CN115137757A - Probiotic composition assisting in reducing blood sugar - Google Patents

Abstract

The invention relates to a probiotic composition for assisting in reducing blood sugar, which mainly comprises composite probiotic freeze-dried powder, wherein the composite probiotic freeze-dried powder comprises a flora consisting of bifidobacterium adolescentis, bifidobacterium longum, bifidobacterium breve, lactobacillus acidophilus, bifidobacterium bifidum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus plantarum, streptococcus thermophilus and lactobacillus delbrueckii subsp. The invention can improve the human body immune response, is beneficial to preventing intestinal diseases, enhancing the immune function, regulating the function of an immune system and inhibiting the growth and the reproduction of pathogenic bacteria.

Description

A kind of probiotic composition for auxiliary hypoglycemic

technical field

The invention relates to the field of probiotics, in particular to a probiotic composition for improving diabetes and assisting in lowering blood sugar.

Background technique

Diabetes mellitus is a heterogeneous metabolic disease characterized by hyperglycemia, which occurs when the pancreas does not produce enough insulin or the body cannot effectively use the insulin it produces. In the past few decades, the incidence of diabetes has increased year by year worldwide, and studies have shown that its prevalence in different regions of China can be as high as 8.3% to 12.7%. Over time, diabetes can also damage many of a patient's body systems, leading to further complications. Therefore, diabetes has become one of the major threats to global health and imposes considerable economic and medical burden on individuals and countries.

There are two main types of diabetes, type 1 and type 2 diabetes (type 2diabetes mellitus, T2DM). Type 1 diabetes occurs when autoreactive T cells attack islet beta cells resulting in insufficient insulin production. Type 2 diabetes, characterized by low-grade cellular failure, is a complex metabolic disorder. Type 2 diabetes, also known as adult-onset diabetes, accounts for more than 90% of all diabetic patients. Research has shown that in addition to genetic factors playing an important role in diabetes susceptibility, there are many determinants that drive the development of type 2 diabetes. Among them, the gut microbiota has been identified as a novel and potential driver in the pathophysiology of type 2 diabetes.

With the application of high-throughput sequencing and metagenomic analysis technologies in recent years, as well as the application of germ-free facilities and germ-free mice, a more comprehensive understanding of the information of the gut microbiota has also been obtained, and the changes in the gut microbiota are related to changes in patients and animal models. Disease development is closely linked. Research shows that each individual's gut microbiome composition is not only unique, but may also be a predictor of disease risk, and that gut microbiota may contribute to the development of metabolic diseases such as obesity and type 2 diabetes, select for specific gut bacteria The regulation of intestinal microbiota balance by bacterial strains is a promising therapeutic approach. For example, patent application 202110878469.5 discloses an intestinal macrogenome feature as a screening marker for the efficacy of fecal bacterial transplantation in patients with type 2 diabetes. The expression levels of microbiota markers can be used to select patients with type 2 diabetes who are suitable for fecal microbiota transplantation. According to this intestinal flora biomarker and different detection technology platforms, corresponding detection kits are designed and developed. This system and product can be used for individualized precision treatment of type 2 diabetes.

However, a single drug treatment method cannot achieve a good therapeutic effect. In the actual treatment process, auxiliary treatment methods are often required. Yes, more or less. Therefore, on the basis of conventional drug treatment, choosing some adjuvant treatments that are easy for patients to accept, such as consumption of probiotics, prebiotics and synbiotics, can improve the balance of intestinal microbes and change the composition of colonic microbes. To improve the symptoms of patients, to achieve the purpose of adjuvant treatment of type 2 diabetes.

SUMMARY OF THE INVENTION

In order to solve the above-mentioned problems, the primary purpose of the present invention is to provide a probiotic composition for assisting hypoglycemia. The probiotic composition selects some edible probiotics, prebiotics and prebiotics that are easy for patients to accept on the basis of conventional medicine treatment. Synbiotics can effectively relieve the symptoms of type 2 diabetes by improving the balance of intestinal microbes and changing the composition of colonic microbes, and achieve the purpose of adjuvant treatment of type 2 diabetes.

For achieving the above object, the technical scheme adopted in the present invention is:

A probiotic composition for auxiliary hypoglycemic, the composite probiotic composition mainly comprises composite probiotic freeze-dried powder, the composite probiotic freeze-dried powder includes Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve Bacillus, Lactobacillus acidophilus, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus plantarum, Streptococcus thermophilus, Lactobacillus delbrueckii subspecies bulgaricus flora.

After in-depth research work, the applicant found that the specific probiotic strains formed by the above-mentioned strains can form a good probiotic environment, have a stimulating effect on intestinal peristalsis, inhibit the growth and reproduction of pathogenic bacteria, thereby improving the immune response of the human body. Helps prevent intestinal diseases.

Further, Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus plantarum, Streptococcus thermophilus, German Lactobacillus bulgaricus subsp. bulgaricus, according to the ratio of 1:0.5:1.5:1:0.5:0.5:1.5:1.5:1:1 to prepare probiotics.

Further, the probiotic composition further includes D-mannitol, resistant dextrin, and magnesium stearate.

Among them, D-mannitol: the only glyconutrient currently used in clinical practice, with low calorie and low sweetness, can replace sugar for special food for diabetic and obese patients, and is also a chewing gum additive.

The probiotic composition further comprises vitamin C and resistant dextrin, so that the stability of the probiotic composition is better.

Among them, vitamin C: is a water-soluble vitamin commonly used in clinical practice. It has antioxidant and immune-enhancing effects, improves the defense function of the intestinal mucosal barrier, and can increase the proportion of beneficial bacteria in the intestinal tract.

Resistant dextrin: It is a representative low-viscosity water-soluble dietary fiber. Because it contains ingredients that are difficult to digest by digestive enzymes, it will not be digested and absorbed by the digestive tract. It can directly enter the intestinal tract and be fermented and utilized by microorganisms in the intestinal tract. It has functions such as regulating blood lipids, adjusting the intestinal environment, controlling body weight, and preventing obesity.

Further, the Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus thermophilus Coccus, Lactobacillus delbrueckii subsp. bulgaricus, according to the ratio of 1: 0.5: 1.5: 1: 0.5: 0.5: 1.5: 1.5: 1: 1 to prepare probiotics, mix them evenly, and then mix with D-mannitol, anti- The synthetic dextrin and magnesium stearate are mixed according to the ratio of D-mannitol: resistant dextrin: magnesium stearate=0.5:2:0.2, and packaged into a product.

The present invention adopts Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus plantarum, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus constructs a probiotic composition, these flora are beneficial bacteria in the human gut, can be used in general food, can stimulate bowel movements, can improve human immune response and help prevent intestinal diseases , enhance immune function, help to regulate immune system function, inhibit the growth and reproduction of pathogenic bacteria.

Description of drawings

Figure 1 is a schematic diagram of the activity change of the accelerated test of the present invention.

Figure 2 is a schematic diagram of the effect of the probiotics of the present invention on STZ mice.

Fig. 3 is a graph showing the abundance of fecal flora in mice of the present invention.

Detailed ways

In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention, but not to limit the present invention.

The probiotic composition for auxiliary hypoglycemic prepared by the present invention mainly comprises freeze-dried composite probiotic powder, and also includes D-mannitol, resistant dextrin and magnesium stearate; Compound probiotic lyophilized powder includes Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus plantarum, The flora composed of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus.

Among them, Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus plantarum, Streptococcus thermophilus, German Lactobacillus bulgaricus subsp. bulgaricus, according to the ratio of 1:0.5:1.5:1:0.5:0.5:1.5:1.5:1:1 to prepare probiotics. Mix it evenly, and then mix with D-mannitol, resistant dextrin, and magnesium stearate according to the ratio of D-mannitol: resistant dextrin: magnesium stearate = 0.5: 2: 0.2, package into a product.

Experimental part (I): Shelf life test.

In order to ensure the preservation of viable bacterial activity of this product, and to explore whether vitamin C and resistant dextrin have a protective effect on bacterial activity, the experiment was divided into two groups. The first group was the group without vitamin C and resistant dextrin. The second group is the product containing vitamin C and resistant dextrin. The two groups were stored in a constant temperature box at 37°C for 4 months, and the humidity was 45-66%. The accelerated test of food storage period (shelf life) (ASLT), and its activity was measured at 14d, 1m, 2m, 3m, and 4m, respectively. The results showed that, as shown in Figure 1, under the accelerated test, the activity of the group without vitamin C and resistant dextrin was 14 days later. It began to decline and reached stability after 1 month, and the viable bacteria rate remained on the order of 10* ⁹ , while the group containing vitamin C and resistant dextrin had better stability, and did not contain vitamin C and resistant dextrin in the 4th month. The activity of dextrin group was 1.69*10 ⁹ , while the activity of vitamin C and resistant dextrin was 6.69*10 ⁹ . There was a very significant difference between the two groups (P<0.01). It may be because the addition of vitamin C and resistant dextrin can reduce the water activity of the product and make the product more stable.

Experimental part (II): The effect of probiotics on diabetic mice.

Twenty-four 7-week-old male Kunming mice were randomly divided into 3 groups with 8 mice in each group. The experiment was divided into a normal group, a model group, and a probiotic treatment group. One-week acclimation was performed in a laboratory in a ventilated cage, with free food and water. After 1 week of adaptive feeding, the model group and probiotic treatment group were given intraperitoneal injection of STZ (40 mg/kg streptozotocin) for 5 consecutive days, and the normal group was intraperitoneally injected with normal saline for 5 consecutive days as the control group. After 5 consecutive days of intraperitoneal injection, when the blood sugar of the mice was stable for a week, blood was collected from the tail vein of the mice to measure their blood sugar. When the blood glucose of the mice was ≥11 mmol/L and stabilized for more than two weeks, the modeling was considered successful. After successful modeling, the treatment group was given 200 μL of the above probiotic complex for treatment every day. Blood glucose, body weight and fecal glucose concentration of mice in each group were recorded daily. Mice feces were collected after 7 weeks, DNA samples were extracted using the QIAamp Fast DNA Stool MiniKIT (QIAGEN) kit, PCR library construction was performed based on the 16S rRNAV3-V4 segment, and high-throughput sequencing was performed by an Illumina HiSeq2500 instrument. Usearch performed OTU cluster analysis to obtain information on flora abundance and composition.

Among them, a 1ml syringe with a 4-gauge needle was used for intraperitoneal injection in mice; during intraperitoneal injection, the syringe was held in the right hand, the little finger and ring finger of the left hand grabbed the tail of the mouse, and the other three fingers grabbed the neck of the mouse, so that the mouse head down. In this way, the organs in the abdominal cavity will naturally fall to the chest, preventing damage to the large intestine, small intestine and other organs when the syringe is pierced. The action of needle insertion should be gentle to avoid puncturing the abdominal organs; during intraperitoneal injection, the needle should pass a short distance under the skin of the abdomen, enter the needle from one side of the abdomen, pass through the midline of the abdomen, and then enter the abdominal cavity on the other side of the abdomen. , slowly withdraw the needle, and rotate the needle slightly to prevent leakage.

Then, bend the new straight gavage needle appropriately, and the angle is similar to the physiological curvature of the rat esophagus; hold the animal with the left hand, hold the syringe with the right hand, and the scale of the syringe faces forward; the gavage needle enters from the corner of the animal's mouth, press the tongue, Hold the upper jaw and push it inward gently; slowly insert the gavage needle into the esophagus along the back wall of the pharynx, and then withdraw the syringe without air backflow before injecting probiotics; observe the reaction of the rat, if there is no excessive struggle, you can try to inject probiotics If the resistance is small, all probiotics can be injected. If the resistance is too large or the animal reacts violently, the breathing is blocked, and the needle is withdrawn and inserted; release the mouse and observe the animal's breathing.

The direct indicators for the evaluation of the therapeutic effect of diabetes are blood sugar level and body weight. Therefore, after the model was successfully started to treat, the blood sugar level, fecal glucose concentration and body weight of the mice in each group were detected at regular intervals. As shown in Figure A in Figure 2, the weight of the mice in the normal control group was relatively stable, reaching 50 g in the seventh week. The weight of mice in the model group (STZ) gradually decreased (p<0.05), and it was 32g in the 7th week. The weight of the mice in the probiotic treatment group (STZ+P) was relieved, and it increased to 41g in the 7th week, and the model group existed. Significant difference; as shown in Figure B, the fecal glucose concentration of the mice in the normal group maintained a normal value of 2 mmol/L, and the fecal glucose concentration of the modeling group (STZ) and the probiotic treatment group (STZ+P) reached the third week. The fecal glucose concentration of mice in the treatment group (STZ+P) began to decrease in the third week, and decreased to the normal concentration in the seventh week. As shown in Figure C, the mice in the normal group maintained normal blood sugar levels, while the blood sugar levels of the mice in the model group (STZ) and the probiotic treatment group (STZ+P) gradually increased, and were maintained at 20 mmol/L from the fourth week. The blood sugar level of the mice in the probiotic treatment group (STZ+P) was relieved after treatment, and gradually decreased, which was significantly different from the model group (STZ). The treatment alleviated the condition of STZ-induced diabetes model mice to a certain extent.

The feces of the mice were collected after 7 weeks, and at the same time combined with high-throughput sequencing, the relative abundance of the flora was further evaluated, according to Figure 3 (in Figure 3, Note: N represents the normal group of mice, DM represents the diabetic model group small group. mice, Treatment represents the probiotic treatment group), it was found that the relative abundance of the genera Sphingomonas and Alistipes in the diabetic group was increased compared with the normal mice, and the relative abundance of the genera Lachnospiraceae and Bifidobacterium was decreased, while the mice in the probiotic-treated group were found to be Sphingomonas and Alistipes compared with the normal mice The relative abundance of mice decreased, while the relative abundance of Bifidobacterium increased. Bifidobacterium, as one of the important members of human and animal intestinal flora, can inhibit the growth of harmful bacteria in the human body, resist the infection of pathogenic bacteria, synthesize vitamins needed by the human body, promote the absorption of minerals by the human body, and produce acetic acid, Organic acids such as acid, butyric acid and lactic acid stimulate intestinal peristalsis, prevent constipation, inhibit intestinal corruption, purify the intestinal environment, stimulate the human immune system, and thus play an important role in improving disease resistance.

In this experiment, there were few bifidobacteria and lactic acid bacteria in the feces of the mice in the diabetes model group, while bifidobacteria and lactic acid bacteria were mainly in this product, so the relative abundance of lactic acid bacteria in the treatment group was higher, and the probiotic bacteria were changed by changing The abundance of intestinal flora, thereby helping to improve diabetes symptoms.

Experimental part (Ⅲ): Probiotics assisted metformin in improving diabetic patients.

Fifty-four diabetic patients were recruited for the study, and their basic characteristics are shown in Table 1. Randomly divided into 2 groups, treatment group and placebo group, for 60 days of probiotics group and placebo intervention, measured fasting blood glucose and glycosylated hemoglobin every 30 days. Experimenters in the probiotic group took the probiotics under the premise of standard metformin treatment. On the basis of standard metformin treatment, the patients in the placebo group were given the corresponding placebo, and the grouped subjects were all given diabetes diet and activity guidance to obtain the basic information and medical history of the patients, collect the patients' height, weight, waist circumference and calculate the BMI. The placebo group received Maltodextrin-only formulation.

Blood collection:

Blood samples were collected from volunteers at 0d, 30d, and 60d before meals into vacuum blood collection tubes, centrifuged at 3000 rpm for 10 min, and serum was collected after centrifugation to collect fasting blood glucose concentration and glycosylated hemoglobin, and use automatic biochemical analysis to determine their indicators.

According to Table 2, after 30 days of probiotic treatment, there was a significant difference in fasting blood glucose and glycosylated hemoglobin compared with the placebo group (P < 0.05), and the average fasting blood glucose of the probiotic treatment group was 0.27/mmol · less than the placebo group. L, and after 60 days of probiotic treatment, there was a significant difference in fasting blood glucose and glycosylated hemoglobin compared with the placebo group (P<0.001), and the gap widened. The fasting blood glucose of the probiotic treatment group was 1.32 mmol lower than that of the placebo group. · L, glycated hemoglobin was 0.48% less in the added probiotic treatment group than in the placebo group. Some studies have found that the role of metformin is mainly in the gut, and intravenous metformin cannot regulate blood sugar in humans. This may be due to the fact that the hypoglycemic effect of metformin is related to the regulation of some species in the intestinal flora, and the probiotic complex can assist the hypoglycemic effect of metformin. These flora are all beneficial bacteria in the human intestinal tract, which can stimulate peristalsis, improve the immune response of the human body, help prevent intestinal diseases, enhance immune function, help regulate immune system function, and inhibit the growth and reproduction of pathogenic bacteria. The above research also found that adding the compound probiotics can significantly enhance the hypoglycemic effect of metformin.

Table 1 Basic information of diabetic patients

Table 2 Main measurement indicators

The experimental data were analyzed by SPSS 22.0 Univariate method, and the factors that reached a significant level by the F test were analyzed by variance analysis and Duncan's multiple comparisons. The experimental data of each group were expressed as (mean ± standard deviation).

The statistical significance level was P<0.05, and the extremely significant level was P<0.01.

The equipment involved in the test includes a high-speed centrifuge, a pipette gun, a biological safety cabinet, a blood biochemical analyzer, an electronic balance, a vortex shaker, a gavage needle, and a 1ml syringe, which are all realized by the existing technology, and will not be repeated here.

It can be seen from the above test that the compound probiotics realized by the present invention can improve the blood sugar index of diabetics by adjusting the intestinal flora and using with metformin.

The above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention shall be included in the protection scope of the present invention. Inside.

Claims (4)

1. an auxiliary hypoglycemic probiotic composition is characterized in that this composite probiotic composition mainly comprises composite probiotic freeze-dried powder, and described composite probiotic freeze-dried powder comprises Bifidobacterium adolescentis, Bifidobacterium longum Bacillus, Bifidobacterium breve, Lactobacillus acidophilus, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus plantarum, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus . 2. The probiotic composition of auxiliary hypoglycemic according to claim 1 is characterized in that Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Bifidobacterium bifidum, Reuteri Lactobacillus, Lactobacillus rhamnosus, Lactobacillus plantarum, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, according to the ratio of 1:0.5:1.5:1:0.5:0.5:1.5:1.5:1:1 to prepare probiotics bacteria. 3. The probiotic composition for auxiliary hypoglycemic according to claim 2, characterized in that the probiotic composition further comprises D-mannitol, resistant dextrin and magnesium stearate. 4. The probiotic composition of claim 3, characterized in that the Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Bifidobacterium bifidum, Bifidobacterium Lactobacillus eische, Lactobacillus rhamnosus, Lactobacillus plantarum, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, according to the ratio 1:0.5:1.5:1:0.5:0.5:1.5:1.5:1:1 Prepare probiotics, mix them evenly, and then mix with D-mannitol, resistant dextrin, and magnesium stearate according to the ratio of D-mannitol: resistant dextrin: magnesium stearate = 0.5:2:0.2 Mixed and packaged into product.

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