Pharmaceutical composition for regulating intestinal microecological balance and reducing blood sugar, preparation and application

Abstract

The invention relates to the technical field of traditional Chinese medicine, and provides a pharmaceutical composition for regulating intestinal microecological balance and reducing blood sugar, a preparation and application, and the pharmaceutical composition comprises a traditional Chinese medicine effective component composition and a probiotic composition, the traditional Chinese medicine effective component composition comprises balsam pear glycoside, purslane phenolic acid, mulberry leaf alkaloid, corn stigma flavone, buckwheat flavone, moringa oleifera leaf flavone, sophora flower flavone, pollen pini flavone, golden camellia flavone, spirulina platensis phycocyanin, opuntia ficus-indica fruit polyphenol, rhizoma tupistrae chinensis total saponin, valeriana jatamansi volatile oil, cucurbit seven sesquiterpenoids and pholidota nameko unsaturated fatty acid; the probiotic composition comprises lactobacillus gasseri, lactobacillus paracasei, lactobacillus plantarum, lactobacillus bulgaricus and clostridium sporogenes; the pharmaceutical preparation comprises the pharmaceutical composition and auxiliary materials. The invention also discloses application of the pharmaceutical composition in preparation of hypoglycemic drugs. The composition can regulate intestinal micro-ecological balance and improve intestinal health and pancreas islet functions, thereby being beneficial to reducing blood sugar.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine compositions, and in particular to a pharmaceutical composition, preparation and application for regulating intestinal microecological balance and lowering blood sugar. Background Art

[0002] As a complex metabolic disease, the core pathology of diabetes lies in insulin deficiency and impaired insulin utilization in the body, with persistent hyperglycemia as a prominent feature. The current treatment status of diabetes covers multi-level intervention measures, mainly including drug therapy and non-drug treatments such as lifestyle intervention, but these treatment strategies have significant shortcomings. While emerging therapies such as cell therapy and artificial pancreas show promise, their technological maturity is limited and they face challenges such as immune rejection, scarce cell sources, and high costs. Therefore, the field of diabetes treatment still needs continuous exploration and innovation to overcome the limitations of existing treatment strategies.

[0003] In the field of traditional Chinese medicine treatment, Chinese patents such as CN111228316A and CN114712400A disclose the potential of probiotics in treating diabetes, but there are still obvious deficiencies. These patents still need to be further studied in terms of the screening of probiotic strains, strain numbering, combination optimization, individual difference adaptability and mechanism of action. Chinese patents with publication number CN104383123A disclose the hypoglycemic effects of eight traditional Chinese medicines, including bitter melon, purslane, mulberry leaves, corn silk, buckwheat, golden camellia, spirulina and cactus, but they only point out the hypoglycemic and lipid-lowering effects of tea polysaccharides, tea polyphenols and flavonoids in these traditional Chinese medicines, and the active ingredients are unclear, which has caused many obstacles to exploring the mechanism of hypoglycemic and thus treating diabetes.

[0004] Therefore, identifying suitable active ingredients for hypoglycemic effects is crucial for exploring deeper mechanisms of hypoglycemic action. Furthermore, combining appropriate active ingredients from traditional Chinese medicines with probiotics to achieve greater efficacy in the treatment of diabetes remains a challenge, requiring further research and optimization to enhance their efficacy and reach. Summary of the Invention

[0005] The purpose of the present invention is to provide a pharmaceutical composition, preparation and application for regulating intestinal microecological balance and lowering blood sugar. It accurately regulates the intestinal microecological balance through the synergistic effect of the Chinese medicine effective ingredient composition and the probiotic composition, significantly improves intestinal health and immunity, and improves pancreatic islet function, thereby showing excellent effects in lowering blood sugar.

[0006] The embodiments of the present invention are achieved through the following technical solutions:

[0007] The present invention provides a pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar, comprising a traditional Chinese medicine effective ingredient composition and a probiotic composition;

[0008] The traditional Chinese medicine effective ingredient composition comprises, by weight, 10-40 parts of momordica charantia glycosides, 10-40 parts of purslane phenolic acid, 5-30 parts of mulberry leaf alkaloids, 10-50 parts of corn silk flavonoids, 10-50 parts of buckwheat flavonoids, 10-50 parts of Moringa oleifera leaf flavonoids, 10-50 parts of sophora japonica flower flavonoids, 10-50 parts of pine pollen flavonoids, 10-50 parts of golden camellia flavonoids, 20-50 parts of Spirulina platensis phycocyanin, 20-40 parts of prickly pear cactus fruit polyphenols, 10-30 parts of total saponins from bamboo root seven, 15-30 parts of spider incense volatile oil, 10-25 parts of cucurbit septetesters, and 25-50 parts of unsaturated fatty acids from euphorbia cerifera.

[0009] The probiotic composition comprises: Lactobacillus gasseri, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus bulgaricus and Clostridium sporogenes, and the ratio thereof is 1-5:1-5:1-5:1-5:1-5:1-5 (unit: CFU / g or CFU / ml).

[0010] The present invention also provides a pharmaceutical preparation for lowering blood sugar, comprising the above-mentioned pharmaceutical composition and pharmaceutically acceptable excipients; wherein the excipients include any one or more of excipients, disintegrants, lubricants, binders, fillers, sweeteners, preservatives or flavorings.

[0011] The present invention also provides an application of the pharmaceutical composition in preparing a drug for treating hypoglycemia.

[0012] The technical solutions of the embodiments of the present invention have at least the following advantages and beneficial effects:

[0013] 1. The present invention combines the active ingredients of the above-mentioned traditional Chinese medicine effective ingredient composition with the probiotic composition, which is beneficial to drug absorption and enhances the blood sugar lowering effect, has the effects of regulating intestinal microecological balance and improving pancreatic islet function, and can be used to treat diabetes.

[0014] 2. The core advantage of this invention is that it can significantly improve intestinal health and immunity, improve pancreatic islet function, and thus show excellent results in lowering blood sugar by precisely regulating the balance of intestinal microecology. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0016] Figure 1 Schematic diagram of the effects of the drugs of Comparative Example 1 and Example 1 on the survival rate of probiotics in a simulated gastric fluid environment with a pH value of 1.5;

[0017] Figure 2 Schematic diagram of the effects of the drugs of Comparative Example 1 and Example 1 on the survival rate of probiotics in a simulated gastric fluid environment with a pH value of 2.5;

[0018] Figure 3 Schematic diagram of the effects of the drugs of Comparative Example 1 and Example 1 on the survival rate of probiotics in a simulated gastric fluid environment with a pH value of 3.5;

[0019] Figure 4 Schematic diagram of the effects of the drugs of Comparative Example 1 and Example 1 on the survival rate of probiotics in a simulated gastric fluid environment with a pH value of 6.8;

[0020] Figure 5 Schematic diagram of the fasting blood glucose results of mice in each group within 4 weeks;

[0021] Figure 6 Schematic diagram of the blood glucose-time curve results under OGTT test for each group of mice;

[0022] Figure 7 For each group of mice Figure 6 Schematic diagram of the area under the curve results;

[0023] Figure 8 Schematic diagram of the changes in liver index of mice in each group;

[0024] Figure 9 Schematic diagram of the changes in glycated hemoglobin (GHb) content in mice in each group;

[0025] Figure 10 Schematic diagram of the changes in total cholesterol (T-CHO) content in each group of mice;

[0026] Figure 11 Schematic diagram of the changes in triglyceride (TG) content in mice in each group;

[0027] Figure 12 Schematic diagram of HE staining results of pancreatic tissues of mice in each group;

[0028] Figure 13 Schematic diagram of HE staining results of liver tissues of mice in each group. DETAILED DESCRIPTION

[0029] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be purchased commercially.

[0030] The following is a detailed description of a pharmaceutical composition, preparation and application for lowering blood sugar provided by the embodiments of the present invention.

[0031] A pharmaceutical composition for lowering blood sugar, comprising a traditional Chinese medicine effective ingredient composition and a probiotic composition;

[0032] The traditional Chinese medicine effective ingredient composition comprises, by weight, 10-40 parts of momordica charantia glycosides, 10-40 parts of purslane phenolic acid, 5-30 parts of mulberry leaf alkaloids, 10-50 parts of corn silk flavonoids, 10-50 parts of buckwheat flavonoids, 10-50 parts of Moringa oleifera leaf flavonoids, 10-50 parts of sophora japonica flower flavonoids, 10-50 parts of pine pollen flavonoids, 10-50 parts of golden camellia flavonoids, 20-50 parts of phycocyanin from Spirulina platensis, 20-40 parts of polyphenols from prickly pear cactus fruit, 10-30 parts of total saponins from rhizoma schoenopsis, 15-30 parts of volatile oil from spider incense, 10-25 parts of cucurbita septetane extract, and 25-50 parts of unsaturated fatty acids from euphorbia milii.

[0033] The probiotic composition comprises: Lactobacillus gasseri, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus bulgaricus and Clostridium sporogenes, and the ratio thereof is 1-5:1-5:1-5:1-5:1-5:1-5 (unit: CFU / g or CFU / ml).

[0034] More specifically, the Chinese medicine effective ingredient composition comprises, by weight: 25 parts of momordica charantia glycosides, 25 parts of purslane phenolic acid, 15 parts of mulberry leaf alkaloids, 30 parts of corn silk flavonoids, 30 parts of buckwheat flavonoids, 30 parts of Moringa oleifera leaf flavonoids, 30 parts of Sophora japonica flower flavonoids, 30 parts of pine pollen flavonoids, 30 parts of Camellia chrysantha flavonoids, 30 parts of Spirulina platensis phycocyanin, 30 parts of opuntia fruit polyphenols, 20 parts of total saponins from bamboo root, 30 parts of spider incense volatile oil, 20 parts of cucurbita septetane extract, and 35 parts of unsaturated fatty acids from euphorbia milii.

[0035] Among them, the extraction methods of the effective ingredients of each Chinese medicine are as follows:

[0036] a. The extraction method of momordica charantia glycosides is as follows: an alcohol solvent (ethanol or methanol) extraction method is used, with a solid-liquid ratio of 1:10-20 (g / mL), ultrasonic-assisted (40-50°C, 30-60 minutes) or reflux extraction (1-3 hours), the crude extract is defatted and extracted, and then separated by column chromatography, and then purified by high-performance liquid chromatography to obtain momordica charantia glycosides.

[0037] b. Extraction of total phenols from Portulaca oleracea: The dried whole herb of Portulaca oleracea is crushed and passed through a 30-50 mesh sieve. Extraction is performed using an alcohol solvent (containing 0.1% HCl or formic acid) at a solid-liquid ratio of 1:10-20 (g / mL). Ultrasonic-assisted extraction (40-50°C, 30-60 min) or reflux extraction (1-3 h) is performed. After rotary evaporation and concentration, phenols are enriched using a macroporous resin to obtain total phenols from Portulaca oleracea.

[0038] c. Extraction of mulberry leaf alkaloids: mulberry leaf powder is extracted using an alcohol solvent (containing 0.015 mol / L HCl) or water, combined with ultrasonic-assisted extraction, with a solid-liquid ratio of 1:10-20 (g / mL), an extraction temperature of 30-50°C, an extraction time of 10-30 min, and at least 2 extractions. The mulberry leaf alkaloids are concentrated and dried.

[0039] d. Extraction of flavonoids from corn silk, Moringa leaves, pine pollen, buckwheat, Sophora japonica flowers, and Camellia chrysantha: The powders of the above raw materials were extracted using an alcohol solvent with a solid-liquid ratio of 1:20-40 (g / mL), a temperature of 40-60°C, an ultrasonic power of 150-250W, and an ultrasonic time of 30-60min. Flavonoid compounds were obtained by centrifugation, filtration, and concentration.

[0040] e. Extraction of phycocyanin from Spirulina platensis: The dried algae of Spirulina platensis were ground into powder using liquid nitrogen, passed through an 80-100 mesh sieve, and the cells were broken by enzymatic hydrolysis. Complex enzyme (papain + saccharifying enzyme, mass ratio 1:1-2) was used to assist in cell wall breaking at a pH of 4-6 and ultrasonication in a water bath at 30-45°C for 30-60 min. Ammonium sulfate was added for graded precipitation (25% precipitated phycoerythrin, 30% precipitated phycocyanin), purified and refined by ion exchange chromatography, desalted and concentrated, and stored by freeze-drying (pre-freezing at -40°C, vacuum drying at -50°C for 24 h) or spray drying (inlet air 180°C, outlet air 80°C).

[0041] f. Extraction of polyphenols from prickly pear cactus fruit: peel and deseed the cactus fruit, cut it into pieces, freeze-dry it, and grind it through a 30-50 mesh sieve to obtain a uniform fruit powder. The fruit powder is added to an alcohol solvent at a solid-liquid ratio of 1:20-40 (g / mL). Ultrasonic extraction is performed for 20-40 min at a temperature of 50-60°C. The mixture is centrifuged at 3000-5000 r / min for 15 min, the supernatant is collected, and concentrated to 1 / 5 of the original volume using a rotary evaporator (50°C). The supernatant is further purified by a macroporous resin, washed with water to remove impurities, and eluted with an alcohol solvent to obtain polyphenols from prickly pear cactus fruit.

[0042] g. Extraction of total saponins from bamboo root seven: dry bamboo root seven, crush it, pass it through a 30-50 mesh sieve, and use a solvent extraction method with an alcohol solvent aqueous solution at a solid-liquid ratio of 1:20-30 (g / mL), a temperature of 70-90°C, and an extraction time of 50-80 min. Recover the solvent to obtain a residue, dissolve the residue in water, and extract it multiple times. Combine the extracts and concentrate them to obtain total saponins from bamboo root seven.

[0043] h. Extraction of spider incense volatile oil: Dry spider incense and crush it, pass it through a 30-50 mesh sieve, use steam distillation method, the material-liquid ratio is 1:10-20 (g / mL), heat distillation for 5-7 hours, collect the volatile oil, and obtain spider incense volatile oil.

[0044] i. Extraction of cucurbitacin heptapeptide terpenes: dried cucurbitacin was crushed and passed through a 30-50 mesh sieve. The cucurbitacin heptapeptide terpenes were extracted by an alcohol solvent method at a solid-liquid ratio of 1:10-30 (g / mL), a temperature of 60-90°C, and an extraction time of 150-200 min. The extraction was repeated multiple times. The extracts were combined and concentrated under reduced pressure to obtain an extract. The extract was further extracted and enriched to obtain cucurbitacin heptapeptide terpenes.

[0045] j. Extraction of unsaturated fatty acids from the ground peach: Dried ground peach is crushed and passed through a 30-50 mesh sieve. The unsaturated fatty acids from the ground peach are extracted using an alcohol solvent at a solid-liquid ratio of 1:10-30 (g / mL), a temperature of 60-90°C, and an extraction time of 50-80 min. The unsaturated fatty acids from the ground peach are obtained by concentration and drying.

[0046] More specifically, the probiotic composition includes Lactobacillus gasseri SHMB 0001, Lactobacillus paracasei DN-173010, Lactobacillus plantarum THIS22, Lactobacillus delbrueckii subsp. bulgaricus GLB 44, and Clostridium sporogenes MIYAIRI 588;

[0047] The present inventors screened the individual efficacy of each strain from various aspects for various lactic acid bacteria isolated from soil, kimchi, and cheese. In addition, various strains were used to carry out efficacy evaluations in various aspects in various combinations. As a result, the present inventors clearly confirmed the unexpected synergistic effect in terms of hypoglycemic function when using a combination of five strains: Lactobacillus gasseri SHMB0001, Lactobacillus paracasei DN-173010, Lactobacillus plantarum THIS22, Lactobacillus delbrueckii subsp. bulgaricus GLB 44, and Clostridium MIYAIRI 588. In addition, the present inventors also confirmed a significant therapeutic effect for diabetes in pharmacological terms. Considering that probiotic properties are strain-dependent even in the same type of bacteria, and it is difficult to prepare a composition that shows excellent performance for all probiotic requirements, the five types of lactic acid bacteria compositions of the present invention and the significant effects confirmed by the present inventors have great technical significance.

[0048] The composition of the present invention is not limited thereto, but can be formulated so that five types of lactic acid bacteria (Lactobacillus gasseri subsp., Lactobacillus paracasei subsp., Lactobacillus plantarum subsp., Lactobacillus bulgaricus subsp., Clostridium megaterium subsp.) are present in one container as a mixture, or, these five types of lactic acid bacteria are packaged in separate containers, but can be formulated so that they are administered simultaneously or sequentially when used.

[0049] In the composition of the present invention, the effective amount of colony forming units (CFU) of each strain can be determined by those skilled in the art according to needs (disease prevention, health or treatment) or necessity, and can also be determined according to the final formulation.

[0050] When preparing the probiotic composition of the five specific strains of the present invention, the recommended amount of bacteria to be added is 1×10 5 to 1×10 15 colony forming units (CFU), where 1×10 6 to 1×10 13 The addition amount of CFU / g (or ml) is taken as the preferred range to ensure that the concentration of probiotics in the product reaches the optimized level, so as to effectively play its potential role in health promotion or disease management.

[0051] The recommended daily intake of a composition containing a specific strain, calculated on a total cell basis, should be maintained at 1 × 10 5 to 1×10 10 CFU / kg body weight to ensure the maximization of physiological benefits. 6 to 1×10 9 CFU / kg body weight is considered a more optimal intake range, aiming to balance adequate probiotic supply and safety considerations. This recommended amount can be used as a guideline, supporting both single-use intake and flexible allocation to multiple daily intakes based on individual needs, thereby enhancing the flexibility and personalization of the intake plan.

[0052] The traditional Chinese medicine composition comprises momordicoside I, trans-Ferulic acid from purslane, 1-deoxynojirimycin from mulberry leaves, luteoline from corn silk flavonoids, rutin from buckwheat, quercetin from Moringa oleifera leaves, rutin from sophora japonica flowers, naringenin from pine pollen, C-phycocyanin from camellia chrysantha flavonoids from Spirulina platensis, condensed tannins from prickly pear cactus fruit, naringenin-7-O-glucoside from camellia chrysantha flavonoids, tupichigenin E from bamboo root seven total saponins, vetivonol from spider incense volatile oil, 1β,10β-epoxyfuranoeremophilan-6β-ol from cucurbita septuprenoids, and oleic acid from unsaturated fatty acids from euphorbia cerifera.

[0053] The inventors discovered through research that the balance of intestinal microbes is closely related to the onset of diabetes. Therefore, the inventors set out to lower blood sugar and treat diabetes by activating pancreatic islet function and regulating the two-way balance between intestinal microbes.

[0054] Specifically, the probiotic composition of the present invention can inhibit the activity of α-amylase and α-glucosidase, limiting the rate at which starch is hydrolyzed into glucose by amylase. In addition, probiotics can promote the metabolism and absorption of glucose by increasing the number of beneficial bacteria in the intestine, thereby achieving the physiological effect of regulating blood sugar homeostasis, namely "sugar control". Moreover, the effective ingredients of traditional Chinese medicine contain water-soluble sugars, mostly in the form of glucosides. After oral administration, glucoside compounds are difficult to absorb in the intestine, have low bioavailability, and have a long retention time in the intestine. After probiotics colonize in the intestine and form a dominant flora, they can produce a rich enzyme system through metabolic activities, including β-glucosidase, β-glucuronidase, β-galactosidase, protease, carbohydrase and cellulase, etc. These enzymes can catalyze the conversion reaction of the effective ingredients of traditional Chinese medicine, decomposing them into more easily absorbed bioactive small molecules, thereby promoting the absorption of drug ingredients in the intestine, allowing them to enter the blood circulation more smoothly and exert their pharmacological effects; at the same time, the various extracellular enzymes secreted by probiotics during the metabolic process can synergistically act on different substrates, converting toxic components in Chinese herbal medicines into pharmacologically active substances or reducing their toxicity, thereby significantly reducing the toxic and side effects of the drugs; therefore, the present invention combines the active ingredients of the above-mentioned traditional Chinese medicine effective ingredient composition with the probiotic composition, which is beneficial to drug absorption while enhancing the hypoglycemic effect, has the effect of regulating intestinal microecological balance and activating pancreatic islet function, and can be used to treat diabetes. The core advantage of this invention is that it significantly improves intestinal health and immunity by precisely regulating the balance of intestinal microecology, and improves pancreatic islet function, thereby showing excellent results in lowering blood sugar.

[0055] The present invention also provides a pharmaceutical preparation for regulating intestinal microecological balance and lowering blood sugar, comprising the above-mentioned pharmaceutical composition and pharmaceutically acceptable excipients; wherein the excipients include any one or more of excipients, disintegrants, lubricants, adhesives, fillers, sweeteners, preservatives, emulsifiers, thickeners or flavorings; specifically, such as: starch, sodium carboxymethyl starch, talc, polyethylene glycol, micropowdered silica gel, starch slurry (paste), cellulose, methylcellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, chitosan, magnesium stearate, povidone, powdered sugar, syrup, lactose, maltodextrin, sucrose, mannitol, benzoic acid, sodium benzoate, glycerin, menthol, polyoxyethylene octanol ether, xanthan gum and natural flavors, etc.

[0056] The pharmaceutical composition of the present invention can be administered to mammals, including humans, by any method. For example, it can be administered orally or parenterally (e.g., by smearing). In addition, the pharmaceutical composition of the present invention can be formulated into a formulation for oral or parenteral administration according to the above-mentioned administration routes.

[0057] Therefore, when preparing preparations for oral administration, the dosage form of the pharmaceutical preparation includes but is not limited to one or more of tablets, capsules, granules, oral solutions, suspensions, lyophilized powders, pills, dragees, gels, syrups and ointments.

[0058] The present invention also provides an application of the pharmaceutical composition in preparing a drug for treating hypoglycemia.

[0059] Example 1

[0060] This embodiment provides a pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar, comprising a Chinese medicine active ingredient composition and a probiotic composition;

[0061] Among them, the effective ingredient composition of traditional Chinese medicine includes, by weight: 25 parts of momordica charantia glycosides, 25 parts of purslane phenolic acid, 15 parts of mulberry leaf alkaloids, 30 parts of corn silk flavonoids, 30 parts of buckwheat flavonoids, 30 parts of Moringa oleifera leaf flavonoids, 30 parts of Sophora japonica flower flavonoids, 30 parts of pine pollen flavonoids, 30 parts of Camellia chrysantha flavonoids, 30 parts of Spirulina platensis phycocyanin, 30 parts of prickly pear cactus fruit polyphenols, 20 parts of total saponins of bamboo root seven, 30 parts of spider incense volatile oil, 20 parts of cucurbitacin heptesters, and 35 parts of unsaturated fatty acids of euphorbia milii.

[0062] The probiotic composition comprises: Lactobacillus gasseri, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus bulgaricus and Clostridium sporogenes, and the ratio thereof is 3:4:5:2:3 (unit: CFU / g or CFU / ml).

[0063] More specifically, the preparation method of the probiotic composition comprises the following steps:

[0064] (1) Preparation of modified MRS medium (mMRS)

[0065] Add 10 g of tryptone, 10 g of beef extract, 5 g of yeast powder, 20 g of glucose, 5 g of sodium acetate, 2 g of diammonium hydrogen citrate, 2 g of dipotassium hydrogen phosphate, 0.5 g of magnesium sulfate heptahydrate, 1 mL of Tween 80, 0.25 g of manganese sulfate monohydrate, and 0.5 g of cysteine ​​hydrochloride to 1 L of water and adjust the pH of the culture medium to 6.8-7.0;

[0066] (2) Preparation of probiotic protective agent for freeze-drying

[0067] Add 120g skim milk powder, 70g sucrose, and 60g trehalose to 1L water;

[0068] (3) Preparation of freeze-dried bacterial powder

[0069] Lactobacillus gasseri SHMB 0001 (isolated from natural soil), Lactobacillus paracasei DN-173010 (isolated from fermented milk samples), Lactobacillus plantarum THIS22 (isolated from fermented soy milk samples), Lactobacillus delbrueckii subsp. bulgaricus GLB 44 (isolated from traditional Bulgarian yogurt), Clostridium MIYAIRI 588 (isolated from a soil sample) was streaked from the bacterium preservation tube on a solid culture medium plate to obtain a single colony, cultured at 36°C in an anaerobic workstation for 40 h, and single colonies were picked and inoculated into modified MRS liquid culture medium, cultured at 36°C in an anaerobic workstation for 20 h, and then inoculated into a larger volume of liquid culture medium at a 3% (v / v) inoculation rate. After cultured at 36°C in an anaerobic workstation for 20 h, the culture sludge was obtained by centrifugation at 5000 rpm for 15 min, washed three times with phosphate buffer at pH 7.2, and resuspended with the above-mentioned freeze-dried protective agent of the same mass as the bacterial sludge to make the bacterial content reach 10 10 CFU / mL or more; then, the suspension was pre-cultured at 37°C under anaerobic conditions for 50 minutes, then pre-frozen at -18°C for 12 hours, and finally vacuum freeze-dried to obtain freeze-dried bacterial powders of Lactobacillus gasseri SHMB 0001, Lactobacillus paracasei DN-173010, Lactobacillus plantarum THIS22, Lactobacillus delbrueckii subsp. bulgaricus GLB 44, and Clostridium sporogenes MIYAIRI 588, respectively;

[0070] (4) Preparation of probiotic composition

[0071] The Lactobacillus gasseri SHMB 0001, Lactobacillus paracasei DN-173010, Lactobacillus plantarum THIS22, Lactobacillus delbrueckii subspecies bulgaricus GLB 44, and Clostridium sporogenes MIYAIRI 588 prepared above are compounded and mixed according to a certain proportion to obtain a probiotic composition.

[0072] The preparation method of the Chinese medicine active ingredient composition is as follows:

[0073] a. Extraction of Momordica charantia: Extraction was performed using 70% ethanol or methanol at a solid-liquid ratio of 1:15 (g / mL), with ultrasound-assisted extraction (40°C, 40 min) or reflux extraction (2 h). The crude extract was defatted with petroleum ether and extracted with ethyl acetate. The extract was then separated by silica gel column chromatography (eluent: chloroform-methanol gradient), and then purified by high-performance liquid chromatography to obtain Momordica charantia.

[0074] b. Extraction of total phenolics from Portulaca oleracea: The dried whole herb was ground and passed through a 40-mesh sieve. Extraction was performed using 70% ethanol (containing 0.1% HCl or formic acid) at a solid-liquid ratio of 1:15 (g / mL). Ultrasonic extraction (40°C, 30 min) or reflux extraction (2 h) was performed. After rotary evaporation and concentration, the extract was concentrated using a macroporous resin to obtain total phenolics from Portulaca oleracea.

[0075] c. Extraction of mulberry leaf alkaloids: Mulberry leaf powder was extracted with 30% ethanol (containing 0.015 mol / L HCl) using ultrasonic-assisted extraction at a solid-liquid ratio of 1:15 (g / mL), at a temperature of 30°C, for 15 min, and three extractions. The mulberry leaf alkaloids were concentrated and dried to obtain the extract.

[0076] d. Extraction of flavonoids from corn silk, Moringa leaves, pine pollen, buckwheat, Sophora japonica, and Camellia chrysantha: The powders of the above raw materials were extracted with 50% ethanol solvent at a solid-liquid ratio of 1:30 (g / mL), at a temperature of 50 ° C, an ultrasonic power of 200 W, and an ultrasonic time of 30 min. Flavonoids were obtained by centrifugation, filtration, and concentration.

[0077] e. Extraction of phycocyanin from Spirulina platensis: The dried algae of Spirulina platensis were ground into powder using liquid nitrogen, passed through a 90-mesh sieve, and the cells were broken by enzymatic hydrolysis using a complex enzyme (papain + saccharifying enzyme, mass ratio of 1:1-2). Ultrasonication was performed in a water bath at pH 5 and 35°C for 45 minutes to assist in cell wall breaking. Ammonium sulfate was added for graded precipitation (25% precipitated phycoerythrin, 30% precipitated phycocyanin). The product was purified and refined by ion exchange chromatography, desalted and concentrated, and stored by freeze-drying (pre-freezing at -40°C and vacuum drying at -50°C for 24 hours).

[0078] f. Extraction of polyphenols from prickly pear cactus fruit: The prickly pear cactus fruit was peeled, seeded, cut into pieces, freeze-dried, and crushed through a 40-mesh sieve to obtain a uniform powder. The fruit powder was added to 70% ethanol at a solid-liquid ratio of 1:30 (g / mL). Ultrasonic extraction was performed for 25 min at a temperature of 55°C and centrifuged at 4000 rpm for 15 min. The supernatant was collected and concentrated to 1 / 5 of its original volume using a rotary evaporator (50°C). The supernatant was further purified using a macroporous resin, washed with water to remove impurities, and eluted with 70% ethanol to obtain prickly pear cactus fruit polyphenols.

[0079] g. Extraction of total saponins from bamboo roots: dried bamboo roots were crushed and passed through a 40-mesh sieve. Solvent extraction was performed using 80% aqueous ethanol at a solid-liquid ratio of 1:20 (g / mL), at a temperature of 80°C, and for 60 min. The solvent was recovered to obtain the residue, which was dissolved in water and extracted three times with water-saturated n-butanol. The n-butanol layers were combined, concentrated, and fixed to volume with methanol to obtain total saponins from bamboo roots.

[0080] h. Extraction of volatile oil from spider incense: The dried spider incense was crushed and passed through a 40-mesh sieve, and steam distilled at a solid-liquid ratio of 1:10 (g / mL), heated and distilled for 6 hours, and the volatile oil was collected to obtain volatile oil from spider incense;

[0081] i. Cucurbita septupletanyl extraction: dried cucurbita 7 crushed, passed through a 40 mesh sieve, and subjected to solvent extraction using 85% ethanol aqueous solution at a solid-liquid ratio of 1:20 (g / mL), temperature 80 ° C, extraction time: 180min, extracted 3 times, the extracts were combined and concentrated under reduced pressure to obtain an extract, which was further extracted with n-butanol or ethyl acetate to enrich the sesquiterpenes to obtain cucurbita septupletanyl extract;

[0082] j. Extraction of unsaturated fatty acids from ground peach: dried ground peach crushed, passed through a 40-mesh sieve, and subjected to solvent extraction using 80% aqueous ethanol at a solid-liquid ratio of 1:20 (g / mL), a temperature of 60 ° C, extraction time: 60 min, and concentrated to dryness to obtain ground peach unsaturated fatty acids;

[0083] h. The above extracts were combined according to the ratio, dissolved in 600 mL of purified water, 0.1 mol / L of citric acid was added to adjust the pH to 5.8, sterilized at 115 ° C for 15 minutes, cooled to below 30 ° C, and the probiotic composition and potassium sorbate were added. The volume was adjusted to 1000 mL and aseptically packaged to prepare an oral solution.

[0084] Example 2

[0085] The difference between this embodiment and Example 1 is that: this embodiment is a tablet, and in step h of its preparation method, each extract is spray-dried (inlet air temperature 180°C, outlet air temperature 80°C) to obtain a dry powder; then the obtained dry powder is evenly mixed with freeze-dried probiotic powder, microcrystalline cellulose, and lactose in a ratio of 3:2:3:2, passed through a 60-mesh sieve, and tableted with magnesium stearate (tablet weight 0.5 g / tablet), and coated with hypromellose to obtain tablets.

[0086] Example 3

[0087] The difference between this embodiment and Example 1 is that the Chinese medicine effective ingredient composition comprises, by weight, 30 parts of momordica charantia glycosides, 20 parts of purslane phenolic acid, 20 parts of mulberry leaf alkaloids, 40 parts of corn silk flavonoids, 20 parts of buckwheat flavonoids, 40 parts of Moringa oleifera leaf flavonoids, 20 parts of Sophora japonica flower flavonoids, 20 parts of pine pollen flavonoids, 40 parts of Camellia chrysantha flavonoids, 40 parts of Spirulina platensis phycocyanin, 25 parts of prickly pear cactus fruit polyphenols, 15 parts of bamboo root seven total saponins, 20 parts of spider incense volatile oil, 15 parts of cucurbita septetane extract, and 30 parts of unsaturated fatty acids from ophiopogon chinensis.

[0088] Example 4

[0089] The difference between this embodiment and Example 1 is that the Chinese medicine effective ingredient composition includes, by weight, 35 parts of momordica charantia glycosides, 25 parts of purslane phenolic acid, 10 parts of mulberry leaf alkaloids, 20 parts of corn silk flavonoids, 40 parts of buckwheat flavonoids, 20 parts of Moringa oleifera leaf flavonoids, 40 parts of Sophora japonica flower flavonoids, 40 parts of pine pollen flavonoids, 20 parts of Camellia chrysantha flavonoids, 35 parts of Spirulina platensis phycocyanin, 35 parts of prickly pear cactus fruit polyphenols, 30 parts of bamboo root seven total saponins, 25 parts of spider incense volatile oil, 10 parts of cucurbita septetane extract, and 40 parts of unsaturated fatty acids from ophiopogon chinensis.

[0090] Comparative Example 1

[0091] The difference between this comparative example and Example 1 is that it does not contain the Chinese medicine effective ingredient composition, that is, it only contains the probiotic composition.

[0092] Experimental Example 1 - Simulating the effect of adding effective ingredients of traditional Chinese medicine to the gastrointestinal tract on the activity of probiotics

[0093] Viable bacteria count: Probiotics are counted according to the Lactobacillus counting method in GB 4789.35-2023 "National Food Safety Standard - Microbiological Examination of Food - Lactic Acid Bacteria". Take 1.0g of the sample to be tested, dilute it with physiological saline, and select 2 to 3 continuous appropriate dilution gradients. For each dilution, use a pipette to take 100μL of the sample into sterilized MRS solid culture medium, spread it evenly with a spreading rod, and then culture it. Test 3 parallel samples for each dilution. Place the culture dish upside down in an anaerobic culture bag and culture it at a constant temperature of 37℃ for 48 hours, and then count the viable bacteria. The calculation formula for the dilution spread plate method is: CFU / mL = C ÷ V × M (where: C is the average number of colonies grown on the plate at a certain dilution; V is the volume of the diluent used when spreading the plate (mL); M is the dilution factor).

[0094] Preparation of in vitro simulated gastrointestinal digestive fluid: The pH value of human gastric fluid varies between 1.5 and 3.5 depending on the type of food consumed and the time of consumption. Take 0.1 mol / L potassium phosphate buffer as the base solution, adjust the pH value to three gradients of 1.5, 2.5, and 3.5 with hydrochloric acid, add an appropriate amount of pepsin according to a concentration of 10 g / L, mix thoroughly, and sterilize by passing through a 0.22 μm sterile membrane to obtain simulated gastric fluid. Take 0.1 mol / L potassium phosphate buffer as the base solution, adjust the pH value to 6.8, add an appropriate amount of pancreatic enzyme according to a concentration of 10 g / L, and add an appropriate amount of porcine bile salt according to a concentration of 3 g / L, mix thoroughly, and sterilize by passing through a 0.22 μm sterile membrane to obtain simulated intestinal fluid.

[0095] In vitro simulated gastrointestinal digestion: The reference human gastric digestion time is about 3 hours, so the time range of the experimental design is 0 to 3 hours. The probiotic composition of comparative example 1 of the present invention and the pharmaceutical composition of embodiment 1 of the present invention (i.e., the effective ingredient composition of traditional Chinese medicine + probiotic composition) were inoculated into 20.0 mL of simulated gastric fluid environment with different pH values ​​(1.5, 2.5, 3.5, 6.8), respectively, and cultured under anaerobic conditions at 37°C and 90rpm to simulate gastric fluid digestion. Samples were taken at 0, 1, 2, and 3 hours respectively, and the live bacteria were counted by the dilution coating plate counting method, and the survival rate was calculated; the results are shown in FIG. Figure 1-4 .

[0096] in, Figure 1 The results of the effects of the drugs of comparative example 1 and example 1 on the survival rate of probiotics in a simulated gastric fluid environment with a pH value of 1.5 are shown in FIG. Figure 1The results show that: at 0 hours, the viable bacterial count was 100%. After 1 hour of digestion, the survival rate of the bare probiotic bacteria in Comparative Example 1 was 55.13%, while the survival rate of the probiotic bacteria added with the Chinese herbal extract in Example 1 was about 89.77%. After 2 hours of digestion, the survival rate of the probiotic bacteria dropped to below 41.78%, but the survival rate of the probiotic bacteria added with the Chinese herbal active ingredient extract in Example 1 reached 73.67%. After 3 hours of digestion, the survival rate of the probiotic bacteria added with the Chinese herbal active ingredient extract in Example 1 reached 69.87%, while the survival rate of the bare probiotic bacteria in Comparative Example 1 was only 22.65%. The results show that when the gastric fluid pH is 1.5, the synergistic effect of the Chinese herbal active ingredient extract of the present invention can significantly improve the tolerance of probiotics to simulated gastric fluid (p < 0.0001).

[0097] Figure 2 The results of the effects of the drugs of Comparative Example 1 and Example 1 on the survival rate of probiotics in a simulated gastric fluid environment with a pH value of 2.5 are shown. Figure 3 The results of the effects of the drugs of Comparative Example 1 and Example 1 on the survival rate of probiotics in a simulated gastric fluid environment with a pH value of 3.5 are shown. Figure 2-3 The results show that: in this experiment, the viable bacterial count was 100% at 0 h. In the simulated gastric fluid with a pH of 2.5 and 3.5, the survival rate of the probiotics in Comparative Example 1 and Example 1 did not decrease significantly. After digestion for 0, 1, 2, and 3 h, the viable bacterial count remained between 85.62% and 99.08%. It can be seen that there was no significant difference in the viable bacterial count between the naked bacteria with and without the addition of the effective component extract of traditional Chinese medicine (p>0.1).

[0098] Figure 4 The results of the effects of the drugs of Comparative Example 1 and Example 1 on the survival rate of probiotics in a simulated gastric fluid environment with a pH value of 6.8 are shown. The reference food stays in the intestine for about 8 hours, so the time range of the experimental design is 0 to 8 hours. Figure 4 The results show that:

[0099] After 8 hours of digestion, the survival rate of the probiotics in Example 1 and Comparative Example 1, that is, whether or not the extract of the effective components of traditional Chinese medicine was added, decreased. After 2 hours of intestinal digestion, the probiotics survival rate of the probiotics added with the extract of the effective components of traditional Chinese medicine in Example 1 was 83.65%, while the survival rate of the naked probiotics in Comparative Example 1 was only 75.37%. As the digestion time increased, after 8 hours, the number of viable probiotics gradually decreased regardless of whether the extract of the effective components of traditional Chinese medicine was added. The survival rate of the probiotics added with the extract of the effective components of traditional Chinese medicine in Example 1 was 72.51%, while the survival rate of the naked probiotics in Comparative Example 1 was 49.36%. The results show that the extract of the effective components of traditional Chinese medicine of the present invention can significantly improve the tolerance of probiotics in intestinal fluid (p < 0.0001).

[0100] In summary, Figure 1-4 The results show that: through the in vitro simulated gastrointestinal fluid screening experiment, the results show that the simulated gastric fluid at pH 2.5 and pH 3.5 has basically no significant effect on the survival rate of probiotics (naked bacteria or added with Chinese herbal medicine active ingredient extracts). However, whether or not to add Chinese herbal medicine active ingredient extracts to the simulated gastric fluid at pH 1.5 has a significant effect on the survival rate of probiotics. After adding Chinese herbal medicine active ingredient extracts, the survival rate of probiotics is significantly higher than that of probiotic naked bacteria, indicating that Chinese herbal medicine active ingredient extracts have a protective effect on the activity of probiotics in simulated gastric fluid at pH 1.5, and in simulated intestinal fluid at pH 6.8, the survival rate of probiotics after adding Chinese herbal medicine active ingredient extracts is also significantly higher than that of probiotic naked bacteria. It can be seen that adding Chinese herbal medicine active ingredient extracts in simulated intestinal fluid can protect the activity of probiotics and improve the tolerance of probiotics to gastric fluid.

[0101] Experimental Example 2 - Comparison of the efficacy of the Chinese medicine preparation of the present invention and the chemical drug metformin in treating diabetes

[0102] The experimental method is as follows:

[0103] (1) Take the probiotic composition prepared in Example 1.

[0104] (2) Experimental animals

[0105] Male Kunming mice (8 weeks old) were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd. Five mice were placed in each cage in an environmentally controlled facility (temperature 22±2°C, humidity 55%±5%) with a 12 / 12 h light / dark cycle.

[0106] All experiments were conducted in the Animal Experiment Center of Northwest Agriculture and Forestry University and all animal experiments complied with relevant IACUC policies and regulations (Animal Ethics Number: IACUC2024-0316).

[0107] Establishment of animal model and feeding and administration: 36 Kunming mice were fed adaptively for 1 week and then randomly divided into 6 groups according to their body weight, with 6 mice in each group, namely: blank group, model group, metformin group, effective part of Chinese medicine group, probiotic group and the Chinese medicine preparation of Example 1, i.e. effective part of Chinese medicine + probiotic group, hereinafter referred to as: Chinese + benefit group. The blank group was fed with ordinary feed, and the other mice were fed with high-fat feed. They were all free to drink water. After 21 days, they were fasted but not watered for 12h and weighed. According to their body weight, they were intraperitoneally injected with streptozotocin (STZ) dissolved in 0.1mol / L sodium citrate buffer at a dose of 50mg / kg for 5 days. After 5 days of STZ injection, the blood glucose levels of the mice were measured for three consecutive days. When the blood glucose concentration of the mice was >11.1mmol / L, it was considered that the type 2 diabetes model was successfully established. The mice were gavage-administered at 9:00-10:00 every day for 4 consecutive weeks. The specific administration content is shown in Table 1. During the oral administration, the mice were kept in an environment with a temperature of 23±1°C and a humidity of 50-60% RH, and were exposed to natural light with a uniform day and night distribution (lighting time: 7:00-19:00). The water was kept fresh. The experimental results are shown in Table 1.

[0108] Table 1-Animal experimental design and drug administration content

[0109] Group Dosage content (4 weeks) Blank group 0.2 mL / kg sodium carboxymethylcellulose solution Model Group 0.2 mL / kg sodium carboxymethylcellulose solution Metformin group 0.075g / kg metformin hydrochloride Effective parts of traditional Chinese medicine group 0.58g / kg Chinese medicine active ingredient composition Probiotic group 0.48g / kg probiotic composition Medium + Benefit Group 1.06g / kg Chinese medicine and probiotic composition

[0110] After the four-week dosing period, mice were fasted for 8 hours without food or water. Blood was then collected from the mice under anesthesia using sterile tubes. The blood was allowed to clot naturally at room temperature for 15 minutes, and serum was collected by centrifugation at 4°C (3000 rpm) for approximately 20 minutes. Serum was then stored at -80°C until further use. Mice were sacrificed by cervical dislocation, and the liver and pancreas were quickly removed, weighed, and fixed in 4% paraformaldehyde.

[0111] 2.1 Changes in blood glucose over four weeks: Fasting blood glucose of each group of mice was measured and recorded weekly. The experimental results are shown in Figure 5 ,Depend on Figure 5 The results showed that compared with the blank group, the fasting blood glucose levels of the model group mice remained at a high level (P<0.0001), and their blood glucose levels were more than twice that of the blank group. After 4 weeks of administration, the blood glucose levels of the treated groups (metformin group, effective Chinese medicine fraction group, probiotic group, and Zhong + Yi group) all decreased to varying degrees compared with the model group. More notably, after 4 weeks of administration, the blood glucose levels of the metformin group mice decreased extremely significantly (P<0.05). The blood glucose levels of the effective Chinese medicine fraction group and probiotic group mice also decreased significantly. The most obvious difference was that the blood glucose levels of the Zhong + Yi group mice decreased significantly (P<0.006), and were lower than those of the metformin group.

[0112] 2.2OGTT test: First, the mice are fasted, usually for 12 to 16 hours to ensure that they are in a fasting state during the experiment. Subsequently, a predetermined dose of anhydrous glucose solution (usually 2g / kg body weight) is given to the mice orally, and the starting time is recorded. At specific time points after the administration of glucose (0min, 30min, 60min and 120min), blood samples are collected from the tail of the mouse using a blood collection needle to determine the blood glucose concentration. The collected blood samples should be processed immediately, and the blood glucose level should be measured using a blood glucose meter to assess the mouse's tolerance to glucose and then analyze its glucose metabolism status. Finally, a blood glucose-time curve is drawn based on the blood glucose values ​​at each time point, and the results are as follows: Figure 6 shown.

[0113] Depend on Figure 6 The results showed that the blood sugar levels of mice in the effective part of Chinese medicine group and probiotic group reached a peak 60 minutes after oral administration of glucose, while the blood sugar levels of mice in other groups reached a peak 30 minutes after oral administration of glucose and then began to decline; among them, the blood sugar levels of mice in the Chinese medicine + probiotic group and metformin group decreased the most and in the shortest time.

[0114] In addition, for Figure 6 The results of the blood glucose-time curve area (AUC value) of different groups of mice are shown in Figure 7 ,Depend on Figure 7 The results show that compared with the metformin group, the effective part of traditional Chinese medicine group and the probiotic group, the area under the OGTT curve (AUC value) of the Zhong+Yi group is the lowest, proving that the synergistic effect of the effective part of traditional Chinese medicine and the probiotic composition of the present invention can optimize the control of blood sugar in mice.

[0115] 2.3 Changes in Mouse Liver Index: The livers of mice in each group were quickly removed after being killed by cervical dislocation, and the results after weighing are shown in Figure 8 ,Depend on Figure 8 The results showed that compared with the blank group, the liver index of the mice in the model group was increased (P < 0.0001). This may be because the high-fat diet for 21 days caused fatty degeneration of the liver of the mice, increased liver weight, and possible inflammatory cell infiltration, accompanied by ballooning of liver cells and liver fibrosis. After 4 weeks of administration, compared with the model group, the liver index of the metformin group, the effective part of Chinese medicine group, the probiotic group and the Chinese + Yi group decreased to varying degrees, indicating that the liver of the mice had recovered to a certain extent. Among them, the best effect was the Chinese medicine and probiotic combination group, with P < 0.0007 compared with the blank group.

[0116] 2.4 Changes in Glycated Hemoglobin (GHb) Content: Glycated hemoglobin (GHb) is the product of the combination of hemoglobin in red blood cells and sugars (mainly glucose) in serum through non-enzymatic reactions. Its content is determined by the blood glucose concentration in the past 2 to 3 months rather than the immediate blood glucose concentration. Glycated hemoglobin is composed of HbA1a, HbA1b, and HbA1c, of which HbA1c accounts for about 70% and has a relatively stable structure. It is often used as a monitoring indicator for diabetes control, and its concentration should be expressed as a percentage of adult hemoglobin. Blood samples were taken from each group of mice to detect glycated hemoglobin, and the results are as follows: Figure 9 As shown by Figure 9 It can be seen that compared with the mice in the blank group, the glycated hemoglobin level of the diabetic mice in the model group was significantly increased (P<0.0001). Compared with the mice in the model group, the GHb levels of the metformin group, the effective part of traditional Chinese medicine group, the probiotics group and the Zhong + Yi group were reduced by 21.6%, 12.2%, 20.3% and 28.4%, respectively. The blood glucose levels of the mice in the metformin group and the Zhong + Yi group were extremely significantly reduced (P<0.0001), and the blood glucose levels of the mice in the Zhong + Yi group were more significantly reduced. It can be seen that the blood glucose lowering effect of the pharmaceutical preparation of the present invention is better than that of the traditional chemical drug metformin.

[0117] 2.5 Changes in serum total cholesterol (T-CHO) content: Blood samples were taken from mice in each group to detect changes in serum total cholesterol (T-CHO) content. The results are as follows: Figure 10 As shown by Figure 10 It can be seen that compared with normal mice, the serum total cholesterol level of diabetic mice in the model group was significantly increased (P<0.0001). After 4 weeks of administration, the serum total cholesterol levels of mice in the treatment groups (metformin group, effective part of Chinese medicine group, probiotic group and Zhong + Yi group) were reduced to varying degrees compared with the model group. After 4 weeks of administration, the serum total cholesterol values ​​of mice in the metformin group were extremely significantly reduced (P<0.003). The serum total cholesterol values ​​of mice in the effective part of Chinese medicine group and probiotic group were also significantly reduced (P<0.03). The most obvious was that the serum total cholesterol values ​​of mice in the Zhong + Yi group were significantly reduced (P<0.0003), and were lower than those in the metformin group.

[0118] 2.6 Changes in serum triglyceride levels: Blood samples were taken from each group of mice to detect changes in serum triglyceride levels. The results are as follows: Figure 11 As shown by Figure 11It can be seen that compared with normal mice, the serum triglyceride levels of diabetic mice in the model group were significantly increased (P<0.0001). After 4 weeks of administration, the triglyceride levels of mice in the treatment groups (metformin group, effective fraction of traditional Chinese medicine group, probiotic group and Zhong + Yi group) were reduced to varying degrees compared with the model group. Among them, the triglyceride levels of mice in the metformin group and Zhong + Yi group were extremely significantly reduced (P<0.0001), and the triglyceride levels of mice in the Zhong + Yi group were more significantly reduced. It can be seen that the therapeutic effect of the pharmaceutical preparation of the present invention on triglyceride levels is better than that of the traditional chemical drug metformin.

[0119] 2.7 Effects on organ pathology in diabetic mice:

[0120] Pancreatic beta cells secrete insulin to regulate the body's blood sugar balance. Pancreatic cell dysfunction or cell damage is the main cause of diabetes.

[0121] 2.7.1 The pathological condition of pancreatic tissue of mice was observed by paraffin tissue sectioning and HE staining. Figure 12 As shown by Figure 12 It can be seen that: the pancreatic tissue acinar cell membrane of blank group mice is intact, islets are round or oval structure, the boundary is clear and the islet mass has more cells and cytoplasm is abundant. Compared with blank group mice, the islet mass of model group mice has obvious cell necrosis, disordered islet structure, fuzzy islet tissue boundary, atrophy and deformation, inflammatory cell infiltration occurs, and the number of islet cells decreases. After administration for 4 weeks, there is regeneration phenomenon in the islet cells of treatment group mice (metformin group, Chinese medicine effective part group, probiotic group and Zhong+Yi group), wherein it is more obvious that metformin group and Zhong+Yi group, and Zhong+Yi group is significantly better than metformin group, the cell arrangement of Zhong+Yi group develops towards neat direction, cytoplasm is more abundant, inflammatory cell infiltration reduces, islet morphology has a trend of becoming better, and it can be seen that the therapeutic effect of pharmaceutical preparation of the present invention to pancreatic cell dysfunction and cell damage is better than traditional chemical drug metformin.

[0122] 2.7.2 Paraffin tissue sections and HE staining were used to observe the pathological condition of mouse liver tissue. Figure 13 As shown by Figure 13 It can be seen that the liver structure of the blank group mice was intact, the hepatocytes were uniform in size, there were no balloons, hyaline degeneration, no interstitial edema, and no inflammatory cell infiltration. Compared with the blank group mice, a large number of glycogen granules and fat vacuoles were observed in the hepatocytes of the model group mice, the hepatocytes were arranged irregularly, the interstitial edema was present, and inflammatory cell infiltration was present. After 4 weeks of administration, the liver structure of the mice in the other groups was significantly improved, among which the most obvious was the Zhong+Yi group, where fat vacuoles and glycogen granules were reduced, the hepatocyte structure gradually became intact, and the inflammatory cell infiltration was reduced. It can be seen that the therapeutic effect of the pharmaceutical preparation of the present invention on the liver tissue of mice is better than that of the traditional chemical drug metformin.

[0123] Experimental Example 3

[0124] The clinical efficacy test of the pharmaceutical preparation prepared in Example 1 of the present invention was carried out, and the results are as follows:

[0125] This experiment selected 144 diabetic cases for clinical trials:

[0126] Inclusion criteria: 1. Fasting blood glucose ≥ 7.0 mmol / L and / or 2-hour postprandial blood glucose ≥ 11.1 mmol / L, as per the "Guidelines for the Prevention and Treatment of Type 2 Diabetes in China" (2020 edition). 2. Ages 25-70 years, regardless of gender.

[0127] Exclusion criteria: 1. Patients in critical or aggravated condition or with complex complications. 2. Patients with allergic reactions to the ingredients in this Chinese medicine and probiotic combination. 3. Pregnant or lactating women.

[0128] Patients drank one bag of the oral liquid prepared in Example 1 of the present invention every day, and their blood sugar levels were tested and recorded. Patients with diabetes under 10 years of age took the liquid for more than 5 months, and patients with diabetes over 10 years of age took the liquid for more than 10 months. When the patient's fasting blood sugar was ≤7.0 mmol / L, 2-hour postprandial blood sugar was ≤11.1 mmol / L, and glycosylated hemoglobin was ≤5.7% for 7 consecutive days, the patient's blood sugar was considered to have returned to normal. The results are shown in Table 2.

[0129] Table 2 - Clinical efficacy testing experiments

[0130]

[0131] As shown in Table 2, the average blood sugar reduction for patients with a history of less than 10 years was 7.1 mmol / L, with a cure rate of over 90%. The average blood sugar reduction for patients with a history of over 10 years was 5.0 mmol / L, with a cure rate of over 81%. In summary, it can be concluded that the Chinese herbal oral solution provided in Example 1 of the present invention has a significant effect on controlling human blood sugar and treating diabetes.

[0132] Typical clinical case data:

[0133] Case 1: Mr. Hu, 58 years old, male, employee, had diabetes for 2 years and came to the clinic on February 16, 2025. Symptoms: slightly thin body, dry mouth, hot flashes, red tongue with scanty fur, and a strong and rapid pulse. The glycosylated hemoglobin concentration was 6.51% (normal range: 4% to 6%). He took one bag of the oral solution prepared in Example 1 daily. After 3 days, his fasting blood glucose level was 5.6 mmol / L (normal range: 3.9 mmol / L to 6.1 mmol / L), and his postprandial blood glucose level was 6.7 mmol / L (normal range: <7.8 mmol / L).

[0134] Case 2: Aunt Wang, 53 years old, female, farmer, has suffered from diabetes for 2 years. She came to the hospital for treatment on February 11, 2024. Her symptoms were: fat body, dry mouth, hot flashes, red tongue with little coating, thin and deep pulse, and blood sugar value of 8.1mmol / L. After taking one bag of oral solution prepared in Example 1 for 2 days, her fasting blood sugar was 6.0mmol / L and her postprandial blood sugar was 7.5mmol / L.

[0135] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. A pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar, characterized in that: include: A composition of effective ingredients of traditional Chinese medicine and a composition of probiotics; The traditional Chinese medicine effective ingredient composition comprises, by weight, 10-40 parts of momordica charantia glycosides, 10-40 parts of purslane phenolic acid, 5-30 parts of mulberry leaf alkaloids, 10-50 parts of corn silk flavonoids, 10-50 parts of buckwheat flavonoids, 10-50 parts of Moringa oleifera leaf flavonoids, 10-50 parts of sophora japonica flower flavonoids, 10-50 parts of pine pollen flavonoids, 10-50 parts of golden camellia flavonoids, 20-50 parts of Spirulina platensis phycocyanin, 20-40 parts of prickly pear cactus fruit polyphenols, 10-30 parts of total saponins from bamboo root seven, 15-30 parts of spider incense volatile oil, 10-25 parts of cucurbit septetesters, and 25-50 parts of unsaturated fatty acids from euphorbia cerifera. The probiotic composition comprises: Lactobacillus gasseri, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus bulgaricus and Clostridium sporogenes, and the ratio thereof is 1-5:1-5:1-5:1-5:1-5:1-5 CFU / g or CFU / ml.

2. The pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar according to claim 1, characterized in that: The extraction method of momordica charantia is as follows: using an alcohol solvent extraction method, according to a solid-liquid ratio of 1:10-20 g / mL, ultrasonic-assisted or reflux extraction for a period of time to obtain a crude extract, the crude extract is defatted and extracted, and then separated by column chromatography, and then purified by high-performance liquid chromatography to obtain momordica charantia; The method for extracting total phenols from purslane is as follows: drying the whole purslane herb, crushing and sieving it, extracting it with an alcohol solvent at a solid-liquid ratio of 1:10-20 g / mL, performing ultrasonic-assisted or reflux extraction for a period of time to obtain a crude extract, concentrating the crude extract, and enriching phenols with a macroporous resin to obtain total phenols from purslane; The extraction method of the mulberry leaf alkaloids is as follows: mulberry leaf powder is extracted by an alcohol solvent method at a solid-liquid ratio of 1:10-20 g / mL, ultrasonic-assisted extraction is performed for a period of time, and the mulberry leaf alkaloids are obtained by concentration and drying.

3. The pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar according to claim 1, characterized in that: The extraction method of flavonoids from corn silk, Moringa leaves, pine pollen, buckwheat, Sophora japonica flowers, and Camellia chrysantha is as follows: the powders of the above raw materials are extracted with an alcohol solvent at a material-liquid ratio of 1:20-40 g / mL, ultrasonic-assisted extraction is performed for a period of time, and flavonoid compounds are obtained by centrifugation, filtration, and concentration.

4. The pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar according to claim 1, characterized in that: The extraction method of the phycocyanin of Spirulina platensis is as follows: dried algae of Spirulina platensis are ground into powder with liquid nitrogen, sieved, cells are broken by enzymatic hydrolysis, and then ammonium sulfate is added for graded precipitation. The phycocyanin of Spirulina platensis is purified and refined by ion exchange chromatography, desalted and concentrated, and freeze-dried for storage or spray-dried to obtain the phycocyanin of Spirulina platensis.

5. The pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar according to claim 1, characterized in that: The method for extracting polyphenols from prickly pear cactus fruit is as follows: peeling, removing seeds, cutting cactus fruit into pieces, freeze-drying, crushing and sieving to obtain uniform fruit powder; adding the fruit powder into an alcohol solvent at a material-liquid ratio of 1:30-60 g / mL; performing ultrasonic extraction for a period of time; collecting the supernatant after centrifugal separation; and obtaining the prickly pear cactus fruit polyphenols after concentration, purification, impurity removal and elution.

6. The pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar according to claim 1, characterized in that: The extraction of the total saponins of bamboo root seven is as follows: dried bamboo root seven is crushed and sieved, and then a solvent extraction method is adopted. The bamboo root seven powder is added to an alcohol solvent according to a material-liquid ratio of 1:20-40g / mL, and extracted for a period of time. The solvent is recovered to obtain a residue, and the residue is dissolved in water and extracted multiple times. The extracts are combined and concentrated to obtain the total saponins of bamboo root seven.

7. The pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar according to claim 1, characterized in that: The extraction method of the spider incense volatile oil is as follows: dried spider incense is crushed and sieved, and then steam distilled with a material-liquid ratio of 1:5-10g / mL, heated and distilled for a period of time, and the volatile oil is collected to obtain the spider incense volatile oil.

8. The pharmaceutical composition for regulating intestinal microecological balance and lowering blood sugar according to claim 1, characterized in that: The extraction method of the cucurbitacin septuplene is as follows: dried cucurbitacin septuplene is crushed and sieved, and then the cucurbitacin septuplene powder is added to the alcohol solvent according to the material-liquid ratio of 1:10-20 g / mL by an alcohol solvent extraction method, and extracted for a period of time. After multiple extractions, the extracts are combined, and the cucurbitacin septuplene is obtained after reduced pressure concentration and extraction enrichment. The method for extracting unsaturated fatty acids from the ground peach is as follows: dried ground peach is crushed and sieved, and then an alcohol solvent extraction method is adopted to add ground peach powder into an alcohol solvent according to a material-liquid ratio of 1:10-30 g / mL, extract for a period of time, and concentrate and dry to obtain the ground peach unsaturated fatty acids.

9. A Chinese medicine preparation for regulating intestinal microecological balance and lowering blood sugar, characterized in that: The invention comprises the pharmaceutical composition according to any one of claims 1 to 8 and pharmaceutically acceptable excipients.

10. Use of the pharmaceutical composition according to any one of claims 1 to 8 in the preparation of a drug for treating hypoglycemia.

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