Composition for improving diabetic encephalopathy and application

Abstract

The invention discloses a composition for improving diabetic encephalopathy and application, the composition comprises intestinal microorganism bacteroides simplex and traditional Chinese medicine component monomer baicalein, and the diabetic encephalopathy is the diabetic encephalopathy with excessive and aggravated selenium; according to the composition, the abundance and proportion of intestinal bacteria are regulated and controlled by utilizing a microbial community-intestinal-brain axis, the composition of intestinal flora is changed, the decomposition capacity of intestinal microorganisms on an amino acid substrate is further changed, short-chain fatty acid metabolism and the neurotransmitter level are regulated and controlled, and neurons are protected from being damaged by oxidative stress; the traditional Chinese medicine composition can effectively relieve high-selenium-aggravated diabetic encephalopathy induced by high-fat diet, improve structural damage of colon tissues and damage of neurons in hippocampus in a diabetic state, and improve the spatial learning and memory ability of patients with diabetic encephalopathy.

Description

Technical Field

[0001] The present invention relates to a composition for improving diabetic encephalopathy and also relates to the application of the composition. Background Art

[0002] Metformin is a traditional drug for the treatment of diabetes. It has been identified as the first-line drug for type 2 diabetes by the International Diabetes Diagnosis and Treatment Guidelines. Since its introduction, it has been the most widely used oral hypoglycemic drug in the world. Metformin can inhibit hepatic glucose output to regulate blood sugar, and has a protective effect on cardiovascular diseases related to type 2 diabetes, including myocardial infarction, hypertrophy and cardiomyopathy. In addition, metformin has been found to have anti-tumor and anti-aging effects, and has a certain therapeutic effect on polycystic ovary or gestational diabetes. Metformin is a classic drug for the treatment of type 2 diabetes. Its treatment of diabetic complications is mainly reflected in improving blood sugar and energy metabolism, thereby playing a role in nerve and cardiovascular protection. It can also indirectly play a metabolic protective role by affecting intestinal microorganisms. However, the current connection between metformin and cognitive function improvement still needs to be verified by large-scale randomized clinical trials. In addition, metformin is prone to cause gastrointestinal disorders, diarrhea, interference with the absorption of vitamin B12 and folic acid, and increased risk of cognitive impairment.

[0003] Bacteroides uniformis (Bu) belongs to the genus Bacteroides. It is a strictly anaerobic Gram-negative bacterium that plays an important ecological role in the intestine and is closely related to diseases such as enteritis, obesity, diabetes, cognitive development, and emotional movement. It can degrade and digest complex carbohydrates and affect the transport of intestinal amino acids. Bu has been shown to have beneficial properties against obesity and metabolic disorders. In animal experiments, Bu can induce the production of anti-inflammatory cytokines in vitro and improve the metabolic and immune dysfunction of obese mice induced by a high-fat diet. As the main next-generation candidate probiotic, Bu also shows positive therapeutic effects on diseases such as cognitive impairment. Bu can significantly reduce CUMS-induced depression and anxiety-like behaviors in rats; in autism spectrum disorders, it can alleviate related phenotypes and the imbalance of the excitation / inhibition ratio in the brain. Bu (Bacteroides uniformis), as a new generation of potential probiotics, has not yet been fully explored in the treatment of diseases. In previous studies, the focus has been on the treatment of metabolic diseases represented by obesity caused by a high-fat diet. B. uniformis is a specific bacterial species associated with the degradation of isoflavone genistein in feces. It usually uses polysaccharides as a carbon source and can produce cellulase and amylase to degrade complex polysaccharides. Studies have shown that B. uniformis is associated with lowering serum cholesterol and triglyceride levels and improving hepatic steatosis. It is also associated with lower serum leptin levels, lower fasting blood glucose concentrations, and improved glucose tolerance. It may play a metabolic regulatory role by regulating intestinal flora, improving intestinal barrier function, reducing chronic low-grade inflammation, and regulating intestinal peptide secretion. In the actual application of probiotics, one of the main challenges is to ensure the vitality and stability of the strain. It is very sensitive to environmental factors such as pH, temperature, oxygen exposure, and processing methods, which will reduce its vitality and shelf life and affect the viability of probiotics. Therefore, it is crucial to choose a suitable processing method to ensure the biological activity of the strain.

[0004] Baicalein is one of the main active ingredients of Scutellaria baicalensis, with pharmacological effects such as anti-oxidation, free radical scavenging, anti-inflammatory, anti-diabetic and neuroprotective, and has shown good effects in controlling metabolic syndrome and related diseases. However, repeated administration of baicalein can cause mild adverse events such as constipation, abdominal distension and peripheral blood leukopenia, which can be relieved without treatment, suggesting that attention should be paid to monitoring liver function changes and erythema when baicalein is used clinically; Coptis chinensis also has some side effects and adverse reactions such as drug rash and allergies, and symptoms such as dizziness, palpitations, shortness of breath and joint pain can also occur, so finding new drug combinations is very important for the treatment of type 2 diabetes.

[0005] Diabetic encephalopathy is a serious complication of diabetes caused by metabolic disorders, including dysfunction of the central nervous system and cognitive decline. High selenium intake can aggravate cognitive impairment in mice with type 2 diabetes mellitus (T2DM) and cause intestinal flora imbalance. Diabetic encephalopathy is a cognitive disorder caused by imbalanced blood sugar levels and energy metabolism disorders in diabetes. It specifically refers to cognitive and motor dysfunction caused by central nervous system lesions such as neurochemical, neurophysiological and structural abnormalities. The clinical manifestations are the decline of learning ability, memory ability, language, understanding, judgment and problem-solving ability. Past epidemiological cross-sectional studies have found that supplementing selenium nutrients for people with normal selenium intake will further aggravate diabetic cognitive impairment. Currently, there is no symptomatic treatment for diabetic encephalopathy aggravated by high selenium. Cognitive impairment complications are usually alleviated by treating type 2 diabetes. The treatment drugs include metformin, anti-oxidative stress drugs, drugs that inhibit aldose reductase activity, drugs that improve microcirculation, etc. There is no teammate drug to treat diabetic encephalopathy aggravated by excessive selenium, and the symptoms can only be slightly improved through existing drugs. In recent years, some herbal medicines and Chinese medicine preparations with blood circulation and stasis-removing effects have also been commonly used to treat diabetic encephalopathy, such as Mudan granules and compound Danshen dripping pills. However, these drugs contain alkaloids or other allergenic ingredients, and allergic reactions may occur during treatment, leading to serious consequences. Therefore, it is urgent to develop a targeted drug for the treatment of diabetic encephalopathy aggravated by excessive selenium. Summary of the invention

[0006] Purpose of the invention: The purpose of the present invention is to provide a composition for improving diabetic encephalopathy, and also to provide the application of the above composition.

[0007] Technical solution: The present invention discloses a composition for improving diabetic encephalopathy, comprising intestinal microorganism Bacteroides monomorpha and a traditional Chinese medicine monomer baicalin.

[0008] Wherein, the diabetic encephalopathy is diabetic encephalopathy aggravated by excessive selenium.

[0009] The composition is 10-150 mg / kg of baicalein monomer and 5×10 7 -5×10 9 CFU / mL combination; preferably baicalein monomer 25mg / kg + Bacteroides monomorpha 5×10 8 CFU / mL.

[0010] The invention also discloses the application of the composition in preparing medicine for treating cognitive impairment complications caused by animals under high-selenium and high-fat diet.

[0011] The composition is orally administered to treat cognitive impairment complications caused by animals on a high-selenium and high-fat diet.

[0012] The application is to mix the composition in sterile physiological saline and administer it orally.

[0013] Among them, the application is to alleviate diabetic encephalopathy induced by a high-fat diet which is aggravated by high selenium, improve the structural damage of colon tissue and the damage of hippocampal neurons in the diabetic state, and improve the spatial learning and memory ability of patients with diabetic encephalopathy.

[0014] Among them, the application is to utilize the microbial community-gut-brain axis to regulate the abundance and proportion of intestinal bacteria, change the composition of intestinal flora, and then change the ability of intestinal microorganisms to decompose amino acid substrates, regulate short-chain fatty acid metabolism and neurotransmitter levels, and protect neurons from damage by oxidative stress.

[0015] Wherein, the dosage form of the drug is capsule, tablet or oral film, which maintains the stability of the composition and improves its bioavailability.

[0016] Wherein, the medicine is an enteric-coated probiotic preparation.

[0017] The preparation method of the enteric probiotic preparation is as follows: inoculating Bacteroides monotheca in an anaerobic fermentation environment, continuing to culture under anaerobic conditions until the late logarithmic growth period or the late stable period of Bacteroides monotheca to obtain a fermentation product; mixing baicalein with the fermentation product, spray-drying the mixture for preservation, and coating the surface with an enteric material to obtain the preparation. The preparation is insoluble in the environment of gastric acid and bile, and begins to dissolve in the intestinal fluid environment, thereby achieving a targeted regulatory effect on the intestine.

[0018] Principle of the invention: The composition of the present invention adopts the traditional Chinese medicine baicalein and the probiotic Bacteroides uniformis in combination, which is beneficial to improve intestinal development and flora structure, thereby affecting the central nervous system through the gut-brain axis. The combination of drugs is commonly used in clinical practice to treat diseases. Berberine + probiotics has a greater blood sugar lowering effect than berberine alone. Glutamine combined with probiotics can inhibit intestinal inflammation and oxidative stress. The combination of B. uniformis and wheat bran extract shows a more superior anti-obesity effect.

[0019] In the mechanism of baicalin in treating T2DM, baicalin can delay the occurrence of diabetic peripheral neuropathy and maintain the normal function of nerves. Studies have reported that intestinal microorganisms have the function of transforming Chinese medicine components, and Chinese medicine can simultaneously regulate the metabolism of intestinal microbiota. The use of baicalin to treat T2DM patients can significantly reduce glucose tolerance and the abundance of bifidobacteria, while increasing the intestinal microbiota of lactic acid bacteria and Akkermansia and improving the structure of intestinal flora. Baicalin can act on different pathways to improve the complications of diabetic cognitive impairment, and supplementation of Bacteroides monocytogenes can target and offset the changes in intestinal flora caused by high selenium intake. Therefore, the combination of baicalin and Bu is a very promising strategy for treating DE.

[0020] In the application experiment of the composition of the present application, the open field data showed that each dosing group showed obvious therapeutic effects, among which the effect of the combined low-dose group was the most significant. In terms of the total moving distance in the open field, compared with HFD+Se, the combined low-, medium- and high-dose groups significantly increased the moving distance of mice (P<0.01, P<0.01, P<0.05) and total moving time (P<0.01, P<0.01, P<0.05). The number of times mice enter the central area can indicate the exploration desire of mice. Compared with HFD+Se, the combined low-dose group significantly increased the number of times mice entered the center and improved the exploration desire of mice (P<0.01).

[0021] In the Y-maze test, after drug treatment, the moving time of mice was prolonged compared with the HFD+Se group, among which the low-dose combined drug treatment group showed significant difference (P < 0.01); for the percentage statistics of diabetic encephalopathy mice entering the novel arm, the percentage of entering the novel arm in the low- and medium-dose combined drug treatment groups increased significantly (P < 0.01; P < 0.01), indicating that after combined drug treatment, the spatial exploration and memory abilities of diabetic encephalopathy mice were significantly improved.

[0022] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: the baicalein combined with Bacteroides monotheca in the composition can have a better therapeutic effect, and can be used to treat diabetic encephalopathy aggravated by excessive selenium, can alleviate diabetic encephalopathy induced by a high-fat diet aggravated by high selenium, improve the structural damage of colon tissue under diabetic conditions, damage to hippocampal neurons, and improve the exploration ability and spatial learning and memory ability of diabetic encephalopathy mice. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The results of the water maze escape latency of mice after treatment with baicalein (BE), Bacteroides uniformis (B.uniformis), and different ratios of the two drugs;

[0024] Figure 2 are the results of behavioral tests, where A is the statistics of the number of platform crossings of each group of mice in the water maze test, B is the statistics of the target quadrant movement time of each group of mice in the water maze, and C is the movement trajectory of each group of mice in the water maze;

[0025] Figure 3 is the result of the open field test, where A is the total moving distance of each group of mice, B is the total moving time, C is the number of times entering the central area, and D is the movement trajectory diagram and heat map of each group of mice in the open field;

[0026] Figure 4is the result of the Y-maze test, where A is the escape latency of mice in each treatment group to enter the novel arm, B is the total moving time of mice in each group in the Y-maze after drug treatment, C is the percentage statistics of mice in each group entering the novel arm, and D is the trajectory diagram and movement heat map of mice in each group in the Y-maze;

[0027] Figure 5 This is a diagram showing a stained section of mouse colon tissue;

[0028] Figure 6 Nissl-stained sections of hippocampus and cortex. DETAILED DESCRIPTION

[0029] The technical scheme of the present invention is further described below in conjunction with the examples. The test materials used in the examples can all be purchased through conventional channels.

[0030] Example 1

[0031] Bacteroides uniformis ATCC 8492 was purchased from Beijing Beina Chuanglian Biotechnology Research Institute, No.: BNCC 240886. The lyophilized bacterial powder was activated on Columbia blood agar plates, and the bacteria were cultured under anaerobic conditions. After 1-2 generations, they were used in the experiment. The first generation of bacteria with strong bacterial viability and stable metabolism were selected and transferred to 5ml liquid culture medium and cultured under anaerobic conditions for 48h. The preparation method of liquid culture medium is as follows: 9.8g GAM culture medium (purchased from Haibo Biotechnology, product number: HB8518-1) was heated and dissolved in 200ml distilled water, autoclaved at 121℃ for 15min, and when cooled to about 50℃, 1 ml hemin (purchased from Haibo Biotechnology, product number: HB0310a) and 100μl vitamin K1 (purchased from Haibo Biotechnology, product number: HB0310b) were added, mixed and set aside, and 1.8% agar was added to pour the plate to make a solid culture medium. After 48 hours, the bacterial solution was diluted gradiently, the absorbance was measured and 100 μl of the bacterial solution was taken for plate counting. The dilution multiples, OD values ​​and colony counts of different gradients were recorded to determine the linear relationship between absorbance and bacterial concentration.

[0032] Every day, take the B.uniformis bacterial suspension with good activity, centrifuge at 4000g for 15min, and resuspend it in physiological saline to adjust the concentration of the bacterial suspension. Weigh an appropriate amount of baicalin and dissolve it in physiological saline. Mix the bacterial suspension and baicalin solution to obtain the B.uniformis+baicalin solution. The ratios are as follows: low-dose group: 25mg / kg baicalin + 5×10 8 CFU / mL B. uniformis, medium dose group: 50mg / kg baicalin + 5×10 8CFU / mL B.uniformis, high-dose group: 100 mg / kg baicalin + 5×10 8 CFU / mL B.uniformis. In addition, the control group, HFD+Se group, Live Bu group and baicalein group were set up, and the dosage was as follows: Live Bu group: 5×10 8 CFU / mL, baicalein group: 100 mg / kg, HFD+Se group and Control group were treated with normal saline in parallel, the administration volume was 200 μL, and the administration lasted for 4 weeks.

[0033] Animal behavioral assessment plays an important role in the evaluation of animal models of cognitive dysfunction-related diseases. Mouse behavioral measurements were performed in the fourth week to evaluate the cognitive function of each group of mice. 24 hours after the last behavioral experiment, 3 mice were randomly selected from each group and decapitated. The brain tissue was carefully removed and fixed in paraformaldehyde, and then paraffin sections were prepared for Nissl staining to observe the morphology of neurons in the hippocampus of each group. The colon area was separated and fixed in paraformaldehyde, then paraffin-embedded and sectioned for HE staining to observe the structural status of the colon area of ​​each group of mice.

[0034] Example 2

[0035] Water maze to evaluate the spatial cognition ability of mice:

[0036] The Morris Water Maze (MWM) experiment is divided into two parts, the first part is a directional navigation experiment, and the second part is a spatial exploration experiment. The water maze device consists of a circular pool with a diameter of 120 cm and a height of 60 cm. A platform with a diameter of 10 cm and a height of 1 cm below the water surface is placed in the pool. The pool wall is marked with obvious signs to provide more visual information related to spatial positioning. The mouse is put into the water with the purpose of finding the platform hidden in the water and successfully escaping from the water. The experiment uses Xeye software to detect the mouse trajectory. Xeye software simulates the pool into four quadrants, in which the platform is placed in the center of a quadrant, which is set as the target quadrant (the quadrant where the platform is located is the target quadrant). Titanium dioxide is added to the water to make the water white, so that the background is deepened and the running trajectory of the black mouse can be clearly captured. In the first part of the directional navigation experiment, the mouse is placed head to the pool wall and randomly selected into a quadrant. The experiment records the escape latency of the mouse, that is, the time it takes for the mouse to find the underwater platform within 90 seconds and stay for 3 seconds. In the experiment, if the mouse failed to find the platform within 90 seconds or found the platform but failed to stay for 3 seconds, the escape latency was recorded as 90 seconds. After the experiment, the mouse was guided to climb onto the platform and allowed to adapt for 15 seconds to enhance its spatial memory ability. In the second part of the spatial exploration experiment, on the second day after the directional navigation training experiment, the platform was removed and a 90-second exploration experiment was conducted. The mouse was placed in the quadrant opposite to the target quadrant, and its movement trajectory was recorded. The target quadrant time / number of platform crossings and escape latency were recorded using Graphpad.

[0037] The results of behavioral tests were Figure 2 As shown in (A) and (B), in the water maze test, the number of crossing the platform in the low-dose group and the medium-dose group was significantly more than that in the HFD+Se group (P<0.05; P<0.05). In terms of the target quadrant activity time, the BE group, the low-dose group, the medium-dose group, and the high-dose group all showed good therapeutic effects (P<0.05; P<0.01; P<0.05; P<0.05). The efficacy of the combined administration of baicalin and Live Bu was significantly better than that of the single administration. In the combined administration, the low-dose group 25 mg / kg baicalin + 5×10 8 The administration of CFU / mL showed better therapeutic effect. During the positioning and navigation process of mice, Figure 1 As shown in the figure, compared with the HFD+Se group, each treatment group showed a good improvement in cognitive ability. Among them, starting from the third day, compared with the HFD+Se group, the time for the low-dose and high-dose combined drug groups to reach the platform was significantly shortened (P<0.05). On the fifth day, the time for the mice in the low-dose combined drug group to find the platform was significantly lower than that in the HFD+Se group (P<0.05), indicating that under this drug administration method, the learning and spatial memory abilities of mice with diabetic encephalopathy were best restored. Figure 2 (C) The movement trajectories of mice in each group in the water maze show that after treatment, the trajectory routes of mice in each group are more inclined to the target quadrant where the platform is located, indicating that the memory ability of mice has been improved after drug treatment.

[0038] Example 3

[0039] Evaluation of the mice's ability to move autonomously by open field testing:

[0040] The open field test (OFT) is used to evaluate the exploratory behavior of animals in a new environment. Four identical open boxes (50cm*50cm*50cm) were placed in a quiet and spacious room. The material was polyvinyl chloride (PVC) and had no odor. The recording camera was fixed above the open box. The ANY-MAZE software was used for the experiment. The software simulated the division of the bottom of the 50cm*50cm open box into 25 small grids of 10cm*10cm. The middle 9 grids (3*3) were assumed to be the central area, and the remaining grids around were assumed to be the edge area. There was no overlap between the areas. ANY-MAZE will record the movement trajectory of the mice in each area. The mice were placed in the room for 1 hour before the experiment to adapt to the indoor environment. At the beginning of the experiment, the mice will explore freely in the open box for 5 minutes. After each experiment, the mouse excrement in the open box was cleaned and wiped thoroughly with 75% alcohol to completely cover up the residual smell of the mice. The total moving distance, total moving time and number of times the mice entered the central area in each group were recorded by Graphpad.

[0041] In the open field test, Figure 3 It can be seen that compared with the HFD+Se group, the Live Bu group, the baicalein group and the combined drug treatment group all showed obvious therapeutic effects, among which the combined drug treatment had the most significant effect. In terms of the total moving distance in the open field, compared with HFD+Se, the low, medium and high doses of combined drug treatment groups significantly increased the moving distance of mice (P<0.01, P<0.01, P<0.05). As shown in (B), compared with the HFD+Se group, the Live Bu group and the low, medium and high doses of combined drug treatment groups also significantly increased the total moving time of diabetic encephalopathy mice (P<0.05, P<0.01, P<0.01, P<0.05). In the open field test, the number of times mice enter the central area can indicate the exploration desire of mice. As shown in (C), compared with HFD+Se, the low, medium and high doses of combined drug treatment groups significantly increased the exploration desire of mice (P<0.01; P<0.05; P<0.05). It can be seen from the movement trajectories of each group of mice that after treatment, the autonomous activity ability and exploration interest of each group of mice were improved. Among them, the movement trajectories of the mice in the combined drug treatment group were more scattered, indicating that the mice with diabetic cognitive impairment received more effective treatment.

[0042] Example 4

[0043] The spontaneous exploration ability of mice was evaluated by Y-maze:

[0044] The Y-maze test is a spontaneous alternating exploration of animals in a Y-shaped structure, which is used to measure the willingness of mice to explore a new environment. The Y-maze apparatus consists of three identical PVC arms spaced 120° apart. The material is opaque, and each arm is 30 cm long, 6 cm wide, and 15 cm high. When exploring, mice usually prefer to explore unfamiliar arms (novel arms) rather than exploring previously explored arms. Therefore, one arm of the Y-maze is randomly blocked, defined as the novel arm, and additional signs are posted on the entrance wall of the novel arm to provide more obvious maze clues. Before the start of the experiment, the mice were placed in the room 1 hour in advance to adapt to the indoor environment. The ANY-MAZE software was used for the experiment. At the beginning of the experiment, mice were randomly placed in the other two arms. The mice were allowed to freely explore the two arms for 10 minutes, and then the mice were put back into their own cages. After an interval of 1 hour, the novel arm was opened and the mice were placed in the Y maze again to freely explore the three arms for 5 minutes. When the four limbs of the mice entered the new arm, it was defined as entering an arm. After each mouse was finished with the experiment, the excrement left in the arm was cleaned and thoroughly cleaned with 75% alcohol. The movement trajectory of the mice was recorded, and the escape latency and residence time of each group entering the novel arm were compared. The percentage of entering the novel arm was calculated as follows:

[0045]

[0046] In the Y-maze test, Figure 4 As shown in (A), the escape latency of mice in each treatment group to enter the novel arm tended to be shortened; Figure 4 As can be seen in (B), after drug treatment, the movement time of mice was prolonged compared with the HFD+Se group, among which the Live group and the low and medium dose groups of BE combination showed significant differences (P<0.05; P<0.01; P<0.01; P<0.05); Figure 4 As shown in (C), the percentage of diabetic encephalopathy mice entering the novel arm was statistically increased in the low and medium dose groups of combined administration (P < 0.01; P < 0.01), indicating that after combined administration, the spatial exploration and memory abilities of diabetic encephalopathy mice were significantly improved. Figure 4 Middle (D) reflects the movement of mice in each group in the Y maze after drug treatment. It can be seen that Live Bu, baicalin, and Live B.u+baicalin combined administration can significantly improve the spatial exploration and learning and memory abilities of diabetic encephalopathy mice.

[0047] Example 5

[0048] Effects on mouse colon tissue:

[0049] Place the colon tissue in 4% paraformaldehyde for 24 hours and then dehydrate it with gradient methanol. Dehydrate it in 75% ethanol, 90% ethanol, 95% ethanol, and anhydrous ethanol for 10 minutes. Soak the tissue in xylene to make it transparent, then soak it in heated molten wax for 2 hours, pour the molten wax into the mold, immediately use the preheated tweezers to clamp the tissue and put it into the mold, with the cross section facing down, and trim the wax block after solidification. Adjust the slicer, cut the wax block into 4μm thick, place it in 42℃ warm water to unfold the wax slice, use a clean slide to pick up the wax slice so that it is located in the center of the slide and wait for drying and staining. Dewax with xylene I and II solutions respectively, and then treat it in gradient anhydrous ethanol for 1 minute in turn, put the slice in hematoxylin staining solution for 10 minutes, then wash off the floating color with water, then put the slice in eosin staining solution for 10 minutes, wash off the skin color with water in turn, and repeat the above steps until the slice is stained blue. Put 2-3 drops of gum on the stained glass slide to cover the tissue, and cover the slide with a clean cover glass at an angle. Then use a slide scanner to scan the slide.

[0050] from Figure 5 It can be seen that the mucosal layer and submucosal layer of the Control group had normal microstructures, regular arrangement of mucosal glands, complete crypts, and a large number of goblet cells; the crypt structure of the colon in the HFD+Se group was severely damaged, with irregular surfaces, fewer goblet cells, and a large number of inflammatory infiltrations; the colon tissue structure of the baicalein group was relatively complete, with a small amount of inflammatory infiltrations scattered among the goblet cells, indicating that drug administration can improve the colon tissue damage caused by a high-selenium and high-fat diet; the crypt structure of the mice in the Live Bu group became more normal, the goblet cells were full and arranged more regularly, and the number of goblet cells was less compared with the control group; for the combined drug administration group, it can be seen that compared with the HFD+Se group, the combined administration of low, medium, and high doses improved the colon villus structure of the mice better, with full goblet cells and glands, neatly arranged, and rare inflammatory infiltrations. This indicates that after the intervention of B. uniformis and baicalein, the colon structure damage of diabetic encephalopathy mice was alleviated.

[0051] Example 6

[0052] Effects on Mouse Brain Neurons:

[0053] After the mice were killed, they were quickly decapitated and the whole brain was carefully removed and placed in 4% paraformaldehyde for tissue fixation. After 24 hours, they were dehydrated, paraffin-embedded, and serially sectioned with a thickness of 5 μm. The same parts were selected for sectioning and attached to slides for Nissl staining to observe the changes in the neurons in the hippocampus of the brain sections of each group of mice.

[0054] Depend on Figure 6 From the sections, it can be seen that the neurons in the hippocampus and cortex of the mice in the Control group were plump and closely arranged, with many Nissl bodies visible; the neurons in the CA3 region of the hippocampus of the mice in the HFD+Se group were disorderly arranged, with obvious neuronal damage, and the Nissl bodies in some neuronal cells dissolved to form translucent areas, and a large number of neurons in the CA3 region and cortex appeared pyknosis, as shown by the arrows; compared with the HFD+Se group, the neuronal cells in the CA3 region of the hippocampus in the LiveB.u group had a more complete structure, large and round nuclei, and a more closely arranged neuronal damage; the neuronal cells in the cortical region had a more regular morphology, plumper cell bodies, and fewer pyknosis and necrotic neurons. The neuronal damage of the LiveBu mice was greatly improved, with neurons in the CA3 region of the hippocampus closely arranged, plump cell bodies, large and round nuclei, more Nissl bodies, and relatively uniform distribution; the neurons in the cortical region were loosely arranged, but the nuclear pyknosis of neurons was reduced. The neuronal damage in the CA3 and cortical regions of the brain in the baicalin group was significantly improved, the nerve cells were arranged relatively neatly, fewer Nissl bodies were lost, and the nuclear pyknosis was improved. The state of the neuronal cells in the mice treated with combined medication was improved to a certain extent. The CA3 cells in the low-dose combined medication group were loosely arranged, but the pyknosis was reduced, and individual nerve cell damage was seen in the cortical region; the CA3 nerve cell damage in the medium-dose group was still relatively large, and the arrangement was loose, and individual pyknosis nerve cells were scattered in the cortical region; the state and arrangement of nerve cells in the CA3 region of the hippocampus in the high-dose combined medication group were well improved, and there were still damaged nerve cells scattered in the cortical region. The experimental results show that LiveB.u, baicalin and the combination of the two can improve the neuronal damage in mice with diabetic encephalopathy complications.

[0055] Therefore, the composition of the intestinal microorganism Bacteroides monotheca and the traditional Chinese medicine ingredient baicalein of the present invention can effectively alleviate diabetic encephalopathy induced by a high-fat diet and aggravated by high selenium, achieve the effect of improving colon tissue structure damage and hippocampal neuron damage under diabetic conditions, and improve the spatial learning and memory ability of patients with diabetic encephalopathy.

Claims

1. A composition for improving diabetic encephalopathy, characterized in that: The composition comprises intestinal microorganism Bacteroides monomorpha and a traditional Chinese medicine component monomer baicalin.

2. The composition according to claim 1, characterized in that The diabetic encephalopathy is diabetic encephalopathy aggravated by excessive selenium.

3. The composition according to claim 1, characterized in that The composition is 10-150 mg / kg of baicalein monomer and 5×10 7 -5×10 9 CFU / mL combination.

4. Use of the composition according to claim 1 in the preparation of a drug for treating cognitive impairment complications caused by a high-selenium and high-fat diet in animals.

5. The use according to claim 4, characterized in that: The composition is orally administered to treat cognitive impairment complications caused by animals fed with a high-selenium and high-fat diet.

6. The use according to claim 5, characterized in that: The application is to mix the composition in sterile physiological saline and administer orally.

7. The use according to claim 4, characterized in that: The application is to alleviate diabetic encephalopathy induced by a high-fat diet and aggravated by high selenium, improve the structural damage of colon tissue and the damage of hippocampal neurons in the diabetic state, and improve the spatial learning and memory ability of patients with diabetic encephalopathy.

8. The use according to claim 4, characterized in that: The application is to utilize the microbial community-gut-brain axis to regulate the abundance and proportion of intestinal bacteria, change the composition of the intestinal flora, and then change the ability of intestinal microorganisms to decompose amino acid substrates, regulate short-chain fatty acid metabolism and neurotransmitter levels, and protect neurons from damage by oxidative stress.

9. The use according to claim 4, characterized in that: The dosage form of the drug is capsule, tablet or oral film.

10. The use according to claim 4, characterized in that: The drug is an enteric-coated probiotic preparation, and the preparation method comprises the following steps: inoculating Bacteroides monotheca in an anaerobic fermentation environment, continuing to culture under anaerobic conditions until the late logarithmic growth stage or the late stable stage of Bacteroides monotheca to obtain a fermentation product; mixing baicalein with the fermentation product, spray-drying the mixture for storage, and coating the surface with an enteric-coated material to obtain the drug.

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