CN110638841A - Probiotics and prebiotics compound preparation, preparation method and application thereof - Google Patents
Probiotics and prebiotics compound preparation, preparation method and application thereof Download PDFInfo
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- CN110638841A CN110638841A CN201910908541.7A CN201910908541A CN110638841A CN 110638841 A CN110638841 A CN 110638841A CN 201910908541 A CN201910908541 A CN 201910908541A CN 110638841 A CN110638841 A CN 110638841A
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Mycology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Microbiology (AREA)
- Diabetes (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Endocrinology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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- Obesity (AREA)
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Abstract
The invention discloses a probiotic and prebiotic composite preparation, a preparation method and application thereof, and belongs to the field of biomedicine. The invention consists of composite probiotics and composite prebiotics, wherein the composite probiotics consist of clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei. The type of the composite probiotics in the invention is obtained by collecting intestinal microorganisms of diabetics and healthy people, researching theories to experiments, and verifying results, so that the probiotic and prebiotics composite preparation which can prevent and treat diabetes in a real sense is obtained; the research result shows that the traditional Chinese medicine composition can effectively reduce blood sugar, improve insulin resistance and simultaneously has a certain protection effect on organs.
Description
Technical Field
The invention relates to the field of biomedicine, and particularly relates to a probiotic and prebiotic composite preparation, a preparation method and application thereof.
Background
It is statistical that about 340 million people die globally in 2004 due to diabetes and its complications. According to the survey results of the international diabetes union, the number of people suffering from diabetes is 2.4 hundred million in 2007 and is increased to 3.47 hundred million in 2011. The international union for diabetes predicts that the number of people suffering from diabetes will reach 6.29 hundred million worldwide in 2045 years. It has been reported that China has become a big-mouth country for diabetes patients, and the prevalence rate is in a continuously rising state. Hyperglycemia is often accompanied with blood lipid metabolism disorder, which causes blood viscosity and blood flow speed to be slow, and chronic cardiovascular diseases such as atherosclerosis, thrombus and the like can be caused after long-term development. Diabetes and complications thereof seriously threaten the physical health of people and reduce the life quality of people.
Clinically, oral hypoglycemic drugs such as sulfonylureas and biguanides are generally adopted to treat diabetes patients. However, the long-term application of the treatment method easily causes side effects, such as diarrhea, dizziness, nausea, inappetence, hypoglycemia, liver function damage and the like, and seriously threatens the health of people. Therefore, the development of a novel natural and safe hypoglycemic agent or adjuvant therapy agent becomes important.
In recent years, probiotics gradually become a research hotspot for preventing and treating diabetes due to the characteristics of safety, easy obtaining, low cost, no side effect and the like.
Disclosure of Invention
In order to make up the defects of the prior art, the invention provides a probiotic and prebiotics compound preparation, a preparation method and application thereof.
The technical scheme of the invention is as follows:
a probiotic and prebiotic compound preparation comprises compound probiotic and compound prebiotic, wherein the compound probiotic comprises clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei.
Preferably, in the composite probiotics, clostridium butyricum is used as the probiotic bacteriaThe number of viable bacteria is 109～1010CFU/g, the viable count of the lactobacillus plantarum is 109～1010CFU/g, viable count of Bifidobacterium longum is 109～1010CFU/g, the viable count of lactobacillus paracasei is 109～1010CFU/g。
Preferably, the composite probiotics comprise 35-55% of clostridium butyricum, 20-45% of lactobacillus plantarum, 15-30% of bifidobacterium longum and 10-25% of lactobacillus paracasei in parts by mass.
Preferably, the complex prebiotics consist of isomaltooligosaccharide and galacto-oligosaccharide.
Furthermore, in the composite prebiotics, the mass percent of isomaltooligosaccharide is 25-85% and the mass percent of galacto-oligosaccharide is 15-75%.
As a preferred scheme, the mass percent of the composite probiotics is 35-85%, and the mass percent of the composite prebiotics is 15-65%.
Most preferably, the mass percent of the composite probiotics is 72 percent, and the mass percent of the composite prebiotics is 28 percent.
The preparation method of the probiotic and prebiotics compound preparation comprises the steps of placing clostridium butyricum bacterium powder, lactobacillus plantarum bacterium powder, bifidobacterium longum bacterium powder and lactobacillus paracasei bacterium powder in an aseptic tank, stirring uniformly, adding the compound prebiotics, and continuing to stir uniformly to obtain the probiotic and prebiotics compound preparation.
As a preferred scheme, the preparation method of the clostridium butyricum bacterial powder, the lactobacillus plantarum bacterial powder, the bifidobacterium longum bacterial powder and the lactobacillus paracasei bacterial powder comprises the following steps:
1) respectively activating clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei by an MRS culture medium, and then placing the activated clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei in a fermentation culture medium for high-density culture;
2) centrifugally separating thallus, adding into freeze-dried protective solution containing skimmed milk powder, glutamic acid and vitamin E, mixing, and freeze-drying to obtain corresponding thallus powder.
Preferably, the probiotic and prebiotic compound preparation is powder, tablet or capsule.
The probiotics and prebiotics compound preparation is used for reducing blood sugar and improving insulin resistance.
The related research of human microbiology and various diseases based on high-throughput sequencing provides an indication for selecting probiotics for people on a large basis. In order to reveal the causal relationship between the diabetic patients and the intestinal flora disorder and search for a suitable probiotic combined drug, the applicant collects a large number of excrement samples of the diabetic patients and healthy people in the early stage, obtains big data of the intestinal microorganisms of the diabetic patients and the healthy people through metagenomic sequencing analysis, integrates the big data with the existing intestinal data of the diabetic patients, and constructs a gene set of the intestinal flora of the diabetic patients. Meanwhile, classification and gene function annotation of intestinal flora species of diabetes and healthy people are completed, and by comparison, species and functional genes specific to diabetes and microbiota with remarkably reduced content relative to healthy people are found. Through a random forest model, four specific microorganisms (clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei) are further accurately obtained, model verification is carried out on the basis of independent people, and the combined change of the four microorganisms can be used for well predicting the early stage of diabetes.
The prebiotics can provide food for the probiotics, can be decomposed and absorbed by beneficial bacteria in intestinal tracts, promote the growth and reproduction of the beneficial bacteria and further have beneficial effects on hosts. The invention adds two prebiotics on the basis of four mixed probiotics, further provides continuous energy for the probiotics, and is a novel compound preparation with the function of obviously reducing blood sugar.
The invention has the beneficial effects that:
1. the probiotic and prebiotic composite preparation is prepared by compounding composite probiotics and composite prebiotics; the type of the composite probiotics in the invention is obtained by collecting intestinal microorganisms of diabetics and healthy people, researching theories to experiments, and verifying results, so that the probiotic and prebiotics composite preparation which can prevent and treat diabetes in a real sense is obtained; the research result shows that the traditional Chinese medicine composition can effectively reduce blood sugar, improve insulin resistance and simultaneously has a certain protection effect on organs.
2. The probiotic and prebiotics compound preparation is easy to be safe and has no toxin effect, can be used as a dietary supplement or a nutritional agent, is used for preventing diabetes, or is used for assisting in treating diabetes and relieving diabetic complications, and greatly relieves the pain of patients with diabetes. 3. The invention not only can play the role of reducing blood sugar of the probiotics, but also can play other probiotic functions of the probiotics, such as regulating immunity, resisting oxidation, delaying aging and the like, and has important significance for human health.
4. The used composite probiotics and composite prebiotics can effectively inhibit the growth of harmful strains, no preservative is required to be additionally added, and the risk of side effects caused by the addition of the preservative is reduced.
5. The composite preparation has the advantages of easily obtained raw materials, simple preparation method, low cost and easy industrial production.
Detailed Description
EXAMPLE 1 preparation of probiotic and prebiotic composite formulations
1) Preparation of probiotic powder
Preparing seed solutions of clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei:
respectively inoculating clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei into an MRS liquid culture medium according to the inoculation amount of 4 percent, and activating for 16 hours at the temperature of 37 ℃. And (3) dipping a small amount of bacterial liquid by using an inoculating loop, carrying out three-zone lineation on an MRS solid culture medium, culturing for 48h at 37 ℃, picking a single colony to a fermentation culture medium, and culturing for 16h at 37 ℃.
Centrifugally separating the thallus, adding into a freeze-drying protective solution containing 10% of skimmed milk powder, 0.8% of glutamic acid and 1% of vitamin E, mixing, and freeze-drying to obtain corresponding bacterial powder. The number of viable bacteria in each bacterial powder is 109-1010CFU/g。
Wherein the content of the first and second substances,
the MRS liquid culture medium comprises the following components in percentage by mass: 2.0% of glucose, 0.2% of ammonium citrate, 0.5% of sodium acetate, 0.03% of manganese sulfate, 0.05% of magnesium sulfate, 1.0% of peptone, 1.0% of beef extract, 0.5% of yeast extract, soil temperature-800.2% and the balance of water; the pH value is 6.0-6.2.
The MRS solid culture medium comprises the following components in percentage by mass: 2.0% of glucose, 0.2% of ammonium citrate, 0.5% of sodium acetate, 0.03% of manganese sulfate, 0.05% of magnesium sulfate, 1.0% of peptone, 1.0% of beef extract, 0.5% of yeast extract, Tween-800.2%, 1.5% of agar powder and the balance of water; the pH value is 6.0-6.2.
The fermentation medium is MRS liquid medium.
2) Ingredients
7.2Kg of composite probiotics and 2.8Kg of composite prebiotics.
Wherein, the composite probiotics contains 3.24Kg of clostridium butyricum bacterial powder, 1.80Kg of lactobacillus plantarum bacterial powder, 1.44Kg of bifidobacterium longum bacterial powder and 0.72Kg of lactobacillus paracasei bacterial powder.
The composite prebiotics contains 1.82Kg of isomaltooligosaccharide and 0.98Kg of galacto-oligosaccharide.
3) Mixing
Sequentially adding the above 10 into a sterile stirring tank9-1010CFU/g clostridium butyricum powder and 109-1010CFU/g lactobacillus plantarum powder, 109-1010CFU/g Bifidobacterium longum powder, 109-1010Stirring for 20 minutes, wherein the CFU/g of lactobacillus paracasei powder is stirred for 20 minutes; and sequentially adding isomaltooligosaccharide and galacto-oligosaccharide, and continuously stirring for 10 minutes to obtain the probiotic and prebiotics compound preparation.
EXAMPLE 2 preparation of probiotic and prebiotic composite formulations
1) Ingredients
6.5Kg of composite probiotics and 3.5Kg of composite prebiotics.
Wherein, the composite probiotics contains 2.275Kg of clostridium butyricum bacterial powder, 1.30Kg of lactobacillus plantarum bacterial powder, 1.95Kg of bifidobacterium longum bacterial powder and 0.975Kg of lactobacillus paracasei bacterial powder. The bacterial powder adopts the steps of example 1
1) And (4) preparing the bacterial powder.
The composite prebiotics contains 1.05Kg of isomaltooligosaccharide and 2.45Kg of galacto-oligosaccharide.
2) Mixing
Sequentially adding the above 10 into a sterile stirring tank9-1010CFU/g clostridium butyricum powder and 109-1010CFU/g lactobacillus plantarum powder, 109-1010CFU/g Bifidobacterium longum powder, 109-1010Stirring for 20 minutes, wherein the CFU/g of lactobacillus paracasei powder is stirred for 20 minutes; and sequentially adding isomaltooligosaccharide and galacto-oligosaccharide, and continuously stirring for 10 minutes to obtain the probiotic and prebiotics compound preparation.
EXAMPLE 3 preparation of probiotic and prebiotic composite formulations
1) Ingredients
4.0Kg of composite probiotics and 6.0Kg of composite prebiotics.
Wherein, the composite probiotics contains 1.60Kg of clostridium butyricum bacterial powder, 1.00Kg of lactobacillus plantarum bacterial powder, 1.00Kg of bifidobacterium longum bacterial powder and 0.40Kg of lactobacillus paracasei bacterial powder. The bacterial powder prepared in the step 1) of the example 1 is adopted.
The composite prebiotics contains 3.30Kg of isomaltooligosaccharide and 2.70Kg of galacto-oligosaccharide.
2) Mixing
Sequentially adding the above 10 into a sterile stirring tank9-1010CFU/g clostridium butyricum powder and 109-1010CFU/g lactobacillus plantarum powder, 109-1010CFU/g Bifidobacterium longum powder, 109-1010Stirring for 20 minutes, wherein the CFU/g of lactobacillus paracasei powder is stirred for 20 minutes; and sequentially adding isomaltooligosaccharide and galacto-oligosaccharide, and continuously stirring for 10 minutes to obtain the probiotic and prebiotics compound preparation.
EXAMPLE 4 preparation of probiotic and prebiotic composite formulations
1) Ingredients
6.0Kg of composite probiotics and 4.0Kg of composite prebiotics.
Wherein, the composite probiotics contains 2.40Kg of clostridium butyricum bacterial powder, 1.20Kg of lactobacillus plantarum bacterial powder, 1.20Kg of bifidobacterium longum bacterial powder and 1.20Kg of lactobacillus paracasei bacterial powder. The bacterial powder prepared in the step 1) of the example 1 is adopted.
The composite prebiotics contains 3.00Kg of isomaltooligosaccharide and 1.00Kg of galacto-oligosaccharide.
2) Mixing
Sequentially adding the above 10 into a sterile stirring tank9-1010CFU/g clostridium butyricum powder and 109-1010CFU/g lactobacillus plantarum powder, 109-1010CFU/g Bifidobacterium longum powder, 109-1010Stirring for 20 minutes, wherein the CFU/g of lactobacillus paracasei powder is stirred for 20 minutes; and sequentially adding isomaltooligosaccharide and galacto-oligosaccharide, and continuously stirring for 10 minutes to obtain the probiotic and prebiotics compound preparation.
Example 5 probiotic and prebiotic Complex formulation
In the ingredients, the composite probiotics account for 80 percent of the mass of the composite preparation, and the composite prebiotics account for 20 percent of the treatment of the composite preparation. The mass ratio of each bacteria powder in the composite probiotics and the mass ratio of the two components in the composite prebiotics are the same as those in example 1.
Example 6 Complex formulation of Probiotics and prebiotics
In the ingredients, the composite probiotics account for 35% of the mass of the composite preparation, and the composite prebiotics account for 65% of the treatment of the composite preparation. The mass ratio of each bacteria powder in the composite probiotics and the mass ratio of the two components in the composite prebiotics are the same as those in example 1.
Example 7 probiotic and prebiotic Complex formulation
The mass ratio of each bacterial powder in the composite probiotics is the same as that in the embodiment 1, and the rest is the same as that in the embodiment 1.
The composite probiotics comprise 35% of clostridium butyricum powder, 25% of lactobacillus plantarum powder, 25% of bifidobacterium longum powder and 15% of lactobacillus paracasei powder by mass.
Example 8 Complex formulation of Probiotics and prebiotics
The mass ratio of each bacterial powder in the composite probiotics is the same as that in the embodiment 1, and the rest is the same as that in the embodiment 1.
The composite probiotics comprise 50% of clostridium butyricum powder, 20% of lactobacillus plantarum powder, 20% of bifidobacterium longum powder and 10% of lactobacillus paracasei powder by mass.
Example 9 Complex formulation of Probiotics and prebiotics
The mass ratio of each bacterial powder in the composite probiotics is the same as that in the embodiment 1, and the rest is the same as that in the embodiment 1.
The composite probiotics comprise 35% of clostridium butyricum powder, 30% of lactobacillus plantarum powder, 25% of bifidobacterium longum powder and 10% of lactobacillus paracasei powder by mass.
Example 10 determination of environmental tolerance of four probiotic bacteria
(1) Preparation of artificial gastrointestinal fluid
An amount of pepsin was dissolved in a PBS buffer solution at pH 3.0 to adjust the final concentration to 3g/L, thereby obtaining an artificial gastric juice.
An amount of trypsin was dissolved in a PBS buffer solution at pH 8.0 to adjust the final concentration to 1g/L, thereby obtaining an artificial intestinal juice.
Both the artificial gastric juice and the artificial intestinal juice are filtered through a 0.22 mu m filter membrane under the aseptic condition.
(2) Artificial gastrointestinal fluid tolerance
Inoculating the target strain into MRS liquid culture medium in an inoculation amount of 3%, continuously activating for three generations at 37 ℃, centrifugally collecting thalli, and preparing into bacterial suspension for later use. Inoculating the prepared bacterial suspension into artificial gastric juice filtered by a 0.22 mu m filter membrane according to the inoculation amount of 3%, culturing at 37 ℃, diluting and coating 100 mu L of bacterial liquid respectively at 0, 1, 2 and 3h, and calculating the survival rate. Then sucking 1mL of bacteria-containing artificial gastric juice cultured for 3h, adding into 9mL of artificial intestinal juice filtered by a 0.22 μm filter membrane, culturing at 37 ℃, sucking 100 μ L of bacterial liquid for dilution and coating at 1, 3, 5 and 8h respectively, and calculating the survival rate.
The survival rate calculation formula is as follows:
wherein N istIs processedNumber of colonies, N0Number of colonies without treatment.
The environmental tolerance results of the four probiotics artificial gastrointestinal fluids are shown in the table 1-table 4.
TABLE 1 results on the resistance of Clostridium butyricum to artificial gastrointestinal fluids
TABLE 2 results on the tolerance of Lactobacillus plantarum to artificial gastrointestinal fluids
TABLE 3 results on the tolerance of Bifidobacterium longum to artificial gastrointestinal fluids
TABLE 4 results on the tolerance of Lactobacillus paracasei to artificial gastrointestinal fluids
As can be seen from tables 1-4, the survival rates of the four strains in the artificial gastric juice are all above 85%, and the survival rates in the artificial intestinal juice are all above 75%, which shows that the four strains have better tolerance capability in the artificial simulated gastrointestinal juice, in particular to lactobacillus paracasei.
Example 11 Effect of probiotic and prebiotic Complex formulations on diabetic mice
Grouping and establishing a diabetes model:
360 male C57BL/6J mice were selected for the experiment, the age of the mice was about 3 weeks, and the mice were randomly divided into 3 groups of 120 mice each: normal group, model group and experimental group. The temperature of the rat room is 22 +/-2 ℃, the humidity is 55 +/-5%, the rat room is alternated day and night for 12 hours, and the rat room is fed with the basic feed adaptively for one week. From week 2 onwards, the model group and the experimental group were fed with high-fat high-sugar diet. After 3 weeks, the model group and the experimental group were administered with 110mg/kg of STZ intraperitoneally to construct a diabetes model, and the normal group was administered with the same amount of citrate buffer as a control. After one week of molding, the random blood sugar of the mice is measured, and the mice with blood sugar value higher than 11.1mmol/L are successful models. Then 9:00 a.m. each day, the experimental group was gavaged with the probiotic and prebiotic composite preparation obtained in gavage example 1, and the normal group and the model group were gavaged with the same dose of physiological saline. The experimental period was 13 weeks.
And (3) measuring blood sugar and insulin: blood glucose levels were measured and recorded before and after the experiment. After the mice were fasted overnight without water deprivation, tail blood was taken and the FBG value was determined. The PBG values of the mice were measured 2 hours after feeding the feed. And insulin was measured after the end of the experiment, and the results are shown in tables 5, 6 and 7.
TABLE 5 Probiotics and prebiotics combination preparation obtained in example 1 susceptibility to blood glucose changes before and after diabetic mice experiments
As can be seen from Table 5, the probiotic and prebiotic composite preparation of the present invention can reduce the hyperglycemia to some extent and delay the onset of diabetes.
TABLE 6 Effect of the probiotic and prebiotic combination formulation obtained in example 1 on insulin levels in diabetic mice
As can be seen from Table 6, the probiotic and prebiotic composite preparation of the present invention can reduce the insulin release amount of diabetic mice and alleviate hyperinsulinemia.
TABLE 7 Effect of the probiotic and prebiotic combination preparation obtained in example 1 on the visceral index of diabetic mice
As shown in Table 7, the probiotic and prebiotics composite preparation of the invention can inhibit the increase of the organ index of the diabetic mice, has a certain protection effect on organs, and relieves diabetic complications.
The above description is only for the purpose of illustrating the present invention, and it should be understood that the present invention is not limited to the above embodiments, and various modifications conforming to the spirit of the present invention are within the scope of the present invention.
Claims (10)
1. A probiotic and prebiotic composite preparation is composed of composite probiotics and composite prebiotics, and is characterized in that: the composite probiotics comprise clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei.
2. The probiotic and prebiotic complex formulation of claim 1, wherein: in the composite probiotics, the number of the live bacteria of the clostridium butyricum is 109~1010 CFU/g, the viable count of the lactobacillus plantarum is 109~1010 CFU/g, viable count of Bifidobacterium longum is 109~1010 CFU/g, the viable count of lactobacillus paracasei is 109~1010 CFU/g。
3. The probiotic and prebiotic complex formulation as claimed in claim 1 or 2, wherein the complex probiotic comprises, in mass fraction, 35% ~ 55% clostridium butyricum, 20% ~ 45% lactobacillus plantarum, 15% ~ 30% bifidobacterium longum, 10% ~ 25% lactobacillus paracasei.
4. The complex formulation of probiotics and prebiotics of claim 1 or 2, characterized by: the composite prebiotics consist of isomaltooligosaccharide and galacto-oligosaccharide.
5. The probiotic and prebiotic complex formulation of claim 4, wherein the complex prebiotic comprises isomaltooligosaccharide 25% ~ 85% by weight and galacto-oligosaccharide 15% ~ 75% by weight.
6. The probiotic and prebiotic composite formulation according to claim 1, wherein the mass percent of the composite probiotic is 35% ~ 85%, and the mass percent of the composite prebiotic is 15% ~ 65%.
7. The method for preparing the probiotic and prebiotic composite formulation of claim 1 wherein: and (3) placing the clostridium butyricum bacterium powder, the lactobacillus plantarum bacterium powder, the bifidobacterium longum bacterium powder and the lactobacillus paracasei bacterium powder in an aseptic tank, adding the composite prebiotics after uniformly stirring, and continuously and uniformly stirring to obtain the probiotic and prebiotic composite preparation.
8. The method for preparing the probiotic and prebiotic composite preparation as described in claim 7, wherein the method for preparing the clostridium butyricum bacterial powder, the lactobacillus plantarum bacterial powder, the bifidobacterium longum bacterial powder and the lactobacillus paracasei bacterial powder comprises the following steps:
1) respectively activating clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei by an MRS culture medium, and then placing the activated clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei in a fermentation culture medium for high-density culture;
2) centrifugally separating thallus, adding into freeze-dried protective solution containing skimmed milk powder, glutamic acid and vitamin E, mixing, and freeze-drying to obtain corresponding thallus powder.
9. The method for preparing the probiotic and prebiotic composite formulation as claimed in claim 7 or 8 wherein: the probiotic and prebiotics compound preparation is powder, tablet or capsule.
10. Use of a probiotic and prebiotic complex formulation according to claim 1 characterised in that: can be used for lowering blood sugar and improving insulin resistance.
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