Composite plant polypeptide composition for reducing blood glucose, repairing cells, resisting oxidation and enhancing immunity as well as preparation method and application of composite plant polypeptide composition

Abstract

The invention provides a composite plant polypeptide composition for reducing blood glucose, repairing cells, resisting oxidation and enhancing immunity as well as a preparation method and application thereof, and belongs to the technical field of biology. The invention provides a composition. Comprising a protein peptide composed of millet protein peptide, soybean protein isolate peptide, whey protein peptide and sea buckthorn protein peptide, dietary fiber composed of chia seed powder and konjaku flour, a probiotic compound composed of bifidobacterium lactis BB-12 and lactobacillus rhamnosus LGG, fructo-oligosaccharide, balsam pear peptide, a cyclocarya paliurus extract, a corn stigma extract, a mulberry leaf extract, a vitamin E extract, a vitamin E extract, a vitamin E extract, a vitamin E extract, a vitamin E extract and a vitamin E extract. The extract mixture is composed of a radix puerariae extract, a rhizoma polygonati polysaccharide extract, a moringa oleifera leaf extract and leech powder, and the vitamin mineral substance mixture is composed of other functional components. The composition provided by the invention is matched with various proteins, dietary fibers, probiotics, prebiotics, plant extracts, vitamins, minerals, other functional components and the like, so that the purpose of effectively and synergistically improving the conditions of diabetes and nephropathy from multiple targets and multiple ways is finally achieved.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a compound plant polypeptide composition with hypoglycemic, cell repair, antioxidant, and immune-enhancing effects, as well as its preparation method and application. Background Technology

[0002] Diabetes mellitus is a prevalent metabolic disease that easily leads to various chronic complications during its progression. Among these, diabetic nephropathy, as one of the most common and serious microvascular complications, has become the leading cause of end-stage renal disease. The pathogenesis of diabetes and nephropathy is extremely complex, not caused by a single pathological step, but rather by the intertwined and synergistic effects of multiple pathological processes, including metabolic disorders, oxidative stress, inflammatory responses, renal fibrosis, and intestinal microecological imbalance. Oxidative stress is the core initiating step, directly inducing oxidative damage and apoptosis in pancreatic β-cells, renal parenchymal cells, and vascular endothelial cells, thereby causing a series of problems such as insufficient insulin secretion, progressive decline in renal function, and microvascular circulatory disorders, further increasing the difficulty of disease management. In addition, diabetic patients often have metabolic syndrome, and the synergistic abnormalities of multiple metabolic indicators such as blood glucose, blood lipids, and blood pressure further aggravate the damage to target cells, increasing the complexity of disease management. Single intervention methods are insufficient to achieve comprehensive disease improvement and organ protection.

[0003] Currently, the prevention and treatment of diabetes and kidney disease mainly fall into two categories: clinical drug therapy and dietary intervention (dietary therapy). Various preventive and treatment compositions (including drug compositions and dietary therapy compositions) play an important role in clinical application and research, but all have significant technical deficiencies and unresolved technical difficulties, making it difficult to meet the clinical needs for multi-faceted protection and synergistic regulation of diabetes and kidney disease. Specifically, current clinical drug compositions used for the prevention and treatment of diabetes and kidney disease often focus on a single pathological target or a single disease process. Interventions targeting oxidative stress are mostly single-component, with limited antioxidant coverage. Furthermore, they do not target the core pathology of diabetic nephropathy—"oxidative stress-induced target cell damage and apoptosis leading to progressive decline in kidney function"—by designing targeted cell repair mechanisms. This fails to achieve the dual intervention of "inhibiting damage at its source and promoting the regeneration of damaged cells," making it difficult to reverse early kidney damage. In recent years, dietary therapy compositions have received widespread attention in the adjunctive prevention and treatment of diabetes and kidney disease due to their high safety and suitability for long-term intervention. However, most existing dietary therapy compositions lack a systematic antioxidant system design and have no targeted formulation plan for cell repair. They can only achieve basic nutritional supplementation or mild blood sugar reduction, and are difficult to effectively alleviate the chronic oxidative stress and target cell damage in patients with diabetic nephropathy.

[0004] In summary, current pharmaceutical and dietary compositions used for the prevention and treatment of diabetes and kidney disease suffer from several drawbacks. Firstly, pharmaceutical compositions often target only a single point, limiting their scope of action, resulting in insufficient long-term safety and hindering comprehensive disease control. Secondly, dietary compositions often lack scientific rigor, have unclear pharmacodynamic bases, and ambiguous mechanisms of action, and lack systematic antioxidant systems and targeted cell repair designs, failing to meet the multi-faceted protection and synergistic regulation needs of diabetic and kidney disease patients. Addressing these shortcomings, and considering the core characteristics of diabetes and kidney disease—"intertwined pathological processes, oxidative stress as the core, target cell damage as the key, and limited effectiveness of single interventions"—developing a scientifically formulated, effective, multi-target, multi-pathway synergistic nutritional formula that combines potent antioxidant effects, targeted cell repair, enhanced immunity, blood glucose regulation, and kidney protection, while also ensuring high safety and suitability for long-term intervention, is a pressing technical challenge in this field. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide a composition with a four-fold synergistic system of "antioxidant-damage inhibition-repair promotion-metabolism regulation", which can synergistically improve the condition of diabetes and kidney disease from multiple targets and multiple pathways, and also has enhanced antioxidant effect, which can achieve targeted repair of pancreatic islet, kidney and vascular target cells, so as to overcome the defects of the prior art.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a composition that combines the functions of enhancing immunity, anti-oxidation, cell repair, blood sugar regulation and kidney protection, including protein peptides, dietary fiber, probiotic and prebiotic mixture, extract mixture, vitamin and mineral mixture and other functional ingredients; The protein peptides include 10g-20g of millet protein peptides, 20g-30g of soybean protein peptide isolate, 10g-20g of whey protein peptides, and 10g-15g of sea buckthorn protein peptides. The dietary fiber includes 10g-20g of chia seed powder and 20g-50g of konjac powder; The probiotic and prebiotic mixture includes 35 billion to 50 billion CFU of probiotic complex and 15g to 30g of fructooligosaccharides. The probiotic complex is composed of Bifidobacterium lactis BB-12 and Lactobacillus rhamnosus LGG. The extract mixture includes 1000mg~1800mg of bitter melon peptide, 1000mg~1500mg of Eucommia ulmoides extract, 1000mg~2000mg of corn silk extract, 1000mg~1500mg of mulberry leaf extract, 200mg~1000mg of kudzu root extract, 200mg~1000mg of Polygonatum sibiricum polysaccharide extract, 1000mg~1500mg of Moringa oleifera leaf extract, and 200mg~400mg of leech powder; The vitamin and mineral mixture comprises B vitamins, calcium 800mg-1000mg, vitamin D3 600IU-800IU, selenium 40μg-50μg, zinc 4mg-5mg, vitamin E 70mg-80mg, vitamin C 70mg-80mg, and β-carotene 400mg-500mg. The B vitamins include vitamin B1 0.8mg-1.2mg, vitamin B6 1.0mg-1.5mg, and vitamin B2. 12 1.8μg~2.4μg; The remaining functional ingredients include taurine 400mg~500mg, yeast chromium 700μg~800μg, α-lipoic acid 700mg~800mg, Omega-3 500mg~600mg, stevioside 400mg~500mg, astaxanthin 700~900mg, and curcumin 700~900mg.

[0007] Preferably, the ratio of viable counts of Bifidobacterium lactis BB-12 to Lactobacillus rhamnosus LGG is 2:3.

[0008] The present invention also provides a method for preparing the above composition, comprising the following steps: Polylactic acid-glycolic acid copolymer, dichloromethane-acetone solution, protein peptides and Tween 80 were mixed to obtain an oil phase; the oil phase was added dropwise to a polyvinyl alcohol aqueous solution with a mass-volume percentage of 4% to 6%, and emulsified by high-speed shearing to obtain an emulsion; the emulsion was poured into a polyvinyl alcohol aqueous solution with a mass-volume percentage of 0.5% to 1.5%, and stirred to obtain a secondary emulsion; the organic solvent in the secondary emulsion was removed, the precipitate was centrifuged, washed, resuspended in an aqueous solution containing a lyophilization protectant, and lyophilized to obtain nanoparticles; Microcapsules were obtained by encapsulating a mixture of probiotics and prebiotics using sodium alginate and chitosan. Medium-chain triglycerides, vitamin E, β-carotene, Omega-3, astaxanthin, curcumin and ascorbyl palmitate were mixed under light-protected conditions to obtain an oil phase; water, Tween 80 and propylene glycol were mixed to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase and stirred, then ultrasonicated and filtered to obtain a nanoemulsion. Water, sodium carboxymethyl cellulose, carbomer 940, leech powder, and glycerin were mixed to obtain an aqueous phase; liquid paraffin and Tween 80 were mixed to obtain an oil phase; the aqueous phase was added dropwise to the oil phase and stirred to emulsify, thus obtaining an emulsion; a CaCl2 aqueous solution of 4%~6% by mass was added dropwise to the emulsion and stirred to solidify; then anhydrous ethanol was added and stirred to break the emulsion, the precipitate was collected, washed, dried, and sieved to obtain microspheres; The kudzu root extract was subjected to high-pressure homogenization at 145MPa~155MPa and 30℃~35℃ to obtain the treated kudzu root extract. Nanoparticles, microcapsules, nanoemulsions, microspheres, treated kudzu root extract, and other raw materials are mixed to obtain a composition.

[0009] Preferably, the volume ratio of dichloromethane to acetone in the dichloromethane-acetone solution is 1:1; the mass-to-volume ratio of polylactic acid-glycolic acid copolymer, dichloromethane-acetone solution, protein peptide, and Tween 80 is 18g~22g:180mL~220mL:8g~12g:1mL~3mL; the volume ratio of 4%~6% polyvinyl alcohol aqueous solution to dichloromethane-acetone solution is 350~450:180~220; and the rotation speed of the high-speed shear emulsification is... Stirring at 11000rpm~13000rpm for 4min~6min; the volume ratio of polyvinyl alcohol aqueous solution (0.5%~1.5% by mass) to dichloromethane acetone solution is 350~450:180~220; the method for removing organic phase solvents from the double emulsion is magnetic stirring at 150rpm~250rpm for 4~6h; the lyophilization protectant is mannitol, and the mass-volume percentage of the lyophilization protectant in the aqueous solution is 4%~6%.

[0010] Preferably, the mass ratio of sodium alginate to chitosan is 2~4:1; the concentration of sodium alginate is 1.5%~2.5%; and the curing time is 25min~35min.

[0011] Preferably, the weight ratio of the medium-chain triglycerides, vitamin E, β-carotene, Omega-3, astaxanthin, curcumin, and ascorbyl palmitate is 100:5:31.25:37.5:4:4:0.5; the weight ratio of water, Tween 80, and propylene glycol is 790~795:25~35:8~12; the dropping rate of the oil phase into the aqueous phase is 4mL / min~6mL / min; the ultrasonic power of the ultrasonic treatment is 250W~350W, the working mode is 5s on, 3s off, and the total time is 12min~18min; the filtration is performed using a 200-mesh filter.

[0012] Preferably, the weight-to-volume ratio of water, sodium carboxymethyl cellulose, carbomer 940, leech powder, and glycerin is 70-90 mL: 13-17 g: 6.5-8.5 g: 15-25 g: 2-4 mL; the volume ratio of liquid paraffin to Tween 80 is 140-160:5; the dropping rate when the aqueous phase is added to the oil phase is 1 mL / min; the stirring and emulsification speed is 500-700 rpm, and the time is 25-35 min; the weight ratio of CaCl2 to sodium carboxymethyl cellulose is 1:2-4; and the particle size of the microspheres is 100 μm-500 μm.

[0013] The present invention also provides the application of the above-described composition or preparation method in the preparation of immune-enhancing products, antioxidant products, cell repair products, or diabetes prevention and treatment products.

[0014] The present invention also provides the use of the above-described composition or preparation method in the preparation of products for the prevention and / or treatment of diabetic nephropathy.

[0015] The present invention also provides the use of the above-described composition or preparation method in the preparation of products that help maintain healthy blood sugar levels.

[0016] The beneficial effects of this invention are: The composition provided by this invention combines various proteins, dietary fiber, probiotics, prebiotics, plant extracts, vitamins, minerals and other functional ingredients to create a four-fold synergistic system of "antioxidant-damage suppression-repair promotion-metabolism regulation". Ultimately, it can effectively improve the condition of diabetes and kidney disease from multiple targets and pathways, and also has a strong antioxidant effect, enabling targeted repair of pancreatic islet, kidney and vascular target cells. Attached Figure Description

[0017] Figure 1 Phenotypic image of fluorescence intensity of neutrophils in zebrafish (magnification 40x) for blank control group. Figure 2 Phenotypic image of fluorescence intensity of zebrafish neutrophils in the sample treatment group (magnification 40x). Figure 3 The results show the statistical analysis of fluorescence intensity in neutrophils from different groups of zebrafish. This indicates that P < 0.05; Figure 4 Phenotypic image of ROS fluorescence intensity in zebrafish (magnification 40x) for blank control group. Figure 5 Phenotypic diagram of ROS fluorescence intensity in zebrafish of the sample treatment group (magnification 40x); Figure 6 Statistical results of ROS fluorescence intensity in zebrafish from different groups. This indicates that P < 0.05; Figure 7 Phenotypic images of the results of zebrafish tail fin regeneration experiments in different groups (magnification 40x). Figure 8 A graph showing the statistical results of tail fin regeneration area in zebrafish from different groups. This indicates that P < 0.001; Figure 9 The graph shows the statistical results of blood glucose levels in different groups of zebrafish. This means P < 0.001. Detailed Implementation

[0018] This invention provides a composition that combines the functions of enhancing immunity, anti-oxidation, cell repair, blood sugar regulation and kidney protection, including protein peptides, dietary fiber, probiotic and prebiotic mixture, extract mixture, vitamin and mineral mixture and other functional ingredients; The protein peptides include 10g-20g of millet protein peptides, 20g-30g of soybean protein peptide isolate, 10g-20g of whey protein peptides, and 10g-15g of sea buckthorn protein peptides. The dietary fiber includes 10g-20g of chia seed powder and 20g-50g of konjac powder; The probiotic and prebiotic mixture includes 35 billion to 50 billion CFU of probiotic complex and 15g to 30g of fructooligosaccharides. The probiotic complex is composed of Bifidobacterium lactis BB-12 and Lactobacillus rhamnosus LGG. The extract mixture includes 1000mg~1800mg of bitter melon peptide, 1000mg~1500mg of Eucommia ulmoides extract, 1000mg~2000mg of corn silk extract, 1000mg~1500mg of mulberry leaf extract, 200mg~1000mg of kudzu root extract, 200mg~1000mg of Polygonatum sibiricum polysaccharide extract, 1000mg~1500mg of Moringa oleifera leaf extract, and 200mg~400mg of leech powder; The vitamin and mineral mixture comprises B vitamins, calcium 800mg-1000mg, vitamin D3 600IU-800IU, selenium 40μg-50μg, zinc 4mg-5mg, vitamin E 70mg-80mg, vitamin C 70mg-80mg, and β-carotene 400mg-500mg. The B vitamins include vitamin B1 0.8mg-1.2mg, vitamin B6 1.0mg-1.5mg, and vitamin B2. 12 1.8μg~2.4μg; The remaining functional ingredients include taurine 400mg~500mg, yeast chromium 700μg~800μg, α-lipoic acid 700mg~800mg, Omega-3 500mg~600mg, stevioside 400mg~500mg, astaxanthin 700~900mg, and curcumin 700~900mg.

[0019] This invention does not specifically limit the source of each raw material in the composition; commercially available raw materials commonly used in the art can be used. In the protein peptides described in this invention, the preferred amount of millet protein peptide is 12g-18g, more preferably 14g-16g; the preferred amount of soybean protein peptide isolate is 22g-28g, more preferably 24g-26g; the preferred amount of whey protein peptide is 12g-18g, more preferably 14g-16g; and the preferred amount of sea buckthorn protein peptide is 11g-14g, more preferably 12g-13g. In this invention, the preferred molecular weight of the millet protein peptide is <1000Da and the purity is ≥90%; the preferred molecular weight of the soybean protein peptide isolate is <1500Da and the purity is ≥90%; the preferred molecular weight of the whey protein peptide is 500-800Da and the purity is ≥95%; and the preferred molecular weight of the sea buckthorn protein peptide is <1200Da and the purity is ≥85%. The four small-molecule protein peptides in the composition of this invention not only serve as a high-quality protein source for patients with diabetic nephropathy, but also as a core raw material supply layer for cell repair. Their low molecular weight and easy absorption characteristics allow them to be directly absorbed by the intestines without liver metabolism, providing sufficient amino acid raw materials for the regeneration and repair of pancreatic β-cells, renal parenchymal cells, and vascular endothelial cells without increasing the excretory burden on the kidneys, thus meeting the organ tolerance requirements of patients with diabetic nephropathy. Simultaneously, each protein peptide also possesses unique physiological activities: millet protein peptides have a mild glycemic regulatory effect; soybean protein peptides' antioxidant and anti-inflammatory properties can reduce renal microvascular damage and inhibit the progression of glomerulosclerosis; whey protein peptides can effectively improve insulin sensitivity and help prevent hyperphosphatemia; and sea buckthorn protein peptides have significant hypoglycemic and renal protective effects. The combination of these four protein peptides achieves the dual value of "raw material supply + active target protection." The proportions of the protein peptides in the composition of this invention can meet the patient's protein needs while reducing the burden on the kidneys. The combination of these four protein peptides not only takes into account the special effects of each component, but also emphasizes the scientific nature of the overall proportions, which can provide a more comprehensive amino acid profile and exert a synergistic effect to achieve multiple goals of blood glucose regulation, kidney protection and nutritional support at the molecular level.

[0020] In the dietary fiber composition of this invention, the preferred amount of chia seed powder is 12g-18g, more preferably 14g-16g; the preferred amount of konjac powder is 25g-45g, more preferably 30g-40g, and the glucomannan content in the konjac powder is preferably ≥80%. The complex system of dietary fiber of this invention exerts a positive impact on diabetes and nephropathy through multiple mechanisms, including regulating intestinal microecology, improving intestinal barrier function, and inhibiting systemic inflammatory responses. This forms a unique gut-kidney axis intervention strategy (referring to an intervention method that protects kidney tissue by regulating intestinal microecological balance, improving intestinal barrier function, reducing uremic toxin production and systemic chronic inflammatory responses), while simultaneously creating a favorable in vivo microenvironment for cell repair. Chia seed powder can form a high-viscosity gel, which significantly slows down gastric emptying and small intestine absorption of glucose, stabilizes the postprandial blood glucose curve, and reduces repeated damage to pancreatic β cells caused by blood glucose fluctuations; konjac powder's glucomannan can selectively promote the proliferation of probiotics such as bifidobacteria and lactic acid bacteria, improve intestinal flora imbalance, reduce the release of intestinal inflammatory factors, and avoid the inhibition of target cell repair by systemic chronic inflammation.

[0021] In the probiotic-prebiotic mixture of the present invention, the amount of the probiotic complex is preferably 40 billion CFU to 45 billion CFU, and the ratio of viable bacteria of Bifidobacterium lactis BB-12 to Lactobacillus rhamnosus LGG in the probiotic complex is preferably 2:3. The prebiotic is preferably fructooligosaccharide, and the amount of fructooligosaccharide is preferably 18g to 27g, more preferably 20g to 25g. The present invention combines dietary fiber and the probiotic-prebiotic mixture. In terms of blood sugar control, intestinal flora regulation can increase the secretion of incretins such as GLP-1 and improve insulin sensitivity; in terms of kidney protection, a healthy intestinal microecology can reduce the production of uremic toxins, alleviate systemic micro-inflammatory states, and delay the progression of kidney disease; in terms of cell repair, the short-chain fatty acids produced by probiotic metabolism can provide energy for intestinal epithelial cells, repair the intestinal barrier, and inhibit the expression of pro-inflammatory factors, avoiding interference of inflammatory factors on the repair of pancreatic, renal, and vascular target cells, thus providing a non-inflammatory in vivo environment for cell repair. The unique feature of this intestinal targeted intervention system is its multi-target mechanism of action and high safety; even with long-term use, it will not produce side effects similar to those of drugs.

[0022] In the extract mixture of the composition of the present invention, the amount of bitter melon peptide is preferably 1200mg~1600mg, more preferably 1300mg~1500mg, and the molecular weight of the bitter melon peptide is preferably <2000Da and the content of bitter melon extract is ≥20%; the amount of Cyclocarya paliurus extract is preferably 1100mg~1400mg, more preferably 1200mg~1300mg, and the total flavonoid content of the Cyclocarya paliurus extract is preferably ≥8%; the amount of corn silk extract is preferably 1200mg~1800mg, more preferably 1400mg~1600mg; the amount of mulberry leaf extract is preferably 1100mg~1400mg, more preferably 1200mg~1300mg; and the amount of kudzu root extract is preferably 1200mg~1400mg, more preferably 1200mg~1300mg. The preferred dosage is 400mg~800mg, more preferably 500mg~600mg, and the puerarin content of the kudzu root extract is preferably ≥40%; the preferred dosage of the polygonatum polysaccharide extract is 400mg~800mg, more preferably 500mg~600mg, and the polysaccharide content of the polygonatum polysaccharide extract is preferably ≥50%; the preferred dosage of the moringa leaf extract is 1100mg~1400mg, more preferably 1200mg~1300mg; the preferred dosage of the leech powder is 250mg~350mg, more preferably 280mg~320mg, and the leech powder is preferably purified leech powder that has been de-allergenic, has a heavy metal content ≤2mg / kg, and whose microbial limits meet the national health food standards. The extract mixture of this invention forms the core active layer for targeted cell repair. Each component precisely repairs key damaged target cells in diabetic nephropathy, while also possessing hypoglycemic, anti-inflammatory, and antioxidant effects: bitter melon peptides have insulin-like effects, inhibiting α-glucosidase activity and reducing the secretory burden on pancreatic β-cells; Eucommia ulmoides extract improves insulin signal transduction, inhibits hepatic gluconeogenesis, and reduces glucose toxicity to pancreatic β-cells; corn silk extract has significant diuretic and anti-inflammatory effects, alleviating glomerular hyperfiltration in the early stages of diabetic nephropathy and protecting renal parenchymal cells; puerarin in kudzu root extract is a key component for glomerular podocyte repair, reducing glomerular podocyte shedding. It inhibits the epithelial-mesenchymal transition of renal parenchymal cells, delays renal fibrosis, and improves microvascular circulation, providing sufficient nutritional supply for vascular endothelial cell repair. Polygonatum polysaccharide extract is the core component for pancreatic β-cell repair, which can activate the pancreatic β-cell proliferation pathway, enhance insulin secretion, and reverse apoptosis caused by glucose toxicity. Moringa leaf extract is rich in isothiocyanates and phenolic substances, which have strong antioxidant and anti-inflammatory capabilities, reducing secondary damage to target cells caused by oxidative stress. Leech powder contains the natural anticoagulant hirudin, which can improve the hypercoagulable state of blood in diabetic patients, prevent microthrombus formation, and create a good blood circulation microenvironment for vascular endothelial cell repair.

[0023] In the vitamin and mineral mixture of the composition of the present invention, the amount of calcium is preferably 850 mg to 950 mg, more preferably 900 mg; the amount of vitamin D3 is preferably 650 IU to 750 IU, more preferably 700 IU; the amount of selenium is preferably 42 μg to 48 μg, more preferably 44 μg to 46 μg; the amount of zinc is preferably 4.2 mg to 4.8 mg; the amount of vitamin E is preferably 72 mg to 78 mg, more preferably 74 mg to 76 mg; the amount of vitamin C is preferably 72 mg to 78 mg, more preferably 74 mg to 76 mg; the amount of β-carotene is preferably 420 mg to 480 mg, more preferably 440 mg to 460 mg; the amount of vitamin B1 is preferably 0.9 mg to 1.1 mg; the amount of vitamin B6 is preferably 1.2 mg to 1.4 mg; and the amount of vitamin B... 12 The preferred dosage is 2.0 μg to 2.2 μg. The vitamin and mineral mixture of this invention serves as a coenzyme support layer for cell repair, not only forming a multi-layered antioxidant defense network but also acting as a core component of cell repair-related enzymes, ensuring the smooth progress of the repair process: B vitamins participate in glucose and energy metabolism, improve insulin sensitivity, and simultaneously act as coenzymes for dehydrogenases and transaminases, participating in material metabolism and energy synthesis during cell repair; zinc participates in insulin synthesis and secretion, maintaining the normal morphology of pancreatic β cells, and simultaneously acts as a coenzyme for DNA polymerase and RNA polymerase, directly participating in DNA replication and repair of damaged cells; selenium, as a component of glutathione peroxidase, strengthens the antioxidant repair capacity of renal cells and vascular endothelial cells, reducing the interference of oxidative stress on the repair process; the combination of calcium and vitamin D3 improves calcium and phosphorus metabolism, regulates the renin-angiotensin system, and protects renal vascular endothelial cells; vitamin E, vitamin C, and β-carotene constitute a complete free radical scavenging system, clearing excess free radicals generated during cell repair, avoiding secondary damage, and ensuring repair efficiency.

[0024] In the remaining functional components of the composition of the present invention, the amount of taurine is preferably 420 mg to 480 mg, more preferably 440 mg to 460 mg; the amount of yeast chromium is preferably 720 μg to 780 μg, more preferably 740 μg to 760 μg; the amount of α-lipoic acid is preferably 720 mg to 780 mg, more preferably 740 mg to 760 mg; the amount of Omega-3 is preferably 520 mg to 580 mg, more preferably 540 mg to 560 mg; the amount of steviol glycoside is preferably 420 mg to 480 mg, more preferably 440 mg to 460 mg, wherein the steviol glycoside content is preferably ≥90%; the amount of astaxanthin is preferably 800 mg, wherein the astaxanthin has a trans structure ≥98% and a purity ≥5%; the amount of curcumin is preferably 800 mg, wherein the total curcumin content is ≥95%. In this invention, the Omega-3 is preferably Omega-3 with a total content of not less than 60% of DHA and EPA. The functional ingredients provided by this invention are meticulously designed to construct a powerful, multi-dimensional antioxidant system and a targeted cell repair system, forming a unique multi-target functional regulation system: Taurine can protect pancreatic β-cells from glucose toxicity and lipotoxicity, and enhance insulin secretion function; at the same time, it regulates calcium ion balance and nerve conduction velocity, improving symptoms of diabetic peripheral neuropathy; Yeast chromium can enhance insulin receptor tyrosine kinase activity, improve insulin sensitivity, and uses a safe and efficient organic chromium form to avoid nephrotoxicity; α-lipoic acid can directly scavenge various oxygen free radicals, regenerate the endogenous antioxidant system, and inhibit aldose reductase activity, reducing osmotic pressure damage caused by sorbitol accumulation and protecting renal parenchymal cells; Omega-3 has significant anti-inflammatory and lipid-regulating effects, inhibits NF-κB nuclear translocation, reduces the production of pro-inflammatory factors such as TNF-α and IL-6, and replenishes cell membrane phospholipids, accelerating the membrane repair of damaged vascular endothelial cells and pancreatic β-cells, restoring normal cellular physiological function; Stevia glycosides, as a natural sweetener, provide a zero-calorie sweetness alternative, avoiding repeated damage to target cells caused by blood sugar fluctuations.

[0025] Astaxanthin is a super fat-soluble antioxidant with an antioxidant capacity more than 500 times that of vitamin E. It can target and eliminate lipid peroxidation free radicals in cell membranes and vascular endothelium, protect microvascular endothelial cells and glomerular podocytes in patients with diabetic nephropathy, inhibit cell oxidative apoptosis, maintain the integrity of the filtration barrier, and improve microcirculatory disorders in diabetic patients, providing sufficient nutrition and oxygen for target cell repair. Curcumin has dual core effects of antioxidation and anti-inflammation. On the one hand, it can directly eliminate water-soluble free radicals, forming a "fat-soluble + water-soluble" antioxidant synergy with astaxanthin. On the other hand, it can inhibit the NF-κB inflammatory pathway, reduce the release of inflammatory factors, delay the process of renal fibrosis, and inhibit renal parenchymal cell apoptosis and pancreatic β-cell inflammatory damage. It also synergistically enhances the target cell repair effect with Polygonatum polysaccharide and Puerarin. Astaxanthin, curcumin, alpha-lipoic acid, vitamin E / C, beta-carotene, and selenium form a comprehensive, multi-target antioxidant network, fundamentally alleviating the core pathological damage caused by oxidative stress in diabetic nephropathy. Simultaneously, these components, along with kudzu root extract, polygonatum polysaccharide extract, four protein peptides, and minerals such as zinc and selenium, form a targeted cell repair system. Astaxanthin specifically protects vascular endothelial cells and glomerular podocytes; curcumin inhibits renal parenchymal cell fibrosis and pancreatic β-cell inflammation; kudzu root extract reduces glomerular podocyte shedding; polygonatum polysaccharide activates pancreatic β-cell proliferation; and small-molecule protein peptides provide easily absorbed amino acid raw materials for the regeneration and repair of all damaged cells. Zinc and selenium act as coenzymes in the synthesis of cell repair-related enzymes, ultimately achieving a quadruple cell repair effect: "antioxidant to prevent damage, active ingredients to promote repair, protein peptides to replenish raw materials, and minerals to provide coenzymes." All components work synergistically without antagonistic effects, and the 800mg dose is a clinically safe and effective dose that will not increase the burden on the liver and kidneys.

[0026] In summary, the composition of this invention differs from traditional single-component interventions. This formulation, through the synergistic effects of multiple targets and pathways, constructs a four-fold synergistic system of "antioxidant-damage inhibition-repair promotion-metabolism regulation," achieving an organic unity of potent antioxidation, targeted cell repair, blood glucose regulation, and kidney protection, providing a new approach for the adjunctive treatment of diabetes and its complications. From a clinical perspective, diabetic nephropathy, as a complex metabolic microvascular complication, faces five major challenges in its treatment: difficulty in blood glucose control, progressive decline in renal function, persistent damage from chronic oxidative stress, irreversible target cell apoptosis, and the coexistence of multiple complications. While traditional drug treatments are effective, they have limitations in terms of long-term safety, overall efficacy, and patient quality of life. The innovation of the composition of this invention lies in its simultaneous targeting of key pathological aspects of diabetic nephropathy: insulin resistance, β-cell dysfunction, oxidative stress, chronic inflammation, renal hyperfiltration, microvascular damage, and target cell apoptosis. The scientific proportions and synergistic relationships of the components in this composition reflect the advanced nature of its design concept: the protein combination provides raw materials for cell repair and active target protection; dietary fiber and probiotics regulate gut-kidney axis metabolism, creating a non-inflammatory and toxin-free in vivo microenvironment for cell repair; plant extracts, as the core active layer targeting cell repair, precisely act on damaged cells in the pancreas, kidneys, and blood vessels; vitamins and minerals are not only important components of the antioxidant system but also coenzyme support layers for cell repair; functional components construct a potent antioxidant system, enhancing metabolic regulation and organ protection. All components in this invention's composition are of natural origin or inherent to the human body and exhibit high safety at recommended dosages. Possible minor adverse reactions include initial gastrointestinal discomfort (bloating, loose stools) and allergic reactions to individual components. Regarding drug interactions, it should be noted that dietary fiber may delay the absorption of certain drugs; it is recommended to take them 2 hours apart. This invention's composition is preferably taken in divided doses between meals, providing nutritional support without causing drastic fluctuations in postprandial blood glucose, and maximizing its postprandial blood glucose regulation and cell repair effects. The design of the composition in this invention represents a shift in the management model of diabetic nephropathy, moving from simple blood glucose control to a combination of antioxidant therapy, cell repair, and overall metabolic regulation; from passive treatment to proactive health management; and from single-target intervention to multi-faceted synergistic protection. This composition is neither a replacement for traditional drug therapy nor a simple nutritional supplement, but rather a multi-target intervention strategy based on systems biology. With the development of precision medicine, it is expected that future biomarker analysis will enable more personalized component adjustments and dosage optimization, bringing greater benefits to patients with diabetes and nephropathy.

[0027] The beneficial effects of the composition of the present invention are as follows: Innovative formulation with significant synergistic effects: This invention is the first to scientifically combine protein peptides, dietary fiber, probiotics and prebiotics, plant extracts, vitamins and minerals, and functional active ingredients to construct a four-fold synergistic system. It targets multiple points to intervene in the core pathological links of diabetic nephropathy, such as oxidative stress, inflammatory response, renal fibrosis, and intestinal microecological imbalance, achieving an organic unity of antioxidation, damage inhibition, repair promotion, and metabolism regulation. At the same time, it also enhances immunity, solving the shortcomings of existing compositions that target only one point. Advanced technology and high bioavailability: Four advanced drug delivery processes are combined to specifically improve the physicochemical properties of each active ingredient, effectively improving the intestinal absorption efficiency, activity retention rate and bioavailability of the ingredients. For example, the survival rate of probiotics in gastric acid is ≥95%, the bioavailability of leech powder is ≥40% higher than that of ordinary powder, and the solubility of fat-soluble components is ≥50% higher. High safety and suitable for long-term intervention: All components of the composition are of natural origin or are substances inherent in the human body. They have been verified by acute toxicity and long-term toxicity tests. At the recommended dose, there are no toxic side effects, and only mild intestinal adaptation reactions may occur. The contraindications and scope of application are clearly defined, and the use is highly safe. With a clear scope of application and strong targeting: the composition is clearly defined in the preparation of products that enhance immunity, provide antioxidant effects, and repair cells, as well as in the prevention and treatment of type 2 diabetes, the treatment of diabetic nephropathy in stages G1-G3, and the maintenance of healthy blood glucose levels. The applicable population and usage methods are precisely defined to avoid misuse and improve the targeting and effectiveness of clinical applications.

[0028] In this invention, if the composition is in powder form, the overall moisture content is preferably ≤5%, and the particle size is preferably 80 mesh to 100 mesh.

[0029] The present invention also provides a method for preparing the above composition, comprising the following steps: Polylactic acid-glycolic acid copolymer, dichloromethane-acetone solution, protein peptides and Tween 80 were mixed to obtain an oil phase; the oil phase was added dropwise to a polyvinyl alcohol aqueous solution with a mass-volume percentage of 4% to 6%, and emulsified by high-speed shearing to obtain an emulsion; the emulsion was poured into a polyvinyl alcohol aqueous solution with a mass-volume percentage of 0.5% to 1.5%, and stirred to obtain a secondary emulsion; the organic solvent in the secondary emulsion was removed, the precipitate was centrifuged, washed, resuspended in an aqueous solution containing a lyophilization protectant, and lyophilized to obtain nanoparticles; Microcapsules were obtained by encapsulating a mixture of probiotics and prebiotics using sodium alginate and chitosan. Medium-chain triglycerides, vitamin E, β-carotene, Omega-3, astaxanthin, curcumin and ascorbyl palmitate were mixed under light-protected conditions to obtain an oil phase; water, Tween 80 and propylene glycol were mixed to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase and stirred, then ultrasonicated and filtered to obtain a nanoemulsion. Water, sodium carboxymethyl cellulose, carbomer 940, leech powder, and glycerin were mixed to obtain an aqueous phase; liquid paraffin and Tween 80 were mixed to obtain an oil phase; the aqueous phase was added dropwise to the oil phase and stirred to emulsify, thus obtaining an emulsion; a CaCl2 aqueous solution of 4%~6% by mass was added dropwise to the emulsion and stirred to solidify; then anhydrous ethanol was added and stirred to break the emulsion, the precipitate was collected, washed, dried, and sieved to obtain microspheres; The kudzu root extract was subjected to high-pressure homogenization at 145 MPa to 155 MPa to obtain the treated kudzu root extract. Nanoparticles, microcapsules, nanoemulsions, microspheres, treated kudzu root extract, and other raw materials are mixed to obtain a composition.

[0030] This invention employs polymer nanoparticle technology, using polylactic-co-glycolic acid copolymer (PLGA), a biodegradable material, as a carrier to encapsulate active ingredients (protein peptides) through an emulsification-solvent evaporation method. These polymer nanoparticles (100~500nm) protect peptides from degradation by gastrointestinal enzymes and enhance absorption through endocytosis by intestinal M cells or epithelial cells. In the emulsification-solvent evaporation method of the present invention, the volume ratio of dichloromethane (DCM) to acetone in the dichloromethane-acetone solution is preferably 1:1; the mass-volume ratio of PLGA, dichloromethane-acetone solution, protein peptides, and Tween 80 is preferably 18g~22g:180mL~220mL:8g~12g:1mL~3mL, more preferably 20g:200mL:10g:2mL; the volume ratio of 4%~6% polyvinyl alcohol aqueous solution to dichloromethane-acetone solution is preferably 350~450:180~220, more preferably 400:200; the rotation speed of the high-speed shear emulsification is preferably 11000rpm. The stirring speed is 13000 rpm, more preferably 12000 rpm, and the time is preferably 4 min to 6 min, more preferably 5 min; the volume ratio of the polyvinyl alcohol aqueous solution (0.5% to 1.5% by mass) to the dichloromethane acetone solution is preferably 350 to 450:180 to 220, more preferably 400:200; the method for removing the organic solvent from the double emulsion is preferably magnetic stirring at 150 rpm to 250 rpm for 4 to 6 h, more preferably magnetic stirring at 200 rpm for 5 h; the freeze-drying protectant is preferably mannitol, and the mass-volume percentage of the freeze-drying protectant in the aqueous solution is preferably 4% to 6%, more preferably 5%.

[0031] This invention utilizes advanced microencapsulation technology to encapsulate high-dose compound probiotics and prebiotics, forming a double-layer protective structure with sodium alginate and chitosan, resulting in a high acid resistance and survival rate of up to 95% for the strains. This technological innovation ensures that a large number of live bacteria can safely pass through the acidic environment of the stomach and reach the intestinal colonization site. The addition of prebiotics further provides a nutrient base for the probiotics, stimulating their growth and metabolic activity. In this invention, the preferred mass ratio of sodium alginate to chitosan is 2-4:1, more preferably 3:1; the preferred concentration of sodium alginate is 1.5%-2.5%, more preferably 2%; and the preferred curing time is 25-35 minutes, more preferably 30 minutes. In this invention, the preferred encapsulation method for preparing the microcapsules is electrostatic dripping, the preferred needle diameter is 0.8 mm, the preferred voltage is 10 kV, and the preferred receiving distance is 15 cm. After treatment with artificial gastric juice for 2 hours, the microcapsules prepared using this method show a probiotic survival rate of ≥95%.

[0032] This invention prepares vitamin E, β-carotene, Omega-3, astaxanthin, and curcumin using a nanoemulsion system. The nanoemulsion particle size is typically between 20 nm and 200 nm, which can significantly improve the dispersibility of fat-soluble components in an aqueous environment, promote bile salt-mediated micelle formation, and increase the absorption rate by 50% to 80%. Astaxanthin and curcumin are fat-soluble active ingredients, and they are prepared in the same pot as the original fat-soluble components, eliminating the need for additional preparation steps. Furthermore, ascorbate palmitate can effectively prevent the oxidative decomposition of astaxanthin and curcumin during the preparation process, ensuring their antioxidant and cell repair activities. In this invention, the preferred weight ratio of the medium-chain triglycerides, vitamin E, β-carotene, Omega-3, astaxanthin, curcumin, and ascorbyl palmitate is 100:5:31.25:37.5:4:4:0.5; the preferred weight ratio of the water, Tween 80, and propylene glycol is 790~795:25~35:8~12, more preferably 792.5:30:10; the preferred dripping rate of the oil phase into the aqueous phase is 4mL / min~6mL / min, more preferably 5mL / min; the preferred ultrasonic power of the ultrasonic treatment is 250W~350W, more preferably 300W, the working mode is 5s on, 3s off, and the total time is preferably 12min~18min, more preferably 14min~16min; the preferred filtration method is a 200-mesh filter.

[0033] This invention prepares leech powder into bioadhesive microspheres, which can prolong the retention time in specific parts of the intestine, thereby ensuring that the leech powder can successfully reach its site of action (proximal small intestine). Animal experiments show that the bioadhesive microspheres can extend the retention time of hirudin in the upper small intestine from the conventional 2-3 hours to 6-8 hours, and increase the absolute bioavailability from 22% to 41%. In this invention, the preferred weight-to-volume ratio of water, sodium carboxymethyl cellulose, carbomer 940, leech powder, and glycerin is 70 mL~90 mL: 13 g~17 g: 6.5 g~8.5 g: 15 g~25 g: 2 mL~4 mL, more preferably 80 mL: 15 g: 7.5 g: 20 g: 3 mL; the preferred volume ratio of liquid paraffin and Tween 80 is 140~160:5, more preferably 150:5; when the aqueous phase is added dropwise to the oil phase, the preferred dropping rate is 1 mL / min; the preferred stirring speed for emulsification is 500 rpm~700 rpm, more preferably 600 rpm, and the preferred time is 25 min~35 min, more preferably 30 min; the preferred weight ratio of CaCl2 to sodium carboxymethyl cellulose is 1:2~4, more preferably 1:3; the preferred particle size of the microspheres is 100 μm~500 μm, more preferably 200 μm~400 μm.

[0034] In this invention, kudzu root extract is treated with high-pressure homogenization technology to increase the bioavailability of its main active ingredient, puerarin, by more than 40%, ensuring the activity of puerarin in targeting and repairing glomerular podocytes and improving microvascular circulation. The preferred pressure for the high-pressure homogenization treatment is 150 MPa, the preferred temperature is 33°C, and the preferred number of high-pressure homogenization treatments is 2. During the high-pressure homogenization treatment, it is preferable to maintain a constant temperature throughout.

[0035] In the preparation method of this invention, after each drug-loaded system is prepared, it is mixed in a specific order of nanoparticles → microcapsules → microspheres → nanoemulsion → treated kudzu root extract → other raw materials. This ensures good compatibility of each component, avoids particle agglomeration and component antagonism, and guarantees the stability and activity of the composition.

[0036] This invention also provides the application of the above-described composition or preparation method in the preparation of products for enhancing immunity, antioxidants, cell repair, or preventing and treating diabetes. Furthermore, it provides the application of the above-described composition or preparation method in the preparation of products for preventing and / or treating diabetes. This invention also provides the application of the above-described composition or preparation method in the preparation of products for preventing and / or treating diabetic nephropathy. In this invention, the diabetic nephropathy is preferably diabetic nephropathy stages G1-G3 (according to the Kidney Disease Outcome Quality Initiative (KDIGO) chronic kidney disease staging criteria, stage G1 eGFR ≥ 90 mL / min / 1.73 m 2 , G2 stage eGFR 60~89mL / min / 1.73m2 , G3 stage eGFR 30~59mL / min / 1.73m 2 All of them had concurrent diabetes as a cause of death. The estimated glomerular filtration rate (eGFR) of the patients with diabetic nephropathy in stages G1-G3 was ≥30 mL / min / 1.73 m³ / min. 2 This invention also provides the application of the above-described composition or preparation method in the preparation of products that help maintain healthy blood sugar levels. In this invention, the type of product preferably includes dietary products or medicines. The effective adult dose of the composition of this invention is 10g / person daily, taken with warm water between meals, 2 hours apart from hypoglycemic drugs and nephropathy medications. The composition of this invention is suitable for patients with type 2 diabetes mellitus and mild kidney damage, patients with diabetic nephropathy in stages G1-G3, excluding patients with end-stage renal disease on dialysis, patients with type 1 diabetic ketoacidosis, those allergic to leeches / seafood, pregnant women, and lactating women.

[0037] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0038] Unless otherwise specified, the following embodiments are all conventional methods.

[0039] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0040] In the following examples, millet protein peptides, soy protein isolate peptides, whey protein peptides, sea buckthorn protein peptides, chia seed powder, konjac powder, Bifidobacterium lactis BB-12, Lactobacillus rhamnosus LGG, fructooligosaccharides, bitter melon peptides, Eucommia ulmoides extract, corn silk extract, mulberry leaf extract, kudzu root extract, Polygonatum sibiricum polysaccharide extract, Moringa oleifera leaf extract, and leech powder were all purchased from Shanxi Nanba Biotechnology Co., Ltd.; calcium, vitamin D3, selenium, zinc, vitamin E, vitamin C, β-carotene, vitamin B1, vitamin B6, and vitamin B1 were also mentioned. 12 Taurine, yeast chromium, α-lipoic acid, Omega-3, stevioside, astaxanthin, and curcumin were all purchased from Shaanxi Baichuan Kangze Biotechnology Co., Ltd.

[0041] The method for determining the safe concentration in zebrafish experiments in the following examples refers to the relevant requirements of the static water method in the national standard GB / T 13267-1991, "Methods for Determining Acute Toxicity of Substances in Water to Freshwater Fish (Zebrafish)". This method is applicable to the toxicity determination of a single chemical substance in water. The results showed that the sample group with a concentration of 2000 μg / mL (the composition obtained in Example 3 of this invention) did not differ significantly from the blank control group; therefore, 2000 μg / mL is the maximum safe concentration.

[0042] Example 1 A composition that helps maintain healthy blood sugar levels, made from protein peptides, dietary fiber, a probiotic and prebiotic blend, an extract blend, a vitamin and mineral blend, and other functional ingredients; The protein peptides consist of 10g of millet protein peptides, 20g of soybean protein peptide isolate, 10g of whey protein peptides, and 10g of sea buckthorn protein peptides. The dietary fiber consists of 10g of chia seed powder and 20g of konjac powder; The probiotic and prebiotic mixture consists of 35 billion CFU of probiotic complex and 15g of fructooligosaccharide. The probiotic complex consists of Bifidobacterium lactis BB-12 and Lactobacillus rhamnosus LGG, with a live bacteria ratio of 2:3. The extract mixture consists of 1000 mg of bitter melon peptide, 1000 mg of Eucommia ulmoides extract, 1000 mg of corn silk extract, 1000 mg of mulberry leaf extract, 200 mg of kudzu root extract, 200 mg of Polygonatum sibiricum polysaccharide extract, 1000 mg of Moringa leaf extract and 200 mg of leech powder. The vitamin and mineral mixture consists of B vitamins, 800 mg of calcium, 600 IU of vitamin D3, 40 μg of selenium, 4 mg of zinc, 70 mg of vitamin E, 70 mg of vitamin C, and 400 mg of beta-carotene. The B vitamins consist of 0.8 mg of vitamin B1, 1.0 mg of vitamin B6, and [missing information - likely a specific vitamin B complex]. 12 Composition: 1.8 μg; The remaining functional components consist of 400 mg taurine, 700 μg yeast chromium, 700 mg α-lipoic acid, 500 mg Omega-3, 400 mg stevioside, 800 mg astaxanthin, and 800 mg curcumin.

[0043] The composition is prepared by: Millet protein peptides, soybean protein peptide isolate, whey protein peptides, and sea buckthorn protein peptides were mixed to obtain protein peptides. The protein peptides were freeze-dried at -50℃ for 24 hours under vacuum to remove moisture, yielding protein peptide powder. Polylactic acid-glycolic acid copolymer (PLGA, LA:GA=50:50, molecular weight 10000Da) was vacuum-dried at 60℃ for 12 hours to remove residual solvent. PLGA was dissolved in a dichloromethane-acetone solution (dichloromethane to acetone volume ratio 1:1) and magnetically stirred at 300 rpm and 25℃ until completely dissolved. Protein peptide powder and Tween 80 were added, and stirring continued for 30 minutes to form a homogeneous oil phase (the oil phase matrix of a W / O type emulsion), wherein the mass-volume ratio of PLGA, dichloromethane-acetone solution, protein peptides, and Tween 80 was 20 g: 200 mL: 10 g: 2 mL. The oil phase was slowly added dropwise to a 5% (w / v) polyvinyl alcohol aqueous solution (PVA aqueous solution), with a volume ratio of 400:200 between the 5% PVA aqueous solution and the dichloromethane-acetone solution. The mixture was emulsified at 12,000 rpm for 5 min to form a W / O emulsion. The emulsion was then slowly poured into a 1% (w / v) PVA aqueous solution, with a volume ratio of 400:200 between the 1% PVA aqueous solution and the dichloromethane-acetone solution. The mixture was magnetically stirred at 500 rpm for 30 min to form a double emulsion (W / O / W type). The double emulsion was magnetically stirred at 200 rpm for 5 h at room temperature to allow complete evaporation of the organic solvent and solidification of the nanoparticles. The nanoparticle suspension was then centrifuged at 10,000 rpm for 20 min, and the supernatant was discarded. The precipitate was washed three times with ultrapure water to remove residual PVA and unencapsulated protein peptides. The precipitate was resuspended in an aqueous solution containing 5% (w / v) mannitol (lyophilization protectant) and lyophilized under vacuum for 24 h to obtain white powdery nanoparticles.

[0044] A mixture of probiotics and prebiotics was encapsulated using sodium alginate solution and chitosan via electrostatic droplet method. The mass ratio of sodium alginate solution to chitosan was 3:1. The concentration of sodium alginate solution was 2%, the needle diameter was 0.8 mm, the voltage was 10 kV, the receiving distance was 15 cm, and the curing time was 30 min. Microcapsules with a particle size of 200 μm to 400 μm and an encapsulation rate of probiotics and prebiotics of ≥90% were obtained.

[0045] Medium-chain triglycerides (MCT) were added to a brown, light-proof beaker, followed by vitamin E, beta-carotene, Omega-3, astaxanthin, curcumin, and ascorbyl palmitate in that weight ratio was 100:5:31.25:37.5:4:4:0.5. The mixture was magnetically stirred at 400 rpm and 25°C for 30 minutes until the beta-carotene, astaxanthin, and curcumin were completely dissolved, forming a transparent oil phase. In another beaker, ultrapure water, Tween 80, and propylene glycol were added in that weight ratio was 792.5:30:10 (in this method, the weight ratio of MCT to water is 100:792.5). The mixture was magnetically stirred at 300 rpm and 25°C for 15 minutes until the emulsifier was completely dissolved, forming an aqueous phase. The oil phase was slowly added dropwise to the aqueous phase at a rate of 5 mL / min, while the mixture was magnetically stirred at 600 rpm. After the addition was complete, stirring was continued for 20 min to form a milky white crude emulsion (O / W type). The crude emulsion was transferred to an ultrasonic cell disruptor, with the probe immersed 1 cm below the liquid surface. The mixture was then subjected to ultrasonic treatment in an ice bath, protected from light. The ultrasonic power was 300 W, and the operating mode was 5 s on / 3 s off, for a total time of 15 min. The ice bath temperature was controlled below 30℃. After ultrasonication, the emulsion was filtered through a 200-mesh filter to remove unemulsified large particles, yielding a nanoemulsion (transferred to a brown bottle and stored protected from light).

[0046] Sodium carboxymethyl cellulose (CMC-Na) and carbomer 940 were added to ultrapure water and magnetically stirred at 300 rpm for 30 min until completely dissolved. Then, leech powder and glycerol were added. The weight-to-volume ratio of ultrapure water, CMC-Na, carbomer 940, leech powder, and glycerol was 80 mL:15 g:7.5 g:20 g:3 mL. The mixture was stirred for another 20 min and then sonicated at 200 W for 5 min to ensure uniform dispersion and prevent particle agglomeration, thus obtaining the aqueous phase. Liquid paraffin was added to Tween 80 at a volume ratio of 150:5 (in this preparation method, the volume ratio of ultrapure water to liquid paraffin is 80:150). The mixture was magnetically stirred at 200 rpm for 10 min to form a homogeneous oil phase, which was then transferred to a three-necked flask. The aqueous phase was slowly added dropwise to the oil phase at a rate of 1 mL / min, and the mixture was mechanically stirred at 600 rpm for 30 min to emulsify, forming a stable W / O emulsion. Prepare a 5% (w / v) CaCl2 aqueous solution (CaCl2 to CMC-Na mass ratio of 1:3), and slowly add it dropwise to the emulsion at a rate of 0.5 mL / min. Continue stirring for 60 min. 2+The carboxyl groups of CMC-Na bind to solidify the aqueous droplets into microspheres. Anhydrous ethanol (1:1 volume ratio to the emulsion) is added to the system, and the mixture is stirred for 10 min to break the emulsion. The mixture is then allowed to stand and separate into layers. The mixture is centrifuged at 3000 rpm for 10 min, and the supernatant is discarded. The precipitate is washed three times with anhydrous ethanol and twice with ultrapure water to remove residual oil phase and uncrosslinked material. The precipitate is spread evenly on a tray and freeze-dried at -50℃ and 10 Pa for 24 h to remove moisture. After drying, the precipitate is passed through 100 μm and 500 μm standard sieves to remove large particles (>500 μm) and fine powder (<100 μm). Microspheres of the appropriate particle size (100 μm~500 μm) with a drug loading of ≥15% are collected and stored in a light-proof, sealed container.

[0047] The kudzu root extract was subjected to high-pressure homogenization at 150 MPa and 30℃ to obtain the treated kudzu root extract. Nanoparticles, microcapsules, nanoemulsions, microspheres, treated kudzu root extract, and other raw materials were added to a three-dimensional motion mixer in the following order: nanoparticles → microcapsules → microspheres → nanoemulsions → treated kudzu root extract → other raw materials. The mixture was then mixed at 150 rpm and 25 ± 2℃ in the dark for 30 minutes to obtain the composition.

[0048] Example 2 A composition that helps maintain healthy blood sugar levels, made from protein peptides, dietary fiber, a probiotic and prebiotic blend, an extract blend, a vitamin and mineral blend, and other functional ingredients; The protein peptides consist of 20g of millet protein peptides, 30g of soybean protein peptide isolate, 20g of whey protein peptides, and 15g of sea buckthorn protein peptides. The dietary fiber consists of 20g of chia seed powder and 50g of konjac powder; The probiotic and prebiotic mixture consists of 50 billion CFU of probiotic complex and 30g of fructooligosaccharide. The probiotic complex consists of Bifidobacterium lactis BB-12 and Lactobacillus rhamnosus LGG, with a live bacteria ratio of 2:3 between Bifidobacterium lactis BB-12 and Lactobacillus rhamnosus LGG. The extract mixture consists of 1800 mg of bitter melon peptide, 1500 mg of Eucommia ulmoides extract, 2000 mg of corn silk extract, 1500 mg of mulberry leaf extract, 1000 mg of kudzu root extract, 1000 mg of Polygonatum sibiricum polysaccharide extract, 1500 mg of Moringa leaf extract and 400 mg of leech powder. The vitamin and mineral mixture consists of B vitamins, 1000 mg of calcium, 800 IU of vitamin D3, 50 μg of selenium, 5 mg of zinc, 80 mg of vitamin E, 80 mg of vitamin C, and 500 mg of β-carotene. The B vitamins consist of 1.2 mg of vitamin B1, 1.5 mg of vitamin B6, and 1 mg of vitamin B2. 12 Composition: 2.4 μg; The remaining functional components consist of 500 mg taurine, 800 μg yeast chromium, 800 mg α-lipoic acid, 600 mg Omega-3, 500 mg stevioside, 800 mg astaxanthin, and 800 mg curcumin.

[0049] The composition is prepared by: Millet protein peptides, soybean protein peptide isolate, whey protein peptides, and sea buckthorn protein peptides were mixed to obtain protein peptides. The protein peptides were freeze-dried at -50℃ for 24 hours under vacuum to remove moisture, yielding protein peptide powder. Polylactic acid-glycolic acid copolymer (PLGA, LA:GA=50:50, molecular weight 10000Da) was vacuum-dried at 60℃ for 12 hours to remove residual solvent. PLGA was dissolved in a dichloromethane-acetone solution (dichloromethane to acetone volume ratio 1:1) and magnetically stirred at 300 rpm and 25℃ until completely dissolved. Protein peptide powder and Tween 80 were added, and stirring continued for 30 minutes to form a homogeneous oil phase (the oil phase matrix of a W / O type emulsion), wherein the mass-volume ratio of PLGA, dichloromethane-acetone solution, protein peptides, and Tween 80 was 22 g:220 mL:12 g:3 mL. The oil phase was slowly added dropwise to a 5% (w / v) polyvinyl alcohol aqueous solution (PVA aqueous solution), with a volume ratio of 5% PVA aqueous solution to dichloromethane-acetone solution of 450:200. The mixture was emulsified at 13000 rpm for 4 min to form a W / O type emulsion. The emulsion was then slowly poured into a 1% (w / v) PVA aqueous solution, with a volume ratio of 1% PVA aqueous solution to dichloromethane-acetone solution of 450:200. The mixture was magnetically stirred at 500 rpm for 30 min to form a double emulsion (W / O / W type). The double emulsion was magnetically stirred at 250 rpm for 4 h at room temperature to allow complete evaporation of the organic solvent and solidification of the nanoparticles. The nanoparticle suspension was then centrifuged at 10000 rpm for 20 min, and the supernatant was discarded. The precipitate was washed three times with ultrapure water to remove residual PVA and unencapsulated protein peptides. The precipitate was resuspended in an aqueous solution containing 6% (w / v) mannitol (lyophilization protectant) and lyophilized under vacuum for 24 h to obtain white powdery nanoparticles.

[0050] A mixture of probiotics and prebiotics was encapsulated using sodium alginate solution and chitosan via electrostatic droplet deposition. The mass ratio of sodium alginate solution to chitosan was 4:1. The concentration of sodium alginate solution was 2.5%, the needle diameter was 0.8 mm, the voltage was 10 kV, the receiving distance was 15 cm, and the curing time was 35 min. Microcapsules with a particle size of 200 μm to 400 μm and an encapsulation rate of probiotics and prebiotics of ≥90% were obtained.

[0051] Medium-chain triglycerides (MCT) were added to a brown, light-proof beaker, followed by vitamin E, beta-carotene, Omega-3, astaxanthin, curcumin, and ascorbyl palmitate in that weight ratio was 100:5:31.25:37.5:4:4:0.5. The mixture was magnetically stirred at 400 rpm and 25°C for 30 minutes until the beta-carotene, astaxanthin, and curcumin were completely dissolved, forming a transparent oil phase. In another beaker, ultrapure water, Tween 80, and propylene glycol were added in that weight ratio was 795:35:12 (in this method, the weight ratio of MCT to water is 100:792.5). The mixture was magnetically stirred at 300 rpm and 25°C for 15 minutes until the emulsifier was completely dissolved, forming an aqueous phase. The oil phase was slowly added dropwise to the aqueous phase at a rate of 6 mL / min, while the mixture was magnetically stirred at 600 rpm. After the addition was complete, stirring was continued for 20 min to form a milky white crude emulsion (O / W type). The crude emulsion was transferred to an ultrasonic cell disruptor, with the probe immersed 1 cm below the liquid surface. The mixture was then subjected to ultrasonic treatment in an ice bath, protected from light. The ultrasonic power was 350 W, and the operating mode was 5 s on / 3 s off, for a total time of 12 min. The ice bath temperature was controlled below 30℃. After ultrasonication, the emulsion was filtered through a 200-mesh filter to remove unemulsified large particles, yielding a nanoemulsion (transferred to a brown bottle and stored protected from light).

[0052] Sodium carboxymethyl cellulose (CMC-Na) and carbomer 940 were added to ultrapure water and magnetically stirred at 300 rpm for 30 min until completely dissolved. Then, leech powder and glycerol were added. The weight-to-volume ratio of ultrapure water, CMC-Na, carbomer 940, leech powder, and glycerol was 90 mL:17 g:8.5 g:25 g:4 mL. The mixture was stirred for another 20 min and then sonicated at 200 W for 5 min to ensure uniform dispersion and prevent particle agglomeration, thus obtaining the aqueous phase. Liquid paraffin was added to Tween 80 at a volume ratio of 160:5 (in this preparation method, the volume ratio of ultrapure water to liquid paraffin is 90:160). The mixture was magnetically stirred at 200 rpm for 10 min to form a homogeneous oil phase, which was then transferred to a three-necked flask. The aqueous phase was slowly added dropwise to the oil phase at a rate of 1 mL / min, and the mixture was mechanically stirred at 700 rpm for 35 min to emulsify, forming a stable W / O emulsion. Prepare a 5% (w / v) CaCl2 aqueous solution (CaCl2 to CMC-Na mass ratio of 1:4), and slowly add it dropwise to the emulsion at a rate of 0.5 mL / min. Continue stirring for 60 min. 2+The carboxyl groups of CMC-Na bind to solidify the aqueous droplets into microspheres. Anhydrous ethanol (1:1 volume ratio to the emulsion) is added to the system, and the mixture is stirred for 10 min to break the emulsion. The mixture is then allowed to stand and separate into layers. The mixture is centrifuged at 3000 rpm for 10 min, and the supernatant is discarded. The precipitate is washed three times with anhydrous ethanol and twice with ultrapure water to remove residual oil phase and uncrosslinked material. The precipitate is spread evenly on a tray and freeze-dried at -50℃ and 10 Pa for 24 h to remove moisture. After drying, the precipitate is passed through 100 μm and 500 μm standard sieves to remove large particles (>500 μm) and fine powder (<100 μm). Microspheres of the appropriate particle size (100 μm~500 μm) are collected and stored in a light-proof, sealed container.

[0053] The kudzu root extract was subjected to high-pressure homogenization at 155 MPa and 33 °C to obtain the treated kudzu root extract. Nanoparticles, microcapsules, nanoemulsions, microspheres, treated kudzu root extract, and other raw materials were added to a three-dimensional motion mixer in the following order: nanoparticles → microcapsules → microspheres → nanoemulsions → treated kudzu root extract → other raw materials. The mixture was then mixed at 150 rpm and 25 ± 2℃ in the dark for 30 minutes to obtain the composition.

[0054] Example 3 A composition that helps maintain healthy blood sugar levels, made from protein peptides, dietary fiber, a probiotic and prebiotic blend, an extract blend, a vitamin and mineral blend, and other functional ingredients; The protein peptides consist of 15g of millet protein peptides, 25g of soybean protein peptide isolate, 15g of whey protein peptides, and 13g of sea buckthorn protein peptides. The dietary fiber consists of 15g of chia seed powder and 35g of konjac powder; The probiotic and prebiotic mixture consists of 40 billion CFU of probiotic complex and 22g of fructooligosaccharide. The probiotic complex consists of Bifidobacterium lactis BB-12 and Lactobacillus rhamnosus LGG, with a live bacteria ratio of 2:3 between Bifidobacterium lactis BB-12 and Lactobacillus rhamnosus LGG. The extract mixture consists of 1500 mg of bitter melon peptide, 1300 mg of Eucommia ulmoides extract, 1500 mg of corn silk extract, 1200 mg of mulberry leaf extract, 600 mg of kudzu root extract, 600 mg of Polygonatum sibiricum polysaccharide extract, 1200 mg of Moringa leaf extract and 300 mg of leech powder. The vitamin and mineral mixture consists of B vitamins, 900 mg of calcium, 700 IU of vitamin D3, 45 μg of selenium, 4 mg of zinc, 75 mg of vitamin E, 75 mg of vitamin C, and 450 mg of β-carotene. The B vitamins consist of 1.0 mg of vitamin B1, 1.2 mg of vitamin B6, and 1.0 mg of vitamin B2. 12 Composition: 2.0 μg; The remaining functional components consist of 450 mg taurine, 750 μg yeast chromium, 750 mg α-lipoic acid, 550 mg Omega-3, 450 mg stevioside, 800 mg astaxanthin, and 800 mg curcumin.

[0055] The preparation method of the composition is the same as in Example 1.

[0056] Experimental Example 1 The composition obtained in Example 3 was used for the zebrafish immune enhancement project - zebrafish neutrophil detection. I. Reference Standards The national standard GB / T 39649-2020, "Quality of Laboratory Animals and Laboratory Fish," specifies the requirements for the rearing environment and feed of zebrafish.

[0057] II. Brief Description of the Method 1. Detection Principle Zebrafish are small tropical freshwater fish native to South Asia. Due to their ease of rearing, high reproductive rate, rapid development, and easily observable transparent fertilized eggs, they have become widely used vertebrate laboratory animals worldwide. As a representative of lower vertebrates, zebrafish share 87% genetic homology with humans and are increasingly used in various fields such as disease model creation, gene function assessment, drug screening, toxicity testing, and environmental health risk assessment. Furthermore, zebrafish experiments have contributed to the research and development of traditional Chinese medicine, natural drugs, health foods, and functional foods. Humans and zebrafish share remarkably similar cellular compositions in their immune systems; the number and fluorescence intensity of neutrophils in zebrafish samples with immunomodulatory effects will show corresponding changes.

[0058] 2. Materials and Methods 2.1 Experimental Instruments and Reagents Stereo fluorescence microscope (SZX7, Olympus, Japan).

[0059] 2.2 Detection system and sample size Experimental system: transgenic fluorescent neutrophil strain of zebrafish.

[0060] Zebrafish age: 3 days after fertilization.

[0061] Sample size per group: 30 samples per well, 1 well per group.

[0062] Adult fish rearing and breeding methods: Follow the standard rearing and breeding methods in this field.

[0063] 3. Experimental Procedure The zebrafish were divided into two groups: a blank control group and a sample treatment group. The test substance stock solution was diluted with buffer water to prepare a series of appropriate concentration gradient solutions (200 μg / mL), with a dilution factor of no less than 10 times, and kept for later use. The blank control group received no treatment, while the sample treatment group received the test sample solution containing the composition obtained in Example 3. After a period of culture, the number of neutrophils or fluorescence intensity in the zebrafish was detected.

[0064] III. Data Processing Methods SPSS 26 software was used for analysis, and GraphPad Prism 8 was used for plotting. Data statistics are expressed as mean ± standard deviation. Experimental results were tested for normality using SPSS software. If the data conformed to normality, the significance was determined by the T-parameter test or analysis of variance (ANOVA). If the data did not conform to normality, nonparametric tests were used (P-value < 0.05 indicates statistical significance).

[0065] IV. Judgment Criteria Based on the fact that the experiment met the validity requirements, compared with the blank control group, the sample treatment group showed a significant increase in the number of neutrophils or fluorescence intensity (P<0.05), indicating that the sample has the effect of enhancing immunity.

[0066] V. Test Results The test results are shown in Tables 1 and 2. Figures 1-3 .

[0067] Table 1 Test Results

[0068] Table 2. Statistical results of fluorescence intensity of zebrafish neutrophils.

[0069] Note: Improvement rate (%) = [(Sample group - Blank group) / Blank group] × 100% The results showed that, in zebrafish experiments, the composition of the present invention had the effect of enhancing immunity, and the fluorescence intensity of zebrafish neutrophils increased by 47.46% compared with the blank group.

[0070] Experimental Example 2 The composition obtained in Example 3 was used to detect the ROS fluorescence intensity in zebrafish for antioxidant purposes. I. Reference Standards Same as the reference standard in Test Example 1.

[0071] II. Brief Description of the Method 1. Detection Principle Oxidative stress is a significant factor contributing to aging. During normal physiological metabolism, substances with oxidative activity, such as reactive oxygen species (ROS), are produced. When the production of ROS exceeds the body's clearance capacity, oxidative stress is triggered. This oxidative stress damages intracellular biomolecules such as lipids, proteins, and DNA. If a test substance can accelerate the synthesis of antioxidants and enhance the activity of antioxidant enzymes, it can rapidly eliminate ROS produced in the body, thereby reducing ROS-induced lipid and protein peroxidation levels and maintaining a healthy balance between ROS production and elimination. The reduction in ROS levels in zebrafish treated with the test substance directly reflects its antioxidant capacity, thus evaluating the sample's antioxidant efficacy.

[0072] 2. Materials and Methods 2.1 Experimental Instruments and Reagents Stereoscopic fluorescence microscope (SZX7, Olympus, Japan), ROS detection kit.

[0073] 2.2 Detection system and sample size Experimental system: wild AB strain zebrafish.

[0074] Zebrafish age: 3 days after fertilization.

[0075] Sample size per group: 30 samples per well, 1 well per group.

[0076] Adult fish rearing and breeding methods: Follow the standard rearing and breeding methods in this field.

[0077] 3. Experimental Procedure The zebrafish were divided into two groups: a blank control group and a sample treatment group. The blank control group received no treatment, while the sample treatment group received the test sample solution (200 μg / mL) of the composition obtained in Example 3. After a period of culture, the zebrafish were stained with ROS and observed and photographed using a fluorescence microscope.

[0078] III. Data Processing Methods SPSS 26 software was used for analysis, and GraphPad Prism 8 was used for plotting. Data statistics are expressed as mean ± standard deviation. Experimental results were tested for normality using SPSS software. If the data conformed to normality, the significance was determined by the T-parameter test or analysis of variance (ANOVA). If the data did not conform to normality, nonparametric tests were used (P-value < 0.05 indicates statistical significance).

[0079] IV. Judgment Criteria Compared with the blank control group, the fluorescence intensity of the sample treatment group was significantly reduced (P<0.05), indicating that the sample has antioxidant effects.

[0080] V. Test Results The results are shown in Tables 3 and 4. Figures 4-6 .

[0081] Table 3 Detection Results

[0082] Table 4. Statistical results of average ROS fluorescence intensity in zebrafish.

[0083] Note: Improvement rate (%) = [(mean of blank group - mean of test group) / mean of blank group] × 100% The results showed that, based on the zebrafish experiment, the composition obtained in Example 3 had antioxidant effects, and the ROS fluorescence intensity increased by 16.39% compared with the blank group.

[0084] Experimental Example 3 The composition obtained in Example 3 was used to detect the regenerated area of ​​zebrafish caudal fin during cell repair and regeneration. I. Reference Standards Same as the reference standard in Test Example 1.

[0085] II. Brief Description of the Method 1. Detection Principle Zebrafish skin, comprising the epidermis, dermis, and subdermal tissue, has a structure highly similar to that of humans. Therefore, zebrafish are used in experiments to test the regenerative efficacy of samples. Mechanical damage causes structural destruction of skin tissue. Zebrafish tail fins have self-regenerative capabilities, and the repair efficacy of test substances can be evaluated by observing the rate of tail fin regeneration.

[0086] 2. Materials and Methods 2.1 Experimental Instruments and Reagents Stereoscopic microscope (SZX7, Olympus, Japan).

[0087] 2.2 Detection system and sample size Experimental system: wild AB strain zebrafish.

[0088] Zebrafish age: five days after fertilization.

[0089] Sample size per group: 58 samples per well, 1 well per group.

[0090] Adult fish rearing and breeding methods: Follow the standard rearing and breeding methods in this field.

[0091] 3. Experimental Procedure The zebrafish were divided into two groups: a model control group and a sample treatment group. The test substance stock solution was diluted with buffer water to prepare a series of suitable concentration gradient solutions (1000 μg / mL), with a dilution factor of no less than 10 times, and kept for later use. In the model control group, tail amputation was performed, while in the sample group, the composition sample obtained in Example 3 was added during tail amputation. After a period of cultivation, the regeneration area of ​​the zebrafish tails was measured.

[0092] III. Data Processing Methods SPSS 26 software was used for analysis, and GraphPad Prism 8 was used for plotting. Data statistics are expressed as mean ± standard deviation. Experimental results were tested for normality using SPSS software. If the data conformed to normality, the significance was determined by the T-parameter test or analysis of variance (ANOVA). If the data did not conform to normality, nonparametric tests were used (P-value < 0.05 indicates statistical significance).

[0093] IV. Judgment Criteria Compared with the model control group, the regeneration area after tail amputation in the sample test group was significantly higher than that in the model control group (P<0.05), indicating that the sample has cell repair and regeneration effects.

[0094] V. Test Results The test results are shown in Tables 5 and 6. Figure 7 and Figure 8 As shown.

[0095] Table 5 Test Results

[0096] Table 6. Statistical Results of Zebrafish Tail Fin Regeneration Area (Pixels)

[0097] Note: Improvement rate (%) = [(Sample group average - Model group average) / Model group average] × 100% The results showed that, in zebrafish experiments, the composition obtained in Example 3 had cell repair and regeneration effects, and the regenerated area of ​​the tail fin increased by 81.54% compared with the model group.

[0098] Test Example 4 The composition obtained in Example 3 was used to test the blood glucose-lowering effect of zebrafish by detecting blood glucose levels. I. Reference Standards Same as the reference standard in Test Example 1.

[0099] II. Brief Description of the Method 1. Detection Principle Zebrafish and humans share certain similarities in glucose metabolism. When zebrafish are exposed to a high-glucose environment, their blood glucose regulation mechanisms face challenges. Excessive glucose intake exceeds their normal glucose uptake and utilization capacity. Normally, cells take up glucose through insulin-mediated signaling pathways and use it for energy production or conversion into glycogen storage. However, under high-glucose conditions, excessive glucose leads to increased insulin secretion, but prolonged high-glucose stimulation may decrease insulin sensitivity, meaning cells respond less to insulin, resulting in reduced glucose uptake. Furthermore, a high-glucose environment may damage pancreatic islet cells, affecting insulin synthesis and secretion, impairing blood glucose regulation, leading to persistently elevated blood glucose levels, and ultimately, a hyperglycemic model.

[0100] 2. Materials and Methods 2.1 Experimental Instruments and Reagents Centrifuge, ultrasonic cell disruptor, Roche Smart Voyager blood glucose meter, glucose.

[0101] 2.2 Detection system and sample size Experimental system: wild AB strain zebrafish Zebrafish age: 1 day after fertilization Sample size per group: 30 samples per well, 1 well per group Adult fish rearing and breeding methods: Follow the standard rearing and breeding methods in this field. 3. Experimental Procedure The zebrafish were divided into three groups: a normal control group, a model control group, and a sample treatment group. The normal control group received no treatment, the model control group was placed in a high-glucose environment, and the sample treatment group received the test sample (2000 μg / mL) while the high-glucose environment was being established. After a period of culture, the zebrafish from all three groups were crushed, centrifuged, and the supernatant was collected. Blood glucose levels were measured, and the data from the three groups were compared and analyzed.

[0102] III. Data Processing Methods SPSS 26 software was used for analysis, and GraphPad Prism 8 was used for plotting. Data statistics are expressed as mean ± standard deviation. Experimental results were tested for normality using SPSS software. If the data conformed to normality, the significance was determined by the T-parameter test or analysis of variance (ANOVA). If the data did not conform to normality, nonparametric tests were used (P-value < 0.05 indicates statistical significance).

[0103] IV. Judgment Criteria Based on the established validity of the experiment, the blood glucose level in the model control group was significantly increased compared to the blank control group (P<0.05), indicating successful modeling. Compared to the model control group, the blood glucose level in the sample treatment group was significantly reduced (P<0.05), indicating that the sample has an auxiliary blood glucose-lowering effect.

[0104] V. Test Results The test results are shown in Tables 7 and 8. Figure 9 This indicates that the composition of the present invention has an auxiliary effect in lowering blood sugar, with blood sugar levels decreasing by 63.49% compared to the model control group.

[0105] Table 7 Test Results

[0106] Table 8. Statistical Results of Blood Glucose Levels (mmol / L) in Zebrafish

[0107] Note: Improvement rate calculation formula = (Average value of model control group - Average value of sample treatment group) / Average value of model control group × 100% The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A composition possessing the combined effects of enhancing immunity, antioxidation, cell repair, blood glucose regulation, and kidney protection, characterized in that, Includes protein peptides, dietary fiber, probiotic and prebiotic mixtures, extract mixtures, vitamin and mineral mixtures, and other functional ingredients; The protein peptides include 10g-20g of millet protein peptides, 20g-30g of soybean protein peptide isolate, 10g-20g of whey protein peptides, and 10g-15g of sea buckthorn protein peptides. The dietary fiber includes 10g-20g of chia seed powder and 20g-50g of konjac powder; The probiotic and prebiotic mixture includes 35 billion to 50 billion CFU of probiotic complex and 15g to 30g of fructooligosaccharides. The probiotic complex is composed of Bifidobacterium lactis BB-12 and Lactobacillus rhamnosus LGG. The extract mixture includes 1000mg~1800mg of bitter melon peptide, 1000mg~1500mg of Eucommia ulmoides extract, 1000mg~2000mg of corn silk extract, 1000mg~1500mg of mulberry leaf extract, 200mg~1000mg of kudzu root extract, 200mg~1000mg of Polygonatum sibiricum polysaccharide extract, 1000mg~1500mg of Moringa oleifera leaf extract, and 200mg~400mg of leech powder; The vitamin and mineral mixture includes B vitamins, calcium 800mg-1000mg, vitamin D3 600IU-800IU, selenium 40μg-50μg, zinc 4mg-5mg, vitamin E 70mg-80mg, vitamin C 70mg-80mg, and β-carotene 400mg-500mg. The B vitamins include vitamin B1 0.8mg-1.2mg, vitamin B6 1.0mg-1.5mg, and vitamin B2. 12 1.8μg~2.4μg; The remaining functional ingredients include taurine 400mg~500mg, yeast chromium 700μg~800μg, α-lipoic acid 700mg~800mg, Omega-3 500mg~600mg, stevioside 400mg~500mg, astaxanthin 700~900mg, and curcumin 700~900mg.

2. The composition according to claim 1, characterized in that, The ratio of viable counts of Bifidobacterium lactis BB-12 and Lactobacillus rhamnosus LGG is 2:

3.

3. A method for preparing the composition according to claim 1 or 2, characterized in that, Includes the following steps: Polylactic acid-glycolic acid copolymer, dichloromethane-acetone solution, protein peptides and Tween 80 were mixed to obtain an oil phase; the oil phase was added dropwise to a polyvinyl alcohol aqueous solution with a mass-volume percentage of 4% to 6%, and emulsified by high-speed shearing to obtain an emulsion; the emulsion was poured into a polyvinyl alcohol aqueous solution with a mass-volume percentage of 0.5% to 1.5%, and stirred to obtain a secondary emulsion; the organic solvent in the secondary emulsion was removed, the precipitate was centrifuged, washed, resuspended in an aqueous solution containing a lyophilization protectant, and lyophilized to obtain nanoparticles; Microcapsules were obtained by encapsulating a mixture of probiotics and prebiotics using sodium alginate and chitosan. Medium-chain triglycerides, vitamin E, β-carotene, Omega-3, astaxanthin, curcumin and ascorbyl palmitate were mixed under light-protected conditions to obtain an oil phase; water, Tween 80 and propylene glycol were mixed to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase and stirred, then ultrasonicated and filtered to obtain a nanoemulsion. Water, sodium carboxymethyl cellulose, carbomer 940, leech powder, and glycerin were mixed to obtain an aqueous phase; liquid paraffin and Tween 80 were mixed to obtain an oil phase; the aqueous phase was added dropwise to the oil phase and stirred to emulsify to obtain an emulsion; a CaCl2 aqueous solution with a mass-volume percentage of 4%~6% was added dropwise to the emulsion and stirred to solidify; then anhydrous ethanol was added and stirred to break the emulsion, the precipitate was collected, washed, dried, and sieved to obtain microspheres; The kudzu root extract was subjected to high-pressure homogenization at 145MPa~155MPa and 30℃~35℃ to obtain the treated kudzu root extract. Nanoparticles, microcapsules, nanoemulsions, microspheres, treated kudzu root extract, and other raw materials are mixed to obtain a composition.

4. The preparation method according to claim 3, characterized in that, The volume ratio of dichloromethane to acetone in the dichloromethane-acetone solution is 1:1; the mass-to-volume ratio of polylactic acid-glycolic acid copolymer, dichloromethane-acetone solution, protein peptide, and Tween 80 is 18g~22g:180mL~220mL:8g~12g:1mL~3mL; the volume ratio of 4%~6% polyvinyl alcohol aqueous solution to dichloromethane-acetone solution is 350~450:180~220; the rotation speed of the high-speed shear emulsification is 11. The stirring speed is 000rpm~13000rpm for 4min~6min; the volume ratio of polyvinyl alcohol aqueous solution (0.5%~1.5%) to dichloromethane acetone solution is 350~450:180~220; the method for removing organic phase solvents from the double emulsion is magnetic stirring at 150rpm~250rpm for 4~6h; the freeze-drying protectant is mannitol, and the mass-volume percentage of the freeze-drying protectant in the aqueous solution is 4%~6%.

5. The preparation method according to claim 3, characterized in that, The mass ratio of sodium alginate to chitosan is 2-4:1; the concentration of sodium alginate is 1.5%-2.5%; and the curing time is 25-35 minutes.

6. The preparation method according to claim 3, characterized in that, The weight ratio of the medium-chain triglycerides, vitamin E, β-carotene, Omega-3, astaxanthin, curcumin, and ascorbyl palmitate is 100:5:31.25:37.5:4:4:0.5; the weight ratio of water, Tween 80, and propylene glycol is 790~795:25~35:8~12; the dripping rate of the oil phase to the aqueous phase is 4mL / min~6mL / min; the ultrasonic power of the ultrasonic treatment is 250W~350W, the working mode is 5s on, 3s off, and the total time is 12min~18min; the filtration is performed using a 200-mesh filter.

7. The preparation method according to claim 3, characterized in that, The weight-to-volume ratio of water, sodium carboxymethyl cellulose, carbomer 940, leech powder, and glycerin is 70-90 mL: 13-17 g: 6.5-8.5 g: 15-25 g: 2 mL-4 mL; the volume ratio of liquid paraffin to Tween 80 is 140-160:5; the dropwise addition rate of the aqueous phase to the oil phase is 1 mL / min; the stirring and emulsification speed is 500-700 rpm, and the time is 25-35 min; the weight ratio of CaCl2 to sodium carboxymethyl cellulose is 1:2-4; and the particle size of the microspheres is 100 μm-500 μm.

8. The use of the composition according to any one of claims 1 to 2 or the preparation method according to any one of claims 3 to 7 in the preparation of products that enhance immunity, antioxidants, cell repair, or prevent and treat diabetes.

9. The use of the composition according to any one of claims 1 to 2 or the preparation method according to any one of claims 3 to 7 in the preparation of products for the prevention and / or treatment of diabetic nephropathy.

10. The use of the composition according to any one of claims 1 to 2 or the preparation method according to any one of claims 3 to 7 in the preparation of products that help maintain healthy blood sugar levels.

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