CN117298141B - Application of polysaccharide in reducing chicken abdomen fat deposition - Google Patents
Application of polysaccharide in reducing chicken abdomen fat deposition Download PDFInfo
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- A—HUMAN NECESSITIES
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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Abstract
The invention provides an application of polysaccharide in reducing fat deposition on chicken abdomen, and belongs to the technical field of livestock breeding. The invention explores the effect of the fructus viticis fruit polysaccharide with different molecular weights on chicken precursor fat cells, and the result shows that when the 0-5kDa fructus viticis fruit polysaccharide reaches 10 mug/mL, the fructus viticis fruit polysaccharide has remarkable effects of inhibiting proliferation, inhibiting differentiation and reducing fat deposition on chicken precursor fat cells. The invention proves that the 0-5kDa fructus viticis fruit polysaccharide is a potential chicken fat regulator, so that the fructus viticis fruit polysaccharide can be used for developing a biological agent for improving excessive fat deposition of broiler chickens.
Description
Technical Field
The invention belongs to the technical field of livestock breeding, and particularly relates to application of polysaccharide in reducing fat deposition on chicken abdomen.
Background
As important economic birds, the current large-scale breeding of broiler chickens has been developed in a great deal worldwide. For many years, through scientific and reasonable selective breeding, the growth speed and the meat yield of broilers are greatly improved. However, while weight gain is being pursued, excessive deposition of fat in broiler chickens is also becoming increasingly serious. When the feed is overabundant, the abdominal fat of the broiler chickens can be deposited rapidly, so that not only is the feed conversion efficiency affected, but also the meat quality is reduced, and the sustainable development of the broiler chickens industry is seriously affected and restricted.
Chicken precursor adipocytes are precursor cells of adipocytes, which are present in adipose tissues and can differentiate into mature adipocytes. When the body takes excessive energy, precursor fat cells can largely differentiate and proliferate to form more fat cells for storing fat. During the growth of broilers, if the feed energy is excessive, precursor fat cells are activated to differentiate, increasing the number of fat cells and fat storage, thereby causing obesity and fat accumulation. Limiting the differentiation of precursor adipocytes can be effective in reducing adipogenesis. Therefore, the proliferation and differentiation of chicken precursor fat cells are effectively regulated, and the method has important significance for improving the feed conversion efficiency of broiler chickens.
Fructus Viticis is a common herb of Chinese traditional medicine for clearing heat, purging fire, detoxicating and detumescence. Fructus Viticis negundo fruit contains abundant vitamin C, pectin, polysaccharide, etc., wherein the polysaccharide is used as one of main active components of fructus Viticis negundo, and has certain medicinal value. However, the research on the fruit polysaccharide of the fructus viticis is relatively weak at present, and particularly, the application research on the aspect of chicken fat deposition regulation is not reported. Therefore, the invention develops more systematic research on the fructus viticis fruit polysaccharide, and aims to develop a new natural plant extract for solving the problem of chicken fat deposition.
Disclosure of Invention
The invention aims to provide an application of polysaccharide in reducing chicken abdominal fat deposition, thereby providing a new effective solution for solving the problem of chicken abdominal fat deposition.
In order to achieve the above purpose, the present invention provides the following technical solutions:
firstly, the invention provides a second aspect, the invention provides an application of polysaccharide in preparing biological agent for reducing chicken abdomen fat deposition, the polysaccharide is 0-5kDa fructus viticis fruit polysaccharide,
preferably, the preparation method of the fructus viticis fruit polysaccharide comprises the following steps:
(a) Grinding the dried fructus Viticis into powder to obtain fructus Viticis powder;
(b) According to sterile water: powder = 20mL: adding sterile water in a proportion of 1g, uniformly stirring, heating to 90 ℃ and soaking and extracting for 3 hours;
(c) Centrifuging to remove precipitate to obtain fructus Viticis negundo fruit extract;
(d) Concentrating fructus Viticis fruit extract by rotary evaporation to 1/5 of the original volume, adding 4 times of anhydrous ethanol, and precipitating with ethanol to obtain fructus Viticis fruit crude polysaccharide;
(e) Dissolving fructus Viticis negundo fruit crude polysaccharide into crude polysaccharide solution by using distilled water, repeating for 8 times by using sevage method, and removing protein in the polysaccharide solution to obtain fructus Viticis negundo fruit polysaccharide solution;
(f) Filtering with 5kDa ultrafilter membrane to obtain 0-5kDa filtrate;
(g) Precipitating the obtained 0-5kDa filtrate with ethanol, and lyophilizing to obtain fructus Vitics Simplicifoliae polysaccharide.
Preferably, the concentration of the fructus viticis fruit polysaccharide is more than or equal to 10 mug/mL.
Preferably, in the biological agent, the concentration of the fructus viticis fruit polysaccharide is 10-200 mug/mL.
Preferably, the biological agent achieves the effect of reducing fat deposition in the abdomen of a chicken by reducing proliferation of chicken precursor fat cells, adipogenic differentiation and lipid droplet formation.
Secondly, the invention provides application of polysaccharide in preparation of biological preparation for inhibiting proliferation of chicken precursor fat cells, wherein the polysaccharide is 0-5kDa fructus viticis fruit polysaccharide.
Preferably, the fructus viticis polysaccharide is prepared by the preparation method of the fructus viticis polysaccharide.
Preferably, the concentration of the fructus viticis fruit polysaccharide in the biological agent is 10-200 mug/mL.
Secondly, the invention provides application of polysaccharide in preparation of biological preparation for inhibiting chicken precursor fat cell differentiation, wherein the polysaccharide is 0-5kDa fructus viticis fruit polysaccharide.
Preferably, the fructus viticis polysaccharide is prepared by the preparation method of the fructus viticis polysaccharide.
Preferably, the concentration of the fructus viticis fruit polysaccharide in the biological agent is 10-200 mug/mL.
Secondly, the invention provides application of polysaccharide in preparation of biological agents for inhibiting lipid drop formation in chicken precursor fat cells, wherein the polysaccharide is 0-5kDa fructus viticis fruit polysaccharide, and the fructus viticis fruit polysaccharide is prepared by the preparation method of the fructus viticis fruit polysaccharide.
Preferably, the concentration of the fructus viticis fruit polysaccharide in the biological agent is 10-200 mug/mL.
Secondly, the invention provides a biological agent for reducing fat deposition in the abdomen of chickens, which is characterized in that the biological agent is prepared by the following steps:
(1) With reference to the preparation method, the fructus viticis fruit polysaccharide is prepared;
(2) Adding fructus Viticis negundo fruit polysaccharide into sterile distilled water to obtain 50 μg/mL fructus Viticis negundo fruit polysaccharide solution;
(3) The biological preparation was obtained after filtration sterilization with a 0.22 μm filter membrane.
The invention has the beneficial effects that:
the invention explores the influence of the fructus viticis fruit polysaccharide with different molecular weights on the proliferation of chicken precursor fat cells, and the result shows that the fructus viticis fruit polysaccharide with 0-5kDa can effectively inhibit the proliferation of chicken precursor fat cells;
further experiments show that the low concentration of the fructus viticis fruit polysaccharide with the molecular weight of 10 mug/mL can effectively inhibit the adipogenic differentiation of chicken precursor fat cells and the formation of intracellular fat drops, thereby effectively reducing the production of chicken abdominal fat and the deposition of chicken abdominal fat. Thus, the 0-5kDa fructus Viticis negundo fruit polysaccharide is a potential natural active molecule, which can be made into biological preparation for improving chicken abdominal fat deposition.
Drawings
FIG. 1 shows the relative expression levels of PPARgamma genes in chicken precursor adipocytes after treatment with different concentrations of polysaccharide;
in fig. 1, P < 0.01 and P < 0.001;
FIG. 2 shows the relative expression levels of the C/EBP alpha gene in chicken precursor adipocytes after treatment with different concentrations of polysaccharide;
in fig. 2, P < 0.01 and P < 0.001;
FIG. 3 shows differences in lipid droplet formation in chicken precursor adipocytes after treatment with different concentrations of polysaccharide;
FIG. 4 shows the difference in absorbance after lipid droplet extraction in chicken precursor adipocytes after treatment with different concentrations of polysaccharide;
in fig. 4, P < 0.05 and P < 0.01.
Detailed Description
The present invention will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
EXAMPLE 1 acquisition and culture of chicken precursor adipocytes
(a) Selecting commercial broilers of 2 weeks old, after sacrifice, aseptically collecting abdominal adipose tissue, placing into a plate which is poured into PBS (phosphate buffer solution) in advance, repeatedly washing, and removing blood vessels and fascia;
(b) Shearing the adipose tissues by using an ophthalmic scissors, and placing the sheared tissues into a test tube added with digestive juice;
(c) After digestion for 60min at 37 ℃ (shaking and mixing uniformly every 5 min), adding DMEM/F12 culture medium to terminate digestion after digestion is finished;
(d) After blowing with a suction pipe, filtering with a stainless steel screen mesh of 100 meshes and 600 meshes;
(e) Subpackaging the filtrate into a centrifuge tube, setting the parameters of the centrifuge to 2000rpm, centrifuging for 10min, removing the culture medium after the centrifugation is finished, and re-suspending cells by using erythrocyte lysate to prepare cell suspension;
(f) After incubation for 10min at room temperature, centrifugation for 10min at 2000rpm, washing the obtained cell pellet with culture medium for 2 times, re-suspending cells with culture medium to obtain chicken precursor fat cells,placing in a cell incubator at 37deg.C with 5% CO 2 Culturing under the condition.
Example 2: preparing fructus Viticis negundo fruit polysaccharide with different molecular weights
(a) Grinding the dried fructus Viticis into powder to obtain fructus Viticis powder;
(b) According to sterile water: powder = 2000mL: adding 100g of sterile water, uniformly stirring, heating to 90 ℃ and soaking and extracting for 3 hours;
(c) Centrifuging at 5000rpm for 25min to remove precipitate to obtain fructus Vitics Simplicifoliae fruit extractive solution;
(d) Concentrating fructus Viticis fruit extract into 400mL by rotary evaporation, adding 1600mL absolute ethanol, and precipitating with ethanol at 4deg.C for 24 hr to obtain fructus Viticis fruit crude polysaccharide;
(e) Dissolving fructus Viticis negundo fruit crude polysaccharide into 1mg/mL crude polysaccharide solution with distilled water, repeating for 8 times with sevage method, and removing protein in polysaccharide solution to obtain fructus Viticis negundo fruit polysaccharide solution;
(f) Filtering with 5kDa ultrafiltration membrane to obtain 0-5kDa filtrate 1, filtering the retentate with 10kDa ultrafiltration membrane to obtain 5-10kDa filtrate 2, and filtering the retentate with 30kDa ultrafiltration membrane to obtain 10-30kDa filtrate 3;
(g) Precipitating the obtained 0-5kDa filtrate 1,5-10kDa filtrate 2 and 10-30kDa filtrate 3 with ethanol, and lyophilizing to obtain fructus Vitics Simplicifoliae polysaccharide 1, fructus Vitics Simplicifoliae polysaccharide 2 and fructus Vitics Simplicifoliae polysaccharide 3.
Example 3: investigation of the Effect of different molecular weight of the fruit polysaccharide of Vitex negundo on proliferation of chicken precursor adipocytes
(a) Washing chicken precursor fat cells in logarithmic growth phase with PBS for 2 times, digesting with pancreatin, and adding DMEM complete medium to terminate digestion;
(b) Collecting the culture medium into a centrifuge tube, centrifuging to remove supernatant, and adjusting cell density to 1×10 with DMEM complete medium 5 /mL；
(c) Cells were seeded at 9 in an amount of 100ul per wellIn a 6-well plate at 37℃in 5% CO 2 Culturing in a cell incubator;
(d) After the cells were completely adherent, the medium was aspirated, the control group was replaced with fresh DMEM medium, the polysaccharide 1 treatment group was replaced with 100 μg/mL of the solution of the fruit polysaccharide 1 of chaste tree (0.22 μm filter membrane degerming after DMEM medium preparation), the polysaccharide 2 treatment group was replaced with 100 μg/mL of the solution of the fruit polysaccharide 2 of chaste tree, the polysaccharide 3 treatment group was replaced with 100 μg/mL of the solution of the fruit polysaccharide 3 of chaste tree, and each treatment was provided with 3 multiplex wells.
(f) After 48h of cultivation, the absorbance at 450nm was measured using CCK-8, and the change of chicken precursor adipocytes after treatment with different fructus Viticis negundo fruit polysaccharides was compared, and the measurement results are shown in Table 1.
TABLE 1 absorbance change of cells after treatment with different fruit polysaccharides of Vitex negundo
From table 1 we can see that the absorbance was significantly reduced and the difference was very significant in the polysaccharide 1 treated group compared to the control group;
compared with the control group, the absorbance of the polysaccharide 2 treatment group is slightly lower than that of the control group, and the difference is obvious;
the absorbance of the polysaccharide 3 treated group is not obviously different from that of the control group;
the 0-5kDa fructus viticis fruit polysaccharide adopted by the polysaccharide 1 treatment group has a remarkable inhibition effect on proliferation of chicken precursor fat cells, and the balance of the molecular weight is poor, so that the 0-5kDa fructus viticis fruit polysaccharide 1 is selected for subsequent research.
Example 4: exploration of suitable concentrations of polysaccharide for inhibition of proliferation of chicken precursor adipocytes
(a) Preparing fructus Viticis negundo fruit polysaccharide 1 into 10 μg/mL, 25 μg/mL,50 μg/mL,100 μg/mL, 200 μg/mL solution, and filtering for later use;
(b) Washing chicken precursor fat cells in logarithmic growth phase with PBS for 2 times, digesting with pancreatin, and adding DMEM complete medium to terminate digestion;
(c) Collecting the culture medium into a centrifuge tube, centrifuging to remove supernatant, and adjusting cell density to 1×10 with DMEM complete medium 5 /mL；
(d) Cells were seeded in 96-well plates at 100ul per well at 37℃with 5% CO 2 Culturing in a cell incubator;
(e) After the cells are completely adhered, the culture medium is sucked, the control group is replaced by a new DMEM culture medium, the polysaccharide 1 treatment groups 1-5 are respectively added with 10-200 mug/mL of the fructus viticis fruit polysaccharide 1 solution, and each treatment is provided with 3 compound holes;
(f) After 48h incubation, absorbance at 450nm was measured using CCK-8 and the change in chicken precursor adipocytes after treatment with different polysaccharides was compared and the results of the measurements are shown in Table 2.
TABLE 2 absorbance changes of cells after treatment with different concentrations of fruit polysaccharide 1 from Vitex negundo
As can be seen from table 2, there was no significant difference in polysaccharide 1 treatment group 1 compared to the control group, with a P value greater than 0.05; the polysaccharide 1 treatment group 2 has a certain degree of difference, and the P value is less than 0.05; the differences among polysaccharide 1 treatment groups 3, 4 and 5 are obvious, and the P value is less than 0.01;
the results show that when the concentration of the solution of the fruit polysaccharide 1 of the fructus viticis is more than or equal to 25 mug/mL, the proliferation of chicken precursor fat cells can be effectively inhibited, when the concentration of the solution of the fruit polysaccharide 1 of the fructus viticis is equal to 50 mug/mL, the optimal concentration is achieved, and the inhibition effect of the fruit polysaccharide 1 of the fructus viticis on the chicken precursor fat cells is not remarkably increased after the concentration is continuously increased. The reason for this is probably that the target point of action of the fructus viticis fruit polysaccharide 1 is saturated, and further inhibition effect is not generated even if the fructus viticis fruit polysaccharide 1 is continuously increased.
Meanwhile, since further increasing the concentration did not lead to a decrease in cell proliferation, it was speculated that the fructus Viticis negundo fruit polysaccharide 1 exerted an inhibitory effect on chicken precursor adipocytes, not by a cytotoxic effect.
Example 5: exploration of Regulation and control of the Equisetum fruit polysaccharide 1 on the chicken precursor adipocyte differentiation related genes PPARgamma and C/EBPalpha
RNA extraction
(a) Washing chicken precursor fat cells in logarithmic growth phase with PBS for 2 times, digesting with pancreatin, and adding DMEM complete medium to terminate digestion;
(b) Collecting the culture medium into a centrifuge tube, centrifuging to remove supernatant, and adjusting cell density to 1×10 with DMEM complete medium 5 /mL；
(c) Inoculating cells into a 6-hole cell culture plate, wherein each hole is 2mL, after the cells are completely adhered, the control group is replaced by a new DMEM complete culture medium, the treatment group 1 is replaced by 10 mug/mL of the solution of the fructus viticis polysaccharide 1, the treatment group 2 is replaced by 25 mug/mL of the solution of the fructus viticis polysaccharide 1, and the treatment group 3 is replaced by 50 mug/mL of the solution of the fructus viticis polysaccharide 1;
(d) Oleic acid is added to induce differentiation, after 48 hours, the culture medium is removed, and after PBS is used for cleaning cells, 1mL of precooled Trizol solution is added into each hole;
(e) Collecting the solution into a centrifuge tube, and standing at room temperature for 5min to completely lyse the cells;
(f) Adding 200ul of precooled chloroform into each centrifuge tube, tightly covering a tube cover, vibrating for 15 seconds, uniformly mixing, and then placing on ice for 15 minutes to fully perform phase separation;
(g) Centrifuging at 12000 rpm and 4 ℃ for 15 minutes, carefully sucking the supernatant without the interface substance by a pipette, and transferring the supernatant into a new tube;
(h) Adding 0.5mL of precooled isopropanol into the supernatant, covering a tube cover, gently reversing and mixing, centrifuging at a temperature of 12000 r/min and a temperature of 4 ℃ for 10 minutes, discarding the supernatant, and reserving RNA sediment at the bottom of the tube;
(i) Adding 1mL of 75% ethanol into the RNA precipitate, gently shaking the tube to wash the RNA precipitate, centrifuging at 8000 rpm and 4 ℃ for 5 minutes, discarding the supernatant, and air-drying the RNA precipitate for 5-10 minutes until the residual ethanol volatilizes;
(j) RNA was reconstituted by adding 30. Mu.L of DEPC water and the quality and concentration of RNA was measured by UV spectrophotometry.
RNA reverse transcription reaction
Reverse transcription of RNA with reference to reverse transcription reagents:
(a) Removal of genomic DNA
The reaction system of the reactant is as follows:
5×gDNA Eraser Buffer2.0μL，
gDNA Eraser1.0μL，
Total RNA1.0μg，
RNase Free dH 2 Oup to 10.0μL；
the reaction conditions of the reactant are as follows: 42 ℃ for 2 minutes at 4 ℃;
(b) Reverse transcription reaction
The reaction system of the reactant is as follows:
10.0. Mu.L of the reaction solution in the step (a),
PrimeScript RT Enzyme Mix I1.0μL，
RT Primer Mix1.0 μL，
5×PrimeScript Buffer 24.0 μL，
RNase Free dH 2 O4.0 μL。
the reaction conditions of the reactant are as follows: 37℃for 15 minutes, 85℃for 5 seconds, 4 ℃.
3. Fluorescent quantitative PCR reaction
The reaction system of the reactant is as follows:
SYBR Green Premix Ex Taq II（2×）10.0μL，
forward primer: 0.4. Mu.L of the total amount,
reverse primer: 0.4. Mu.L of the total amount,
2.0. Mu.L of cDNA template,
ddH 2 O7.2μL。
the reaction conditions of the reactant are as follows: 95 ℃ for 5min;95 ℃ 60s,62 ℃ 40s 35 cycles; and at 72℃for 5min.
The primer sequences of the chicken PPARgamma gene are as follows:
an upstream primer: CATCATGCCATGCAGTCTGC;
a downstream primer: CACACAAAGCGACGTAAGCC;
the primer sequences of the chicken C/EBP alpha gene are as follows:
an upstream primer: CGAGCACTCCATCGACATCA;
a downstream primer: TTGTGCTTCTCCTGCTGCTT;
4. the obtained data is adopted to adopt 2 -△△Ct The relative expression levels of the chicken PPARgamma gene and the chicken C/EBP alpha gene are calculated by the method, and the obtained results are shown in figures 1 and 2.
In FIG. 1, the relative expression level of PPARgamma gene in treatment group 1 was 0.733.+ -. 0.045, the relative expression level of PPARgamma gene in treatment group 2 was 0.537.+ -. 0.060, and the relative expression level of PPARgamma gene in treatment group 3 was 0.501.+ -. 0.044, as compared with the control group;
in FIG. 2, the relative expression level of C/EBP.alpha.in treatment group 1 was 0.702.+ -. 0.044, in treatment group 2 was 0.497.+ -. 0.042, and in treatment group 3 was 0.436.+ -. 0.041, compared to the control group.
It can be seen that when the concentration of the solution of the fructus viticis fruit polysaccharide 1 reaches 10 mug/mL, obvious inhibition effect can be generated on the expression of PPARgamma genes and C/EBP alpha in the adipogenic differentiation process of chicken precursor fat cells, so that the adipogenic differentiation of chicken precursor fat cells can be effectively inhibited.
Meanwhile, as the concentration of the solution of the fruit polysaccharide 1 of the fructus viticis continuously increases, the solution can generate a dose-dependent inhibition effect on the expression of the adipogenic differentiation PPARgamma gene and the C/EBPalpha of chicken precursor adipocytes.
Example 6: exploration of the Regulation of fat deposition in chicken precursor fat cells by the fruit polysaccharide 1 of Vitex negundo
(a) Washing chicken precursor fat cells in logarithmic growth phase with PBS for 2 times, digesting with pancreatin, and adding DMEM complete medium to terminate digestion;
(b) Collecting the culture medium into a centrifuge tube, centrifuging to remove supernatant, and adjusting cell density to 1×10 with DMEM complete medium 5 /mL；
(c) Inoculating cells into a 6-hole cell culture plate, wherein each hole is 2mL, after the cells are completely adhered, the control group is replaced by a new DMEM complete culture medium, the treatment group 1 is replaced by 10 mug/mL of the solution of the fructus viticis polysaccharide 1, the treatment group 2 is replaced by 25 mug/mL of the solution of the fructus viticis polysaccharide 1, and the treatment group 3 is replaced by 50 mug/mL of the solution of the fructus viticis polysaccharide 1;
(d) Adding oleic acid to induce differentiation, removing culture medium after 48h, washing cells for 2 times by using PBS, and adding 4% paraformaldehyde to fix the cells for 30min;
(e) Sucking paraformaldehyde, and adding 500 mu L of oil red O staining solution into each hole for staining;
(f) After staining for 30min, the cells were washed 3 times with PBS, photographed under a microscope and recorded, and the results obtained are shown in fig. 3;
(g) Oil red O in the cells was extracted by adding 500. Mu.L of isopropyl alcohol, and after 20 minutes, each group of the extracted isopropyl alcohol was collected and added to a 96-well plate, and the mixture was subjected to detection of absorbance at 510nm in a microplate reader, and the results were shown in FIG. 4.
As can be seen from the results of fig. 3 and 4, as the treatment concentration of the fructus viticis fruit polysaccharide 1 increases, the number of lipid droplets dyed by the red oil O in the chicken precursor fat cells gradually decreases (fig. 3), and the absorbance value also gradually decreases (fig. 4), which indicates that the fat deposition of the chicken precursor fat cells can be effectively reduced along with the fructus viticis fruit.
Meanwhile, it can be seen that when the concentration of the solution of the fructus viticis fruit polysaccharide 1 reaches 10 mug/mL, a remarkable effect can be generated, which shows that the fructus viticis fruit polysaccharide 1 provided by the invention has a remarkable inhibiting effect on fat deposition of chicken precursor fat cells, and can realize an excellent inhibiting effect at a low concentration.
Example 7: biological agent 1 for reducing fat deposition in abdomen of chicken
(1) With reference to the preparation method of example 2, fructus viticis fruit polysaccharide 1 is prepared;
(2) Adding the fructus viticis fruit polysaccharide 1 into sterile distilled water to prepare 10 mug/mL fructus viticis fruit polysaccharide solution;
(3) After filtration sterilization with a 0.22 μm filter membrane, biological preparation 1 was obtained.
Example 8: biological agent 2 for reducing fat deposition in abdomen of chicken
(1) With reference to the preparation method of example 2, fructus viticis fruit polysaccharide 1 is prepared;
(2) Adding fructus Viticis negundo fruit polysaccharide 1 into sterile distilled water to obtain 25 μg/mL fructus Viticis negundo fruit polysaccharide solution;
(3) After filtration sterilization with a 0.22 μm filter membrane, biological preparation 2 was obtained.
Example 9: biological agent 3 for reducing fat deposition in abdomen of chicken
(1) With reference to the preparation method of example 2, fructus viticis fruit polysaccharide 1 is prepared;
(2) Adding fructus Viticis negundo fruit polysaccharide 1 into sterile distilled water to obtain 50 μg/mL fructus Viticis negundo fruit polysaccharide solution;
(3) After filtration sterilization with a 0.22 μm filter membrane, biological preparation 3 was obtained.
Claims (2)
1. Use of a polysaccharide in the preparation of a preparation for reducing fat deposition in chicken abdomen, characterized in that the polysaccharide is a 0-5kDa fructus viticis fruit polysaccharide;
the preparation method of the fructus viticis polysaccharide comprises the following steps:
(a) Grinding the dried fructus Viticis into powder to obtain fructus Viticis powder;
(b) According to sterile water: powder = 20mL: adding sterile water in a proportion of 1g, uniformly stirring, heating to 90 ℃ and soaking and extracting for 3 hours;
(c) Centrifuging to remove precipitate to obtain fructus Viticis negundo fruit extract;
(d) Concentrating fructus Viticis fruit extract by rotary evaporation to 1/5 of the original volume, adding 4 times of anhydrous ethanol, and precipitating with ethanol to obtain fructus Viticis fruit crude polysaccharide;
(e) Dissolving fructus Viticis negundo fruit crude polysaccharide into crude polysaccharide solution by using distilled water, repeating for 8 times by using sevage method, and removing protein in the polysaccharide solution to obtain fructus Viticis negundo fruit polysaccharide solution;
(f) Filtering with 5kDa ultrafilter membrane to obtain 0-5kDa filtrate;
(g) Precipitating the obtained 0-5kDa filtrate with ethanol, and lyophilizing to obtain fructus Vitics Simplicifoliae polysaccharide;
the concentration of the fructus viticis fruit polysaccharide is 50 mug/mL.
2. Use of a polysaccharide according to claim 1 for the preparation of a formulation for reducing abdominal fat deposition in chickens, wherein the formulation achieves the effect of reducing abdominal fat deposition in chickens by reducing proliferation of chicken precursor adipocytes, adipogenic differentiation and lipid droplet formation.
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| CN116019165A (en) * | 2023-02-23 | 2023-04-28 | 广西大学 | Plant extract for reducing abdominal fat deposition of broiler chickens and improving meat quality and application thereof |
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