CN115137757A - Probiotic composition assisting in reducing blood sugar - Google Patents
Probiotic composition assisting in reducing blood sugar Download PDFInfo
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- CN115137757A CN115137757A CN202210903020.4A CN202210903020A CN115137757A CN 115137757 A CN115137757 A CN 115137757A CN 202210903020 A CN202210903020 A CN 202210903020A CN 115137757 A CN115137757 A CN 115137757A
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- lactobacillus
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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
Technical Field
The invention relates to the field of probiotics, in particular to a probiotic composition for improving diabetes and assisting in reducing blood sugar.
Background
Diabetes is a heterogeneous metabolic disease characterized by hyperglycemia, which results when the pancreas fails to produce enough insulin or the body fails to use the insulin it produces effectively. Over the past several decades, the incidence of diabetes has increased worldwide year by year, with studies showing that it can reach 8.3% to 12.7% prevalence in different regions of china. Over time, diabetes also damages many of the patient's body systems, further causing various complications. Therefore, diabetes has become one of the major threats facing global health, and imposes considerable economic and medical burdens on individuals and countries.
Diabetes mainly includes two types, type 1 diabetes and type 2diabetes (T2dm), and type 1 diabetes occurs when autoreactive T cells attack islet β cells to cause insufficient insulin production; type 2diabetes, which is characterized by low-grade inflammation, insulin resistance and low-grade cell failure, is a complex metabolic disorder. Among them, type 2diabetes is also called adult onset diabetes, and accounts for more than 90% of all diabetic patients. Studies have shown that, in addition to genetic factors playing an important role in diabetes susceptibility, there are a number of determinants that can drive the development of type 2 diabetes. Among them, intestinal microorganisms are considered as a new, potential driver in the pathophysiology of type 2 diabetes.
With the application of high-throughput sequencing and metagenomic analysis technology, sterile facilities and sterile mice in recent years, the information of the intestinal microbiota is more comprehensively known, and the change of the intestinal microbiota is closely related to the disease development of patients and animal models. Studies have shown that the gut microbiota composition of each individual is not only unique, but may also be a predictor of disease risk, that gut microbiota may contribute to the development of metabolic diseases such as obesity and type 2diabetes, and that selection of specific gut bacterial strains to regulate the gut microbiota balance is a promising treatment. For example, patent application 202110878469.5 discloses a macrogenomic feature of intestinal tract as a marker for screening the curative effect of fecal bacteria transplantation in type two diabetic patients, wherein the macrogenomic feature of intestinal tract microorganism is characterized by intestinal flora of Rinkenellaceae and Anaerotruncus, and type two diabetic patients suitable for fecal bacteria transplantation treatment are selected according to the expression level of the markers of the specific intestinal flora. According to the intestinal flora biomarker and different detection technology platforms, corresponding detection kits are designed and developed. The system and product can be used for the precise treatment of type II diabetes based on individuation.
However, a single treatment means cannot achieve a good treatment effect, and an auxiliary treatment means is often needed in an actual treatment process. Therefore, on the basis of the conventional medicine treatment, some auxiliary treatment means which are easily accepted by patients, such as edible probiotics, prebiotics and synbiotics, are selected, the symptoms of the patients can be improved by improving the balance of intestinal microorganisms and changing the composition of colon microorganisms, and the aim of auxiliary treatment of the type 2diabetes is fulfilled.
Disclosure of Invention
In order to solve the problems, the primary object of the present invention is to provide a probiotic composition for assisting in decreasing blood sugar, wherein the probiotic composition is prepared by selecting edible probiotics, prebiotics and synbiotics which are easily accepted by some patients on the basis of conventional medicine treatment, and can effectively alleviate the symptoms of type 2diabetes by improving intestinal microbial balance and changing the composition of colon microorganisms, so as to achieve the purpose of assisting in the treatment of type 2 diabetes.
In order to achieve the purpose, the invention adopts the technical scheme that:
the composite probiotic composition 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.
After intensive research work, the applicant finds that the specific probiotic strains formed by the strains can form a good probiotic environment, have a stimulation effect on intestinal peristalsis, and inhibit the growth and reproduction of pathogenic bacteria, so that the immune response of a human body is improved, and intestinal diseases are prevented.
Further, bifidobacterium adolescentis, bifidobacterium longum, bifidobacterium breve, lactobacillus acidophilus, bifidobacterium bifidum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus plantarum, streptococcus thermophilus and lactobacillus delbrueckii subspecies bulgaricus according to the mixture ratio of 1:0.5:1.5:1:0.5:0.5:1.5:1.5:1:1 configuration of probiotics.
Further, the probiotic composition also comprises D-mannitol, resistant dextrin and magnesium stearate.
Wherein, D-mannitol: at present, the sugar nutrient is only used for clinical sugar nutrients, has low calorie and low sweetness, can replace sugar to be used as special food for patients with diabetes and obesity, 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.
Wherein, the ratio of vitamin C: is a clinically common water-soluble vitamin, has the functions of resisting oxidation and enhancing the immunity of the organism, improves the barrier defense function of intestinal mucosa, and can improve the proportion of beneficial bacteria in the intestinal tract.
Resistant dextrin: the dietary fiber is representative low-viscosity water-soluble dietary fiber, contains components which are difficult to digest by digestive enzymes, cannot be digested and absorbed by the digestive tract, can directly enter the intestinal tract to be fermented and utilized by microorganisms in the intestinal tract, and has the functions of reducing blood sugar and regulating blood fat, adjusting the environment of the intestinal tract, controlling weight, preventing obesity and the like.
Furthermore, the bifidobacterium adolescentis, the bifidobacterium longum, the bifidobacterium breve, the lactobacillus acidophilus, the bifidobacterium bifidum, the lactobacillus reuteri, the lactobacillus rhamnosus, the lactobacillus plantarum, the streptococcus thermophilus and the lactobacillus delbrueckii subsp bulgaricus are mixed according to the mixture ratio of 1:0.5:1.5:1:0.5:0.5:1.5:1.5:1:1 preparing probiotics, mixing the probiotics evenly, and then mixing the probiotics with D-mannitol, resistant dextrin and magnesium stearate according to the proportion that D-mannitol: resistant dextrin: magnesium stearate =0.5:2: mixing at a ratio of 0.2, and packaging to obtain the final product.
The invention adopts bifidobacterium adolescentis, bifidobacterium longum, bifidobacterium breve, lactobacillus acidophilus, bifidobacterium bifidum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus plantarum, streptococcus thermophilus and lactobacillus delbrueckii subsp bulgaricus to construct the probiotic composition, and the flora are all beneficial bacteria in human intestinal tracts, can be used for common foods, has the functions of stimulating intestinal peristalsis, improving human immune response, helping to prevent intestinal diseases, enhancing immune function, helping to regulate immune system function and inhibiting the growth and reproduction of pathogenic bacteria.
Drawings
FIG. 1 is a schematic diagram showing the activity change of the accelerated test according to the present invention.
FIG. 2 is a schematic representation of the effect of 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 Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The compound probiotic composition prepared by the invention for assisting in reducing blood sugar mainly comprises compound probiotic freeze-dried powder, and also comprises D-mannitol, resistant dextrin and magnesium stearate; 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 bulgaricus.
Wherein, the bifidobacterium adolescentis, the bifidobacterium longum, the bifidobacterium breve, the lactobacillus acidophilus, the bifidobacterium bifidum, the lactobacillus reuteri, the lactobacillus rhamnosus, the lactobacillus plantarum, the streptococcus thermophilus and the lactobacillus delbrueckii subsp bulgaricus are mixed according to the mixture ratio of 1:0.5:1.5:1:0.5:0.5:1.5:1.5:1:1 preparing probiotics. Mixing them uniformly, mixing them with D-mannitol, resistant dextrin and magnesium stearate according to D-mannitol: resistant dextrin: magnesium stearate =0.5:2: mixing at a ratio of 0.2, and packaging to obtain the final product.
Experimental part (i): and (4) shelf life test.
In order to ensure the preservation of the viable bacteria activity of the product and to investigate whether the vitamin C and the resistant dextrin have the protection effect on the flora activity, the experiment is divided into two groups, the first group is a group without the addition of the vitamin C and the resistant dextrin, the second group is a group with the vitamin C and the resistant dextrin, the two groups are placed in a constant temperature box at 37 ℃ for preservation for 4 months, the food storage period (quality guarantee period) acceleration test (ASLT) is carried out under the condition that the humidity is 45-66%, the activity of the groups is measured at 14d, 1m, 2m, 3m and 4m respectively, and the result shows that the activity of the groups without the vitamin C and the resistant dextrin begins to decline after 14 days under the acceleration test and reaches the stability after 1 month, and the viable bacteria rate is kept at 10 ×, as shown in figure 1 9 Of the order of magnitude of (1.69 x 10), while the group containing vitamin C and resistant dextrins is more stable and free of vitamin C and resistant dextrins at month 4 9 And vitamin C and resistant dextrin of 6.69 x 10 9 There was a very significant difference (P) between the two groups<0.01). Probably because the water activity of the product can be reduced after the vitamin C and the resistant dextrin are added, the product is more stable.
Experimental part (ii): probiotic bacteria have an effect on diabetic mice.
24 male Kunming mice with age of 7 weeks are randomly divided into 3 groups of 8 mice, and the experiment is divided into a normal group, a modeling group and a probiotic treatment group. Acclimation for one week, free diet and water drinking were performed in a laboratory in ventilated cages. After adaptive feeding for 1 week, the model group and the probiotic treated group were given treatment by intraperitoneal injection of STZ (streptozotocin 40 mg/kg) for 5 consecutive days, and the normal group was given treatment by intraperitoneal injection of physiological saline for 5 consecutive days as a control group. After 5 consecutive days of intraperitoneal injection, when the blood sugar of the mouse is stable for one week, tail vein blood taking is carried out on the mouse to measure the blood sugar. When the blood sugar of the mouse is more than or equal to 11mmol/L and is stable for more than two weeks, the molding is considered to be successful. After successful modeling, the treatment group was treated with 200 μ L of the probiotic compound per day. Blood glucose, body weight and fecal glucose concentrations were recorded daily for each group of mice. Mouse feces were collected after 7 weeks, DNA samples were extracted using QIAamp Fast DNA pool Mini KIT (QIAGEN) KIT, PCR library construction was performed based on 16965 s rRNAV3-V4 segment, high throughput sequencing was performed by Illumina HiSeq2500 instrument, OTU cluster analysis was performed on sequencing data by Usearch to obtain information on flora abundance and composition.
Wherein, a 1ml syringe for intraperitoneal injection of mice is matched with a No. 4 needle; when the injection is performed in the abdominal cavity, the injector is held by the right hand, the tail of the mouse is held by the little finger and the ring finger of the left hand, the neck of the mouse is held by the other three fingers, and the head of the mouse is downward. Therefore, organs in the abdominal cavity can naturally fall to the chest, and the large intestine, the small intestine and other organs are prevented from being damaged when the injector is punctured. The needle insertion is gentle, so as to prevent the abdominal organs from being punctured; the needle head passes through the abdomen for a short distance subcutaneously in the abdominal cavity injection, the needle is inserted from one side of the abdomen, enters the abdominal cavity from the other side of the abdomen after passing through the abdominal midline, and after the medicine is injected, the needle head is slowly pulled out, and is slightly rotated to prevent leakage.
Then, bending the new straight-line gastric lavage needle head by a proper amount, wherein the angle is similar to the physiological curvature of the mouse esophagus; holding the animal with the left hand, holding the injector with the right hand, and enabling the scale of the injector to face forwards; the stomach irrigation needle head enters from the mouth corner of an animal, presses the tongue, props against the palate and slightly pushes inwards; slowly inserting the stomach filling needle into the esophagus along the posterior pharyngeal wall, and injecting probiotics by a back-pumping injector in an air-free countercurrent way; observing the reaction of the mouse, if struggle without excess, trying to inject probiotics, if the resistance is small, injecting all probiotics, if the resistance is too large or the animal has violent reaction, the breathing is blocked, and the probiotics is inserted after withdrawing the needle; loosening the mouse, observing the respiration of the animal, and determining the success of the gavage if no abnormal respiration exists.
Since blood glucose and body weight are direct indicators for evaluating the therapeutic effect on diabetes, blood glucose, fecal glucose concentration, and body weight of each group of mice were measured at intervals after the start of therapy successfully modeled. As shown in FIG. 2, the body weight of the mice in the normal control group was relatively stable and reached 50g at week 7. The weight of the model building group (STZ) mice is gradually reduced (P is less than 0.05), 32g is obtained in 7 weeks, the weight of the probiotic treatment group (STZ + P) mice is relieved, and the weight of the probiotic treatment group (STZ + P) mice is increased to 41g in 7 weeks, which is obviously different from that of the model building group; as shown in FIG. B, the fecal glucose concentration of the normal group mice was maintained at a normal value of 2mmol/L, the fecal glucose concentration of the model group (STZ) and the probiotic treated group (STZ + P) reached 4mmol/L in the third week, the model group (STZ) had a tendency to gradually increase, the fecal glucose concentration of the treated group (STZ + P) mice began to decrease in the third week, and the fecal glucose concentration decreased to the normal concentration in the seventh week. As shown in the graph C, the blood glucose values of the mice in the normal group were maintained at normal blood glucose level, while the blood glucose values of the mice in the model group (STZ) and the mice in the probiotic treatment group (STZ + P) were gradually increased and maintained at 20mmol/L in the fourth week, but the blood glucose values of the mice in the probiotic treatment group (STZ + P) were relieved after the treatment and gradually decreased, and the results were significantly different from those of the mice in the model group (STZ), which indicates that the treatment with the probiotic compound relieved the conditions of the diabetes-induced mice in the model group (STZ) to some extent.
After 7 weeks the feces of the mice were collected and further evaluated from the relative abundance of the flora in combination with high throughput sequencing, according to fig. 3 (in fig. 3, note: N for normal group mice, DM for diabetic model group mice, and trement for probiotic treated group), it was found that the relative abundance of diabetic group sphingans and Alistipes was increased compared to normal mice, the relative divisions of lachnospiacee and bifidobacteria were decreased, while the relative abundance of mice sphingans and Alistipes subjected to probiotic treated group was decreased compared to normal mice, and the relative abundance of bifidobacteria was increased. The bacteria of the genus Bifidobacterium, as one of the important constituents of the human and animal intestinal flora, can inhibit the growth of harmful bacteria of the human body, resist the infection of pathogenic bacteria, synthesize vitamins required by the human body, promote the absorption of mineral substances by the human body, produce organic acids such as acetic acid, propionic acid, butyric acid and lactic acid to stimulate the intestinal peristalsis, prevent constipation and inhibit the intestinal putrefaction, purify the intestinal environment, stimulate the immune system of the human body, thus have important effects in the aspects of improving the disease resistance, etc.
In the experiment, the diabetes mellitus model group mouse excrement contains fewer bifidobacteria and lactic acid bacteria, while the product mainly contains the bifidobacteria and lactic acid bacteria, so that the relative abundance of the lactic acid bacteria in the treatment group is more, and the probiotics change the abundance of intestinal flora, thereby assisting in improving the symptoms of diabetes mellitus.
Experimental part (iii): the probiotics assist metformin to improve the diabetic patients.
54 diabetic patients were recruited for the study and their basic characteristics are shown in Table 1. Randomized into 2 groups, treatment group and placebo group, with probiotic and placebo intervention at 60d, fasting glucose and glycated hemoglobin measured every 30 d. The probiotic group experimenters take the probiotics under the premise of standard metformin therapy. Patients in the placebo group take corresponding placebo on the basis of standard metformin therapy, all the grouped subjects give diabetic diet and activity guide, know basic information and medical history of the patients, collect height, weight and girth of the patients and calculate BMI, and the placebo group receives a preparation only containing maltodextrin.
Blood collection:
blood samples of volunteers at 0d, 30d and 60d before meals are collected into a vacuum blood collection tube, centrifuged at 3000rpm for 10min, and serum is taken after centrifugation to collect fasting plasma glucose concentration and glycosylated hemoglobin, and indexes of the fasting plasma glucose concentration and the glycosylated hemoglobin are measured by using full-automatic biochemical analysis.
As can be seen from Table 2, after 30 days of probiotic treatment, there was a significant difference in fasting plasma glucose and glycated hemoglobin (P < 0.05) from the placebo group, the probiotic treated group had a 0.27/mmol.L less mean fasting plasma glucose than the placebo group, and after 60 days of probiotic treatment, there was a significant difference in fasting plasma glucose and glycated hemoglobin (P < 0.001) from the placebo group, and the difference was increased, with the addition of probiotic treated group having 1.32 mmol.L less fasting plasma glucose than the placebo group, and the addition of glycated hemoglobin of probiotic treated group having 0.48% less glycated hemoglobin than the placebo group. The research shows that the action site of metformin is mainly in the intestinal tract, and intravenous injection of metformin can not regulate the blood sugar of human beings. This is probably due to the hypoglycemic effect of metformin, which is associated with the regulation of some species in the intestinal flora, the probiotic complex can assist metformin in its hypoglycemic action. The flora is beneficial bacteria in human intestinal tract, and has effects of stimulating intestinal peristalsis, improving immune response, preventing intestinal tract diseases, enhancing immunity, regulating immune system function, and inhibiting growth and reproduction of pathogenic bacteria. The research also finds that the hypoglycemic effect of the metformin can be obviously enhanced by adding the composite probiotics.
TABLE 1 basic information of diabetic patients
TABLE 2 Main measurement indices
The experimental data were analyzed by SPSS 22.0Univariate method for single factor analysis, for factors that achieved significant levels for the F test, and by analysis of variance and Duncan's multiple comparisons, the experimental data for each group are expressed as (mean. + -. Standard deviation).
Statistical significance level was P <0.05, and very significance level was P <0.01.
The equipment involved in the test comprises a high-speed centrifuge, a pipette, a biological safety cabinet, a blood biochemical instrument, an electronic balance, a vortex oscillator, a gastric lavage needle and a 1ml syringe which are all realized by adopting the prior art, and are not described again.
The tests show that the composite probiotics realized by the invention can improve the blood sugar index of the diabetes patients by regulating intestinal flora and matching with metformin.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (4)
1. The probiotic composition is characterized by mainly comprising 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 bulgaricus.
2. The probiotic composition for assisting in reducing blood glucose according to claim 1, wherein the ratio of bifidobacterium adolescentis, bifidobacterium longum, bifidobacterium breve, lactobacillus acidophilus, bifidobacterium bifidum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus plantarum, streptococcus thermophilus, lactobacillus delbrueckii subsp. 0.5:1.5:1:0.5:0.5:1.5:1.5:1:1 preparing probiotics.
3. Probiotic composition for the adjuvant reduction of blood glucose according to claim 2, characterized in that it further comprises D-mannitol, resistant dextrin, magnesium stearate.
4. The probiotic composition for assisting in reducing blood glucose according to claim 3, wherein the ratio of Bifidobacterium adolescentis, bifidobacterium longum, bifidobacterium breve, lactobacillus acidophilus, bifidobacterium bifidum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus plantarum, streptococcus thermophilus, lactobacillus delbrueckii subsp. 0.5:1.5:1:0.5:0.5:1.5:1.5:1:1 preparing probiotics, mixing the probiotics evenly, and then mixing the probiotics with D-mannitol, resistant dextrin and magnesium stearate according to the proportion that D-mannitol: resistant dextrin: magnesium stearate =0.5:2: mixing at a ratio of 0.2, and packaging to obtain the final product.
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