Application of composition containing cold region berry and cold region medicinal and edible Chinese medicine in improving intestinal flora function of alzheimer disease

Abstract

The application discloses application of a composition containing cold land berries and cold land medicinal and edible Chinese herbs in improving intestinal flora function of Alzheimer's disease. The cold land berries contain blueberries, wild cranberries and blue honeysuckle fruits; the cold land medicinal and edible Chinese herbs include acanthopanax powder, schisandra extract and astragalus extract. Preferably, the composition further contains grape seed extract and vitamin C powder. Experiments prove that the composition containing the cold land berries and the cold land medicinal and edible Chinese herbs can improve richness and uniformity of intestinal flora, and can significantly down-regulate six genera of Staphylococcus (Staphylococcus), Vibrio (Vibrio), Collinsella (Collinsella), Actinomycetales (Actinomycetales), Aeromonas (Aeromonas) and Holdemanella (Holdemanella) which are closely related to Alzheimer's disease, so as to achieve the purpose of improving intestinal flora function of Alzheimer's disease.

Description

TECHNICAL FIELD

[0001] The present application relates to a composition containing cold region berries and cold region medicinal and edible Chinese herbs, and also relates to the application of the composition in improving the intestinal flora function of Alzheimer's disease. The present application belongs to the technical field of cold region berry and Chinese herbal preparation. BACKGROUND

[0002] Alzheimer's disease (AD) is an age-related neurodegenerative disease, and there is currently no effective clinical treatment method. The clinical features are the gradual decline of memory and cognitive function caused by a series of factors. More and more evidence shows that the imbalance of human intestinal microbiota is related to neurodegenerative diseases such as Alzheimer's disease through the neuroinflammatory process of the microbiota-gut-brain axis. Intestinal microbiota affects intestinal health by secreting toxins and short-chain fatty acids, thereby regulating intestinal permeability and various immune functions. Intestinal microbiota imbalance can lead to increased permeability of the intestinal epithelial barrier, thereby releasing pro-inflammatory cytokines and promoting neuroinflammatory reactions. Studies have shown that the diversity of intestinal microorganisms in patients with Alzheimer's disease is reduced, which may contribute to the study of the pathogenesis and treatment of the disease. The brain / gut axis may play a certain role in the pathogenesis of AD. AD model mice have excessive Aβ deposition in intestinal epithelial cells, accompanied by increased intestinal permeability, inflammatory changes, and reduced expression of tight junction proteins. These changes in intestinal barrier lead to increased proliferation of goblet cells and Paneth cells and increased mucus synthesis. AD occurs intestinal barrier dysfunction, which may be one of its causes. Future treatment strategies to reverse AD pathology may involve early manipulation of intestinal physiology and its microbiota.

[0003] The regulation of intestinal flora is heavily dependent on external factors, which provides an opportunity to advance the treatment of neurodegenerative diseases. Diet plays a special role, as it provides metabolites that directly affect the body and microbiota. Therefore, a holistic view of the human organism is essential. A healthy microbiota is a state in which most potentially beneficial bacterial species coexist in harmony in the gastrointestinal tract. Although a healthy person's microbiota is usually stable in a healthy state, food and lifestyle choices can affect the dynamic balance of the composition of the intestinal microbiota. Unhealthy lifestyle habits can lead to dysbiosis, which is defined as a disturbance of the intestinal microbiota, which disrupts intestinal homeostasis, triggers inflammation, and has a negative impact on intestinal permeability.

[0004] Berrys and cold region traditional Chinese medicines have broad application prospects in the prevention and treatment of Alzheimer's disease. Traditional Chinese medicines and berrys have multiple active ingredients and multiple targets, and are increasingly valued by the international community. Traditional Chinese medicine adopts holistic therapy to restore the overall functional balance of the human body. These interventions show good results in correcting metabolic imbalances, which is a common phenomenon in Alzheimer's disease. Wild berrys in cold regions, such as Vaccinium uliginosum, Lonicera caerulea and blueberries, are wild berrys exposed to cold highland environments. They produce more polyphenolic compounds to enhance their defense functions and protect the plants themselves from external factors. Therefore, cold region berrys have more bioactive compounds with health benefits. Cold region berrys are also rich in a variety of secondary metabolites with medicinal value, including major components such as phenolic acids and flavonoids, which have strong antioxidant and cell protection properties. Regular consumption of berrys throughout the year plays an important role in preventing or reducing neurodegenerative diseases. Cold region medicines with the same origin as food, such as Acanthopanax senticosus, Schisandra chinensis and Astragalus membranaceus, play an important role in resisting the pathological mechanism of Alzheimer's disease.

[0005] Therefore, the present application provides a composition composed of cold region berrys and cold region medicines with the same origin as food, which can improve the intestinal flora function of Alzheimer's disease patients and achieve the purpose of preventing and treating Alzheimer's disease. SUMMARY

[0006] The present application aims to overcome the technical deficiencies of existing Alzheimer's disease treatment drugs and provide a composition containing cold region berrys and medicines with the same origin as food to improve the intestinal flora of Alzheimer's disease and the application of the composition in improving the intestinal flora function of Alzheimer's disease.

[0007] In order to achieve the above-mentioned purpose, the present application adopts the following technical means:

[0008] The present application provides the application of a composition containing cold region berrys and cold region medicines with the same origin as food in improving the intestinal flora function of Alzheimer's disease. The cold region berrys include blueberries, wild cranberries and blueberries. The cold region medicines with the same origin as food include Acanthopanax senticosus powder, Schisandra chinensis extract and Astragalus membranaceus extract.

[0009] Preferably, the composition further contains grape seed extract and vitamin C powder.

[0010] Preferably, the composition contains 800-900 ml of wild cranberry juice, 4000-4500 ml of blueberry juice, 100-200 ml of blueberry juice, 30-40 ml of Schisandra extract, 100-200 ml of Astragalus extract, 400-500 mg of Acanthopanax powder, 16000-17000 mg of grape seed extract, 25000-30000 mg of vitamin C powder, and distilled water, with the distilled water making up the volume to 10 L. More preferably, the composition contains 833 ml of wild cranberry juice, 4163 ml of blueberry juice, 167 ml of blueberry juice, 33 ml of Schisandra extract, 167 ml of Astragalus extract, 433 mg of Acanthopanax powder, 16650 mg of grape seed extract, 29304 mg of vitamin C powder, and distilled water, with the distilled water making up the volume to 10 L.

[0011] Preferably, the wild cranberry juice, blueberry juice, and blueberry juice are prepared from wild cranberries, blueberries, and blueberries in Daxing'anling, respectively, and the preparation method is as follows:

[0012] (1) The quick-frozen wild cranberries, blueberries, or blueberries are manually selected and then slowly thawed at room temperature for 12-24 h;

[0013] (2) The thawed berries are crushed in a crusher, and the crushed juice is pumped into a plate heat exchanger, heated, and then pumped into an enzymatic hydrolysis tank. Pectinase is added at a proportion of 1 / 10,000 of the weight of the juice, and the juice is stirred and enzymatically hydrolyzed at a temperature of 40-50°C for 3.5-4 h to obtain enzymatically hydrolyzed juice;

[0014] (3) The enzymatically hydrolyzed juice is pumped into a press for 6 min at a pressure of less than 200 bar and a temperature of less than 20°C, and then pumped into a 120-mesh vibrating screen for separation. The separated juice is pumped into a temporary storage tank;

[0015] (4) The separated juice is heated in a plate heat exchanger, and then pumped into a separator for negative pressure enzyme inactivation treatment. The enzyme inactivation process requires a vacuum negative pressure of 0 to -0.5 bar and an outlet material temperature of 50-70°C. After enzyme inactivation, the juice is pumped into a temporary storage tank;

[0016] (5) Pump the juice after enzyme inactivation into a vacuum rotary filter, diatomite filter and paper filter, and perform three filtering, that is, first, prepare the filtering medium by mixing diatomite, expanded perlite and water according to a ratio of 1:9:90, pump into the storage tank below the vacuum rotary filter, start the vacuum rotary filter, and wait until 3-5 cm thick filtering medium is adhered to the surface of the machine, adjust the distance of the scraper so that the thickness of the medium is kept at 2.5-3.5 cm, control the negative pressure of the vacuum rotary filter at-50 to-70 cmHg, the pressure of the diatomite filter at 4-5 bar, and the pressure of the paper filter at 20 bar or less;

[0017] (6) Pump the juice after three filtering into a tank for storage, and obtain wild cranberry juice, blueberry juice and blueberry juice.

[0018] Preferably, the preparation method of the Schisandra extract is as follows: freeze the fresh Schisandra fruit, uniformly place the frozen fruit in a baking tray, set the temperature of the oven to 120 DEG C and bake for 5-6 hours, reduce the temperature to 60-65 DEG C and bake for 18-20 hours, dry the fruit, grind the dried fruit, add 750 kg of water to 50 kg of the ground fruit, heat to boiling, boil for 4 hours, place for 12 hours, filter the residue, perform tubular centrifugal filtration, and then use the finished product.

[0019] Preferably, the preparation method of the Astragalus extract is as follows: boil the stems and leaves of Astragalus with water according to a weight ratio of 1:10, boil for 4-5 hours, centrifugally filter the boiled liquid, then concentrate 10 times, and use the finished product.

[0020] Preferably, the composition containing cold land berries and cold land medicine-food homologous Chinese medicines can improve the richness and uniformity of intestinal flora, and can significantly down-regulate six bacteria genera, Staphylococcus (Staphylococcus), Vibrio (Vibrio), Collinsella (Collinsella), Actinomycetales (Actinomycetales), Aeromonas (Aeromonas) and Holdemanella (Holdemanella), which are closely related to Alzheimer's disease, so as to achieve the purpose of improving the intestinal flora function of Alzheimer's disease.

[0021] Compared with the prior art, the present application has the following advantages:

[0022] 1. The present application provides a composition containing cold land berries and cold land medicinal and edible similar traditional Chinese medicines. Experiments have proved that the composition can reduce the abundance of Staphylococcus (Staphylococcus), Vibrio (Vibrio), Collinsella (Collinsella), Actinomycetales (Actinomycetales), Aeromonas (Aeromonas), Holdemanella (Holdemanella) and other closely related bacterial genera of Alzheimer's disease, improve the richness of intestinal flora, and make the composition of intestinal flora closer to the healthy control group, thereby improving the intestinal flora function of Alzheimer's disease patients, and further achieving the purpose of preventing and treating Alzheimer's disease through brain-gut axis.

[0023] 2. The composition of the present application is composed of cold land berries and cold land medicinal and edible similar traditional Chinese medicines, grape seed extract and vitamin C powder. The raw materials are safe and have no side effects, are rich in nutrients, have good taste, and are suitable for long-term consumption. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 Chao1 value of intestinal flora in each group;

[0025] Figure 2 Shannon plot of intestinal flora in each group;

[0026] Figure 3 Jaccard_pcoa plot of intestinal flora in each group;

[0027] Figure 4 Jaccard_NMDS plot of intestinal flora in each group;

[0028] Figure 5 Genus_abund_Group_cluster plot of intestinal flora in each group;

[0029] Figure 6 Species_abund_Group_cluster plot of intestinal flora in each group;

[0030] Figure 7A -C is the abundance difference analysis of 14 common differential genera (the vertical coordinate in the figure is the relative abundance x 100000). DETAILED DESCRIPTION

[0031] The advantages and characteristics of the present application will be more apparent with the description of the specific examples. However, the examples are only exemplary and do not constitute any limitation on the scope of the present application. Those skilled in the art should understand that the details and forms of the technical solutions of the present application can be modified or replaced without departing from the spirit and scope of the present application, and such modifications and replacements all fall within the protection scope of the present application.

[0032] Application of the composition of wild berry and wild medicinal food in cold region in improving the intestinal flora function of Alzheimer's disease

[0033] 1. Preparation of raw materials

[0034] Cranberry juice, blueberry juice, blueberry juice, Schisandra chinensis extract, Astragalus extract, Acanthopanax powder, grape seed extract, vitamin C powder. Among them, vitamin C is purchased from Xintaiyang Biological Engineering Co., Ltd.; grape seed extract is purchased from Shaanxi KeHong Health Industry Co., Ltd.; Acanthopanax powder is purchased from Shaanxi Lituo Plant Chemical Industry Co., Ltd.

[0035] Cranberry juice, blueberry juice, blueberry juice are prepared from wild cranberry, blueberry and blueberry in Daxing'anling, the specific preparation method is as follows: first, the quick-frozen berries are selected by hand and then slowly thawed at room temperature for 12-24 hours. The thawed berries are crushed in a crusher, and the crushed juice is pumped into a plate heat exchanger for heating and then into an enzymolysis tank. Pectinase is added at a proportion of 1 / 10,000 of the weight of the juice, and the juice is stirred for enzymolysis. The enzymolysis reaction temperature is required to be 40-50℃, and the time is 3.5-4 hours. The enzymolysis juice is pumped into a press for 6 minutes, with a pressure less than 200 bar and a temperature less than 20℃. The pressed juice is pumped into a 120-mesh vibrating screen for separation, and the separated juice is pumped into a storage tank for temporary storage. The pressed juice is heated in a plate heat exchanger and then pumped into a separator for negative pressure enzyme inactivation treatment. The process requires a vacuum negative pressure of 0 to -0.5 bar and an outlet material temperature of 50-70℃. After enzyme inactivation, it is pumped into a clarification tank for temporary storage. The enzyme-inactivated juice is pumped into a vacuum rotary filter, diatomite filter and paper filter for three-stage filtration. First, the filter medium is prepared by mixing diatomite, expanded perlite and water at a ratio of 1:9:90, and then pumped into the lower storage tank of the vacuum rotary filter. After the machine surface is adhered with a 3-5 cm thick filter medium, the scraper distance is adjusted to maintain a medium thickness of 2.5-3.5 cm. The vacuum degree of the vacuum rotary filter is controlled at -50 to -70 cmHg, the diatomite filter pressure is 4-5 bar, and the paper filter pressure is within 20 bar. The three-stage filtered juice is pumped into a tank for storage.

[0036] The preparation method of Schisandra chinensis extract is as follows: the fresh Schisandra chinensis fruit is frozen and then taken out of the freezer, evenly placed in a baking tray, and baked in an oven set at a temperature of 120℃ for 5-6 hours, and then baked at a temperature of 60℃-65℃ for 18-20 hours. After drying, the Schisandra chinensis powder is crushed, 50 kg of the crushed powder is added with 750 kg of water, heated to boiling, boiled for 4 hours, and then filtered after standing for 12 hours. The residue is then filtered by a tubular centrifuge, and the finished product is packaged for use.

[0037] The preparation method of the Astragalus extract solution is as follows: 400 kg of Astragalus stems and leaves are boiled with 4000 kg of water for 4-5 hours, the boiled liquid is centrifuged and filtered, and then 10 times concentration is performed, and the finished product is packaged and used.

[0038] Vitamin C is dissolved in deionized water with an initial concentration of 0.88 g / 5 mL. The grape seed extract is dissolved in methanol with an initial concentration of 20 mg / mL, and then diluted with PBS to the required concentration. The Acanthopanax powder is dissolved in water to obtain an Acanthopanax water solution with an initial concentration of 0.51 mg / mL, and then diluted with PBS by a factor of 5.

[0039] 2. The cytotoxicity and antioxidant damage protection of each raw material are determined respectively

[0040] The determination index is the survival rate of the human neuron cell line SY-HY5Y after administration of the raw materials. Then four dose levels are selected for each raw material, and the final concentrations of the first six (wild cranberry juice, blueberry juice, blueberry juice, Schisandra extract solution, Astragalus extract solution, and Acanthopanax water solution) are 1 / 5 2 , 1 / 5 3 , 1 / 5 4 , and 1 / 5 5 of the initial concentration, respectively, and the orthogonal table corresponds to "1", "2", "3", and "4", respectively. The final concentration levels of the grape seed extract and the vitamin C water solution are 1 / 5 3 , 1 / 5 4 , 1 / 5 5 , and 1 / 5 6 of the initial concentration, respectively, and the orthogonal table corresponds to "1", "2", "3", and "4", respectively. An orthogonal test table of 8 factors and 4 levels is established, and a total of 32 test groups are obtained (Table 1).

[0041] The normalization value calculation method is as follows: 1) The survival rate is calculated using the cell culture medium without cells as a blank well, and the formula is (OD 实验组 -OD 空白 ) / (OD 对照组 -OD 空白 ); 2) The normalization value = survival rate 实验组 / survival rate 损伤组 .

[0042] Table 1 Orthogonal design table

[0043]

[0044]

[0045]

[0046] The first round of experiments by the above results: according to the range R value arrangement of primary and secondary factors, namely: wild cranberry / blueberry / grape seed / Chinese milk vetch / acanthopanax senticosus / indigo fruit / VC / schisandra, according to the average k value of each raw material, the best level, according to the best level, the best combination, namely: wild cranberry(4) indigo fruit(1) / blueberry(1) / schisandra(3 / ) Chinese milk vetch(2) / acanthopanax senticosus(1) / grape seed extract(4) / VC(2).

[0047] According to the repeated experiments, the following primary and secondary factors are obtained, namely: wild cranberry / VC / indigo fruit / Chinese milk vetch / schisandra / blueberry / grape seed / acanthopanax senticosus is the best combination, namely: wild cranberry(3) / indigo fruit(1) / blueberry(3) / schisandra(1) / Chinese milk vetch(4) / acanthopanax senticosus(1) / grape seed extract(3) / VC(2).

[0048] After multiple rounds of determination, the optimal level of each raw material is determined by sorting method, range method and orthogonal experiment variance analysis, and then the optimal combination is combined for testing.

[0049] 3. Establishment of composition of cold region berry and cold region medicine-food homologous traditional Chinese medicine

[0050] Raw materials: wild cranberry juice, indigo fruit juice, blueberry juice, schisandra extract, Chinese milk vetch extract, acanthopanax senticosus powder, grape seed extract, vitamin C powder, distilled water.

[0051] The final selected formula for cell experiment is: wild cranberry(2)+indigo fruit(1)+blueberry(3)+schisandra(4)+Chinese milk vetch(2)+grape seed extract(1)+vitamin C(2). According to the calculation, the composition of cold region berry and cold region medicine-food homologous traditional Chinese medicine is configured as follows: the total volume of the configured formula is 10000ml(10L). Among them, wild cranberry juice 833ml, indigo fruit juice 4163ml, blueberry juice 167ml, schisandra extract 33ml, Chinese milk vetch extract 167ml, acanthopanax senticosus powder 433mg, grape seed extract 16650mg, vitamin C powder 29304mg, distilled water 4639ml. The reagent required for mouse experiment should be 260 times the final concentration of cell experiment.

[0052] 4. Animal experiment

[0053] Feeding method and animal grouping: 5-month-old male APP / PS1 double transgenic mice and littermate negative wild-type mice were selected for 8w gavage administration. The APP / PS1 mice were randomly divided into the composition (referred to as "composition") treatment group (medium-dose group and high-dose group) of cold region berry and cold region medicine-food homologous traditional Chinese medicine combination, positive control drug treatment group, placebo group, and were respectively given 6ml / kg / d of the composition (medium-dose treatment group), 12ml / kg / d of the composition (high-dose treatment group), 0.9mg / kg / d of donepezil (positive control drug treatment group), 6ml / kg / d of 0.9% wild cranberry, and WT group mice were also given 6ml / kg / d of 0.9% saline. Before gavage, 5-month-old male APP / PS1 double transgenic mice (AD_0W) and littermate negative wild-type mice (WT_0W) were subjected to intestinal flora analysis by taking feces; after 8 weeks (8W) of gavage feeding using the above gavage method, the feces were again taken for intestinal flora experiment, and the improvement of the composition on intestinal flora was compared and analyzed.

[0054] Note: WT is wild-type mice, WT_0w is wild-type mice without gavage, WT_veh is wild-type mice gavaged with saline for 8W; AD is an Alzheimer's disease model mouse APP / PS1 mouse, AD_0w is an APP / PS1 mouse without gavage, AD_M is an APP / PS1 mouse gavaged with a medium-dose composition for 8W, AD_H is an APP / PS1 mouse gavaged with a high-dose composition for 8W, AD_DPZ is an APP / PS1 mouse gavaged with donepezil for 8W, and AD_cran is an APP / PS1 mouse gavaged with wild cranberry for 8W.

[0055] 5. Intestinal flora experiment results:

[0056] 5.1 Alpha analysis results

[0057] 1) Chao1 value analysis of each group of intestinal flora richness

[0058] According to the chao1 value of each group of intestinal flora, the fecal sample process of different drug treatment groups in the experiment was analyzed to compare the richness of intestinal flora. The results are shown in Figure 1 As can be seen from the experimental results, the richness of intestinal flora in the donepezil treatment group and the control treatment group is the lowest, followed by the medium-dose composition treatment experimental group (AD_M), the disease model group and the healthy control group are close, and the high-dose composition treatment experimental group (AD_H) is slightly higher.

[0059] 2) OUT number value analysis of each group of intestinal flora richness

[0060] The abundance of gut microbiota was analyzed by comparing fecal samples from different drug treatment groups based on the number of Out-of-Use (OTU) counts in each group. The results showed that the donepezil treatment group and the control group had the lowest gut microbiota abundance, followed by the medium-dose combination therapy group (AD_M). The disease model group and the healthy control group were similar, while the high-dose combination therapy group (AD_H) had a slightly higher abundance. The trend of OTU counts in each group was consistent with the trend of gut microbiota abundance analysis based on chao1 values. Figure 1 )resemblance.

[0061] 3) Shannon value analysis of gut microbiota evenness in each group

[0062] The uniformity of gut microbiota was analyzed by comparing the Shannon values ​​of fecal samples from different drug treatment groups during the experiment. Results are as follows: Figure 2 As shown in the results, the Shannon values ​​indicate that the uniformity of the high-dose combination treatment group (AD_H) is slightly higher than that of the other groups, even higher than that of the wild-type mouse gavage saline group (WT_veh), while the donepezil treatment group has the lowest uniformity.

[0063] The abundance and relative abundance of gut microbiota in different drug treatment groups were compared based on the Shannon values ​​of each group. The Shannon values ​​indicated that the abundance and relative abundance of microbiota were slightly higher in the high-dose group and slightly lower in the untreated disease model group.

[0064] 5.2 β Analysis Results

[0065] 1) Shannon value method for analyzing the similarity of bacterial species in samples

[0066] jaccard_pcoa image as shown Figure 3 As shown in the principal coordinate analysis of fecal microbiota, at 5 months of age, the transgenic model mice were very similar to the healthy control mice, but significantly different from the six groups of 7-month-old mice after two months of feeding. Among them, the high-dose combination group (AD_H) was similar to the healthy group (WT_8w), while the wild cranberry single ingredient group (AD_Cran) was similar to AD_8w.

[0067] 2) Nonmetric Multidimensional Permutation (NMDS) method for analyzing the similarity of bacterial species in samples.

[0068] Depend on Figure 4 The results show that the nonmetric multidimensional permutation (NMDS) analysis results ( Figure 4 ) and PCoA ( Figure 3 )resemblance.

[0069] 5.3 Results of Taxonomic Analysis

[0070] Genera abundance clustering results of fecal microbiota taxonomy analysis (Figure 5 The results showed that the gut microbiota of mice in the high-dose group and the healthy control group were similar at the genus level, followed by the medium-dose group, while the difference from the disease model group was significant. Furthermore, untreated 5-month-old mice were similar to each other, but showed significant differences from the 7-month-old groups.

[0071] Cluster analysis results at various levels, such as Figure 6 As shown, the results indicate that the cluster analysis at the species level is similar to that at the genus level, suggesting that at 7 months of age, there were differences in the gut microbiota between the model group and the healthy control group mice. The high-dose composition altered the gut microbiota of the model mice, making them more similar to those of the healthy control mice.

[0072] 5.4 Abundance Difference Analysis

[0073] Differential analysis showed that 20 genera in the fecal microbiota of the healthy control group (WT_8W) differed significantly from those of the model group (AD_8W), and the abundance of 27 genera in the high-dose group (AD_H) differed significantly from that of the disease model group (AD_veh). Among these, 14 common differentially expressed genera were found, and the abundance of these 14 common differentially expressed genera was lower in both the high-dose group (AD_H) and the healthy control group (WT_8W) than in the model group. See Table 2 for details. Figure 7A -C (The vertical axis in the figure represents relative abundance × 100000).

[0074] Table 2 List of fungal genera with significant differences

[0075]

[0076] Among the 14 differentially expressed bacterial genera that were significantly downregulated after intervention, evidence suggests that six genera—Staphylococcus, Vibrio, Collinsella, Actinomycetales, Aeromonas, and Holdemanella—are closely associated with Alzheimer's disease. These are described in detail below:

[0077] The cell wall of *Staphylococcus* contains lipoteichoic acid (LTA), a major component of which is a TLR2 agonist and a receptor for Aβ-induced microglial activation. LTA mediates Aβ uptake by microglia, and the neurotoxic activation of microglia may be important in the pathogenesis of Aβ and neurodegenerative diseases. This bacterium is more abundant in the central nervous system of AD patients than in healthy controls, and it is associated with a higher risk of death from AD.

[0078] Vibrio mediates changes in gut tight junctions, thereby exacerbating the development of Alzheimer's disease (AD).

[0079] Collinsella (Collinsella sp.) is widely recognized as a risk factor for AD and has pro-inflammatory effects.

[0080] Actinomycetales (Actinomyces sp.) was significantly higher in the AD group than in the control group. Actinomyces sp. was also increased in the mild cognitive impairment group.

[0081] Aeromonas (Aeromonas sp.) belongs to the phylum Proteobacteria and was more abundant in the 3-month-old transgenic AD model mice than in the control group.

[0082] Holdemanella (Holdemania sp.) was the dominant genus in the AD group and was significantly higher than in the healthy control group and the mild cognitive impairment group.

Claims

1. The use of a composition containing cold region berry and cold region medicinal and edible Chinese medicine in the preparation of a drug for improving the intestinal flora function of Alzheimer's disease, characterized in that, The cold region berry includes blueberry, wild cranberry and blue chokeberry; the cold region medicinal and edible Chinese medicine includes acanthopanax powder, schisandra extract and astragalus extract; The composition also contains grape seed extract and vitamin C powder; In 10L of the composition, 800-900 ml of wild cranberry juice, 4000-4500 ml of blue chokeberry juice, 100-200 ml of blueberry juice, 30-40 ml of schisandra extract, 100-200 ml of astragalus extract, 400-500 mg of acanthopanax powder, 16000-17000 mg of grape seed extract, 25000-30000 mg of vitamin C powder and distilled water are contained, and the distilled water is supplemented to 10L.

2. Use according to claim 1, wherein In 10L of the composition, 833 ml of wild cranberry juice, 4163 ml of blue chokeberry juice, 167 ml of blueberry juice, 33 ml of schisandra extract, 167 ml of astragalus extract, 433 mg of acanthopanax powder, 16650 mg of grape seed extract, 29304 mg of vitamin C powder and distilled water are contained, and the distilled water is supplemented to 10L.

3. Use according to claim 1 or 2, characterized in that, The wild cranberry juice, the blue chokeberry juice and the blueberry juice are prepared from wild cranberry, blue chokeberry and blueberry in Daxing'anling, and the specific preparation method is as follows: (1) The quick-frozen wild cranberry, blue chokeberry or blueberry berry is manually selected and then slowly thawed at room temperature for 12-24 hours; (2) The berry thawed in step (1) is crushed in a crusher, the crushed juice is pumped into a plate heat exchanger, heated and then pumped into an enzymolysis tank, pectinase is added in a proportion of 1:10,000 of the weight of the juice, the juice is stirred for enzymolysis, the enzymolysis reaction temperature is 40-50℃, the time is 3.5-4 hours, and the juice after enzymolysis is obtained; (3) The juice after enzymolysis is pumped into a press for pressing for 6 minutes, the pressure is less than 200 bar and the temperature is less than 20℃, the pressed juice is pumped into a 120-mesh vibrating screen for separation, and the separated juice is pumped into a storage tank for temporary storage; (4) The separated juice is heated through a plate heat exchanger, then pumped into a separator for negative pressure enzyme inactivation treatment, the vacuum negative pressure is 0-0.5 bar, the outlet material temperature is 50-70℃, and the juice after enzyme inactivation is pumped into a clarification tank for temporary storage; (5) The juice after enzyme inactivation is pumped into a vacuum rotary filter, a diatomite filter and a paper filter for three filtrations, that is, the diatomite, the expanded perlite and water are mixed in a proportion of 1:9:90 to prepare a filter medium, the filter medium is pumped into a storage tank below the vacuum rotary filter, the vacuum rotary filter is started and then 3-5 cm of the filter medium is adhered to the surface of the machine, the scraper distance is adjusted to keep the thickness of the medium at 2.5-3.5 cm, the negative pressure of the vacuum rotary filter is controlled to be-50--70 cmHg, the pressure of the diatomite filter is 4-5 bar, and the pressure of the paper filter is within 20 bar; (6) The juice after three filtrations is pumped into a tank for storage, and the wild cranberry juice, the blue chokeberry juice and the blueberry juice are obtained.

4. Use according to claim 1 or 2, wherein the compound is ###0002### The preparation method of the Schisandra chinensis extract is as follows: the fresh Schisandra chinensis fruits are frozen, taken out of the freezer, uniformly placed in a baking tray, and baked in an oven with a temperature of 120 DEG C for 5-6 hours, and then baked at a temperature of 60 DEG C-65 DEG C for 18-20 hours; after baking, the Schisandra chinensis fruits are crushed; 50 kg of the crushed Schisandra chinensis fruits are added with 750 kg of water, heated to boiling, boiled for 4 hours, and then filtered after being placed for 12 hours; then the filtered residue is subjected to tubular centrifugal filtration, and the finished product is used after being packaged.

5. Use according to claim 1 or 2, wherein the compound is ###0002### The preparation method of the Astragalus extract is as follows: the stems and leaves of Astragalus are boiled with water at a weight ratio of 1:10 for 4-5 hours, the boiled liquid is subjected to centrifugal filtration, and then 10-fold concentration is performed, and the finished product is used after being packaged.

6. Use according to any one of claims 1 to 5, wherein The composition containing the cold land berry and the cold land medicine-food homologous traditional Chinese medicine can improve the richness and uniformity of intestinal flora, and can significantly down-regulate six bacteria closely related to Alzheimer's disease, i.e., Staphylococcus, Vibrio, Collinsella, Actinomyces, Aeromonas, and Holdemania, so as to achieve the purpose of improving the intestinal flora function of Alzheimer's disease.

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