Application of medium-chain fatty acid in promoting fructose absorption and glycogen supplement

Abstract

The invention discloses application of medium-chain fatty acid in promoting fructose absorption and glycogen supplement. Expression of fructose transporter GLUT5 in intestinal cells is up-regulated through octanoic acid, absorption of fructose and supplement of hepatic glycogen are promoted, a scientific basis is provided for basic research of relieving or preventing gastrointestinal discomfort, malnutrition and rapid energy supplement caused by fructose absorption and product development, and the pharmaceutical composition has great application value.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of medium-chain fatty acids in promoting fructose absorption and glycogen replenishment. Background Technology

[0002] Energy is essential for maintaining life. The main sources of energy for the human body are carbohydrates, fats, and proteins from food. Fructose is a major monosaccharide among carbohydrate energy substances, widely found in foods such as fruits and honey, where it combines with glucose to form sucrose, which we consume daily. After ingestion, fructose is transported and absorbed in the small intestine via the fructose transporter GLUT5. Therefore, the expression level of GLUT5 determines the absorption rate and bioavailability of fructose. As an energy supplement, fructose is widely used for rapid energy replenishment and physical recovery in various situations (such as after exercise or during periods of hunger). However, in some individuals, low GLUT5 expression or impaired function in the small intestine hinders fructose absorption, causing gastrointestinal discomfort and potentially leading to malnutrition in the long term. Some individuals may even experience a range of gastrointestinal symptoms, clinically termed fructose malabsorption or fructose intolerance. Given the important role of GLUT5 in fructose absorption, energy replenishment, and gastrointestinal health, it is of great significance to develop bioactive substances that can upregulate GLUT5 expression and thus promote fructose absorption and energy replenishment.

[0003] Octanoic acid (OA) is an 8-carbon medium-chain fatty acid. Compared with long-chain fatty acids, medium-chain fatty acids are digested and absorbed more quickly, and can be directly transported to the liver via portal vein blood flow for rapid metabolism and energy production, making it a rapid energy source among fat-based energy substances. Octanoic acid may inhibit hepatic fat deposition, improve insulin sensitivity, reduce inflammation, and improve metabolic disorders, thereby intervening in metabolic syndrome (MS) in mice (Chen Weimin, Cao Hui, Chen Hongbiao, et al. Interventional effect of octanoic acid on metabolic syndrome in mice [J]. Journal of Mathematical Medicine, 2025, 38(2):98-109.). In addition, medium-chain fatty acids also have functions such as regulating metabolism, improving gut microbiota, and enhancing cognition.

[0004] There are currently no research reports on the upregulation of GULT5 expression and the promotion of fructose absorption by medium-chain fatty acids such as caprylic acid. Summary of the Invention

[0005] To better promote fructose absorption, meet the body's need for rapid energy replenishment and physical recovery, and fully leverage the benefits of medium-chain fatty acids, this invention provides the following technical solutions.

[0006] In a first aspect, the present invention provides the use of medium-chain fatty acids in the preparation of products that promote fructose absorption and / or glycogen replenishment.

[0007] Preferably, the medium-chain fatty acid is selected from at least one of caprylic acid, capric acid, or lauric acid, and more preferably caprylic acid.

[0008] Preferably, the application includes at least one of the following: (1) Application in the preparation of products that promote the absorption of fructose by intestinal cells; (2) Application in the preparation of products that upregulate the expression level of fructose transporter protein in intestinal tissue; (3) Application in the preparation of products that upregulate the expression level of fructokinase in intestinal tissue; (4) Application in the preparation of products that restore the body’s liver glycogen levels.

[0009] Preferably, the liver glycogen level is the liver glycogen level under starvation conditions.

[0010] Preferably, the concentration of the medium-chain fatty acid used is 100~500 μM, for example: 100 μM, 150 μM, 200 μM, 250 μM, 300 μM, 350 μM, 400 μM, 450 μM, 500 μM.

[0011] Furthermore, the concentration of the medium-chain fatty acid used is 100~200 μM.

[0012] Preferably, the product is a pharmaceutical, feed, health food, emergency food, or sports food.

[0013] Preferably, the product also includes excipients permitted to be added to pharmaceuticals, feed, health foods, emergency foods, or sports foods.

[0014] Preferably, the drug is an oral preparation or an injection.

[0015] Furthermore, the oral preparation is an energy gel, powder, granule, capsule, powder, pill, tablet or oral liquid.

[0016] In a second aspect, the present invention provides a medicament for preventing and / or treating fructose malabsorption, wherein the active ingredient of the medicament includes caprylic acid.

[0017] Preferably, the caprylic acid is the sole active ingredient.

[0018] Furthermore, the drug also includes pharmaceutically or physiologically acceptable excipients, which include one or more of the following: diluents, binders, wetting agents, lubricants, disintegrants, solvents, emulsifiers, cosolvents, preservatives, pH adjusters, osmotic pressure adjusters, surfactants, coating materials, antioxidants, or buffers.

[0019] Furthermore, the drug can be administered via at least one of the following routes: intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, or sublingual administration.

[0020] Preferably, the drug further includes a pharmaceutically acceptable carrier, and more particularly, the carrier is a solid or a liquid.

[0021] Furthermore, the solid carrier can be one or more substances that also function as a diluent, flavoring agent, solubilizer, lubricant, suspending agent, binder, preservative, tablet disintegrant, or encapsulating material.

[0022] Furthermore, the carrier includes, but is not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth gum, methylcellulose, sodium carboxymethyl cellulose, low-melting-point waxes, and cocoa butter.

[0023] Preferably, the drug is a solid or liquid formulation.

[0024] Furthermore, the solid dosage form includes powders, granules, capsules, powders, pills, and tablets.

[0025] Furthermore, the liquid formulation includes oral liquids and injections, wherein the oral liquid is preferably a solution, suspension, or emulsion.

[0026] Thirdly, the present invention provides a method for preventing and / or treating fructose malabsorption, the method comprising the step of administering the drug described in the second aspect to a patient.

[0027] The beneficial effects of this invention are: This invention promotes fructose absorption and liver glycogen replenishment by upregulating the expression of the fructose transporter GLUT5 in intestinal cells with caprylic acid. It provides a scientific basis for basic research and product development to alleviate or prevent gastrointestinal discomfort and malnutrition caused by fructose malabsorption and to provide rapid energy replenishment. It has significant application value. Attached Figure Description

[0028] Figure 1 The results show the effect of caprylic acid on the expression of GLUT5 protein in intestinal cells; Figure 2 The results show the effect of caprylic acid on fructose absorption by intestinal cells; Figure 3The results show the effects of caprylic acid on the expression of GLUT5 protein and its fructokinase (KHK) in intestinal tissue; Figure 4 The results show the effect of caprylic acid on the recovery level of liver glycogen after fructose supplementation under starvation conditions. Detailed Implementation

[0029] The technical solution of the present invention will be further described below with reference to embodiments and accompanying drawings. The advantages and features of the present invention will become clearer as the description unfolds. However, it should be understood that the embodiments are merely exemplary and do not constitute a limitation on the scope of the present invention.

[0030] It should be noted that, unless otherwise specified, the experimental methods used in the following embodiments are conventional methods in the art. Unless otherwise defined, all scientific and technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art.

[0031] Example 1: Effects of caprylic acid on fructose uptake and GLUT5 protein expression in intestinal cells 1.1 Cell Culture: IEC-18 cells were seeded into custom DMEM medium containing 10% fetal bovine serum (FBS, purchased from Biological Industries; DMEM medium, purchased from Wuhan Pronosai Co., Ltd.). The glucose-to-fructose ratio in the medium was 1:1. The cells were cultured at 37°C with 5% CO2. When the confluence reached 80-90%, the cells were passaged or plated proportionally. After plated cells reached approximately 30-40% confluence, IEC-18 cells were treated with different concentrations of OA (100 μM, 150 μM, 200 μM) for 24 hours. Protein was collected and sonicated 4-5 times. After balancing the centrifuge tubes, the cells were centrifuged at 15000 rpm for 30 min at 4°C, and the supernatant was collected. Protein concentration was determined using a BCA protein assay kit. The samples were adjusted to the same protein concentration according to the standard curve, and loading buffer was added proportionally. The mixture was thoroughly mixed and subjected to denaturation in a boiling water bath for 5 min. After centrifugation, the cells were stored at -80°C for later use.

[0032] 1.2 Western Blotting: Prepare stacking and separating gels, as well as running and transfer buffers, according to the specified ratios. Load samples according to their protein concentrations. After loading, begin gel transfer. After transfer, block with 5% skim milk for 1 h. After blocking, wash three times with 1×TBST and incubate with primary antibody overnight at 4°C. After incubation, wash the membrane three times with 1×TBST buffer for 8 min each time. Then add working solution of secondary antibody from the appropriate species (rabbit or mouse) (secondary antibody dilution ratio 1:5000, diluent is 1×TBST, and 1% skim milk is added), and incubate with secondary antibody at room temperature for 1 h; then wash three times with 1×TBST buffer for 8 min each time to remove unbound secondary antibody. Prepare ECL chemiluminescence solution according to the ratio of solution A: solution B = 1:1 for development. In a dark room, ECL luminescent solution is evenly spread on the target band. The target band emits fluorescence through a chemical reaction. X-ray film is then used for exposure, development, and fixing. The X-ray film shows bands of varying shades of color, and the intensity of the color indicates the level of GLUT5 protein expression in the sample.

[0033] 1.3 Determination of fructose uptake: The effects of different concentrations of OA (100 μM, 150 μM, 200 μM) on fructose uptake in IEC-18 cells were determined by 1-NBDF fluorescent probe method and flow cytometry, respectively.

[0034] like Figure 1 As shown, compared with the control group, the OA treatment groups of 100 μM, 150 μM and 200 μM (P<0.01) can all upregulate the expression of fructose transporter GLUT5 in IEC-18 cells, with the effect of 150 μM reaching a significant level (P<0.05) and 200 μM reaching a significant level (P<0.01). Compared to the control group, OA treatments at 100 μM (P<0.01), 150 μM (P<0.001), and 200 μM (P<0.0001) significantly upregulated the expression of the related metabolic enzyme KHK, with P values ​​of 100 μM <0.01, 150 μM <0.001, and 200 μM <0.0001. The expression levels of both proteins were highest at an OA concentration of 200 μM.

[0035] like Figure 2 As shown, flow cytometry analysis of fructose uptake in IEC-18 cells showed that, compared with the control group, the fructose uptake rates of the 100 μM, 150 μM, and 200 μM OA treatment groups increased by 13.81% (P<0.01), 19.28% (P<0.001), and 22.65% (P<0.001), respectively.

[0036] The results above indicate that caprylic acid can effectively upregulate the expression of GLUT5 and KHK proteins in intestinal cells and promote fructose absorption in intestinal cells.

[0037] Example 2: Effects of caprylic acid on the expression of GLUT5 protein and its related metabolic enzyme KHK in intestinal tissue Tricaprylic acid glyceride is a precursor of caprylic acid and is the form in which caprylic acid exists in food. In humans and animals, tricaprylic acid glyceride is rapidly metabolized into caprylic acid (Nagao KY, T. Bioactive lipids in metabolic syndrome [J]. Prog Lipid Res, 2008, 47(2): 127-46.). ] Therefore, the main active ingredient that ultimately exerts its effect in the body is caprylic acid. Caprylic acid has a certain degree of volatility and corrosiveness, as well as a slightly unpleasant odor. Tricaprylic acid glyceride is often used in animal experiments, and this esterified form is more consistent with the properties of its food source (Watanabe ST, S. Applications of Medium-Chain Triglycerides in Foods [J]. Front Nutr, 2022, 9(5): 802-805.). Therefore, this example uses tricaprylic acid glyceride for the experiment.

[0038] 2.1 Grouping of experimental animals: Forty male C57BL / 6J mice, aged 8-10 weeks, were randomly divided into 5 groups of 8 mice each: normal diet control group (NC), control group (Con), tricaprylic acid glyceride group (Tri), fructose group (Fru), and "tricaprylic acid glyceride + fructose" group (Tri+Fru).

[0039] 2.2 Experimental Methods: Mice were administered the food via gavage. After a week of acclimatization, gavage began and continued for one day. Except for the normal diet control group (NC), which did not require fasting, the other four groups fasted for 12 hours before the end of the experiment. Fasting began at 1:30 AM. At 9:30 AM, each mouse in the Tri group and the “Tri+Fru” group was administered 70 μL of tricaprylic acid glyceride via gavage. At 1:30 PM, each mouse in the Fru group and the “Tri+Fru” group was administered 1 g / kg of fructose solution via gavage, while the control group (Con) was administered an equal volume of physiological saline. Two hours later, at 3:30 PM, mice were euthanized and samples were collected. Liver and intestinal tissues were collected, placed in liquid nitrogen, and then stored at -80°C for analysis.

[0040] 2.3 The expression levels of GLUT5 and fructokinase (KHK) were determined according to the Western blotting method in 1.2.

[0041] like Figure 3 As shown, compared with the control group, the tricaprylate treatment group (Tri+Fru) significantly upregulated the expression of fructose transporter 5 (GLUT5) and fructose kinase (KHK) in intestinal tissue (P<0.001).

[0042] Example 3: Effect of caprylic acid on liver glycogen recovery levels after fructose supplementation under starvation conditions. The grouping of experimental animals and the experimental methods are the same as in Example 2. Liver glycogen content determination: The liver glycogen content of mice was determined using the liver / muscle glycogen kit from Nanjing Jiancheng Bioengineering Institute.

[0043] like Figure 4 As shown, compared with the normal diet control group (NC), the liver glycogen content in the fasting control group (Con) decreased by 82.47% after 12 hours of fasting (P<0.0001), indicating that starvation has a significant impact on liver glycogen content in animals. The liver glycogen level in mice treated with oral tricaprylyl glycerol (Tri) recovered by 11.46%, and in mice treated with oral fructose (Fru) by 14.25%, with no significant differences among the Con, Tri, and Fru groups. However, compared with the fasting control group (Con), the liver glycogen level in mice treated with the combined tricaprylyl glycerol and fructose treatment (Tri+Fru) recovered by 49.96% (P<0.0001).

[0044] The above results indicate that caprylic acid promotes the recovery of liver glycogen after fructose supplementation during starvation.

[0045] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

Claims

1. Application of medium-chain fatty acids in the preparation of products that promote fructose absorption and / or glycogen replenishment.

2. The application according to claim 1, characterized in that, The medium-chain fatty acid is caprylic acid.

3. The application according to claim 1 or 2, characterized in that, The application includes at least one of the following: (1) Application in the preparation of products that promote the absorption of fructose by intestinal cells; (2) Application in the preparation of products that upregulate the expression level of fructose transporter protein in intestinal tissue; (3) Application in the preparation of products that upregulate the expression level of fructokinase in intestinal tissue; (4) Application in the preparation of products that restore the body’s liver glycogen levels.

4. The application according to claim 3, characterized in that, The liver glycogen level mentioned refers to the liver glycogen level under starvation conditions.

5. The application according to claim 1, characterized in that, The concentration of the medium-chain fatty acids used is 100~500 μM.

6. The application according to claim 1, characterized in that, The products mentioned are pharmaceuticals, animal feed, health foods, emergency foods, or sports foods.

7. The application according to claim 6, characterized in that, The products also include excipients permitted to be added to pharmaceuticals, feed, health foods, emergency foods, or sports foods.

8. The application according to claim 6, characterized in that, The drug is an oral preparation or an injection.

9. The application according to claim 8, characterized in that, The oral preparations are energy gels, powders, granules, capsules, powders, pills, tablets, or oral liquids.

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