CN112006169A - Application of composition containing plant exosome freeze-dried powder in preparation of intestinal tract conditioning feed - Google Patents
Application of composition containing plant exosome freeze-dried powder in preparation of intestinal tract conditioning feed Download PDFInfo
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Abstract
The invention relates to the field of pet foods, in particular to application of a composition containing plant exosome freeze-dried powder in preparing a diet for conditioning intestinal tracts. The invention provides application of a composition containing plant exosome freeze-dried powder in preparing a feed for conditioning intestinal tracts, wherein the plant exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin. The composition for conditioning the pet intestinal tract can effectively condition the pet intestinal flora.
Description
Technical Field
The invention relates to the field of pet foods, in particular to application of a composition containing plant exosome freeze-dried powder in preparing a diet for conditioning intestinal tracts.
Background
The living mode of people is greatly changed due to the rapid development of the society, more and more people are raised for pets, the raised pets are natural emotion regulators for people, and the loneliness can be resolved and the stress can be relieved when the raised pets are combined with the pets. As the role of pets in pet-raising households becomes more and more important, people are increasingly turning their attention to pet health, where gut health is of particular importance.
The intestinal tract is the main site for the animals to digest and absorb nutrients and is the first barrier to directly contact with the outside, and the intestinal health is concerned with the growth and normal life activities of the animals. Whether the intestinal tract is healthy or not is generally judged, whether the digestion and the absorption of food are normal or not, whether the intestinal tract is diseased or not, whether a normal and stable intestinal microbial environment, an intestinal immune barrier and the like exist or not are judged, and if a certain characteristic appears, the normal life activity of an organism is influenced. The pet food intake has certain passivity, and certain obstacle exists in the communication with people, so when the pet owner finds that the intestinal health of the pet is in a problem, the life health of the pet is often influenced to a certain extent. Among them, the role of the intestinal flora in maintaining the host's physiology is particularly important, and disturbances of the intestinal flora can cause many physiological diseases, including metabolic diseases.
At present, the food for improving the intestinal health of pets on the market is prepared by adding probiotics into daily ration, and the probiotics are only used as an auxiliary treatment or health care product for regulating gastrointestinal tracts or patients with young and old digestive tract diseases or preventing external bacterial infection after operation, and cannot promote the proliferation of intestinal beneficial flora and restore the intestinal microecological balance.
Exosome is a small cup vesicle with a double-layer membrane structure and a diameter of about 30-100nm, secreted by various living cells, has a lipid bilayer structure, can carry a large amount of protein, lipid, mRNA and siRNA, is easy to fuse with receptor cells, and researches report that exosome can participate in processes such as cell communication, cell migration, angiogenesis, immunoreaction and the like.
At present, animal-derived exosomes are researched and applied more, but the animal cell-derived exosomes are complex in extraction process, high in cost, low in yield, easy to cause side reactions in the application process and have certain safety problems. The exosome from plant sources is extracted and applied, the source is reliable and stable, the yield is relatively high, and the biocompatibility and the safety are more reliable. However, the activity of the plant exosomes is easily affected by temperature preservation conditions and the like, and the application of the plant exosomes and the efficacy of the product are limited, so that the research and the application of the plant exosomes are less, and particularly, the research on regulating intestinal tracts by the plant exosomes is less.
Disclosure of Invention
The invention aims to overcome the defects of the background art and provides a composition for conditioning the intestinal tract of pets.
The purpose of the invention is realized by the following technical scheme:
in a first aspect, the invention provides the use of a composition comprising a lyophilized powder of plant exosomes in the preparation of a diet for conditioning the intestinal tract, wherein the lyophilized powder of plant exosomes is sequentially embedded in mannitol and bovine serum albumin.
In a preferred embodiment, the plant exosome lyophilized powder is not ginger exosome lyophilized powder or turmeric exosome lyophilized powder.
In a preferred embodiment, the plant exosome lyophilized powder is an onion exosome lyophilized powder.
In a preferred embodiment, the plant exosome lyophilized powder is 0.5-5 parts by weight.
In a preferred embodiment, the plant exosome lyophilized powder is 0.5-2 parts by weight.
In a preferred embodiment, the plant exosome lyophilized powder is 1-3 parts by weight.
In a preferred embodiment, the plant exosome lyophilized powder is 1.5-4 parts by weight.
In a preferred embodiment, the plant exosome lyophilized powder is 3-5 parts by weight.
In a preferred embodiment, the method for preparing the plant exosome freeze-dried powder comprises the following steps:
s1, extraction of plant exosomes
Weighing 100-; centrifuging the solution in the middle layer at 10000-12000 Xg and 4 ℃ for 50-60 min; taking the supernatant, ultracentrifuging at 120000-150000 Xg and 4 ℃ for 50-70min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000-150000 Xg and 4 ℃ for 50-70 min; after centrifugation, resuspending the precipitate with 1-2mL sterile PBS to obtain the plant exosome extract;
s2. embedding of plant exosomes
First embedding of plant exosomes
1) Preparing a mannitol solution, uniformly mixing 15-30mL of the prepared mannitol solution with 6-12mL of the plant exosome extracting solution, adding 40-60mL of edible oil, and stirring at 200-300r/min for 15-25 min;
2) dropwise adding 20-30mL of edible oil containing 0.5-1g of glacial acetic acid, and stirring for 15-25min at 300r/min under 200-;
3) adding 100-150mL acetate buffer solution, standing for 3-5h, centrifuging, and removing an oil layer;
second embedding of plant exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20-30mL of prepared bovine serum albumin solution;
3) stirring at 300r/min for 15-25min at 200-;
s3, freeze-drying of plant exosomes
Freezing the extracted plant exosomes at about-40 to-35 ℃ for 1 to 3 hours until the central temperature of the material is-35 to-25 ℃, then placing the material on a heating plate for heating, raising the temperature of the heating plate to 90 to 100 ℃ within 0.5 to 1.5 hours, and maintaining the temperature for 4 to 6 hours; reducing the temperature of the heating plate to 80-90 ℃ within 0.5-1.5h, and maintaining the temperature for 4-6 h; reducing the temperature of the heating plate to 55-65 ℃ within 0.5-1.5h, and maintaining the temperature for 4-6 h; and (3) reducing the temperature of the heating plate to 40-50 ℃ within 0.5-1.5h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out the material to obtain the plant exosome freeze-dried powder.
In a preferred embodiment, the composition further comprises flour, corn flour, bone flour, pumpkin flour, shrimp shell flour, seaweed flour, milk powder and water.
In a preferred embodiment, the composition further comprises 10-20 parts of flour, 20-30 parts of corn flour, 2-10 parts of bone meal, 5-15 parts of pumpkin powder, 2-5 parts of shrimp shell powder, 3-8 parts of seaweed powder, 5-10 parts of milk powder and 40-60 parts of water according to parts by weight.
In a preferred embodiment, the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; and (3) reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out the material to obtain the onion exosome freeze-dried powder.
In a preferred embodiment, the mass ratio of the mannitol to the bovine serum albumin is 2-5: 6-8.
In a more preferred embodiment, the mass ratio of the mannitol to the bovine serum albumin is 3: 7.
In a second aspect, there is provided the use of a composition comprising a lyophilized powder of plant exosomes, in the preparation of a pharmaceutical composition, wherein the lyophilized powder of plant exosomes is embedded in mannitol and bovine serum albumin sequentially.
In a preferred embodiment, the plant exosome lyophilized powder is not ginger exosome lyophilized powder or turmeric exosome lyophilized powder.
In a preferred embodiment, the plant exosome lyophilized powder is an onion exosome lyophilized powder.
In a preferred embodiment, the plant exosome lyophilized powder is 0.5-5 parts by weight.
In a preferred embodiment, the plant exosome lyophilized powder is 0.5-2 parts by weight.
In a preferred embodiment, the plant exosome lyophilized powder is 1-3 parts by weight.
In a preferred embodiment, the plant exosome lyophilized powder is 1.5-4 parts by weight.
In a preferred embodiment, the plant exosome lyophilized powder is 3-5 parts by weight.
In a preferred embodiment, the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; and (3) reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out the material to obtain the onion exosome freeze-dried powder.
In a preferred embodiment, the mass ratio of the mannitol to the bovine serum albumin is 2-5: 6-8.
In a more preferred embodiment, the mass ratio of the mannitol to the bovine serum albumin is 3: 7.
Compared with the prior art, the invention has the beneficial effects that:
the invention optimizes the extraction and separation technology of the plant exosomes to obtain the plant exosome extracting solution with high concentration and high activity, and then selects a proper embedding agent and an embedding method to embed the plant exosomes, thereby improving the concentration, the activity, the tolerance, the targeting property and the like of the plant exosomes, improving the flavor taste and the stability of the product, in particular to onion exosomes with bad flavor.
The composition containing the plant exosome freeze-dried powder enhances the intestinal probiotic flora through the targeting property of the onion exosome freeze-dried powder, inhibits the proliferation of harmful flora, remarkably enhances the intestinal rhamnosus and bifidobacteria, and can improve the immunity of the organism at the same time.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention. It is to be understood that the following description is only illustrative of the present invention and is not to be construed as limiting the present invention.
The terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
The conjunction "consisting of …" excludes any unspecified elements, steps or components. If used in a claim, the phrase is intended to claim as closed, meaning that it does not contain materials other than those described, except for the conventional impurities associated therewith. When the phrase "consisting of …" appears in a clause of the subject matter of the claims rather than immediately after the subject matter, it defines only the elements described in the clause; other elements are not excluded from the claims as a whole.
When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted to include the ranges "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.
The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. "optional" or "any" means that the subsequently described event or events may or may not occur, and that the description includes instances where the event occurs and instances where it does not.
Approximating language, as used herein throughout the specification and claims, is intended to modify a quantity, such that the invention is not limited to the specific quantity, but includes portions that are literally received for modification without substantial change in the basic function to which the invention is related. Accordingly, the use of "about" to modify a numerical value means that the invention is not limited to the precise value. In some instances, the approximating language may correspond to the precision of an instrument for measuring the value. In the present description and claims, range limitations may be combined and/or interchanged, including all sub-ranges contained therein if not otherwise stated.
The indefinite articles "a" and "an" preceding an element or component of the invention are not intended to limit the number requirement (i.e., the number of occurrences) of the element or component. Thus, "a" or "an" should be read to include one or at least one, and the singular form of an element or component also includes the plural unless the number clearly indicates the singular.
Furthermore, the description below of the terms "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily for the same embodiment or example. Further, the technical features of the embodiments of the present invention may be combined with each other as long as they do not conflict with each other.
Example 1
The present example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 0.5 in parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 2: 6;
the composition also comprises 10 parts of flour, 20 parts of corn flour, 2 parts of bone meal, 5 parts of pumpkin meal, 2 parts of shrimp shell meal, 3 parts of seaweed meal, 5 parts of milk powder and 40 parts of water.
Example 2
The present example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 5 parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 5: 8;
the composition also comprises 20 parts of flour, 30 parts of corn flour, 10 parts of bone meal, 15 parts of pumpkin meal, 5 parts of shrimp shell meal, 8 parts of seaweed meal, 10 parts of milk powder and 60 parts of water.
Example 3
The present example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 2 parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 3: 7;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone meal, 7 parts of pumpkin meal, 3 parts of shrimp shell meal, 6 parts of seaweed meal, 8 parts of milk powder and 45 parts of water.
Example 4
This example provides a method of preparing a composition comprising a plant exosome lyophilized powder, comprising the steps of:
s1, weighing flour, corn flour, bone powder, pumpkin powder, shrimp shell powder, seaweed powder, cow milk powder, plant exosome freeze-dried powder and water;
s2, mixing the components weighed in the step S1, and then pouring the mixture into a feed granulator for granulation;
and S3, packaging the particles obtained in the step S2.
Comparative example 1
The comparative example provides use of a composition comprising ginger exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the ginger exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the weight part of the ginger exosome freeze-dried powder is 2;
the preparation method of the ginger exosome freeze-dried powder comprises the following steps:
s1, extracting ginger exosomes
Weighing 150g of fresh ginger, pouring the fresh ginger into a crusher, adding 150mL of sterilized PBS into the fresh ginger, crushing and homogenizing the ginger, and then centrifuging the ginger for 10min at 3000 Xg and 4 ℃ to remove larger residues; centrifuging the yellow solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain ginger exosome extract;
s2, embedding of ginger exosomes
Firstly, the first embedding of ginger exosome
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of ginger exosome extracting solution, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
second embedding of ginger exosome
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying ginger exosomes
Freezing the extracted ginger exosome at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, then placing the material on a heating plate for heating, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out the material to obtain the ginger exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 3: 7;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone meal, 7 parts of pumpkin meal, 3 parts of shrimp shell meal, 6 parts of seaweed meal, 8 parts of milk powder and 45 parts of water.
Comparative example 2
The comparative example provides use of a composition comprising turmeric exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the turmeric exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the weight part of the turmeric exosome freeze-dried powder is 2;
the preparation method of the turmeric exosome freeze-dried powder comprises the following steps:
s1, extracting curcuma exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the yellow solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain the turmeric exosome extract;
s2, embedding of turmeric exosomes
Firstly, the first embedding of the exosomes of turmeric
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of a turmeric exosome extracting solution, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of turmeric exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying of turmeric exosomes
Freezing the extracted Curcuma rhizome exosome at about-35 deg.C for 2h until the central temperature of the material is-25 deg.C, heating the material on a heating plate, heating the heating plate to 95 deg.C within 1h, and maintaining at the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out the material to obtain turmeric exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 3: 7;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone meal, 7 parts of pumpkin meal, 3 parts of shrimp shell meal, 6 parts of seaweed meal, 8 parts of milk powder and 45 parts of water.
Comparative example 3
The comparative example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 0.2 in parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 3: 7;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone meal, 7 parts of pumpkin meal, 3 parts of shrimp shell meal, 6 parts of seaweed meal, 8 parts of milk powder and 45 parts of water.
Comparative example 4
The comparative example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 6 parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 3: 7;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone meal, 7 parts of pumpkin meal, 3 parts of shrimp shell meal, 6 parts of seaweed meal, 8 parts of milk powder and 45 parts of water.
Comparative example 5
The comparative example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 2 parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 5: 10;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone meal, 7 parts of pumpkin meal, 3 parts of shrimp shell meal, 6 parts of seaweed meal, 8 parts of milk powder and 45 parts of water.
Comparative example 6
The comparative example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 2 parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a sodium alginate solution, uniformly mixing 15mL of the prepared sodium alginate solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the sodium alginate to the bovine serum albumin is 3: 7;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone meal, 7 parts of pumpkin meal, 3 parts of shrimp shell meal, 6 parts of seaweed meal, 8 parts of milk powder and 45 parts of water.
Comparative example 7
The comparative example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 2 parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing egg white protein solution;
2) dissolving the substance obtained in the step I into 20mL of prepared egg white protein solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the mannitol to the egg white protein is 3: 7;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone meal, 7 parts of pumpkin meal, 3 parts of shrimp shell meal, 6 parts of seaweed meal, 8 parts of milk powder and 45 parts of water.
Comparative example 8
The comparative example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 2 parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 3: 7;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone meal, 7 parts of pumpkin meal, 3 parts of shrimp shell meal, 8 parts of milk powder and 45 parts of water.
Comparative example 9
The comparative example provides the use of a composition comprising onion exosome lyophilized powder in the preparation of a diet for conditioning the intestinal tract, wherein,
the onion exosome freeze-dried powder is sequentially embedded in mannitol and bovine serum albumin;
the onion exosome freeze-dried powder is 2 parts by weight;
the preparation method of the onion exosome freeze-dried powder comprises the following steps:
s1, extraction of onion exosomes
Weighing 150g of fresh onion, pouring into a crusher, adding 150mL of sterilized PBS, crushing and homogenizing, centrifuging at 3000 Xg and 4 ℃ for 10min, and removing larger residues; centrifuging the purple solution in the middle layer at 11000 Xg at 4 deg.C for 50 min; taking the supernatant, ultracentrifuging at 120000 Xg and 4 ℃ for 60min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 130000 Xg and 4 ℃ for 70 min; after centrifugation, resuspending the precipitate with 2mL sterile PBS to obtain onion exosome extract;
s2, embedding of onion exosomes
First embedding of onion exosomes
1) Preparing a mannitol solution, uniformly mixing 15mL of the prepared mannitol solution with 10mL of onion exosome extract, adding 50mL of edible oil, and stirring at 200r/min for 20 min;
2) dropwise adding 20mL of edible oil containing 1g of glacial acetic acid, and stirring at 200r/min for 20 min;
3) adding 100mL of acetate buffer solution, standing for 3h, centrifuging, and removing an oil layer;
② second embedding of onion exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20mL of prepared bovine serum albumin solution;
3) stirring for 20min at a speed of 200 r/min;
s3, freeze-drying onion exosomes
Freezing the extracted onion exosomes at about-35 ℃ for 2h until the central temperature of the material is-25 ℃, heating the material on a heating plate, raising the temperature of the heating plate to 95 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 85 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 60 ℃ within 1h, and maintaining the temperature for 4 h; reducing the temperature of the heating plate to 50 ℃ within 1h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out to obtain onion exosome freeze-dried powder;
the mass ratio of the mannitol to the bovine serum albumin is 3: 7;
the composition also comprises 15 parts of flour, 25 parts of corn flour, 3 parts of bone flour, 7 parts of pumpkin flour, 3 parts of shrimp shell powder, 6 parts of lotus leaf powder, 8 parts of milk powder and 45 parts of water.
Effect verification: experiment for regulating intestinal tract
And (3) testing a sample: inventive examples 1-3, comparative examples 1-9 samples prepared according to the preparation method of example 4
Subject: selecting 120 pet dogs of the same breed, wherein the pet dogs are about 3 months old and have the same number of males and females, the pet dogs are healthy before the experiment, the pet dogs are similar in weight and are randomly divided into twelve groups, and each group contains 10 pet dogs
The experimental method comprises the following steps: the pet dog is subjected to intragastric administration, the pet dog to be tested is continuously given for one week, excrement of the pet dog is aseptically collected before and after the pet dog to be tested is tested for one week, bifidobacteria and lactobacillus rhamnosus are detected, no obvious difference exists between each group of data, the average value of each group of data is taken as a final detection result, and the final detection result is shown in the following table 1.
TABLE 1 results of intestinal conditioning experiments
Likewise, to further illustrate the beneficial effects of the present invention, the following comparative examples are provided:
comparative example 2.1 is provided, which is distinguished from example 3 by the following: replacing onion exosomes with turmeric powder;
comparative example 2.2 is provided, which is distinguished from example 3 by the following: replacing onion exosomes with curcumin;
comparative example 2.3 is provided, which comparative example 2.3 differs from example 3 in that: replacing onion exosomes with turmeric straw powder;
the composition for conditioning the pet intestinal tract is prepared by combining the preparation method disclosed by the invention, the effect of the composition in conditioning the pet intestinal tract is verified, and the composition is tested according to an experimental method of a conditioning intestinal tract experiment. The pet intestine conditioning effect of the pet intestine conditioning compositions prepared in comparative example 2.1, comparative example 2.2 and comparative example 2.3 was observed. The results were similar to those in comparative example 2 described above.
Likewise, to further illustrate the beneficial effects of the present invention, the following comparative examples are provided:
comparative example 3.1 is provided, which comparative example 3.1 differs from example 3 in that: adjusting the weight part of the seaweed powder to 2 parts;
comparative example 3.2 is provided, which comparative example 3.2 differs from example 3 in that: adjusting the weight part of the corn flour to 18 parts;
comparative example 3.3 is provided, which comparative example 3.3 differs from example 3 in that: adjusting the weight part of the bone powder to 1 part;
comparative example 3.4 is provided, which comparative example 3.4 differs from example 3 in that: adjusting the weight part of the shrimp shell powder to 1 part;
the composition for conditioning the pet intestinal tract is prepared by combining the preparation method disclosed by the invention, the effect of the composition in conditioning the pet intestinal tract is verified, and the composition is tested according to an experimental method of a conditioning intestinal tract experiment. The pet intestine conditioning effect of the pet intestine conditioning compositions prepared in comparative example 3.1, comparative example 3.2, comparative example 3.3 and comparative example 3.4 was observed. The results were similar to those in comparative example 3 above.
Likewise, to further illustrate the beneficial effects of the present invention, the following comparative examples are provided:
comparative example 4.1 is provided, which is distinguished from example 3 by the following: adjusting the weight part of the seaweed powder to 9 parts;
comparative example 4.2 is provided, which is distinguished from example 3 by the following: adjusting the weight part of the corn flour to 32 parts;
comparative example 4.3 is provided, which comparative example 4.3 differs from example 3 in that: adjusting the weight part of the bone powder to 12 parts;
comparative example 4.4 is provided, which is distinguished from example 3 by the following: adjusting the weight part of the shrimp shell powder to 6 parts;
the composition for conditioning the pet intestinal tract is prepared by combining the preparation method disclosed by the invention, the effect of the composition in conditioning the pet intestinal tract is verified, and the composition is tested according to an experimental method of a conditioning intestinal tract experiment. The pet intestine conditioning effect of the pet intestine conditioning compositions prepared in comparative example 4.1, comparative example 4.2, comparative example 4.3 and comparative example 4.4 was observed. The results were similar to those in comparative example 4 above.
Likewise, to further illustrate the beneficial effects of the present invention, the following comparative examples are provided:
comparative example 8.1 is provided, which comparative example 8.1 differs from example 3 in that: no corn flour is added;
comparative example 8.2 is provided, which comparative example 8.2 differs from example 3 in that: no bone powder is added;
comparative example 8.3 is provided, which comparative example 8.3 differs from example 3 in that: shrimp shell powder is not added;
comparative example 8.4 is provided, which comparative example 8.4 differs from example 3 in that: no pumpkin powder is added;
the composition for conditioning the pet intestinal tract is prepared by combining the preparation method disclosed by the invention, the effect of the composition in conditioning the pet intestinal tract is verified, and the composition is tested according to an experimental method of a conditioning intestinal tract experiment. The pet intestine conditioning effect of the pet intestine conditioning compositions prepared in comparative example 8.1, comparative example 8.2, comparative example 8.3 and comparative example 8.4 was observed. The results were similar to those in comparative example 8 described above.
Likewise, to further illustrate the beneficial effects of the present invention, the following comparative examples are provided:
comparative example 9.1 is provided, which is distinguished from example 3 by the following: replacing the pumpkin powder with fruit and vegetable powder;
comparative example 9.2 is provided, which comparative example 9.2 differs from example 3 in that: replacing the shrimp shell powder with inulin;
the composition for conditioning the pet intestinal tract is prepared by combining the preparation method disclosed by the invention, the effect of the composition in conditioning the pet intestinal tract is verified, and the composition is tested according to an experimental method of a conditioning intestinal tract experiment. The pet intestine conditioning effect of the pet intestine conditioning compositions prepared in comparative example 9.1 and comparative example 9.2 was observed. The results were similar to those in comparative example 9 described above.
From the above results, it can be seen that:
the composition for conditioning the intestinal tract of the pet provided by the invention is administrated to the pet dog by intragastric administration, and after the pet dog is intragastric administered for one week, both bifidobacterium and lactobacillus rhamnosus in the intestinal tract of the pet dog are obviously increased. And the composition for conditioning the intestinal tract of the pet, which is not prepared by the formula system, is administered to the pet dog by gavage, and the increase of the bifidobacteria and the lactobacillus rhamnosus in the intestinal tract of the pet dog is not obvious after the pet dog is fed with the composition for conditioning the intestinal tract of the pet dog by gavage for one week.
It is to be understood that the invention disclosed is not limited to the particular methodology, protocols, and materials described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.
It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. Use of a composition comprising a lyophilized powder of a plant exosome in the preparation of a diet for conditioning the intestinal tract, wherein the lyophilized powder of a plant exosome is sequentially embedded in mannitol and bovine serum albumin.
2. The use of claim 1, wherein the plant exosome lyophilized powder is not ginger exosome lyophilized powder or turmeric exosome lyophilized powder.
3. The use according to claim 1, wherein the plant exosome lyophilized powder is onion exosome lyophilized powder.
4. The use according to claim 1, wherein the mass ratio of the mannitol to the bovine serum albumin is 2-5: 6-8.
5. The use according to claim 4, wherein the mass ratio of the mannitol to the bovine serum albumin is 3: 7.
6. The use according to claim 1, wherein the plant exosome lyophilized powder is 0.5-5 parts by weight; for example, it may be 0.5-2, 1-3, 1.5-4 or 3-5.
7. Use according to any one of claims 1-6, wherein the method of preparing the plant exosome lyophilized powder comprises the steps of:
s1, extraction of plant exosomes
Weighing 100-; centrifuging the solution in the middle layer at 10000-12000 Xg and 4 ℃ for 50-60 min; taking the supernatant, ultracentrifuging at 120000-150000 Xg and 4 ℃ for 50-70min, then using 1mL sterile PBS to resuspend the precipitate, and then ultracentrifuging at 120000-150000 Xg and 4 ℃ for 50-70 min; after centrifugation, resuspending the precipitate with 1-2mL sterile PBS to obtain the plant exosome extract;
s2. embedding of plant exosomes
First embedding of plant exosomes
1) Preparing a mannitol solution, uniformly mixing 15-30mL of the prepared mannitol solution with 6-12mL of the plant exosome extracting solution, adding 40-60mL of edible oil, and stirring at 200-300r/min for 15-25 min;
2) dropwise adding 20-30mL of edible oil containing 0.5-1g of glacial acetic acid, and stirring for 15-25min at 300r/min under 200-;
3) adding 100-150mL acetate buffer solution, standing for 3-5h, centrifuging, and removing an oil layer;
second embedding of plant exosomes
1) Preparing a bovine serum albumin solution;
2) dissolving the substance obtained in the step I in 20-30mL of prepared bovine serum albumin solution;
3) stirring at 300r/min for 15-25min at 200-;
s3, freeze-drying of plant exosomes
Freezing the extracted plant exosomes at about-40 to-35 ℃ for 1 to 3 hours until the central temperature of the material is-35 to-25 ℃, then placing the material on a heating plate for heating, raising the temperature of the heating plate to 90 to 100 ℃ within 0.5 to 1.5 hours, and maintaining the temperature for 4 to 6 hours; reducing the temperature of the heating plate to 80-90 ℃ within 0.5-1.5h, and maintaining the temperature for 4-6 h; reducing the temperature of the heating plate to 55-65 ℃ within 0.5-1.5h, and maintaining the temperature for 4-6 h; and (3) reducing the temperature of the heating plate to 40-50 ℃ within 0.5-1.5h, and maintaining the temperature for 4-5h until the temperature of the material is not changed any more, and taking out the material to obtain the plant exosome freeze-dried powder.
8. The use according to any one of claims 1-6, wherein the composition further comprises flour, corn flour, bone flour, pumpkin flour, shrimp shell flour, seaweed flour, milk powder and water.
9. The use of claim 8, wherein the composition further comprises 10-20 parts of flour, 20-30 parts of corn flour, 2-10 parts of bone meal, 5-15 parts of pumpkin meal, 2-5 parts of shrimp shell meal, 3-8 parts of seaweed meal, 5-10 parts of milk powder and 40-60 parts of water by weight.
10. Use of a composition comprising a lyophilized powder of a plant exosome in the preparation of a pharmaceutical composition, wherein the lyophilized powder of a plant exosome is embedded in mannitol and bovine serum albumin in sequence.
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| CN202010922517.1A CN112006169A (en) | 2020-09-04 | 2020-09-04 | Application of composition containing plant exosome freeze-dried powder in preparation of intestinal tract conditioning feed |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112451488A (en) * | 2020-12-15 | 2021-03-09 | 南京中医药大学 | Preparation method and application of plant source exosome product |
| CN114642700A (en) * | 2022-03-22 | 2022-06-21 | 成都市第三人民医院 | Application of tea exosome in preparation of medicine for protecting intestinal barrier and treating irritable bowel syndrome |
-
2020
- 2020-09-04 CN CN202010922517.1A patent/CN112006169A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112451488A (en) * | 2020-12-15 | 2021-03-09 | 南京中医药大学 | Preparation method and application of plant source exosome product |
| CN114642700A (en) * | 2022-03-22 | 2022-06-21 | 成都市第三人民医院 | Application of tea exosome in preparation of medicine for protecting intestinal barrier and treating irritable bowel syndrome |
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