieved caused by vomitoxin
Intestinal tract injury, and it is without side-effects to body or side effect is smaller, and can be carried out large-scale production, feed or food can be added to
In preparation.
Summary of the invention
The purpose of the invention is to overcome the above-mentioned deficiency of the prior art, provides glutamic acid binding peptide and alleviate in preparation
Animal vomitoxin induction intestinal tract injury preparation in application, the glutamic acid binding peptide by nutrition regulation means to by
The damage of mouse intestinal caused by vomitoxin has apparent remission effect.
Another object of the present invention is to provide a kind of preparations of the intestinal tract injury of alleviation animal vomitoxin induction.
Another object of the present invention is to provide glutamic acid binding peptides to promote Wnt/ β-catenin signal path egg in preparation
Application in the preparation of white matter expression.
Another object of the present invention is to provide glutamic acid binding peptides in preparation enhancing intestinal stem cell activity and/or to promote
Application in the preparation of expansion of stem cells.
Another object of the present invention is to provide glutamic acid binding peptides to promote Intestinal epitheliual cell proliferation and/or differentiation in preparation
Preparation in application.
Another object of the present invention is to provide glutamic acid binding peptides to increase goblet cell and/or paneth's cell number in preparation
Application in the preparation of amount.
Another object of the present invention is to provide glutamic acid binding peptides in the preparation that preparation maintains enteron aisle structural integrity
Using.
To achieve the goals above, the present invention is achieved by following scheme:
The present invention passes through the study found that gavaging mouse jejunum weight, jejunum villi height and the diamine oxidase of vomitoxin
Vigor significantly reduces；And after gavaging glutamic acid binding peptide, compared with vomitoxin group, can dramatically increase intestines weight, height of naps and
Diamine oxidase vigor, show glutamic acid binding peptide under vomitoxin degree of impairment to maintain enteron aisle structural integrity have compared with
Good protective effect can quickly repair intestinal mucosa, alleviate and damage caused by vomitoxin.
Therefore, the present invention is claimed glutamic acid binding peptide and is preparing the intestinal tract injury for alleviating the induction of animal vomitoxin
Application in preparation.
The present invention is also claimed glutamic acid binding peptide and increases in enteron aisle in the preparation of diamine oxidase vigor in preparation
Using.
A kind of preparation of the intestinal tract injury of alleviation animal vomitoxin induction is also claimed in the present invention, contains glutamic acid knot
Close peptide；The glutamic acid binding peptide includes glutamic acid and glutamine.
Preferably, the glutamic acid binding peptide gavage or additive capacity be 500~2000mg/kg BW.
It is highly preferred that the glutamic acid binding peptide gavage or additive capacity be 1000mg/kg BW.
Preferably, the animal is mouse.It is highly preferred that the animal is C57BL/6 mouse.
The present inventor further study show that, vomitoxin inhibit Jejunum Crypt cell amplification be class intestines group efficiency and speed
Degree reduces jejunum, Wnt/ β-catenin signal path key protein β-catenin and TCF4 in crypts and class intestines group, active
The expression of type intestinal stem cell mark Lgr5, cell Proliferation mark Ki67 and cell terminal differentiation mark KRT20；And gavage paddy ammonia
After sour binding peptide, the formation efficiency of class intestines group is dramatically increased, Wnt/ β-catenin signal path, Lgr5, Ki67 and KRT20 quilt
Significant up-regulation.Since Wnt/ β-catenin signal path determines intestinal stem cell proliferation and differentiation destiny, it is known that, glutamic acid knot
Peptide is closed by the expression of up-regulation Wnt/ β-catenin signaling pathway protein matter, promotes the proliferation and differentiation of intestinal stem cell, is reversed
Vomitoxin is damaged caused by gut epithelium.
In addition, Claudin-1 is tight junction protein, expression quantity level can partially represent enteron aisle mechanical barrier function just
Whether often；MUC2, LYZ are respectively the mark and paneth's cell mark of goblet cell, and wherein MUC2 is the weight of enteron aisle chemical barrier
Constituent is wanted, LYZ is a part of intestinal immunological barrier.And vomitoxin can inhibit Claudin-1 expression, reduce MUC2 and
LYZ positive cell quantity；After gavaging glutamic acid binding peptide, vomitoxin group is compared, tight junction protein is significantly raised, MUC2
It is dramatically increased with LYZ positive cell quantity.Show that glutamic acid binding peptide helps to enhance gut barrier function.
Therefore, glutamic acid binding peptide is also claimed in preparation promotion Wnt/ β-catenin signaling pathway protein in the present invention
Application in the preparation of matter expression.
The present invention is also claimed glutamic acid binding peptide in preparation enhancing intestinal stem cell activity and/or stem cell is promoted to expand
Application in the preparation of increasing.
Glutamic acid binding peptide is also claimed in the preparation that preparation promotes Intestinal epitheliual cell proliferation and/or differentiation in the present invention
Application.
The preparation that glutamic acid binding peptide increases goblet cell and/or paneth's cell quantity in preparation is also claimed in the present invention
In application.
Application of the glutamic acid binding peptide in the preparation that preparation maintains enteron aisle structural integrity is also claimed in the present invention.
Compared with prior art, the invention has the following advantages:
(1) preparation provided by the present invention contains glutamic acid binding peptide, and main component is glutamic acid and glutamine, energy
Enough when enteron aisle is damaged by vomitoxin, up-regulation Wnt/ β-catenin signaling pathway protein is horizontal, improves intestinal stem cell and increases
It grows and differentiation capability, reinforcement gut barrier function safeguards gut integrity so that enteric epithelium be promoted to regenerate, alleviate vomitoxin
Caused by damage.
(2) preparation provided by the present invention has the damage of the mouse intestinal as caused by vomitoxin by nutrition regulation means
Apparent remission effect, and add relatively low-dose glutamic acid binding peptide and can be obtained better effects, reduce the poison to mouse intestinal
Property reaction.
(3) preparation provided by the present invention is applicable not only to the property diarrhea prevention and control of animal feed source, is also mankind's intestinal health
The exploitation of adjusting control agent provides new approach.
Detailed description of the invention
Fig. 1 is embodiment experiment process figure.
Fig. 2 is mouse jejunum HE colored graph under 100 times of optical microscopies in embodiment.
Fig. 3 is mouse jejunum fluff structures figure under 400 times of scanning electron microscope in embodiment.
Fig. 4 is the influence that embodiment Glutamic Acid binding peptide expands vomitoxin inducing mouse Jejunum Crypt cell.
Fig. 5 is embodiment Glutamic Acid binding peptide to Wnt/ β-in vomitoxin inducing mouse jejunum, crypts and class intestines group
The influence of catenin signal path key protein expression.
Fig. 6 be embodiment Glutamic Acid binding peptide to vomitoxin inducing mouse jejunum, crypts and class intestines group in Ki67 and
The influence of PCNA protein expression.
Fig. 7 is embodiment Glutamic Acid binding peptide to KRT20 egg in vomitoxin inducing mouse jejunum, crypts and class intestines group
The influence of white matter expression.
Fig. 8 is the influence that embodiment Glutamic Acid binding peptide expresses MUC2 in vomitoxin inducing mouse jejunum and LYZ.
Fig. 9 is embodiment Glutamic Acid binding peptide in vomitoxin inducing mouse jejunum, crypts and class intestines group
The influence of Claudin-1 protein expression.
Specific embodiment
With reference to the accompanying drawings of the specification and specific embodiment is made the present invention and is further elaborated, the embodiment
It is served only for explaining the present invention, be not intended to limit the scope of the present invention.Test method as used in the following examples is such as without spy
Different explanation, is conventional method；Used material, reagent etc., unless otherwise specified, for the reagent commercially obtained
And material.
Embodiment
One, test method
1, test material
Vomitoxin: commercially available, purity 100%.
Glutamic acid binding peptide: from Gluten hydrolysis, wherein free amino acid is about 4.59%, and dipeptides~tetrapeptide is about
33%, pentapeptide~octapeptide is about 18%, and octapeptide~ten hexapeptides are about 10%.Glutamic acid and the glutamine accounting in peptides composition
Up to 34%, proline 11.54%.
2, experimental animal
4~5 week old C57BL/6 male and healthy mouse 72 similar in weight is chosen, 6 processing, each place are randomly divided into
Manage 12 repetitions, 1 mouse of each repetition.
The test grouping of table 1
| Group | Gavage scheme |
| Control group | Physiological saline |
| 1 group of adjusting control agent | 1000mg/kg body weight (BW) glutamic acid binding peptide |
| 2 groups of adjusting control agent | 2000mg/kg BW glutamic acid binding peptide |
| Vomitoxin group | 2mg/kg BW vomitoxin |
| 1 group of vomitoxin+adjusting control agent | 2mg/kg BW vomitoxin+1000mg/kg BW glutamic acid binding peptide |
| 2 groups of vomitoxin+adjusting control agent | 2mg/kg BW vomitoxin+2000mg/kg BW glutamic acid binding peptide |
3, process is tested
(1) first 3 days, every morning 8:00, to the nutrient solution or physiological saline of mouse gavaging equal volume.Wherein control group
Physiological saline is gavaged with vomitoxin group, 1 group of adjusting control agent and 1 group of vomitoxin+adjusting control agent gavage 1000mg/kg BW glutamic acid
Binding peptide, 2 groups of adjusting control agent and 2 groups of vomitoxin+adjusting control agent gavage 2000mg/kg BW glutamic acid binding peptide.
(2) 7 days intermediate, control group gavages physiological saline, and adjusting control agent group gavages glutamic acid binding peptide, and vomitoxin group gavages
Vomitoxin, vomitoxin+adjusting control agent group gavage vomitoxin+glutamic acid binding peptide.
(3) the last 3 days, it was identical as first 3 days to gavage situation, gavaged to off-test.Experiment process is as shown in Figure 1.
4, testing index and method
(1) mouse feed intake, amount of drinking water and weight are counted, and weighs duodenum, jejunum and ileum weight.
(2) Senile Mouse
By paraffin section and HE dyeing and scanning electron microscope, intestinal tissue form is observed, intestinal mucosa injury degree is evaluated.
(3) gut integrity is evaluated
Enteron aisle and blood serum sample detect diamine oxidase (diamine oxidase, DAO) vigor.
(4) intestinal stem cell activity rating
By separating and cultivating crypts, the formation efficiency and area of statistics class intestines group.
(5) protein expression
Enteron aisle, crypts and class intestines group sample detection Wnt/ β-catenin signal path key protein and Ki67,
The variation of KRT20 and Claudin-1 protein expression；
Jejunal tissue detects MUC2 and LYZ expression and distribution.
5, data processing and statistical method
Test data is indicated with average value ± standard error (mean ± SEM), is carried out with 19.0 statistical software of SPSS to data
One-way analysis of variance, P < 0.05 indicate that significant difference, P < 0.10 indicate variant significant trend.
Two, test result
1, influence of the glutamic acid binding peptide to vomitoxin inducing mouse growth performance
2 mouse growth performance (g) of table
| Group | Average daily gain | Average daily drink amount | Average daily gain |
| Control group | 2.83±0.06 | 4.43±0.18ab | 0.19±0.02bc |
| 1 group of adjusting control agent | 2.82±0.04 | 4.72±0.21a | 0.23±0.03ab |
| 2 groups of adjusting control agent | 2.78±0.07 | 4.30±0.12ab | 0.28±0.03a |
| Vomitoxin group | 2.72±0.05 | 4.12±0.13b | 0.06±0.02d |
| 1 group of vomitoxin+adjusting control agent | 2.83±0.07 | 4.71±0.19a | 0.19±0.02bc |
| 2 groups of vomitoxin+adjusting control agent | 2.82±0.05 | 4.83±0.17a | 0.21±0.03ab |
Note: having an identical lowercase or do not mark alphabetical person with all after column data, indicates the significant trend of indifference
(P>0.05,Duncan’s)。
As shown in Table 2, compared with control group and adjusting control agent group, vomitoxin group mouse, which is averaged, daily drink amount and averagely to increase day by day
Significantly reduce (P < 0.05) again.And after gavaging glutamic acid binding peptide, compared with vomitoxin group, average daily drink amount and average day
Weight gain dramatically increases (P < 0.05).And (P > 0.05) is had no significant effect to average daily gain.
2, influence of the glutamic acid binding peptide to vomitoxin inducing mouse small intestine weight
3 mouse small intestine of table weight (mg/cm)
| Group | Duodenum | Jejunum | Ileum |
| Control group | 39.48±2.49 | 41.03±1.29ab | 27.30±1.41 |
| 1 group of adjusting control agent | 41.52±2.28 | 43.34±2.06a | 26.90±0.85 |
| 2 groups of adjusting control agent | 36.53±2.13 | 39.76±1.29ab | 26.84±2.76 |
| Vomitoxin group | 35.81±4.18 | 29.15±1.00c | 25.77±2.19 |
| 1 group of vomitoxin+adjusting control agent | 39.49±1.97 | 38.29±1.74b | 30.24±2.07 |
| 2 groups of vomitoxin+adjusting control agent | 38.69±2.05 | 39.40±1.98ab | 28.00±3.87 |
Note: having an identical lowercase or do not mark alphabetical person with all after column data, indicate difference it is not significant (P >
0.05,Duncan’s)。
As shown in Table 3, compared with control group and adjusting control agent group, vomitoxin group jejunum significantly reduces (P < 0.05) again.And
After gavaging glutamic acid binding peptide, compared with vomitoxin group, jejunum dramatically increases (P < 0.05) again.In addition, vomitoxin reduces
Mouse duodenal weight (P < 0.10), glutamic acid binding peptide can increase duodenum weight (P < 0.10) to a certain extent, and right
Ileum weight is without influence.
3, influence of the glutamic acid binding peptide to vomitoxin inducing mouse jejunum morphosis
4 mouse jejunum height of naps of table and height of naps/Crypt depth ratio
| Group | Height of naps (μm) | Height of naps/Crypt depth |
| Control group | 189±5b | 3.4±0.2bc |
| 1 group of adjusting control agent | 215±4a | 3.5±0.2bc |
| 2 groups of adjusting control agent | 149±2d | 2.8±0.1cd |
| Vomitoxin group | 84±4e | 2.4±0.2d |
| 1 group of vomitoxin+adjusting control agent | 210±5a | 5.1±0.3a |
| 2 groups of vomitoxin+adjusting control agent | 178±4c | 4.0±0.2b |
Note: having an identical lowercase or do not mark alphabetical person with all after column data, indicate difference it is not significant (P >
0.05,Duncan’s)。
By Fig. 2,3 and table 4 it is found that compared with control group and 1 group of adjusting control agent, vomitoxin group jejunum villi height and villus
Highly/Crypt depth ratio significantly reduces (P < 0.05).And after gavaging glutamic acid binding peptide, compared with vomitoxin group, villus
Height and height of naps/Crypt depth ratio dramatically increase (P < 0.05).
4, influence of the glutamic acid binding peptide to diamine oxidase vigor in vomitoxin inducing mouse jejunum and serum
Diamine oxidase vigor in 5 mouse jejunum of table and serum
| Group | Jejunum (U/mg) | Serum (U/mL) |
| Control group | 7.23±0.21bc | 13.20±3.09b |
| 1 group of adjusting control agent | 7.25±0.21bc | 18.16±1.84ab |
| 2 groups of adjusting control agent | 8.68±0.49a | 23.98±5.14a |
| Vomitoxin group | 6.47±0.14c | 26.37±2.97a |
| 1 group of vomitoxin+adjusting control agent | 8.32±0.26a | 13.10±2.03b |
| 2 groups of vomitoxin+adjusting control agent | 7.19±0.35bc | 17.67±3.29ab |
Note: having an identical lowercase or do not mark alphabetical person with all after column data, indicate difference it is not significant (P >
0.05,Duncan’s)。
As shown in Table 5, compared with control group and 1 group of adjusting control agent, the reduction of vomitoxin group jejunum diamine oxidase vigor (P <
0.10), serum diamine oxidase vigor significantly increases (P < 0.05), compared with 2 groups of adjusting control agent, vomitoxin group jejunum diamines oxygen
Changing enzyme activity significantly reduces (P<0.05), and serum diamine oxidase vigor is without significant change (P>0.05).And gavage 1000mg/kg
After BW glutamic acid binding peptide, compared with vomitoxin group, jejunum and serum diamine oxidase vigor significantly raise and reduce respectively
(P<0.05).After gavaging 2000mg/kg BW glutamic acid binding peptide, have the tendency that significantly raising and reducing (P < 0.1) respectively.
5, the influence that glutamic acid binding peptide expands vomitoxin inducing mouse Jejunum Crypt cell
As shown in Figure 4, vomitoxin can inhibit the amplification of Jejunum Crypt cell, reduce class intestines group's formation efficiency and area (P
< 0.05) after, gavaging glutamic acid binding peptide, class intestines group's formation efficiency and area dramatically increase (P < 0.05).Show glutamic acid knot
Peptide is closed to increase intestinal stem cell activity, promote expansion of stem cells.
6, influence of the glutamic acid binding peptide to vomitoxin inducing mouse jejunal tissue Wnt/ β-catenin signal path
As shown in Figure 5, vomitoxin can inhibit Wnt/ β-catenin signal path (β-catenin, TCF4 and Lgr5) (P
< 0.05) after, gavaging glutamic acid binding peptide, Wnt/ β-catenin signal path is significantly raised (P < 0.05) or is had on significant
The trend (P < 0.1) of tune.Show that glutamic acid binding peptide by the expression of up-regulation Wnt/ β-catenin signal path, increases Lgr5
Intestinal stem cell activity, reverses vomitoxin to damage caused by gut epithelium.
7, influence of the glutamic acid binding peptide to vomitoxin inducing mouse jejunum Ki67 and KRT20 protein expression
By Fig. 6 and Fig. 7 it is found that vomitoxin proliferation mark Ki67, PCNA and terminal differentiation mark KRT20 capable of inhibiting cell
Expression (P < 0.05), and after gavaging glutamic acid binding peptide, compare vomitoxin group, cell Proliferation mark and terminal differentiation mark
It is significantly raised (P < 0.05) or has the tendency that significantly raising (P < 0.1).Show that glutamic acid binding peptide helps to reinforce enteron aisle
Stem cells hyperplasia and differentiation capability promote the regeneration of epithelium.
8, influence of the glutamic acid binding peptide to vomitoxin inducing mouse jejunum barrier function
By Fig. 8,9 it is found that vomitoxin can be reduced MUC2 and LYZ positive cell quantity (P < 0.05), inhibition is close to be connected
PROTEIN C laudin-1 expresses (P < 0.05), and after gavaging glutamic acid binding peptide, vomitoxin group is compared, MUC2 and LYZ are positive thin
Born of the same parents' quantity dramatically increases (P < 0.05), and tight junction protein is significantly raised (P < 0.05).Show that the enhancing of glutamic acid binding peptide is vomitted
It spits toxin and damages lower mouse intestinal barrier function.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than protects to the present invention
The limitation of shield range can also be made on the basis of above description and thinking for those of ordinary skill in the art
Other various forms of variations or variation, there is no necessity and possibility to exhaust all the enbodiments.It is all of the invention
Made any modifications, equivalent replacements, and improvements etc., should be included in the protection of the claims in the present invention within spirit and principle
Within the scope of.
Claims (10)
1. application of the glutamic acid binding peptide in the preparation that the intestinal tract injury of animal vomitoxin induction is alleviated in preparation.
2. a kind of preparation for the intestinal tract injury for alleviating the induction of animal vomitoxin, which is characterized in that contain glutamic acid binding peptide；Institute
Stating glutamic acid binding peptide includes glutamic acid and glutamine.
3. preparation according to claim 2, which is characterized in that the glutamic acid binding peptide gavage or additive capacity be 500
~2000mg/kg BW.
4. preparation according to claim 3, which is characterized in that the glutamic acid binding peptide gavage or additive capacity is
1000mg/kg BW。
5. preparation according to claim 2, which is characterized in that the animal is mouse.
6. application of the glutamic acid binding peptide in the preparation that preparation promotes the expression of Wnt/ β-catenin signaling pathway protein matter.
7. application of the glutamic acid binding peptide in preparation enhancing intestinal stem cell activity and/or the preparation for promoting expansion of stem cells.
8. application of the glutamic acid binding peptide in the preparation that preparation promotes Intestinal epitheliual cell proliferation and/or differentiation.
9. application of the glutamic acid binding peptide in the preparation that preparation increases goblet cell and/or paneth's cell quantity.
10. application of the glutamic acid binding peptide in the preparation that preparation maintains enteron aisle structural integrity.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111849860A (en) * | 2020-06-01 | 2020-10-30 | 浙江大学 | Method for regulating and controlling intestinal stem cell differentiation by using iron element and application |
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| CN1031025A (en) * | 1987-05-01 | 1989-02-15 | 大制药株式会社 | Nutritive emulsion preparation |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111849860A (en) * | 2020-06-01 | 2020-10-30 | 浙江大学 | Method for regulating and controlling intestinal stem cell differentiation by using iron element and application |
| CN111849860B (en) * | 2020-06-01 | 2021-12-21 | 浙江大学 | Method for regulating and controlling intestinal stem cell differentiation by using iron element and application |
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Application publication date: 20190604 |