Synergistic hypoglycemic sargassum fusiforme-inulin compound as well as preparation method, product and application thereof

Abstract

The invention provides a sargassum fusiforme-inulin compound capable of synergistically reducing blood glucose as well as a preparation method, a product and application thereof, and relates to the technical field of functional foods. The sargassum fusiforme-inulin compound is prepared from the following raw materials: a sargassum fusiforme extract, inulin and metabiotics, the invention discloses an animal bifidobacterium subsp. Lactis metaplast, which is characterized in that the metaplast comprises animal bifidobacterium subsp. Lactis BA-C9, and the preservation number of the animal bifidobacterium subsp. Lactis BA-C9 is CCTCC (China Center For Type Culture Collection) NO: M2024457. The compound prepared by the invention has synergistic interaction in the aspect of playing a role in reducing blood sugar, and the effect can be remarkably improved.

Description

Technical Field

[0001] The present invention relates to the technical field of functional foods, and in particular to a synergistically hypoglycemic Sargassum-inulin complex, a preparation method, a product and an application thereof. Background Art

[0002] Diabetes mellitus (DM) is a group of metabolic diseases characterized by chronically elevated blood glucose (glucose) levels, caused by defects in insulin secretion and / or action. This disease can be complicated by chronic damage to multiple organs, including the eyes, kidneys, nerves, and cardiovascular system. Acute metabolic disturbances, such as ketoacidosis and hyperosmolar coma, can occur in severe cases or during periods of stress. For patients with diabetes, particularly type 2 diabetes, postprandial hyperglycemia poses far greater risks than fasting hyperglycemia. Postprandial hyperglycemia is not only highly susceptible to complications, such as cardiovascular, brain, kidney, eye, and nervous system disorders, but can also lead to ketoacidosis and coma, which can be life-threatening in severe cases. Therefore, lowering postprandial blood glucose is a key measure for preventing diabetes, reducing complications, and lowering mortality. α-glucosidase inhibitors are a class of drugs that can lower postprandial hyperglycemia. Numerous clinical studies have demonstrated that α-glucosidase inhibitors are significantly effective in lowering fasting and postprandial blood glucose and glycated hemoglobin, and also significantly improve body weight, insulin levels, cholesterol, triglycerides, and high-density lipoprotein cholesterol in patients with diabetes. The α-glucosidase inhibitors' ability to lower glycated hemoglobin suggests they may be beneficial in preventing diabetic complications.

[0003] Chinese patent CN104784694A discloses bifidobacteria for treating diabetes and related conditions. The patent discloses the use of a combination of (i) bacteria of the genus Bifidobacterium or a mixture thereof and (ii) an antidiabetic drug in the preparation of a food, dietary supplement, or medicine, wherein the food, dietary supplement, or medicine is used to treat one or more of the following diseases and conditions (a) to (g) in mammals: (a) treating diabetes; (b) treating metabolic syndrome; (c) treating glucose intolerance; (d) normalizing insulin sensitivity; (e) improving insulin secretion after eating; (f) reducing fasting insulin secretion; and (g) treating obesity, controlling weight gain, inducing weight loss, reducing body fat mass, and / or reducing mesenteric fat mass. However, the patent only discloses that bacteria of the animal Bifidobacterium lactis subspecies and inulin can treat diabetes in mammals, and does not consider the synergistic effect between the components.

[0004] The article "He Weibin, Gao Liang, Ai Yuting, et al. Research progress on the chemical composition and pharmacological activities of Sargassum fusiformis [J]. Chinese Herbal Medicine, 2023. DOI: 10.7501 / j.issn.0253-2670.2023.07.033" discloses that Jia et al. established a diabetic mouse model on a high-fat diet and found that the Sargassum fusiformis polysaccharide group was better at maintaining blood glucose homeostasis than the metformin group when monitoring glycosylated hemoglobin. Symptoms such as dyslipidemia, oxidative stress, and renal damage in the mice were alleviated, and the polysaccharide promoted glycogen synthesis in the liver and skeletal muscle. Based on this, Wu et al. and Li et al. found that the combination of Sargassum fusiformis polysaccharide and relatively low doses of metformin or acarbose showed good pharmacological effects in controlling fasting blood glucose, improving insulin resistance, and alleviating renal damage in type 2 diabetic mice. In summary, Sargassum fusiforme polysaccharides have demonstrated promising hypoglycemic activity in both in vitro and in vivo studies, acting primarily as α-glucosidase inhibitors and adjunctive hypoglycemic agents, and have the potential to be used in the treatment of type 2 diabetes. However, this literature only discloses that the chemical components of Sargassum fusiforme can treat type 2 diabetes, without considering the synergistic effects of Sargassum fusiforme with other substances.

[0005] In view of this, in order to address the deficiencies of the prior art, the present invention provides a synergistic hypoglycemic Sargassum-inulin complex and a preparation method and application thereof. Summary of the Invention

[0006] The present invention aims to provide a synergistic hypoglycemic Hijiki-inulin complex and its preparation method and application, which overcomes the problem of insufficient hypoglycemic activity of Hijiki extract / Bifidobacterium animalis subsp. lactis BA-C9 and inulin.

[0007] To achieve the above-mentioned object, the present invention is as follows: In a first aspect, the present invention provides a synergistic hypoglycemic sea lettuce-inulin complex, the raw materials of which are composed of sea lettuce extract, inulin and postbiotics; the postbiotics contain animal Bifidobacterium lactis subspecies BA-C9, and the preservation number of animal Bifidobacterium lactis subspecies BA-C9 is CCTCC NO: M2024457.

[0008] Preferably, the mass ratio of the Hijiki extract, inulin and postbiotics is 5-18:5-24:1.

[0009] Further preferably, the mass ratio of the hijiki extract, inulin and postbiotics is 5-10:5-10:1.

[0010] More preferably, the mass ratio of the Hijiki extract, inulin and postbiotics is 5:5:1.

[0011] Preferably, the content of animal Bifidobacterium lactis subspecies BA-C9 in the postbiotics is ≥1.0×1011 pcs / g.

[0012] Preferably, the preparation method of the Sargassum fusiformis extract is: Step (1): crushing the sea cucumber and mixing it with water for extraction, filtering, collecting the filtrate, performing secondary extraction on the filter residue, filtering, and combining and concentrating the filtrates; Step (2): the concentrated solution is centrifuged, ethanol is added to the supernatant, and the extract is concentrated, dialyzed, and freeze-dried to obtain the product.

[0013] Further preferably, the preparation method of the Sargassum fusiformis extract is: Step (1): crushing the sea cucumber and mixing it with water for extraction, filtering, collecting the filtrate, performing secondary extraction on the filter residue, filtering, collecting the filter residue for later use, and combining the filtrates and concentrating them; Step (2): The concentrated solution is centrifuged, and ethanol is added to the supernatant so that the final concentration of ethanol in the extracted solution is 35-45% (v / v). After extraction at 0-5°C for 10-14 hours, the solution is concentrated, and after removing the ethanol, the supernatant is centrifuged and dialyzed with distilled water. The liquid in the dialysis bag is collected and concentrated, and then freeze-dried to obtain crude polysaccharide of Sargassum fusiformis.

[0014] In a second aspect, the present invention provides a method for preparing the aforementioned Sargassum fusiformis-inulin complex, comprising the following steps: mixing raw materials of the Sargassum fusiformis-inulin complex to obtain.

[0015] In a third aspect, the present invention provides a product, which is a food or health product, comprising the aforementioned Sargassum fusiformis-inulin complex and auxiliary materials accepted in the field of food / health products.

[0016] Preferably, the auxiliary materials include but are not limited to flavor enhancers, sweeteners, emulsifiers, leavening agents, stabilizers, thickeners, antioxidants, acidity regulators, preservatives, and colorants.

[0017] Preferably, the flavor enhancer is selected from at least one of 5'-flavor nucleotide disodium, 5'-inosinic acid disodium, L-alanine, glycine, sodium glutamate and disodium succinate.

[0018] Preferably, the sweetener is selected from at least one of D-mannitol, alitame, aspartame, advantame, erythrosine, xylitol and sucralose.

[0019] Preferably, the emulsifier is selected from at least one of D-mannitol sugar, glycerol, modified soybean lecithin, succinic acid monoglyceride, carrageenan, soluble soybean polysaccharide, citric acid fatty acid glyceride and hydroxypropyl starch.

[0020] Preferably, the leavening agent is selected from at least one of D-mannitol sugar, potassium hydrogen tartrate, polydextrose, sodium pyrophosphate, sodium tripolyphosphate, calcium hydrogen phosphate, hydroxypropyl starch and sodium lactate.

[0021] Preferably, the stabilizer is selected from at least one of D-mannitol sugar, α-cyclodextrin, γ-cyclodextrin, xanthan gum, chitin, curdlan, hydroxypropyl starch and sodium lactate.

[0022] Preferably, the thickener is at least one selected from the group consisting of D-mannitol sugar, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, gum arabic, locust bean gum, xanthan gum, methylcellulose, chitin, gellan gum and agar.

[0023] Preferably, the antioxidant is at least one selected from D-isoascorbic acid, tea polyphenols, theaflavins, butylated hydroxyanisole, butylated hydroxytoluene, ascorbic acid, calcium ascorbate, sodium ascorbate and propyl gallate.

[0024] The acidity regulator is selected from at least one of DL-malic acid, dl-tartaric acid, L-malic acid, sodium L-malate, glacial acetic acid, fumaric acid and lactic acid.

[0025] The preservative is selected from at least one of benzoic acid and its sodium salt, parahydroxybenzoates and its sodium salt, and nisin.

[0026] Preferably, the colorant is selected from at least one of erythrosine, lycopene, lycopene, citrus yellow, sorghum red, monascus yellow pigment, red yeast rice, turmeric and caramel color.

[0027] In a fourth aspect, the present invention provides the use of the aforementioned Sargassum fusiformis-inulin complex or the aforementioned product in the preparation of a product for preventing or treating metabolic diseases characterized by hyperglycemia and insulin resistance.

[0028] Preferably, the metabolic disease is diabetes.

[0029] Further preferably, the metabolic disease is borderline diabetes, spontaneous type I diabetes, induced type I diabetes, spontaneous type II diabetes and / or induced type II diabetes.

[0030] More preferably, the metabolic disease is idiopathic type II diabetes and / or induced type II diabetes.

[0031] The beneficial effects of the present invention are: The present invention has found in experiments that the extract of Hijiki, inulin and postbiotics have synergistic effects in lowering blood sugar, protecting pancreatic islet cells, improving blood lipid metabolism and / or enhancing the body's antioxidant function to play a hypoglycemic effect, which can significantly improve the effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The fasting blood glucose graphs of Examples 1-3 and Comparative Examples 1-4 of the present invention after 14 days of gavage and 4 hours of fasting, wherein the model control group is compared with the blank control group. **** p<0.0001; compared with the model group, # p<0.05, ## p<0.01, ### p<0.001, #### p<0.0001. DETAILED DESCRIPTION

[0033] The following non-limiting examples are provided to enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way. The following is merely an illustrative description of the scope of the present invention, and those skilled in the art may make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.

[0034] The present invention will be further described below by way of specific examples. The various chemical reagents used in the examples of the present invention were obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are all by weight. Unless otherwise specified, it is understood that the experiments were conducted at room temperature.

[0035] The preparation method of the Sargassum fusiformis extract is as follows: Weigh 60 g of Sargassum fusiformis powder and dissolve the powder in 3000 mL of distilled water in a beaker. Mix well and stir in a water bath at 90°C for 3 h. After the extract cools to room temperature, filter it through three layers of gauze, collect the filtrate, and then repeat the extraction once with the resulting residue. The residue is set aside.

[0036] The filtrates from the two extractions were combined and concentrated using a rotary evaporator at 60°C. The concentrate was then collected and centrifuged at 10,000 rpm for 5 min.

[0037] Alcohol was added to the supernatant to make the final concentration of the extracted solution 40% (v / v). The solution was concentrated at 4°C overnight. After removing the ethanol, the supernatant was centrifuged and dialyzed with distilled water for 24 h. The liquid in the dialysis bag was collected and concentrated to a certain volume, and then freeze-dried to obtain the Sargassum fusiformis extract.

[0038] Inulin: Manufacturer: Shaanxi Senfu Natural Products Co., Ltd., batch number SFJY240223.

[0039] Postbiotics: Manufacturer: Zhongchuang Yike (Shanghai) Health Technology Co., Ltd., batch number 2025041703, content of Bifidobacterium animalis subspecies lactis BA-C9 is 1.8×1011 pcs / g.

[0040] High-fat feed: Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd.

[0041] Example 1 A synergistic hypoglycemic Sargassum-inulin complex has the following formula: 200 parts of Sargassum fusiformis extract, 200 parts of inulin, and 20 parts of postbiotics.

[0042] Preparation method: Mix the above raw materials evenly to obtain the product.

[0043] Example 2 A synergistic hypoglycemic Sargassum-inulin complex has the following formula: 100 parts of Sargassum fusiformis extract, 100 parts of inulin, and 20 parts of postbiotics.

[0044] Preparation method: Mix the above raw materials evenly to obtain the product.

[0045] Example 3 A synergistic hypoglycemic Sargassum-inulin complex has the following formula: 180 parts of Sargassum fusiformis extract, 240 parts of inulin, and 10 parts of postbiotics.

[0046] Preparation method: Mix the above raw materials evenly to obtain the product.

[0047] Comparative Example 1 A hypoglycemic substance having the following formula: hijiki extract.

[0048] Comparative Example 2 A hypoglycemic substance having the following formula: 350 parts of inulin and 70 parts of postbiotics.

[0049] Comparative Example 3 A synergistic hypoglycemic Sargassum-inulin complex has the following formula: 262.5 parts of Sargassum fusiformis extract, 87.5 parts of inulin, and 70 parts of postbiotics.

[0050] Comparative Example 4 A synergistic hypoglycemic hijiki-inulin complex has the following formula: 175 parts of hijiki extract, 175 parts of inulin, 35 parts of postbiotics, and 35 parts of corn silk powder.

[0051] Preparation method: Mix the above raw materials evenly to obtain the product.

[0052] Test Example 1 Improvement of hyperglycemia model mouse experiments 1. Animals Seventy-two SPF-grade C57BL6 / J male mice, 6-8 weeks old and weighing 18-20 g, were purchased from Shanghai Slack Laboratory Animal Co., Ltd. They were housed in an SPF-grade animal room maintained at room temperature between 20-22°C, humidity between 45-55%, and a 12-hour daylight period. Except for modeling, fasting blood glucose testing, and prior to euthanasia, mice had free access to food and water, and bedding was changed daily.

[0053] 2. Establishment of a mouse hyperglycemia model All mice had free access to food and water and were acclimated for one week. Eight mice were randomly selected from the 72 mice as a blank control group and maintained on a standard diet and distilled water throughout the experiment. The remaining 64 mice were fed a high-fat diet for 12 weeks. Except for the 8 mice in the blank control group, the remaining 64 mice were treated with a fasting blood glucose level ≥10 mmol / L as the model criterion; mice with blood glucose levels <10 mmol / L were excluded. The mice were divided into seven groups of eight mice each based on their blood glucose levels.

[0054] 3. Grouping of experimental animals Based on the fasting blood glucose value, the mice with successful modeling were divided into: model control group, embodiment 1 group, embodiment 2 group, embodiment 3 group, comparative example 1 group, comparative example 2 group, comparative example 3 group, comparative example 4 group, with 8 mice in each group.

[0055] The oral doses are as follows: Example 1 group: daily gavage of Example 1 (420 mg / kg) for 14 days; Example 2 group: daily gavage of Example 2 (420 mg / kg) for 14 days; Example 3 group: daily gavage of Example 3 (420 mg / kg) for 14 days; Comparative Example 1 group: daily gavage of Comparative Example 1 (420 mg / kg) for 14 days; Comparative Example 2 group: daily gavage of comparative example 2 (420 mg / kg) for 14 days; Comparative Example 3 group: daily gavage of comparative example 3 (420 mg / kg) for 14 days; Comparative Example 4 group: daily gavage of Comparative Example 4 (420 mg / kg) for 14 days; Blank control group: gavage with the same volume of blank solvent daily for 14 days; Model control group: The same volume of blank solvent was gavaged daily for 14 days.

[0056] There was no statistically significant difference in fasting blood glucose and body weight between the groups in the model after variance analysis, and the groups were equally comparable (see Table 1).

[0057] Table 1. Body weight and fasting blood glucose values ​​of mice before oral gavage ( ±SD)

[0058] 3.2 Experimental Results (1) Changes in food intake and body weight of mice Each group was gavaged for 14 consecutive days, and the food intake and body weight of the mice were measured every week.

[0059] Table 2. Changes in food intake and body weight of mice ( ±SD)

[0060] (2) Hypoglycemic effect in hyperglycemic model mice On the 14th day after oral administration, mice were fasted for 4 hours and tail blood glucose was measured.

[0061] Table 3. Fasting blood glucose values ​​of mice after oral gavage ( ±SD)

[0062] Note: Compared with the blank control group, **** p<0.0001; Comparison of the embodiment, comparative example and model group, # p<0.05, ## p<0.01, ### p<0.001, #### p<0.0001.

[0063] It can be found that the Sargassum fusiformis-inulin complex prepared by the present invention significantly reduces the fasting blood glucose of mice after oral administration without substantially changing the weight and food intake of the mice, and the three components have significant synergistic synergy under a specific ratio.

[0064] 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, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A synergistic hypoglycemic Sargassum-inulin complex, characterized in that: The raw materials consist of hijiki extract, inulin and postbiotics; the postbiotics contain animal Bifidobacterium lactis subspecies BA-C9, and the preservation number of the animal Bifidobacterium lactis subspecies BA-C9 is CCTCC NO: M2024457.

2. The Sargassum fusiformis-inulin complex according to claim 1, characterized in that The mass ratio of the hijiki extract, inulin and postbiotics is 5-18:5-24:

1.

3. The Sargassum fusiformis-inulin complex according to claim 2, characterized in that The mass ratio of the hijiki extract, inulin and postbiotics is 5-10:5-10:

1.

4. The Sargassum fusiformis-inulin complex according to claim 1, characterized in that The preparation method of the Sargassum fusiformis extract comprises the following steps: step (1): crushing the Sargassum fusiformis and mixing it with water for extraction, filtering, collecting the filtrate, performing secondary extraction on the filter residue, filtering, and combining and concentrating the filtrates; Step (2): the concentrated solution is centrifuged, ethanol is added to the supernatant, and the extract is concentrated, dialyzed, and freeze-dried to obtain the product.

5. The Sargassum fusiformis-inulin complex according to claim 4, characterized in that The preparation method of the Sargassum fusiformis extract is as follows: Step (1): crushing the sea cucumber and mixing it with water for extraction, filtering, collecting the filtrate, performing secondary extraction on the filter residue, filtering, collecting the filter residue for later use, and combining the filtrates and concentrating them; Step (2): The concentrated solution is centrifuged, and ethanol is added to the supernatant so that the final concentration of ethanol in the extracted solution is 35-45% (v / v). After extraction at 0-5°C for 10-14 hours, the concentrated solution is concentrated. After removing the ethanol, the supernatant is centrifuged and dialyzed. The liquid in the dialysis bag is collected and concentrated, and then freeze-dried to obtain the product.

6. The method for preparing the Sargassum fusiformis-inulin complex according to any one of claims 1 to 5, characterized in that: The method comprises the following steps: mixing raw materials of the hijiki-inulin complex to obtain the product.

7. A product, characterized in that The invention is a food or health product, comprising the hijiki-inulin complex according to any one of claims 1 to 5 and auxiliary materials accepted in the field of food or health products.

8. Use of the Sargassum fusiformis-inulin complex according to any one of claims 1 to 5 or the product according to claim 7 in the preparation of a product for preventing or treating metabolic diseases characterized by hyperglycemia and insulin resistance.

9. The use according to claim 8, characterized in that The metabolic disease is diabetes.

10. The use according to claim 9, characterized in that The metabolic disease is borderline diabetes, idiopathic type I diabetes, induced type I diabetes, idiopathic type II diabetes and / or induced type II diabetes.

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