A medicine-food homologous complex based on probiotic fermentation synergism and application thereof in blood glucose regulation

Abstract

The application discloses a kind of based on probiotic fermentation synergism of homoeopathy complex and its application in blood sugar regulation, belong to microbial and fermentation engineering technical field.The application utilizes Saccharomyces cerevisiae Saccharomyces cerevisiae )BLCC4-0032 and plant lactobacillus ( Lactiplantibacillus plantarum )DFR-1 respectively to aloe, mulberry leaf and licorice carry out two-stage fermentation, obtain fermentation mixture;Then add bitter gourd peptide to fermentation mixture, obtain homoeopathy complex, form metabolic closed loop, synergistically regulate "blood sugar-liver-intestinal flora axis", provide new solution for diabetes intervention, more for the prevention of non-alcoholic fatty liver and intestinal inflammation caused by glycolipid metabolism disorder open new path.

Description

Technical Field

[0001] This invention relates to the fields of microbial and fermentation engineering technology, specifically to a food-medicine homology complex based on probiotic fermentation enhancement and its application in blood glucose regulation. Background Technology

[0002] Diabetes mellitus is a metabolic disease characterized by chronically elevated blood glucose levels. Its global prevalence continues to rise, posing a serious threat to human health. The liver, as a major metabolic organ, participates in the body's energy and metabolite homeostasis, and is closely linked to liver diseases such as diabetes, non-alcoholic fatty liver disease, and cirrhosis. Insulin resistance and glucose metabolism disorders can lead to liver dysfunction; hyperglycemia can trigger liver inflammation and oxidative stress through multiple signaling pathways, thereby accelerating the progression of diabetes.

[0003] Current blood sugar lowering strategies face three major bottlenecks: First, chemical drugs have significant limitations. Mainstream drugs such as metformin are prone to causing gastrointestinal reactions, insulin drugs pose risks of hypoglycemia and drug resistance, and while α-glucosidase inhibitors such as acarbose can delay glucose absorption, they cannot improve insulin sensitivity. Second, the development of natural blood sugar lowering products is insufficient. Although plants and their extracts with blood sugar lowering activity have been widely reported, traditional extraction processes result in low bioavailability, bitter taste, and weak clinical evidence. Third, multi-target synergistic mechanisms have not yet been systematically integrated. Hyperglycemic patients often experience side effects such as liver damage and gut microbiota imbalance. Current products mostly focus on single pathways (such as promoting insulin secretion or inhibiting glucose absorption), neglecting the systemic regulatory value of the "blood sugar-liver-gut microbiota axis," making it difficult to achieve long-term maintenance of blood sugar homeostasis. Summary of the Invention

[0004] To address the problems existing in the prior art, the purpose of this invention is to provide a food-medicine homology complex based on probiotic fermentation and its application in blood glucose regulation. This invention utilizes *Saccharomyces cerevisiae* (Saccharomyces cerevisiae). Saccharomyces cerevisiae BLCC4-0032 and Lactobacillus plantarum ( Lactiplantibacillus plantarum DFR-1 was used to perform two-stage fermentation of aloe vera, mulberry leaves and licorice to obtain a fermentation mixture. Bitter melon peptide was then added to the fermentation mixture to obtain a food-medicine homology complex, which forms a metabolic closed loop and synergistically regulates the "blood glucose-liver-gut microbiota axis". This provides a new solution for diabetes intervention and opens up a new path for preventing non-alcoholic fatty liver and enterogenic inflammation caused by glucose and lipid metabolism disorders.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] In a first aspect, the present invention provides a food-medicine homology complex based on probiotic fermentation enhancement, wherein the food-medicine homology complex is composed of aloe vera gel fermentation liquid, licorice extract fermentation liquid, mulberry leaf extract fermentation liquid and bitter melon peptide in a weight ratio of (60-80):(10-15):(3-7):1.

[0007] The aloe vera gel fermentation liquid is prepared by the following method:

[0008] Dissolve aloe vera gel powder in water to obtain aloe vera gel liquid, and sterilize it; add brewer's yeast to aloe vera gel liquid for the first stage of fermentation, sterilize it, and obtain the first stage aloe vera gel fermentation substrate.

[0009] Then, *Lactobacillus plantarum* was inoculated into the first-stage aloe vera gel fermentation substrate for the second-stage fermentation. After pasteurization, the fermentation supernatant was collected by centrifugation to prepare the aloe vera gel fermentation broth.

[0010] The licorice extract fermentation broth is prepared by the following method:

[0011] Licorice was extracted by heating with water to obtain licorice extract, which was then sterilized. Saccharin yeast was added to the licorice extract for the first stage of fermentation, which was then sterilized to obtain the first stage licorice fermentation substrate.

[0012] Then, Bacillus plantarum was introduced into the licorice fermentation substrate of the first stage for the second stage of fermentation. After pasteurization, the fermentation supernatant was collected by centrifugation to prepare the licorice extract fermentation broth.

[0013] The mulberry leaf extract fermentation broth is prepared by the following method:

[0014] Mulberry leaves were extracted by heating water to obtain mulberry leaf extract, which was then sterilized. Saccharin yeast was added to the mulberry leaf extract for the first stage of fermentation, which was then sterilized to obtain the first stage mulberry leaf fermentation substrate.

[0015] Then, Bacillus plantarum was introduced into the mulberry leaf fermentation substrate of the first stage for the second stage of fermentation. After pasteurization, the fermentation supernatant was collected by centrifugation to prepare the mulberry leaf extract fermentation broth.

[0016] Preferably, the brewing yeast is selected from brewing yeast with preservation number CCTCC NO: M 2015123 ( Saccharomyces cerevisiae )BLCC4-0032; The *Lactobacillus plantarum* is selected from *Lactobacillus plantarum* with the preservation number CCTCC NO:M 20251450 ( Lactiplantibacillus plantarum DFR-1.

[0017] Preferably, the temperature for the first stage of fermentation is 26-30℃, and the fermentation is allowed to stand for 20-30 hours; the temperature for the second stage of fermentation is 35-40℃, and the fermentation is allowed to stand for 10-30 hours.

[0018] This invention performs sterilization after the first stage of fermentation to prevent the growth of brewing yeast from affecting the subsequent fermentation of Lactobacillus plantarum.

[0019] Preferably, the food-medicine homology complex is composed of aloe vera gel fermentation liquid, licorice extract fermentation liquid, mulberry leaf extract fermentation liquid and bitter melon peptide in a weight ratio of 70:12:5:1.

[0020] The medicinal and edible homologous complex of this invention selects three medicinal and edible homologous substances—aloe vera, mulberry leaf, and licorice—as core components, and utilizes brewer's yeast ( Saccharomyces cerevisiae The first stage of fermentation was carried out using BLCC4-0032 to achieve cell wall disruption and enhanced efficiency; *Lactobacillus plantarum* (…) was utilized. Lactiplantibacillus plantarum DFR-1 undergoes a second-stage fermentation process to achieve transformation and toxicity reduction. Through this two-stage fermentation, the effective substances in aloe vera, mulberry leaves, and licorice are deeply transformed, and their toxicity is reduced while their efficacy is enhanced, achieving cross-scale bioactivity that is unattainable with unfermented extracts. The inclusion of bitter melon peptides creates a metabolic closed loop in this composition: mulberry leaves and probiotics block intestinal glucose supply → bitter melon peptides activate peripheral tissue glucose consumption → licorice and aloe vera protect the liver and maintain gut microbiota balance → gut microbiota metabolites (such as SCFAs) further assist bitter melon peptides in improving insulin sensitivity. This simultaneously targets four pathways: "inhibition of glucose absorption - enhancement of insulin sensitivity - regulation of hepatic glucose output - gut microbiota immune homeostasis."

[0021] A second aspect of the present invention provides the use of the above-mentioned food-medicine homologous complex in at least one of the following (1)-(3):

[0022] (1) Preparation of hypoglycemic products;

[0023] (2) To prepare products for the prevention and / or treatment of liver injury;

[0024] (3) Prepare products that regulate gut microbiota.

[0025] In the above applications, the food-medicine homology complex lowers blood sugar by inhibiting sugar absorption and enhancing insulin sensitivity.

[0026] In the above application, the liver damage is caused by hyperglycemia.

[0027] In the above applications, the regulation of intestinal flora involves increasing the number of beneficial bacteria and inhibiting the growth of harmful bacteria. Preferably, the beneficial bacteria are Lactobacillus and Bifidobacterium, and the harmful bacteria are Escherichia coli.

[0028] In a third aspect, the present invention provides a drug for the prevention and treatment of diabetes, wherein the drug uses the above-mentioned food-medicine homology complex as its active ingredient.

[0029] Furthermore, the drug also includes a pharmaceutically acceptable carrier. In some preferred embodiments, the carrier may be selected from one or more of lactose, glucose, sucrose, starch, carboxymethyl cellulose, gelatin, talc, peanut oil, glycerin, sorbitol, and mannitol.

[0030] More preferably, the dosage form of the drug is a solution, suspension, emulsion, pill, tablet or capsule.

[0031] The beneficial effects of this invention are:

[0032] This invention pioneers a two-stage yeast-lactic acid bacteria fermentation process for a ternary complex of aloe vera, mulberry leaf, and licorice, overcoming the triple bottlenecks of traditional hypoglycemic therapies and offering the following advantages:

[0033] (1) Enhanced process efficiency and reduced toxicity: brewing yeast ( Saccharomyces cerevisiae BLCC4-0032 and Lactobacillus plantarum DFR-1 co-fermented, significantly reducing the substrate sugar content and solving the risk of glycemic index of natural products; at the same time, it improved the α-glucosidase inhibition rate (inhibition rate after fermentation of aloe / mulberry leaf / licorice >94%), optimizing taste and bioavailability.

[0034] (2) Multi-target synergistic mechanism: By embedding bitter melon peptides, a metabolic closed loop of “intestinal glucose blocking → relief of insulin resistance → liver protection → microbial regulation” is formed, which systematically integrates the “blood glucose-liver-intestinal microbial axis”.

[0035] (3) Blood sugar reduction: The food-medicine homology complex based on probiotic fermentation of the present invention reduced fasting blood glucose in diabetic mice by 29.83%, which was better than acarbose by 16.49%.

[0036] (4) Liver protection: The liver index returned to near normal levels, down 12.57% compared to the model group.

[0037] (5) Regulation of gut microbiota: The number of Bifidobacteria / Lactobacillus increases by 2-3 times, reversing the imbalance of gut microbiota related to hyperglycemia.

[0038] (6) Synergistic effect verification: The fermented composition was significantly better than the single component and the unfermented composition (e.g., compared with the unfermented composition group, the fermented composition group increased GLP-1 secretion by 19.32% and decreased insulin resistance index by 49.72%), confirming the synergistic enhancement effect of fermentation process on the activity of multiple components. Attached Figure Description

[0039] Figure 1 pH values ​​measured before and after fermentation of raw materials.

[0040] Figure 2 Results of the inhibition rate of α-glucosidase before and after raw material fermentation.

[0041] Figure 3 Blood glucose levels in mice during the intervention period: Results of blood glucose measurements in each group.

[0042] Figure 4 Comparison of livers of mice in the model group and mice in the blank group; the left image shows the liver of mice in the model group, and the right image shows the liver of mice in the blank group.

[0043] Figure 5 Results of liver index measurement in mice of each group.

[0044] Figure 6 Results of kidney index measurement in mice of each group.

[0045] Figure 7 Results of liver glycogen assay in mice in each group.

[0046] Figure 8 : Results of lactobacillus count in the intestines of mice in each group after intervention.

[0047] Figure 9 : Results of Bifidobacterium count in the intestines of mice in each group after intervention.

[0048] Figure 10 : Results of Escherichia coli count in the intestines of mice in each group after intervention.

[0049] Figure 11 : Enterococcus count results in the intestines of mice in each group after intervention. Detailed implementation method:

[0050] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of this application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0051] To enable those skilled in the art to more clearly understand the technical solution of this application, the technical solution of this application will be described in detail below with reference to specific embodiments. If specific experimental conditions are not specified in the embodiments, they are generally performed under conventional conditions or according to the conditions recommended by the reagent company; the reagents, consumables, etc. used in the following embodiments and comparative examples, unless otherwise specified, can be obtained commercially. Wherein:

[0052] Lactobacillus plantarum ( Lactiplantibacillus plantarum DFR-1, this strain was deposited on June 23, 2025 at the China Center for Type Culture Collection (CCTCC, Wuhan University, Wuhan, China), with accession number CCTCC NO: M 20251450.

[0053] Lactobacillus plantarum DFR-1 exhibits excellent α-glucosidase inhibitory activity. Using 4-nitrophenol-α-D-glucopyranoside (PNPG) as a substrate, the fermentation supernatant of Lactobacillus plantarum DFR-1 showed an inhibition rate of up to 87.75% against α-glucosidase, demonstrating significant hypoglycemic activity.

[0054] Lactobacillus plantarum DFR-1 also has bile salt hydrolase activity, which can degrade cholesterol. The fermentation supernatant of Lactobacillus plantarum DFR-1 has a cholesterol degradation rate of 81.2%, which can synergistically improve dyslipidemia caused by diabetes.

[0055] Lactobacillus plantarum DFR-1 also has the ability to inhibit xanthine oxidase. According to the test, the fermentation supernatant of Lactobacillus plantarum DFR-1 has an inhibition rate of 57.72% against xanthine oxidase, which can synergistically improve hyperuricemia caused by diabetes.

[0056] Lactobacillus plantarum DFR-1 can also produce high levels of polyphenol oxidase and has in vitro antioxidant properties, which can reduce oxidative stress, protect pancreatic β cells, and improve insulin secretion.

[0057] In summary, using *Lactobacillus plantarum* DFR-1 as a fermentation strain can inhibit glucose metabolism, reduce oxidative stress, lower blood lipids, and indirectly improve insulin resistance, thereby effectively improving blood glucose regulation.

[0058] brewing yeast ( Saccharomyces cerevisiae The accession number of BLCC4-0032 is CCTCC NO: M2015123, and it is recorded in patent CN 105055458 B.

[0059] Kudzu root, galangal, mulberry leaves, mulberries, licorice, ginseng, bitter melon, and lotus leaves are all commercially available raw materials that are both food and medicine.

[0060] Bitter melon peptide, purchased from Dezhou Lanli Biotechnology Co., Ltd., product batch number: 20241013630.

[0061] Aloe vera gel powder was purchased from Xinjiang Bafubafu Agricultural Co., Ltd.; α-glucosidase, 4-nitrophenol-α-D-glucopyranoside (PNPG), and acarbose were all purchased from Shanghai Yuanye Biotechnology Co., Ltd.; streptozotocin (STZ) was purchased from Sigma.

[0062] MRS medium: peptone 10.0 g / L, beef extract 8.0 g / L, yeast extract 4.0 g / L, glucose 20.0 g / L, dipotassium hydrogen phosphate 2.0 g / L, diammonium hydrogen citrate 2.0 g / L, sodium acetate 5.0 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.04 g / L, agar 14.0 g / L, Tween-80 1.0 g / L, pH 6.5±0.2, sterilized at 121℃ for 15 min.

[0063] 0.2 U / mL α-glucosidase: Dissolve 100 U of α-glucosidase in 1 mL of PBS buffer (pH 7.0, 200 mM) to prepare a stock solution. If not used completely at once, aliquot and store at -20°C. When using, dilute the stock solution with PBS buffer to 0.2 U / mL.

[0064] 4-Nitrophenol-α-D-glucopyranoside (PNPG): Dissolve 20 mg PNPG in 4 mL of pure water and sonicate for 10 min to dissolve. Prepare fresh before use.

[0065] 0.2M Sodium Carbonate Solution: Accurately weigh 5.72 g of anhydrous sodium carbonate and dissolve it in pure water. Dilute to 100 mL with pure water in a volumetric flask and set aside.

[0066] Preparation method of citric acid-sodium citrate buffer (pH=4.2-4.5): First, prepare a stock solution, and prepare 0.1 mol / L citric acid solution (solution A) and 0.1 mol / L sodium citrate solution (solution B) separately. Then adjust the pH by mixing solutions A and B in proportion (e.g., 16.5 mL of solution A + 13.5 mL of solution B, which can be finely adjusted according to the pH meter) to stabilize the pH of the final mixture at 4.2-4.5.

[0067] STZ solution: Weigh 90mg of STZ powder (weighing should be done in the dark) and quickly add it to 15mL of citrate-sodium citrate buffer solution. Mix gently until completely dissolved. After preparation, immediately wrap the solution in aluminum foil to protect it from light and place it in an ice bath. Use within 30 minutes.

[0068] Citric acid and sodium citrate were purchased from Sinopharm Chemical Reagent Co., Ltd. (China); the blood glucose meter and blood glucose test strips used for testing were purchased from Sinocare Biosensor Co., Ltd.

[0069] Example 1: Screening of raw materials with hypoglycemic effects

[0070] 1. Experimental Methods

[0071] 1.1 Raw material preparation

[0072] 1.1.1 Preparation of raw material extract before fermentation:

[0073] Weigh 20g each of the following medicinal and edible raw materials: kudzu root, galangal, mulberry leaf, mulberry fruit, licorice root, ginseng, bitter melon, and lotus leaf. Soak each raw material in purified water at room temperature for 2 hours (5 times its weight) and extract at 100℃ for 1 hour. Filter the extract through an 80-mesh standard sieve and collect the supernatant. Add 4 times its weight of purified water to the residue and extract a second time at 100℃ for 1 hour. Filter the residue through an 80-mesh standard sieve and collect the supernatant. Combine the two supernatants and concentrate them to 100 mL. Prepare kudzu root extract, galangal extract, mulberry leaf extract, mulberry fruit extract, licorice root extract, ginseng extract, bitter melon extract, and lotus leaf extract.

[0074] Accurately weigh 1g of aloe vera gel powder and dissolve it in 100mL of purified water to prepare aloe vera gel solution.

[0075] 1.1.2 Preparation of fermentation supernatant:

[0076] The prepared kudzu root extract, galangal extract, mulberry leaf extract, mulberry extract, licorice root extract, ginseng extract, bitter melon extract, lotus leaf extract, and aloe vera gel were sterilized at 121°C under high temperature and pressure for 20 minutes and then cooled to room temperature. Activated brewer's yeast BLCC4-0032 seed culture (10 μL) was then inoculated into the sterilized extracts. 8 The seed culture was inoculated at a concentration of 5% (v / v) with CFU / mL, and the first stage of fermentation was carried out at 28℃ for 24 hours. After fermentation, the mixture was sterilized at 121℃ under high pressure for 20 minutes to obtain the first stage fermentation substrate.

[0077] A suspension of *Lactobacillus plantarum* DFR-1 bacteria activated on MRS medium (concentration 1×10⁻⁶) 9 The fermentation medium (CFU / mL) was inoculated at a rate of 5% (v / v) into the first-stage fermentation substrate for the second-stage fermentation. The second-stage fermentation was carried out at 37°C for 18 hours. After fermentation, pasteurization was performed under the following conditions: the fermentation broth was incubated in an 80°C water bath for 15 minutes, followed immediately by ice bath cooling to inactivate the cells. After pasteurization, the mixture was centrifuged at 5000×g for 10 minutes, and the fermentation supernatant was collected.

[0078] 1.2 Indicator Testing

[0079] 1.2.1 pH measurement before and after fermentation:

[0080] Take 1.0 mL each of the pre-fermentation raw material extract and fermentation supernatant prepared in 1.1 and place them in sample cups. Immerse the pH electrode 1 cm below the liquid surface and record the pH value after the reading stabilizes. Repeat the measurement 3 times for each sample.

[0081] 1.2.2 Determination of sugar content before and after fermentation:

[0082] If the raw materials contain monosaccharides (glucose, fructose) and disaccharides (sucrose, maltose) that can be rapidly absorbed by the human body, they may have an adverse effect on blood sugar levels for people who need to control their blood sugar. Therefore, the sugar content of the extract before fermentation and the supernatant after fermentation of the raw materials to be screened was determined using the following method:

[0083] Accurately weigh 0.01g of glucose standard, place it in a 50 mL beaker, dissolve it in ultrapure water, and dilute to a 100 mL volumetric flask to obtain glucose standard solution.

[0084] Accurately pipette 0.2, 0.4, 0.6, 0.8, and 1.0 mL of glucose standard solution into stoppered test tubes, respectively. Add ultrapure water to a final volume of 1 mL, then add 1 mL of 5% phenol and 4 mL of 98% concentrated sulfuric acid, respectively. Shake well and react for 10 min. Then, rapidly cool to room temperature using an ice-water bath. Zero the tube using a blank tube (1 mL ultrapure water + 1 mL 5% phenol + 4 mL 98% sulfuric acid) and measure the absorbance (OD) at 490 nm. Take 1 mL of the pre-fermentation extract and the post-fermentation supernatant, respectively, and measure the absorbance at 490 nm using the same method (1 mL sample + 1 mL 5% phenol + 4 mL 98% sulfuric acid). Repeat the measurement three times for each sample.

[0085] 1.2.3 Determination of α-glucosidase inhibition rate before and after fermentation:

[0086] The reaction was performed using a 96-well plate. The α-glucosidase activity assay consisted of a 100 μL reaction mixture. First, 25 μL of 20 mmol / L substrate 4-nitrophenol-α-D-glucopyranoside (PNPG) and 25 μL of sample were mixed and incubated at 37°C for 10 min. Then, 50 μL of 0.2 U / mL α-glucosidase was added, mixed, and incubated at 37°C for 30 min. The reaction was then terminated by rapidly adding 100 μL of 0.2 mol / L Na₂CO₃ solution. The absorbance at 405 nm was measured using a microplate reader. Each experiment was performed in triplicate, and the experiment was divided into four groups (sample group, sample blank control group, negative control group, and negative blank control group). The order of sample addition for each group was as shown in Table 1.

[0087] Table 1. Dosage and order of reactant addition (unit: μL)

[0088]

[0089] Note: The samples used in the sample group were the raw material extract before fermentation and the fermentation supernatant, respectively.

[0090] The formula for calculating the inhibition rate is as follows:

[0091] α-glucosidase inhibition rate = [1 - (AB) / (CD)] × 100%

[0092] In the formula: A is the absorbance of the sample at 405 nm; B is the absorbance of the blank control at 405 nm; C is the absorbance of the negative control at 405 nm; D is the absorbance of the negative blank control at 405 nm.

[0093] 1.2.4 Detection of polysaccharide and anthraquinone content in aloe vera gel before and after fermentation

[0094] Unlike monosaccharides in fermentation broth, aloe polysaccharides exert their hypoglycemic effect through multi-target synergistic action. The core mechanism lies in improving insulin resistance, enhancing glucose uptake and utilization by muscle and fat cells, and inhibiting hepatic gluconeogenesis. Simultaneously, aloe polysaccharides can delay carbohydrate absorption and regulate gut microbiota in the intestines. The short-chain fatty acids (SCFAs) produced by the gut microbiota help further improve insulin sensitivity and systemic inflammation, thereby comprehensively stabilizing blood glucose levels. Anthraquinones are another major and important active ingredient in aloe, besides aloe polysaccharides. Studies have shown that lipid-lowering probiotics combined with total anthraquinones from cassia seeds can promote cholesterol absorption and excretion, inhibit cholesterol production and efflux, thereby regulating lipid metabolism, reducing insulin resistance levels, and improving non-alcoholic fatty liver disease. Therefore, the polysaccharide and anthraquinone content of aloe gel before and after fermentation was tested, and samples were sent to Qingdao Yuanxin Testing Technology Co., Ltd. for analysis.

[0095] 2. Experimental Results

[0096] 2.1 pH measurement results before and after fermentation:

[0097] The results are as follows Figure 1 As shown, the pH value of each raw material decreased after fermentation with Saccharomyces cerevisiae BLCC4-0032 and Lactobacillus plantarum DFR-1.

[0098] 2.2 Results of sugar content determination before and after fermentation:

[0099] The sugar content of the selected raw materials with hypoglycemic effects was determined before and after fermentation. The absorbance was measured at 490 nm. A standard curve was plotted with absorbance A as the ordinate and standard solution concentration X as the abscissa, yielding the regression equation: A = 0.015X + 0.1345, R. 2 =0.998, and the sugar content was calculated by substituting the absorbance of each sample. The results are shown in Table 2.

[0100] Table 2. Raw material extract before fermentation, supernatant after fermentation of *Lactobacillus plantarum* DFR-1 single strain, and...

[0101] Determination of sugar content in the supernatant after yeast-lactic acid bacteria two-stage fermentation

[0102]

[0103] Mulberry and ginseng extracts maintained high sugar content before and after fermentation. Before fermentation, the sugar content of mulberry and ginseng extracts was 5.10 g / 100 mL and 3.58 g / 100 mL, respectively. After two stages of fermentation, the sugar content of their supernatants was 3.97 g / 100 mL and 2.14 g / 100 mL, respectively. Kudzu root, aloe vera gel, galangal, mulberry leaf, licorice, bitter melon, and lotus leaf extracts had lower sugar content, with a significant decrease in sugar content in their supernatants after fermentation. The sugar contents of these extracts were 0.58 g / 100 mL, 1.40 g / 100 mL, 0.69 g / 100 mL, 1.48 g / 100 mL, 1.72 g / 100 mL, 1.62 g / 100 mL, and 0.54 g / 100 mL, respectively. After two-stage fermentation using *Saccharomyces cerevisiae* BLCC4-0032 and *Lactobacillus plantarum* DFR-1, the sugar content was significantly reduced and superior to that of single-strain fermentation using *Lactobacillus plantarum* DFR-1 (1×10⁻⁶ cells were inoculated at a rate of 5% (v / v) into the sterilized raw material extract). 9 A CFU / mL *Lactobacillus plantarum* DFR-1 bacterial suspension was fermented at 37℃ for 18 h. The fermentation broth was then incubated in an 80℃ water bath for 15 min, followed immediately by ice bath cooling to inactivate the bacteria. After centrifugation at 5000×g for 10 min, the fermentation supernatant was collected. The sugar contents of the fermentation supernatant after the two-stage fermentation were 0.10 g / 100 mL, 0.41 g / 100 mL, 0.59 g / 100 mL, 0.10 g / 100 mL, 0.50 g / 100 mL, 0.12 g / 100 mL, and 0.11 g / 100 mL, respectively. The results indicate that two-stage fermentation of yeast BLCC4-0032 and *Lactobacillus plantarum* DFR-1 can effectively reduce the content of free sugars that can rapidly raise blood sugar in the extract of the selected hypoglycemic raw material, thus reducing the risk for glycemic-sensitive individuals.

[0104] 2.3 Results of α-glucosidase inhibition rate determination before and after fermentation

[0105] The inhibition rates of α-glucosidase on the extracts and fermentation supernatants of selected raw materials with hypoglycemic effects were determined. PNPG was used as a substrate for screening α-glucosidase inhibitory activity. The results are as follows: Figure 2As shown, the results indicate that the inhibition rate of each raw material on α-glucosidase was increased to varying degrees after fermentation, and it had significant inhibitory activity on the production of p-nitrophenol (pNP) from PNPG. Among them, aloe vera gel, mulberry leaves, and licorice were the most significant. After fermentation, the inhibition rates of α-glucosidase in the fermentation supernatant were 99.72%, 94.84%, and 94.16%, respectively.

[0106] In summary, considering the results of pH, sugar content, and α-glucosidase inhibition rate measurements, aloe vera gel showed the best overall performance, with a sugar content of 0.41 g / 100 mL in the supernatant after fermentation, a high α-glucosidase inhibition rate of 99.72%, and a post-fermentation pH of 3.98. Licorice was second best, with a sugar content of 0.50 g / 100 mL in the supernatant after fermentation, a α-glucosidase inhibition rate of 94.16%, and a pH of 3.78. Mulberry leaf fermentation showed a sugar content of 0.10 g / 100 mL in the supernatant, a α-glucosidase inhibition rate of 94.84%, and a pH of 3.83. Therefore, these three were selected as the core components of the hypoglycemic composition.

[0107] 2.4 Results of determination of polysaccharides and anthraquinones in aloe vera gel before and after fermentation

[0108] The polysaccharide and anthraquinone contents of aloe vera gel before and after fermentation were determined. The polysaccharide content before fermentation was 692 mg / kg, and after fermentation it was 928 mg / kg, representing a 34.10% increase after two stages of fermentation. The anthraquinone content before fermentation was 0.004 mg / kg, and after fermentation it was 0.006 mg / kg, representing a 50% increase after two stages of fermentation. While some literature reports that anthraquinones have a stimulating laxative effect, the anthraquinone content in aloe vera gel before and after fermentation both met the requirement that the anthraquinone content in aloe vera gel products should not exceed 0.1 mg / kg, indicating safe use.

[0109] Example 2: Preparation of a food-medicine homology complex based on probiotic fermentation synergistic effect

[0110] 1. Preparation of aloe vera gel fermentation broth:

[0111] Dissolve 1g of Aloe vera gel powder in 100mL of purified water to obtain an aloe vera gel solution with a concentration of 1g / 100mL. Sterilize at 121℃ for 20min and cool to room temperature.

[0112] Inoculate the sterilized aloe vera gel solution with a seed culture of Saccharomyces cerevisiae BLCC4-0032 at a rate of 5% (v / v). 8 The first stage of fermentation was carried out at 28℃ for 24 hours. After fermentation, the aloe vera gel fermentation substrate was obtained by high temperature and high pressure sterilization at 121℃ for 20 minutes.

[0113] A bacterial suspension of *Lactobacillus plantarum* DFR-1 (concentration 1×10⁻⁶) was prepared. 9 (CFU / mL) was inoculated into the first-stage aloe vera gel fermentation substrate at an inoculation rate of 5% (v / v) for the second-stage fermentation;

[0114] The second stage of fermentation was carried out at 37℃ for 18 hours. After fermentation was completed, the mixture was pasteurized (80℃ water bath for 15 minutes, followed by immediate ice bath cooling to inactivate the bacteria), centrifuged at 5000×g for 10 minutes, and the supernatant was collected to prepare the aloe vera gel fermentation broth.

[0115] 2. Preparation of licorice extract fermentation broth:

[0116] Weigh 20g of licorice, add 100g of purified water, soak for 2 hours, extract at 100℃ for 1 hour, filter, and collect the supernatant; add 80g of purified water to the filter residue, extract a second time at 100℃ for 1 hour, filter, and collect the supernatant; combine the two supernatants, concentrate to 100 mL, and prepare licorice extract; sterilize at 121℃ for 20 minutes under high temperature and high pressure, and cool to room temperature;

[0117] Inoculate the sterilized licorice extract with a seed culture of Saccharomyces cerevisiae BLCC4-0032 at a rate of 5% (v / v). 8 The first stage of fermentation was carried out at 28℃ for 24 hours (CFU / mL). After fermentation, the first stage of fermentation was carried out at 121℃ for 20 minutes to obtain the first stage licorice fermentation substrate.

[0118] A bacterial suspension of *Lactobacillus plantarum* DFR-1 (concentration 1×10⁻⁶) was prepared. 9 (CFU / mL) was inoculated into the first-stage licorice fermentation substrate at an inoculation rate of 5% (v / v) for the second-stage fermentation;

[0119] The second stage of fermentation was carried out at 37℃ for 18 hours. After fermentation was completed, the mixture was pasteurized (80℃ water bath for 15 minutes, followed by immediate ice bath cooling to inactivate the bacteria), centrifuged at 5000×g for 10 minutes, and the supernatant was collected to prepare the licorice extract fermentation broth.

[0120] 3. Preparation of mulberry leaf extract fermentation broth:

[0121] Weigh 20g of mulberry leaves, add 100g of purified water, soak for 2 hours, extract at 100℃ for 1 hour, filter, and collect the supernatant; add 80g of water to the filter residue, extract a second time at 100℃ for 1 hour, filter, and collect the supernatant; combine the two supernatants, concentrate to 100 mL, and prepare mulberry leaf extract; sterilize at 121℃ for 20 minutes under high temperature and high pressure, and cool to room temperature;

[0122] Inoculate sterilized mulberry leaf extract with a seed culture of Saccharomyces cerevisiae BLCC4-0032 at an inoculation rate of 5% (v / v). 8 The first stage of fermentation was carried out at 28℃ for 24 hours (CFU / mL). After fermentation, the mixture was sterilized at 121℃ for 20 minutes to obtain the first stage mulberry leaf fermentation substrate.

[0123] A bacterial suspension of *Lactobacillus plantarum* DFR-1 (concentration 1×10⁻⁶) was prepared. 9 (CFU / mL) was inoculated into the first-stage mulberry leaf fermentation substrate at an inoculation rate of 5% (v / v) for the second-stage fermentation;

[0124] The second stage of fermentation was carried out at 37℃ for 18 hours. After fermentation was completed, the mixture was pasteurized (80℃ water bath for 15 minutes, followed by immediate ice bath cooling to inactivate the bacteria), centrifuged at 5000×g for 10 minutes, and the supernatant was collected to prepare the mulberry leaf extract fermentation broth.

[0125] 4. Preparation of food-medicine homologous complexes based on probiotic fermentation synergy:

[0126] The aloe vera gel fermentation liquid, licorice extract fermentation liquid, mulberry leaf extract fermentation liquid and bitter melon peptide prepared above were mixed in a weight ratio of 70:12:5:1 to prepare a food-medicine homology complex based on probiotic fermentation enhancement.

[0127] Example 3: Preparation of a food-medicine homology complex based on probiotic fermentation synergistic effect

[0128] The preparation methods for aloe vera gel fermentation liquid, licorice extract fermentation liquid, and mulberry leaf extract fermentation liquid are the same as in Example 2.

[0129] Aloe vera gel fermentation liquid, licorice extract fermentation liquid, mulberry leaf extract fermentation liquid and bitter melon peptide were mixed in a weight ratio of 60:10:3:1 to prepare a food-medicine homologous complex based on probiotic fermentation enhancement.

[0130] Example 4: Preparation of a food-medicine homology complex based on probiotic fermentation synergy

[0131] The preparation methods for aloe vera gel fermentation liquid, licorice extract fermentation liquid, and mulberry leaf extract fermentation liquid are the same as in Example 2.

[0132] Aloe vera gel fermentation liquid, licorice extract fermentation liquid, mulberry leaf extract fermentation liquid and bitter melon peptide were mixed in a weight ratio of 80:15:7:1 to prepare a food-medicine homologous complex based on probiotic fermentation enhancement.

[0133] Comparative Example 1:

[0134] The preparation method of aloe vera gel fermentation broth is the same as in Example 2;

[0135] Composition A was prepared by mixing aloe vera gel fermentation broth and bitter melon peptide at a weight ratio of 70:1.

[0136] Comparative Example 2:

[0137] The preparation method of licorice extract fermentation broth is the same as in Example 2;

[0138] Composition B was prepared by mixing licorice extract fermentation broth and bitter melon peptide at a weight ratio of 12:1.

[0139] Comparative Example 3:

[0140] The preparation method of mulberry leaf extract fermentation broth is the same as in Example 2;

[0141] Composition C was prepared by mixing mulberry leaf extract fermentation broth and bitter melon peptide at a weight ratio of 5:1.

[0142] Comparative Example 4:

[0143] Dissolve 1g of aloe vera gel powder in 100mL of purified water to obtain an aloe vera gel solution with a concentration of 1g / 100mL.

[0144] Weigh 20g of licorice, add 100g of purified water, soak for 2 hours, extract at 100℃ for 1 hour, filter, and collect the supernatant; add 80g of purified water to the residue, extract a second time at 100℃ for 1 hour, filter, and collect the supernatant; combine the two supernatants, concentrate to 100 mL, and prepare licorice extract.

[0145] Weigh 20g of mulberry leaves, add 100g of purified water, soak for 2 hours, extract at 100℃ for 1 hour, filter, and collect the supernatant; add 80g of purified water to the filter residue, extract a second time at 100℃ for 1 hour, filter, and collect the supernatant; combine the two supernatants, concentrate to 100 mL, and prepare mulberry leaf extract.

[0146] The aloe vera gel, licorice extract, mulberry leaf extract and bitter melon peptide prepared above were mixed in a weight ratio of 70:12:5:1 to prepare composition D.

[0147] Experimental example:

[0148] 1. Experimental Methods

[0149] 1.1 Construction of a type 2 diabetic mouse model:

[0150] Ninety Kunming mice weighing approximately 25g (purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd.) were pre-fed for 3 days, weighed, and 80 Kunming mice were randomly selected to construct a type 2 diabetic mouse model. They were injected intraperitoneally with 50mg / kg STZ for 7 consecutive days. The remaining 10 Kunming mice served as blank controls and were injected intraperitoneally with an equal amount of physiological saline for 7 consecutive days. During the experiment, the mice had free access to water and food.

[0151] After the model was established, mice were fasted for 12 hours, and their fasting blood glucose was measured. The blank control group was used as the baseline blood glucose value for this batch of animals. Mice with blood glucose values ​​of 10-25 mmol / L were considered to have successfully developed the hyperglycemic model.

[0152] 1.2 Experimental grouping and treatment:

[0153] Seventy mice of similar weight were selected from Kunming mice that had successfully established a type 2 diabetes mouse model and randomly divided into four groups: model group, positive control group, fermented mulberry leaf group, fermented licorice group, fermented aloe vera group, unfermented combination group, and fermented combination group, with 10 mice in each group. The Kunming mice used as a blank control during the model establishment process were used as the control group. The intervention methods for each group are as follows:

[0154] Control group: Oral administration of normal saline, 0.3 mL / day;

[0155] Model group: administered physiological saline by gavage, 0.3 mL / day;

[0156] Positive group: Acarbose, 20 mg / kg, administered by gavage;

[0157] Fermented aloe vera group: Composition A prepared in Comparative Example 1 was administered by gavage at a rate of 0.3 mL / day;

[0158] Fermented licorice group: Composition B prepared by gavage in Comparative Example 2, 0.3 mL / day;

[0159] Fermented mulberry leaf group: Composition C prepared by gavage in Comparative Example 3, 0.3 mL / day;

[0160] Unfermented composition group: Composition D prepared by gavage in Comparative Example 4, 0.3 mL / day;

[0161] Fermentation composition group: the food-medicine homology complex prepared by gavage in Example 2, 0.3 mL / day.

[0162] During the experiment, mice in each group had free access to water and food, were weighed weekly, and were continuously administered gavage for 28 days, with samples taken to measure indicators.

[0163] 1.3 Indicator Testing

[0164] The detection indicators included: weekly weight change, weekly blood glucose level change (blood collected from mouse tail tips), and the area under the blood glucose curve (AUC) after the intervention (one day before the end of the experiment, mice underwent an oral glucose tolerance test, receiving 2 g / kg BW of glucose orally after an 8-hour fast, and then blood glucose levels were measured at 0h, 0.5h, 1h, and 2h; the AUC was calculated using Graph Pad Prism software); after the intervention, changes in liver index, kidney index, liver glycogen, serum indicators (insulin, glucagon-like peptide-1), insulin resistance index, and gut microbiota (Escherichia coli, Enterococcus, Lactobacillus, Bifidobacterium) were measured. Among these:

[0165] The methods for measuring liver and kidney indices are as follows:

[0166] After blood was collected from the eyeballs of mice in each group, they were euthanized by cervical dislocation under carbon dioxide anesthesia and their weight was measured. Following weighing, the mice were dissected; the abdominal cavity was cut along the midline, and the intact liver and both kidneys were removed. Surface bloodstains were rinsed off with pre-cooled physiological saline, and moisture was absorbed with filter paper. The total weight of the liver (including all lobes) and the total weight of both kidneys (left kidney + right kidney) were accurately measured. Liver-kidney indices and kidney indices were calculated after weighing.

[0167] Liver index (g / 100g) = Liver weight (g) / Body weight (g) × 100; Kidney index (g / 100g) = Kidney weight (g) / Body weight (g) × 100

[0168] The liver glycogen assay kit was purchased from Nanjing Jiancheng Biotechnology Co., Ltd., and the liver glycogen levels of mice in each group were measured according to the kit instructions.

[0169] Insulin ELISA kits were purchased from Shanghai Enzyme-Linked Biotechnology Co., Ltd. Insulin levels in the serum of mice in each group were measured according to the kit instructions. Fasting blood glucose was measured using a glucometer, and the insulin resistance index was calculated using the following formula:

[0170] Insulin resistance index ≈ fasting blood glucose × fasting insulin / 22.5; fasting blood glucose unit: mmol / L; insulin unit: µIU / mL.

[0171] The glucagon-like peptide-1 (GLP-1) ELISA kit was purchased from Shanghai Enzyme-Link Biotechnology Co., Ltd. GLP-1 levels in the serum of mice in each group were measured according to the kit instructions.

[0172] Methods for analyzing the quantity of gut microbiota in mice:

[0173] 1g of cecal contents from each group of mice was added to a 99 mL Erlenmeyer flask containing glass beads and sterile physiological saline. The mixture was shaken for 20 min until fully mixed. A 10-fold serial dilution was performed using sterile physiological saline, with each dilution using a different pipette tip. Each dilution was performed in triplicate. After plating the serial dilutions onto plates, the plates were incubated at 37°C for 48 hours before counting.

[0174] 2. Experimental Results

[0175] 2.1 Results of establishing a type 2 diabetic mouse model:

[0176] After seven consecutive days of low-dose STZ injection, mice showed obvious symptoms, with weight gain significantly lower than the control group. They exhibited polydipsia and polyuria, reduced activity, lethargy, sluggishness, and a state of mental depression such as curling up. Their exploratory behavior and spontaneous activity may have been weakened, consistent with the characteristics of type 2 diabetes.

[0177] All model mice exhibited high fasting blood glucose (0 hours) and elevated AUC. The blood glucose levels of all model mice were significantly higher than those of the blank control, and all indicators reached the level of diabetes (2-hour fasting blood glucose >11.1 mmol / L), which is consistent with the characteristics of type 2 diabetes. Based on the mouse weight indicators, polydipsia and polyuria, the type 2 diabetes mouse model was successfully constructed.

[0178] 2.2 Changes in body weight of mice in each treatment group:

[0179] As shown in Table 3, during the intervention period, there were significant differences in body weight changes between the treatment groups and the control group. The main reason was that body weight was suppressed in mice under hyperglycemic conditions. The initial body weight of the model group (31.7g) was significantly lower than that of the control group (35.8g), with a difference of 4.1g. After 4 weeks, the weight difference widened to 4.6g, and the slow weight gain was consistent with the "weight loss" characteristic of type 2 diabetes. Moreover, the weight gain in all treatment groups was higher than that in the model group. The optimal intervention group was the fermented composition group, which had the highest final body weight of 37.1g, which was significantly higher than that of the unfermented composition group (35.3g).

[0180] Table 3. Results of mouse body weight measurement during intervention treatment

[0181]

[0182] 2.3 Blood glucose levels and AUC measurements in mice of each treatment group:

[0183] Changes in blood glucose levels in mice during the intervention period are as follows: Figure 3As shown in Table 4, the AUC results after four weeks of intervention are as follows. The results indicate that the fasting blood glucose in the model group remained consistently high (14.70-15.25 mmol / L) for nearly four weeks, with an AUC as high as 36.86, significantly higher than the control group. Furthermore, the fasting blood glucose in the model group decreased by only 0.55 mmol / L after four weeks, indicating that the uninterrupted diabetic mice showed no tendency for spontaneous remission of blood glucose. The optimal intervention group was the fermented composition group, where fasting blood glucose decreased from 13.31 mmol / L to 9.34 mmol / L, a reduction of 29.83%, with a cumulative reduction of 3.97 mmol / L over four weeks, representing the largest reduction. The endpoint blood glucose was below 10 mmol / L (close to the normal value of 7 mmol / L), and the AUC value was 18.21, a 50.60% reduction compared to the model group, demonstrating superior efficacy compared to the positive control group. The positive control group showed significant efficacy, with fasting blood glucose decreasing from 13.10 mmol / L to 10.94 mmol / L, a reduction of 16.49%, and a cumulative reduction of 2.16 mmol / L over four weeks, which met the expected efficacy, but the effect was not as good as that of the fermented combination group.

[0184] Table 4. AUC measurement results after 4 weeks of intervention

[0185]

[0186] 2.4 Results of liver and kidney index measurements in mice of each treatment group:

[0187] After the intervention, the liver and kidney function of mice in each group were analyzed. The results showed that, compared with the blank group, the liver of the model group was significantly enlarged, and fatty liver had appeared. Figure 4 The liver index was 5.33, an increase of 29.06% compared to the control group. The reason for this was attributed to hyperglycemia inducing hepatic steatosis and glycogen deposition (typical diabetic liver disease). The group with the best liver protection was the fermented composition group, with a liver index of 4.66, closest to the control group, and a decrease of 12.57% compared to the model group. The fermented aloe vera group had a liver index of 4.75, a decrease of 10.88% compared to the model group. Based on the results, the liver protection effect was ranked as follows: fermented composition group > fermented aloe vera group > fermented licorice group / positive group > fermented mulberry leaf group > unfermented composition group (see...). Figure 5 ).

[0188] After the intervention, the kidney index of mice in each group remained stable between 1.31 and 1.34 (see...). Figure 6 The values ​​were within the normal range, indicating that the gavage intervention drug posed no risk of kidney damage in mice.

[0189] 2.5 Results of liver glycogen assay in mice of each treatment group:

[0190] Hyperglycemia can cause disordered liver glycogen metabolism in mice, and glycogen deposition can lead to typical diabetic liver lesions. Experimental results showed that the liver glycogen content in the model group was abnormally elevated, indicating that the liver of the mice in the model group had already been damaged under hyperglycemic conditions. Liver glycogen levels were reduced in the positive control group, fermented mulberry leaf group, fermented licorice group, fermented aloe vera group, unfermented combination group, and fermented combination group, and were close to those in the control group, thus reducing the risk of fatty liver (see...). Figure 7 ).

[0191] 2.6 Results of insulin resistance index measurement in mice of each treatment group:

[0192] The results of insulin resistance index measurement in mice of each treatment group after intervention are shown in Table 5.

[0193] Table 5 Results of insulin resistance index measurement in mice of each treatment group

[0194]

[0195] The results of the HOMA-IR measurement are shown in Table 5. The model group mice showed severe insulin resistance, with a HOMA-IR of 11.37 (>10 times that of the blank group), consistent with the characteristics of insulin resistance in diabetes. The fermented composition showed the best intervention effect, with a reduction of 76.43% compared to the model group. Mechanism speculation: The natural product group activating the insulin signaling pathway (such as Akt phosphorylation) showed outstanding performance. Specifically, the effects of the unfermented composition group and the fermented composition group significantly outperformed the positive control drug acarbose, and the fermented composition intervention effect was 49.72% better than the unfermented composition group. Compared with the fermented mulberry leaf group, fermented licorice group, and fermented aloe vera group, the reduction in insulin resistance index of the fermented composition group was superior to the sum of the reductions of the three groups, achieving a synergistic effect greater than the sum of the two. Therefore, the fermented composition has a significant effect in alleviating insulin resistance, providing a core intervention strategy for preventing diabetic complications.

[0196] 2.7 Results of glucagon-like peptide-1 (GLP-1) assay in mice of each treatment group:

[0197] GLP-1 acts on specific receptors on pancreatic β cells, enhancing glucose-stimulated insulin secretion. When blood glucose levels rise, GLP-1 significantly promotes insulin secretion from β cells. Hyperglycemia causes gut microbiota dysbiosis, impairing the function of intestinal L cells (endocrine cells distributed in the gastrointestinal mucosal epithelium) that secrete glucagon-like peptide. The experimental results are shown in Table 6.

[0198] Table 6. Results of GLP-1 assay in mice of each treatment group

[0199]

[0200] Table 6 shows that GLP-1 levels were significantly reduced in the model group, and all intervention groups partially or completely restored GLP-1 secretion. The unfermented combination group had a 21.62% higher level than the model group, which was superior to the single fermented herbal medicine group, indicating a synergistic effect of the combination of mulberry leaf, licorice, and aloe vera. The fermented combination group had a 45.11% higher level than the model group and a 19.32% higher level than the unfermented combination group, indicating that the fermentation process significantly enhanced the efficacy of the combination.

[0201] The intervention effects were as follows: fermented composition group > unfermented composition group > fermented aloe vera group > fermented licorice group > fermented mulberry leaf group > positive group > model group, and the fermented composition group was close to the blank group.

[0202] 2.8 Results of gut microbiota analysis in mice of each treatment group:

[0203] After the intervention, the counts of lactobacilli, bifidobacteria, Escherichia coli, and enterococci in the mouse gut were as follows: Figures 8-11 As shown (live bacteria count unit: CFU / mL), the results indicate that the model group confirmed the imbalance of gut microbiota, characterized by a decrease in beneficial bacteria (Lactobacillus, Bifidobacterium) and an increase in harmful bacteria (Escherichia coli) and conditionally pathogenic bacteria (Enterococcus), which may be a factor in diabetic complications.

[0204] Effectiveness of the intervention: The fermented composition group showed the most significant effect, substantially increasing beneficial bacteria (Lactobacillus and Bifidobacterium) and inhibiting harmful bacteria (Escherichia coli). The fermented composition group performed exceptionally well, especially in promoting Bifidobacterium growth, outperforming the positive control group and the unfermented composition group, indicating that the fermentation process may have enhanced the prebiotic effect of the composition.

[0205] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A medicinal and edible homologous complex based on probiotic fermentation enhancement, characterized in that, The food-medicine homology complex is composed of aloe vera gel fermentation liquid, licorice extract fermentation liquid, mulberry leaf extract fermentation liquid and bitter melon peptide in a weight ratio of (60-80):(10-15):(3-7):

1. The aloe vera gel fermentation liquid is prepared by the following method: Aloe vera gel powder was dissolved in water to obtain aloe vera gel liquid, which was then sterilized. Saccharomyces cerevisiae was added to the aloe vera gel liquid for the first stage of fermentation, which was then sterilized to obtain the first stage aloe vera gel fermentation substrate. Lactobacillus plantarum was then added to the first stage aloe vera gel fermentation substrate for the second stage of fermentation, which was then pasteurized. The fermentation supernatant was collected by centrifugation to prepare the aloe vera gel fermentation liquid. The licorice extract fermentation broth is prepared by the following method: Licorice was extracted by heating with water to obtain licorice extract, which was then sterilized. Saccharin yeast was added to the licorice extract for the first stage of fermentation, which was then sterilized to obtain the first stage licorice fermentation substrate. Lactobacillus plantarum was then added to the first stage licorice fermentation substrate for the second stage of fermentation, which was then pasteurized. The fermentation supernatant was collected by centrifugation to prepare the licorice extract fermentation broth. The mulberry leaf extract fermentation broth is prepared by the following method: Mulberry leaves were heated with water to extract mulberry leaf extract, which was then sterilized. Saccharomyces cerevisiae was added to the mulberry leaf extract for the first stage of fermentation, which was then sterilized to obtain the first stage mulberry leaf fermentation substrate. Lactobacillus plantarum was then added to the first stage mulberry leaf fermentation substrate for the second stage of fermentation, which was then pasteurized. The fermentation supernatant was collected by centrifugation to prepare the mulberry leaf extract fermentation broth. The brewing yeast was selected from brewing yeast with preservation number CCTCC NO: M 2015123 ( Saccharomycescerevisiae )BLCC4-0032; The *Lactobacillus plantarum* is selected from *Lactobacillus plantarum* with the preservation number CCTCC NO: M20251450 ( Lactiplantibacillus plantarum DFR-1; The bitter melon peptide was purchased from Dezhou Lanli Biotechnology Co., Ltd., product batch number: 20241013630.

2. The medicinal and edible homologous complex according to claim 1, characterized in that, The first stage of fermentation is carried out at a temperature of 26-30℃ for 20-30 hours; the second stage of fermentation is carried out at a temperature of 35-40℃ for 10-30 hours.

3. The food-medicine homologous complex according to claim 1 or 2, characterized in that, The medicinal and edible homologous complex is composed of aloe vera gel fermentation liquid, licorice extract fermentation liquid, mulberry leaf extract fermentation liquid and bitter melon peptide in a weight ratio of 70:12:5:

1.

4. The use of the medicinal and edible homologous complex according to any one of claims 1-3 in at least one of the following (1)-(3): (1) Preparation of hypoglycemic products; (2) To prepare products for the prevention and / or treatment of liver injury; (3) Prepare products that regulate gut microbiota; The liver damage was caused by hyperglycemia.

5. The application according to claim 4, characterized in that, The medicinal and edible homologous complex lowers blood sugar by inhibiting sugar absorption and improving insulin sensitivity.

6. The application according to claim 4, characterized in that, The liver damage was caused by hyperglycemia.

7. The application according to claim 4, characterized in that, The regulation of intestinal flora involves increasing the number of beneficial bacteria and inhibiting the growth of harmful bacteria; the beneficial bacteria are lactobacilli and bifidobacteria, and the harmful bacteria are Escherichia coli.

8. A drug for the prevention and treatment of diabetes, characterized in that, The drug uses the food-medicine homology complex as described in any one of claims 1-3 as its active ingredient.

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