A 4D printing swallowing-friendly nutraceutical gel for simultaneously regulating glycolipid comorbidity and a preparation method thereof

Abstract

This invention discloses a 4D-printed, swallow-friendly, food-grade medicinal gel that simultaneously regulates comorbid glucose and lipid disorders, and its preparation method, belonging to the technical field of food-grade medicinal products. Starting from the complex pathological mechanism of "comorbid glucose and lipid disorders," this invention, based on the theory of "food and medicine homology" and modern systems biology, selects six food-grade medicinal raw materials (Astragalus membranaceus, Pueraria lobata, Morus alba leaf, corn silk, Dioscorea opposita, and Perilla frutescens) and combines them in a specific ratio to exert a synergistic effect of lowering blood sugar and lipids through multiple targets and pathways. Furthermore, it clarifies that its core active ingredients are quercetin, luteolin, and kaempferol, constructing a 4D-printed protective system from raw material combination to active ingredients.

Description

Technical Field

[0001] This invention belongs to the technical field of food and medicine homology products, specifically relating to a 4D-printed swallow-friendly food and medicine homology gel that simultaneously regulates comorbidities of glucose and lipids and its preparation method. Background Technology

[0002] Hyperglycemia and hyperlipidemia are the most common metabolic diseases in clinical practice. They are closely linked pathophysiologically and often coexist. Epidemiological data shows that approximately 70% of patients with type 2 diabetes also have dyslipidemia. This interaction of glucose and lipid metabolism disorders is a major risk factor for serious complications such as atherosclerotic cardiovascular disease (ASCVD), diabetic nephropathy, and retinopathy. Currently, the global prevalence of metabolic diseases continues to rise. According to the International Diabetes Federation (IDF), there were 537 million adults with diabetes worldwide in 2021, and this number is projected to increase to 783 million by 2045, a significant proportion of whom also have dyslipidemia (International Diabetes Federation. IDF Diabetes Atlas, 10th edn. Brussels, Belgium: International Diabetes Federation, 2021.). Therefore, developing safe and effective intervention programs that can simultaneously address glucose and lipid metabolism disorders has significant clinical and social value.

[0003] Currently, the main chemical drugs used clinically to treat hyperglycemia and hyperlipidemia include metformin, sulfonylureas, thiazolidinediones, and other hypoglycemic agents, as well as statins, fibrates, ezetimibe, and other lipid-lowering agents. While these drugs have definite single-target therapeutic effects, they have significant limitations: First, their single target makes it difficult to comprehensively regulate the complex metabolic network—the pathogenesis of hyperglycemia and hyperlipidemia involves multiple aspects such as insulin secretion and sensitivity regulation, lipid metabolism, inflammatory response, and oxidative stress, which single-target drugs cannot achieve synergistic regulation. Second, long-term use can easily lead to side effects such as liver and kidney damage, muscle toxicity, gastrointestinal reactions, and drug tolerance (Lillich FF, Imig JD, Proschak E. Multi-Target Approaches in Metabolic Syndrome. Front Pharmacol. 2020;11:554761.). Third, some patients experience drug intolerance or poor response; for example, the incidence of myalgia and elevated muscle enzymes caused by statins can reach 5%–10%. Therefore, developing intervention programs that are safe for long-term consumption and can synergistically regulate glucose and lipid metabolism through multiple targets has become a research hotspot in this field.

[0004] Substances derived from traditional Chinese medicine theory, often referred to as "food and medicine homology," possess both nutritional and therapeutic functions, with few adverse reactions and suitability for long-term consumption, demonstrating unique advantages in the daily management of metabolic diseases. However, existing metabolic intervention technologies based on food and medicine homology still have several technical shortcomings, specifically as follows: First, existing medicinal and edible compound formulas are mostly small combinations with limited scope of action. Among the published patent literature, medicinal and edible compositions involving hypoglycemia and lipid reduction are mostly simple combinations of 2-5 ingredients, which are insufficient to cover the complex pathological mechanisms of comorbid diabetes and lipid disorders. For example, patent CN113318139A discloses a traditional Chinese medicine composition for lowering blood sugar and lipids composed of chicory and poria cocos. Although it has a certain synergistic effect, it consists of only two ingredients, resulting in a relatively singular target. Patent CN109646579A discloses a composition containing mulberry leaves, bitter melon, and yam, but its focus is primarily on hypoglycemic effects, lacking targeted regulation of lipid metabolism, making it difficult to achieve comprehensive synergistic intervention in glucose and lipid metabolism.

[0005] Second, existing technologies lack a systematic and in-depth explanation of the mechanism of action of the compositions. Many studies on food-medicine homology products only focus on efficacy verification, often using simple animal experiments to confirm their hypoglycemic and lipid-lowering effects. However, they lack systematic research on core scientific issues such as the basis of their active ingredients, key targets, and involved signaling pathways (Besada PM, Abdelaziz AM, Rabie AR, et al. Synergistic impact of polyherbal formulations on metabolic syndrome: a comprehensive review of mechanisms and therapeutic potential. Adv Metab Dis Res. 2025;5(2):112-129). This "black box" research model not only limits the scientific understanding and quality improvement of products but also makes it difficult for related patents to meet inventiveness requirements.

[0006] Third, current technologies lack research on the synergistic regulation of "glycolipid comorbidity." Hyperglycemia and hyperlipidemia interact in their pathological mechanisms: insulin resistance is both a core pathological feature of type 2 diabetes and a significant driver of dyslipidemia; conversely, dyslipidemia can further exacerbate insulin resistance through lipotoxicity. However, most current studies focus only on the single effect of lowering blood glucose or lipids, lacking targeted research on the synergistic regulation of glucose and lipid metabolism; even the few combination therapies involving both have failed to clarify the material basis and molecular mechanism of their synergistic effect.

[0007] Meanwhile, with the development of functional food formulation technology, 3D / 4D food printing technology provides a new approach for the personalized molding of foods for special medical purposes. Dysphagia is a common problem among the elderly and patients with neurological diseases. The International Dietary Standardization Association for Dysphagia (IDDSI) classifies food texture into levels 0-7, and different texture properties are crucial to swallowing safety. Sodium alginate and low-methoxyl pectin can form texture-tunable hydrogels under the cross-linking effect of calcium ions, which has been proven to be able to prepare textured foods that meet IDDSI standards through 3D printing. However, there is currently no functional hydrogel system that combines 4D printing technology with functional extracts of medicinal and edible origin and incorporates pH-responsive anthocyanins as a digestive visualization indicator. Introducing butterfly pea flower anthocyanin (Clitoria ternatea anthocyanin) into a hydrogel system allows for the intuitive visualization of the digestive process by utilizing its reversible color-changing properties under different pH conditions in the oral cavity (purple under alkaline / neutral conditions), stomach (purple-red under acidic conditions), and intestines (blue-green / dark green under alkaline conditions), providing a convenient means for monitoring product digestion and absorption.

[0008] In summary, there is an urgent need to develop a gel composed of food and medicine homologous raw materials that can synergistically regulate hyperglycemia and hyperlipidemia, clarify its active ingredient basis and mechanism of action, so as to solve the technical problems of existing intervention programs such as single target, unclear mechanism, and lack of synergistic regulation research on "glycolipid comorbidity". Summary of the Invention

[0009] Technical Problem Solved: To address the aforementioned technical problems, this invention provides a 4D-printed, swallow-friendly, food-medicine homologous gel for the simultaneous regulation of glucose-lipid comorbidity and its preparation method. Starting from the complex pathological mechanism of "glucose-lipid comorbidity," and based on the theory of "food and medicine homology" and modern systems biology, six food-medicine homologous raw materials (Astragalus membranaceus, Pueraria lobata, Morus alba leaves, corn silk, Dioscorea opposita, and Perilla frutescens) are selected and combined in a specific ratio to exert a synergistic hypoglycemic and lipid-lowering effect through multiple targets and pathways. Furthermore, its core active ingredients are identified as quercetin, luteolin, and kaempferol, constructing a multi-level protection system from raw material combination to active ingredients. This effectively solves the shortcomings of existing methods, such as single target, unclear mechanism, and lack of research on the synergistic regulation of "glucose-lipid comorbidity."

[0010] Technical solution: In a first aspect, the present invention provides a 4D-printed swallowing-friendly food-medicine homologous gel that simultaneously regulates comorbidities of glucose and lipids. The gel comprises sodium alginate, low-methoxyl pectin, butterfly pea pollen, and a composition thereof. The composition is made from the following raw materials in parts by weight: Astragalus membranaceus 40-100 parts, Pueraria lobata 30-80 parts, mulberry leaf 10-40 parts, corn silk 10-40 parts, Dioscorea opposita 5-20 parts, and Perilla frutescens 5-20 parts.

[0011] Preferably, the composition is made from the following raw materials in parts by weight: 80 parts Astragalus membranaceus, 60 parts Pueraria lobata, 20 parts Mulberry leaves, 20 parts Corn silk, 10 parts Dioscorea opposita, and 10 parts Perilla frutescens.

[0012] Among the above-mentioned drugs: Astragalus: As the principal ingredient, astragalus polysaccharides and astragaloside A in astragalus can improve insulin resistance and promote glucose uptake and utilization by activating the AMPK signaling pathway; it also has anti-inflammatory and antioxidant effects, which can reduce vascular endothelial damage. In this invention, astragalus plays a core role in insulin sensitization and vascular protection.

[0013] Kudzu root: As an auxiliary ingredient, puerarin in kudzu root can improve insulin resistance and lower serum total cholesterol and low-density lipoprotein levels; it also has the effects of dilating blood vessels and improving microcirculation. In this invention, kudzu root and astragalus synergistically enhance the insulin-sensitizing effect and exert a lipid-lowering effect.

[0014] Mulberry leaves: As an adjuvant, 1-deoxynojirimycin (DNJ) in mulberry leaves is a potent α-glucosidase inhibitor, which can delay the absorption of carbohydrates in the intestine and effectively control postprandial blood glucose; mulberry leaf flavonoids also have antioxidant and lipid-lowering effects. In this invention, mulberry leaves are mainly responsible for inhibiting intestinal sugar absorption and exerting a rapid hypoglycemic effect.

[0015] Corn silk: As an adjuvant ingredient, the flavonoids and polysaccharides in corn silk have diuretic, anti-edema, hypoglycemic, and lipid-lowering effects; they can promote the excretion of sodium and water, help lower blood pressure, and inhibit fat synthesis. In this invention, corn silk and mulberry leaves synergistically enhance the regulation of glucose metabolism and exert diuretic and lipid-lowering effects.

[0016] Yam: As a medicinal ingredient, yam polysaccharides can regulate the intestinal flora structure and improve glucose and lipid metabolism; it also has the effects of strengthening the spleen and stomach, nourishing yin and replenishing qi, and can moderate the overall medicinal properties, making it suitable for long-term use. In this invention, yam mainly plays the role of harmonizing the medicinal properties and protecting the intestines.

[0017] Perilla: As a medicinal ingredient, perilla contains rosmarinic acid, perillaldehyde, and flavonoids, which have significant anti-inflammatory and antioxidant effects; they can inhibit the release of inflammatory factors and reduce oxidative stress damage. In this invention, perilla is mainly responsible for anti-inflammatory and antioxidant effects, synergistically improving glucose and lipid metabolism from a pathological perspective.

[0018] Furthermore, the active ingredients in the composition include quercetin, luteolin, and kaempferol.

[0019] In a second aspect, the present invention provides a method for preparing a 4D-printed swallowing-friendly food-derived gel that simultaneously regulates comorbidities of glucose and lipids as described in the first aspect, comprising the following steps: S1. Mix and pulverize the astragalus, kudzu root, mulberry leaf, corn silk, yam and perilla according to the weight proportions to obtain the mixture; S2. Add a solvent to the mixture at a ratio of 1g:10-20mL, wherein the solvent is water or 70% ethanol; S3. Ultrasonic extraction at 30–50℃ for 60–100 min yields the extract; S4. Filter the extract and dry the filtrate to obtain the composition; S5. Dissolve sodium alginate, low-methoxyl pectin and butterfly pea pollen in water, add the mixture, and prepare gel ink. S6. Use an extrusion 3D printer to print the gel ink into a shape to obtain the printed product; S7. Immerse the printed product in CaCl2 crosslinking solution for crosslinking and curing to obtain a 4D printed swallow-friendly food-medicine homologous gel.

[0020] Preferably, the material-to-liquid ratio in step S2 is 1g:15mL.

[0021] Preferably, in step S3, extraction is performed at 40°C for 80 min.

[0022] Preferably, the drying process in step S4 is spray drying, vacuum drying, or freeze drying.

[0023] Preferably, in step S5, the mass-volume percentage concentration of sodium alginate is 1.5-2.5%, the mass-volume percentage concentration of low-methoxyl pectin is 0.5-1.5%, the mass-volume percentage concentration of butterfly pea pollen is 0.5-1.0%, and the amount of the composition added is 1-10 mg / mL.

[0024] Preferably, in step S7, the mass-volume percentage concentration of the CaCl2 crosslinking liquid is 0.2-0.8%, and the volume ratio of the printed product to the crosslinking liquid is 1:5-1:20.

[0025] Thirdly, the present invention provides the application of the 4D-printed swallowing-friendly food-medicine homologous gel for simultaneously regulating comorbidities of glucose and lipids as described in the first aspect in the preparation of drugs or functional foods for regulating metabolic disorders.

[0026] Preferably, the regulation of metabolic disorders includes any one or more of the following: (1) Inhibits α-glucosidase activity; (2) Inhibits cholesterol esterase activity; (3) It can exert its antioxidant effect by scavenging free radicals or reducing iron ions; (4) Protect cells from damage induced by glucose and / or oleic acid; (5) Regulate signaling pathways associated with insulin resistance or atherosclerosis, which involve targets such as CASP8, RELA, mTOR, STAT1, or GSK3B.

[0027] Beneficial effects: 1) Synergistic effect on glucose and lipid regulation: The composition of this invention achieves integrated glucose and lipid reduction through multi-target synergy, specifically: α-Glucosidase inhibition: The inhibition rate of the 5 mg / mL water extract was significantly higher than that of the alcohol extract, effectively delaying the rise in postprandial blood glucose; Cholesterol esterase inhibition: The inhibition rates of both the 5 mg / mL water extract and the alcohol extract reached significant levels, reducing intestinal cholesterol absorption; Overall synergistic mechanism: It adopts a multi-linked approach of "intestinal absorption - improvement of insulin resistance - anti-inflammatory and antioxidant", which is superior to single-target drugs and simple formula combinations. 2) Clearly elucidating the mechanism of action: Based on network pharmacology and molecular docking, the core mechanism and scientific basis were clarified, specifically: 59 active compounds and 88 common targets were identified, and 5 core targets were identified: CASP8, RELA, mTOR, STAT1, and GSK3B; it mainly regulates key pathways such as insulin resistance, lipids and atherosclerosis, fluid shear stress and atherosclerosis, and sphingolipid signaling; the core components such as kaempferol have good affinity for binding to the targets, providing structural biological evidence for the mechanism of action; 3) Antioxidant and cell-protective effects: It possesses both antioxidant activity and cell-protective effects, enhancing metabolic stress tolerance, specifically: Antioxidant capacity: ABTS water extract scavenges free radicals (IC). 50 =3.005 mg / mL, DPPH IC 50 =45.54 mg / mL; the ethanol extract has stronger FRAP reducing ability; cell protection: in the high sugar and high fat model, the composition significantly reduced LDH release in a dose-dependent manner, protected cell membrane integrity, and helped delay complications; 4) Excellent biocompatibility: The raw materials are substances that are both medicinal and edible, resulting in high overall safety, specifically: Low cytotoxicity: At concentrations of 1–8 μg / mL, the survival rate of RAW264.7 cells was approximately 100%, with no obvious toxicity observed; suitable for long-term use: high safety and few side effects, suitable as a functional food for long-term conditioning; This invention constructs a full-chain protection system from raw materials to components, with strong anti-circumvention capabilities; the preparation process is advanced and controllable; the 4D printed gel integrates functions, giving the product personalized texture and intelligent digestion regulation. Attached Figure Description

[0028] Figure 1 This is a technical roadmap for the composition of the present invention to regulate blood sugar and blood lipids; Figure 2This is a network diagram of the active compounds and potential targets in the composition of this invention; Figure 3 This is a Venn diagram of the intersection analysis between the target of the composition of the present invention and the target of hyperglycemia and hyperlipidemia. In Figure a), the Venn diagram of the intersection of the potential target of the composition, the target related to hyperglycemia, and the target related to hyperlipidemia shows 88 core targets that are in common intersection. Figure b) is a graph showing the ranking of the Degree values ​​of the core active ingredients in the composition that are most related to the disease targets. Figure 4 This is a network diagram of protein-protein interactions (PPIs) of the composition of the present invention acting on targets related to hyperglycemia and hyperlipidemia; Figure 5 Figure 1 shows the results of GO functional enrichment analysis and KEGG pathway enrichment analysis of the relevant targets of the composition of the present invention. Figure 2 shows the results of biological process (BP) enrichment analysis of GO, Figure 3 shows the results of cellular component (CC) enrichment analysis of GO, Figure 4 shows the results of molecular function (MF) enrichment analysis of GO, Figure 5 shows the bubble diagram of KEGG pathway enrichment analysis, and Figure 6 shows the signaling pathway of fluid shear stress and atherosclerosis. Figure 6 This is a schematic diagram of the molecular docking between the core active ingredient (kaempferol) of the composition of the present invention and the key target (RelA, mTOR); Figure 7 This is a comparison of the in vitro antioxidant activity and enzyme inhibitory activity of the aqueous extract and ethanol extract of the composition of the present invention. In Figure a), the results of the ABTS free radical scavenging activity assay are shown; in Figure b), the results of the DPPH free radical scavenging activity assay are shown; in Figure c), the results of the ferric ion reducing antioxidant capacity (FRAP) assay are shown; and in Figure d), the results of the inhibition rate against α-glucosidase and cholesterol esterase are shown. Figure 8 Figure 1 shows the effect of the composition of the present invention on the survival rate of RAW264.7 cells and the inhibitory effect on LDH release induced by high glucose and high lipid. Figure 2 shows the inhibitory effect of the composition on LDH release induced by glucose and oleic acid, and Figure 3 shows the effect of different concentrations of the composition extract on cell survival rate. Figure 9 This study presents the IDDSI textural characterization and simulated digestion behavior of 4D-printed hydrogels. Figure a) shows the spoon tilting, fork pressing, and fork dripping tests of the hydrogels under different CaCl2 conditions (corresponding to Levels 3-5). Figure b) shows the morphological comparison of the butterfly-shaped 4D-printed hydrogels before and after solidification. Figure c) shows the pH-responsive color changes and structural integrity evolution of anthocyanins during simulated digestion using INFOOGEST 2.0. Detailed Implementation

[0029] The present invention will be described in detail below with reference to specific embodiments. The overall technical route of the composition of the present invention for regulating blood glucose and blood lipids is as follows: Figure 1 As shown.

[0030] Example 1: Preparation of HGSYSZ water extract Weigh out 80 parts of Astragalus membranaceus, 60 parts of Pueraria lobata, 20 parts of mulberry leaves, 20 parts of corn silk, 10 parts of Dioscorea opposita and 10 parts of Perilla frutescens, mix them and grind them to obtain a mixture. Add distilled water at a material-to-liquid ratio of 1g:15mL, place the mixture in an ultrasonic cleaner, and perform ultrasonic-assisted extraction at 40℃ for 80min. The principle of ultrasonic-assisted extraction is to use the cavitation effect, mechanical effect and thermal effect of ultrasound to destroy the plant cell wall, promote solvent penetration and dissolution of active ingredients. Compared with traditional decoction and reflux extraction methods, ultrasonic-assisted extraction has the following advantages: (1) low extraction temperature (30-50℃), which can avoid the destruction of heat-sensitive components; (2) short extraction time (60-100 min), low energy consumption; (3) high extraction efficiency, and significantly improved yield of active ingredients. After extraction, the extract was filtered, and the filtrate was collected. The filtrate was then freeze-dried under vacuum at -50°C for 72 hours to obtain the HGSYSZ water extract powder of the food-medicine homology composition of this invention, which was then sealed and stored for later use.

[0031] Example 2: Preparation of HGSYSZ ethanol extract The preparation method is the same as in Example 1, except that distilled water is replaced with 70% (v / v) ethanol, and HGSYSZ ethanol extract dry powder is finally obtained and sealed for later use.

[0032] Example 3: Network pharmacological analysis of the mechanism of action of the composition Using network pharmacology methods, this study systematically explored the potential mechanisms by which the composition (HGSYSZ) of this invention regulates blood glucose and blood lipids. First, the active chemical components (oral bioavailability OB ≥ 30%, drug-likeness DL ≥ 0.18) and their corresponding potential targets were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). Figure 2 Simultaneously, disease targets related to "hyperglycemia" and "hyperlipidemia" were collected from multiple databases such as GeneCards and DrugBank. The intersection of drug targets and disease targets was used to obtain core targets. A protein-protein interaction (PPI) network was constructed using the STRING database and imported into Cytoscape software for visualization analysis and module mining. Finally, the DAVID database was used to perform Gene Ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses on the core targets.

[0033] The results showed that a total of 59 active compounds and 88 common disease-related targets were identified ( Figure 3 ); PPI network analysis and module mining identified 5 core targets: CASP8, RELA, MTOR, STAT1, and GSK3B ( Figure 4 ); GO enrichment analysis showed that these targets were mainly involved in biological processes such as insulin response, cholesterol metabolic process, and regulation of insulin secretion ( Figure 5 a)-c) in); KEGG pathway analysis showed that the targets were mainly enriched in signaling pathways such as "fluid shear stress and atherosclerosis", "lipids and atherosclerosis", and "insulin resistance" ( Figure 5 d)-e) in).

[0034] To further verify the binding ability of the core active components to the key targets, molecular docking technology was used to perform docking analysis on kaempferol with the core targets RelA and mTOR. The results are as Figure 6 shown: Kaempferol can stably bind to the active pockets of RelA and mTOR proteins through hydrogen bonds and hydrophobic interactions, and the binding free energy is lower than -5.0 kcal / mol, indicating good binding affinity between the two, providing verification at the structural biology level for the mechanism of action predicted by network pharmacology.

[0035] Example 4: Determination of the in vitro antioxidant activity of the combination The aqueous extracts and ethanol extracts prepared in Example 1 and Example 2 were respectively formulated into solutions with a series of concentrations of 1, 2, 5, 8, and 10 mg / mL, and their antioxidant activities were evaluated by ABTS method, DPPH method, and FRAP method.

[0036] Determination of ABTS radical scavenging activity: Operate according to the microplate method of the total antioxidant capacity (T-AOC) detection kit (ABTS method) (Nanjing Jiancheng Bioengineering Institute, product number A015-2-1). The results are as Figure 7 shown in a) below. The half-maximal inhibitory concentration (IC 50 ) of the aqueous extract was 3.005 mg / mL, and the IC 50 of the ethanol extract was 7.772 mg / mL. The results showed that the ability of the aqueous extract to scavenge ABTS radicals was significantly stronger than that of the ethanol extract.

[0037] Determination of DPPH radical scavenging activity: Operate according to the DPPH radical scavenging ability kit (Nanjing Jiancheng Bioengineering Institute, product number A153-1-1). The results are as Figure 7 shown in b) below. The IC 50 of the aqueous extract was 45.54 mg / mL, and the IC 50The concentration was 55.09 mg / mL. The results indicate that both extracts possess DPPH free radical scavenging ability, with the water extract showing slightly better performance.

[0038] Ferrous Reduction Antioxidant Capacity (FRAP) Determination: The procedure was performed according to the instructions of the Total Antioxidant Capacity (T-AOC) Assay Kit (FRAP Method) microplate method (Nanjing Jiancheng Bioengineering Institute, catalog number A015-3-1). Results are as follows: Figure 7 As shown in c), the ethanol extract exhibits a significantly higher iron-reducing capacity than the water extract, with absorbance values ​​increasing significantly with increasing concentration, showing a clear dose-dependent effect. This indicates that the ethanol extract is rich in highly reducing, fat-soluble components such as polyphenols and flavonoids.

[0039] Example 5: Determination of the in vitro enzyme inhibitory activity of the composition The aqueous and ethanol extracts prepared in Examples 1 and 2 were prepared into 5 mg / mL solutions, and their inhibitory activities on α-glucosidase and cholesterol esterase were determined, respectively.

[0040] α-Glucosidase inhibitory activity assay: The procedure was performed according to the instructions for the α-glucosidase (α-Glu) kit (Nanjing Jiancheng Bioengineering Institute, catalog number H419-1-1). Results are as follows: Figure 7 As shown in d), at a concentration of 5 mg / mL, both the water extract and the ethanol extract exhibited significant inhibitory effects, and their inhibition rates were comparable. This indicates that both the water-soluble and alcohol-soluble components in the composition of this invention can effectively delay the digestion and absorption of carbohydrates, thus helping to control postprandial blood glucose.

[0041] Cholesterol esterase inhibitory activity assay: The method described in the literature (Zhao S, Wu Y, Hu L. Identification and synthesis of selective cholesterol esterase inhibitor using dynamic combinatorial chemistry. Bioorganic Chemistry, 2022; 119: 105520.) was adapted accordingly. The results are as follows: Figure 7 As shown in d), at a concentration of 5 mg / mL, the water extract exhibited significantly higher inhibitory activity than the ethanol extract. The results are as follows... Figure 7 As shown in d), at a concentration of 5 mg / mL, the water extract exhibited significantly higher inhibitory activity than the ethanol extract. This suggests that the water-soluble components in the water extract may play a more significant role in inhibiting intestinal cholesterol absorption and regulating blood lipid levels.

[0042] Example 6: Cellular safety evaluation of the composition (MTT assay) RAW264.7 macrophages were used at a rate of 5 × 10⁻⁶. 4 Cells were seeded at a density of 100 cells / well in 96-well plates. After cell attachment, HGSYSZ aqueous extract was added at final concentrations of 1, 2, 4, and 8 μg / mL, respectively. MTT assay was performed after 2 hours of incubation. Results are shown below. Figure 8 As shown in b), within the concentration range of 1–8 μg / mL, the cell viability of each group remained at approximately 100%, and no obvious cytotoxicity was observed, indicating that the composition of the present invention has good biosafety.

[0043] Example 7: Evaluation of the cell protective effect of the composition (LDH release assay) RAW264.7 cells were loaded at a rate of 2 × 10⁻⁶. 5 Cells were seeded at a density of 100 cells / well in 24-well plates, with control, model, low-dose HGSYSZ (2 μg / mL), and high-dose HGSYSZ (4 μg / mL) groups. Except for the control group, all other groups were co-treated with glucose (final concentration 30 mmol / L) and oleic acid (final concentration 0.5 mmol / L) to establish a high-glucose, high-lipidemia cell injury model. Cell culture supernatants were collected, and LDH activity was detected using an LDH release assay kit.

[0044] The results are as follows Figure 8 As shown in Figure a), compared with the control group, the LDH release in the model group was significantly increased; compared with the model group, the LDH release in both the low-dose and high-dose HGSYSZ groups was significantly decreased in a dose-dependent manner. These results indicate that the composition of the present invention can effectively protect the integrity of the cell membrane and alleviate cell damage caused by high-glucose and high-lipid metabolic stress.

[0045] Example 8: Preparation of 4D Printed Hydrogels Sodium alginate and low-methoxyl pectin were weighed and dissolved in distilled water, and stirred in a water bath at 60°C until completely dissolved. The final concentration of sodium alginate was 2.0% w / v, and the final concentration of low-methoxyl pectin was 1.0% w / v. HGSYSZ water extract powder (final concentration 5 mg / mL) prepared in Example 1 and butterfly pea pollen (final concentration 1% w / v) were added, and the mixture was thoroughly mixed and degassed to obtain the 4D printing hydrogel ink, which was stored at 4°C for later use.

[0046] The aforementioned 4D printing hydrogel ink was loaded into an extrusion 3D printer (needle orifice diameter 0.8 mm, printing speed 20 mm / s, air pressure 0.2 MPa) and printed into a preset shape (butterfly shape, 5 layers stacked). Figure 9(b)). After printing, the products were immersed in 0.2%, 0.5%, and 0.8% w / v CaCl2 solutions for cross-linking and curing for 10 min, respectively. After removal, they were rinsed with distilled water to obtain 4D printing hydrogels with different textures. 0.2% CaCl2 concentration corresponds to IDDSI Level 3, 0.5% corresponds to Level 4, and 0.8% corresponds to Level 5. Figure 9 (a)

[0047] INFORGEEST simulation of digestion and observation of color changes in 4D-printed hydrogels: Following the INFORGEEST 2.0 standard method, the 4D-printed hydrogels were sequentially treated in the oral phase (pH 7, 2 min), gastric phase (pH 3, 2 h), and intestinal phase (pH 7, 2 h). Color changes and structural integrity of the hydrogels were recorded after each stage. Results are as follows: Figure 9 As shown in (c): the hydrogel changes from blue-purple to blue-green during the oral phase; it returns to purple during the gastric phase while maintaining its structural integrity; and it turns dark green during the intestinal phase, undergoing progressive disintegration within 60 min until the structure completely collapses, indicating that the structure-dependent digestive behavior and the pH-responsive color change function of anthocyanins are realized simultaneously.

[0048] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A 4D-printed, swallow-friendly, food-grade gel that simultaneously regulates comorbidities of glucose and lipids, characterized in that: The gel comprises sodium alginate, low-methoxyl pectin, and butterfly pea pollen, as well as a composition made from the following raw materials in parts by weight: 40-100 parts of Astragalus membranaceus, 30-80 parts of Pueraria lobata, 10-40 parts of mulberry leaves, 10-40 parts of corn silk, 5-20 parts of Dioscorea opposita, and 5-20 parts of Perilla frutescens.

2. The 4D-printed swallowing-friendly food-grade gel for simultaneously regulating comorbid glucose and lipid disorders according to claim 1, characterized in that: The composition is made from the following raw materials in parts by weight: 80 parts Astragalus membranaceus, 60 parts Pueraria lobata, 20 parts Mulberry leaves, 20 parts Corn silk, 10 parts Dioscorea opposita, and 10 parts Perilla frutescens.

3. The 4D-printed swallowing-friendly food-grade gel for simultaneously regulating comorbidities of glucose and lipids according to claim 1, characterized in that: The active ingredients in the composition include quercetin, luteolin, and kaempferol.

4. A method for preparing a 4D-printed swallowing-friendly food-medicine homologous gel for simultaneously regulating comorbidities of glucose and lipids, as described in any one of claims 1-3, characterized in that, Includes the following steps: S1. Mix and pulverize the astragalus, kudzu root, mulberry leaf, corn silk, yam and perilla according to the weight proportions to obtain the mixture; S2. Add a solvent to the mixture at a ratio of 1g:10-20mL, wherein the solvent is water or 70% ethanol; S3. Ultrasonic extraction at 30–50℃ for 60–100 min yields the extract; S4. Filter the extract and dry the filtrate to obtain the composition; S5. Dissolve sodium alginate, low-methoxyl pectin and butterfly pea pollen in water, add the mixture, and prepare gel ink. S6. Use an extrusion 3D printer to print the gel ink into a shape to obtain the printed product; S7. Immerse the printed product in CaCl2 crosslinking solution for crosslinking and curing to obtain a 4D printed swallow-friendly food-medicine homologous gel.

5. The preparation method according to claim 4, characterized in that: In step S2, the material-to-liquid ratio is 1g:15mL; in step S3, extraction is performed at 40℃ for 80 min; and in step S4, the drying is performed by spray drying, vacuum drying, or freeze drying.

6. The preparation method according to claim 4, characterized in that: In step S5, the mass-volume percentage concentration of sodium alginate is 1.5–2.5%, the mass-volume percentage concentration of low-methoxyl pectin is 0.5–1.5%, the mass-volume percentage concentration of butterfly pea pollen is 0.5–1.0%, and the amount of the composition added is 1–10 mg / mL.

7. The preparation method according to claim 4, characterized in that: In step S7, the mass-volume percentage concentration of the CaCl2 crosslinking solution is 0.2-0.8%, and the volume ratio of the printed product to the crosslinking solution is 1:5-1:

20.

8. The use of the 4D-printed swallowing-friendly food-grade gel for simultaneously regulating comorbidities of glucose and lipids as described in any one of claims 1-3 in the preparation of drugs or functional foods for regulating metabolic disorders.

9. The application according to claim 8, characterized in that, The metabolic disorders mentioned include any one or more of the following: (1) Inhibits α-glucosidase activity; (2) Inhibits cholesterol esterase activity; (3) It can exert its antioxidant effect by scavenging free radicals or reducing iron ions; (4) Protect cells from damage induced by glucose and / or oleic acid; (5) Regulate signaling pathways associated with insulin resistance or atherosclerosis, which involve targets such as CASP8, RELA, mTOR, STAT1, or GSK3B.

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