Application of OPO and composition thereof in promoting intestinal development

Abstract

The invention provides an application of OPO and a composition thereof in promoting intestinal development. The 1, 3-dioleoyl-2-palmitin disclosed by the invention can be used for up-regulating the transcriptional level of a coding gene gpc1 of phospholipid globulin glycan, a coding gene has1 of hyaluronic acid synthetase 1, a coding gene has2 of hyaluronic acid synthetase 2, a coding gene has3 of hyaluronic acid synthetase 3, a coding gene ext1 of exosome glycosyltransferase 1 and a coding gene ext2 of exosome glycosyltransferase 2; therefore, the development of a protein skeleton of the glycocalyx layer, the synthesis of hyaluronic acid and the synthesis of heparan sulfate are promoted, the development of the glycocalyx layer is further promoted, and the effects of improving the intestinal barrier, regulating the intestinal flora and improving the intestinal health are achieved.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and more particularly to the application of OPO and its compositions in promoting intestinal development. Background Technology

[0002] In the physiological structure of the intestine, the glycocalyx is a specialized and intricate structure covering the surface of intestinal epithelial cells, primarily composed of glycans and proteins. A well-developed intestinal glycocalyx plays a crucial role in intestinal health, preventing pathogen adhesion and invasion, reducing the risk of intestinal infection, and providing specific binding sites for symbiotic microorganisms in the intestine. This helps maintain the balance and stability of the intestinal microbiota, which is essential for the normal functioning of the intestinal barrier and the homeostasis of the overall intestinal ecosystem. For example, during neonatal intestinal development, a well-developed glycocalyx provides binding sites for symbiotic microorganisms, prevents pathogen adhesion, and acts as a barrier against luminal toxins and enzymes.

[0003] Currently, although gut health has received considerable attention and numerous products on the market exist to improve gut function, such as probiotic preparations, prebiotic products, and some gut repair drugs, products specifically targeting the development of the intestinal epithelial glycocalyx are extremely scarce. Most existing gut health products indirectly affect gut health by regulating the quantity and types of gut microbiota, with unclear direct effects on this important structure. The few products that involve intestinal barrier repair mainly focus on strengthening the tight junctions between intestinal epithelial cells, lacking effective means to promote glycocalyx development. Therefore, developing a product that can effectively promote intestinal glycocalyx development is significantly necessary and has broad market prospects, playing a crucial role in improving gut health maintenance. Summary of the Invention

[0004] This invention provides the application of SN-2 palmitic acid triglyceride in promoting the development of the intestinal glycocalyx layer.

[0005] In the applications described above, SN-2 palmitate triglycerides include at least one of 1,3-dioleo-2-palmitoyl glyceride, 1-oleo-2-palmito-3-linoleic acid triglyceride, and lipoprotein lipase;

[0006] Preferably, SN-2 palmitic acid triglyceride is 1,3-dioleoyl-2-palmitoyl glyceride;

[0007] Preferably, the application concentration of 1,3-dioleoyl-2-palmitoylglycerol is 3-15 mg / mL.

[0008] The applications described above, wherein promoting the development of the intestinal glycocalyx layer include:

[0009] Upregulate the transcriptional level of gpc1, the gene encoding phosphoglobulin; and / or,

[0010] Upregulate the transcriptional level of the hyaluronic acid synthase 1 encoding gene has1; and / or,

[0011] Upregulate the transcriptional level of the hyaluronic acid synthase 2 encoding gene has2; and / or,

[0012] Upregulate the transcriptional level of the hyaluronic acid synthase 3 encoding gene has3; and / or,

[0013] Upregulate the transcriptional level of ext1, the gene encoding exochrome glycosyltransferase 1; and / or,

[0014] Upregulate the transcriptional level of ext2, the gene encoding exochrome glycosyltransferase 2; and / or,

[0015] Promotes the development of the protein backbone of the intestinal glycocalyx; and / or,

[0016] Promotes hyaluronic acid synthesis in the glycocalyx layer of the intestine; and / or,

[0017] Promotes the synthesis of heparan sulfate in the glycocalyx layer of the intestine.

[0018] This invention provides the application of a composition of 1,3-dioleoyl-2-palmitoylglycerol and casein phosphopeptide in promoting the development of the intestinal glycocalyx.

[0019] As described above, the application concentration of 1,3-dioleoyl-2-palmitoylglycerol is 3-15 mg / mL; and / or,

[0020] The application concentration of casein phosphopeptide is 0.05-1.5 mg / mL.

[0021] In the application described above, the concentration ratio of 1,3-dioleoyl-2-palmitoyl glyceride to casein phosphopeptide is (10-70):1.

[0022] The applications described above, wherein promoting the development of the intestinal glycocalyx layer include:

[0023] Upregulate the transcriptional level of gpc1, the gene encoding phosphoglobulin; and / or,

[0024] Upregulate the transcriptional level of the hyaluronic acid synthase 1 encoding gene has1; and / or,

[0025] Upregulate the transcriptional level of the hyaluronic acid synthase 2 encoding gene has2; and / or,

[0026] Upregulate the transcriptional level of the hyaluronic acid synthase 3 encoding gene has3; and / or,

[0027] Upregulate the transcriptional level of ext1, the gene encoding exochrome glycosyltransferase 1; and / or,

[0028] Upregulate the transcriptional level of ext2, the gene encoding exochrome glycosyltransferase 2; and / or,

[0029] Promotes the development of the protein backbone of the intestinal glycocalyx; and / or,

[0030] Promotes hyaluronic acid synthesis in the glycocalyx layer of the intestine; and / or,

[0031] Promotes the synthesis of heparan sulfate in the glycocalyx layer of the intestine.

[0032] This invention provides the use of a composition in the preparation of products related to promoting the development of the intestinal glycocalyx, wherein the composition comprises glyceryl 1,3-dioleoyl-2-palmitoylglycerol and optionally casein phosphopeptide.

[0033] The applications described above include at least one of food, health products, and pharmaceuticals;

[0034] Preferably, the food includes at least one of the following: infant formula, children's formula, adolescent formula, adult formula, dietary supplements, snacks, complementary foods, nutritional products, liquid milk, non-alcoholic beverages, and foods for special medical purposes;

[0035] Preferably, the health products include at least one of the following: health foods, functional foods, functional beverages, functional powders, functional capsules, and functional powders, which improve the intestinal barrier, regulate the intestinal flora, or improve intestinal health.

[0036] Preferably, the drug includes at least one of the following: prevention or treatment of gastrointestinal infection caused by glycocalyx damage, prevention or treatment of inflammatory bowel disease caused by glycocalyx damage, prevention or treatment of intestinal barrier impairment caused by glycocalyx damage, and prevention or treatment of intestinal flora imbalance caused by glycocalyx damage.

[0037] In the applications described above, the product also includes a carrier and / or physiologically acceptable excipients;

[0038] The carrier includes at least one of microcapsules, microspheres, nanoparticles, and liposomes;

[0039] Physiologically acceptable excipients include at least one of the following: fillers, flavoring agents, diluents, wetting agents, dispersants, binders, disintegrants, lubricants, color, flavor and aroma modifiers, solvents, solubilizers, co-solvents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, local analgesics, pH adjusters, and isotonic or isotropic modifiers.

[0040] This invention, through long-term research and extensive experiments, has discovered that 1,3-dioleoyl-2-palmitoylglycerol can upregulate the transcriptional level of the gene encoding phospholipoglobulin gpc1, thereby promoting the development of the glycocalyx protein backbone; it can upregulate the transcriptional levels of the genes encoding hyaluronic acid synthase 1 (has1), hyaluronic acid synthase 2 (has2), and hyaluronic acid synthase 3 (has3), thereby promoting the synthesis of hyaluronic acid in the glycocalyx; and it can also upregulate the transcriptional levels of the genes encoding exochrome glycosyltransferase 1 (ext1) and exochrome glycosyltransferase 2 (ext2), thereby promoting the synthesis of heparan sulfate in the glycocalyx. Therefore, 1,3-dioleoyl-2-palmitoylglycerol can promote the development of the glycocalyx by promoting the production of important components in the intestinal glycocalyx (glycocalyx protein backbone, hyaluronic acid, and heparan sulfate), and further improve the intestinal barrier, regulate the intestinal flora, and enhance intestinal health. The novel applications of 1,3-dioleoyl-2-palmitoylglycerol provided by this invention not only fill existing market gaps and overcome the limitations of existing products, but also offer new approaches to regulating gut microbiota, improving gut health, preventing or treating gastrointestinal infections caused by glycocalyx damage, preventing or treating inflammatory bowel disease caused by glycocalyx damage, preventing or treating intestinal barrier damage caused by glycocalyx damage, and preventing or treating gut microbiota dysbiosis caused by glycocalyx damage. This invention has significant necessity and broad market prospects, and is of great importance for improving the level of gut health maintenance. Attached Figure Description

[0041] Figure 1 This is a diagram showing the relative transcriptional level of the gpc1 gene in one embodiment of the present invention;

[0042] Figure 2 This is a diagram showing the relative transcriptional level of the has1 gene in one embodiment of the present invention;

[0043] Figure 3 This is a diagram showing the relative transcriptional level of the has2 gene in one embodiment of the present invention;

[0044] Figure 4 This is a diagram showing the relative transcriptional level of the has3 gene in one embodiment of the present invention;

[0045] Figure 5 This is a diagram showing the relative transcriptional level of the ext1 gene in one embodiment of the present invention;

[0046] Figure 6 This is a diagram showing the relative transcriptional level of the ext2 gene in one embodiment of the present invention;

[0047] Figure 7 This is a diagram showing the relative transcriptional levels of the gpc1 gene in the OPO and CPP groups in one embodiment of the present invention.

[0048] Figure 8 This is a diagram showing the relative transcriptional levels of the has1 gene in the OPO and CPP groups in one embodiment of the present invention;

[0049] Figure 9 This is a diagram showing the relative transcriptional levels of the has2 gene in the OPO and CPP groups in one embodiment of the present invention;

[0050] Figure 10 This is a diagram showing the relative transcriptional levels of the has3 gene in the OPO and CPP groups in one embodiment of the present invention;

[0051] Figure 11 This is a diagram showing the relative transcriptional levels of the ext1 gene in the OPO and CPP groups in one embodiment of the present invention;

[0052] Figure 12 This is a diagram showing the relative transcriptional levels of the ext2 gene in the OPO and CPP groups in one embodiment of the present invention. Detailed Implementation

[0053] To enable those skilled in the art to better understand the present invention, the present invention will be further described in detail below. The specific embodiments listed below are merely descriptions of the principles and features of the present invention, and the examples are only for explaining the present invention and are not intended to limit the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0054] The intestinal glycocalyx is a complex network structure covering the surface of intestinal epithelial cells. It serves as a core hub for the intestinal physical barrier and functional regulation, primarily composed of mucins, proteoglycans, glycoproteins, glycolipids, and free polysaccharides assembled through covalent or non-covalent interactions to form a dynamic functional layer rich in carbohydrates. Among these, phosphoglobulin 1 (Glypican 1) is a key membrane-bound proteoglycan in the glycocalyx. Its core protein is anchored to the intestinal epithelial cell membrane surface via glycosylphosphatidylinositol (GPI), and covalently linked to glycosaminoglycan chains such as heparan sulfate. As a protein backbone component of the glycocalyx, Glypican 1 not only maintains the spatial stability of the glycocalyx through its own structure, but its heparan sulfate chains also bind water, ions, and small molecule nutrients through charge interactions, participating in the osmotic regulation and substance exchange functions of the glycocalyx. Heparan sulfate (HS) is a linear sulfated polysaccharide. Besides being a glycan component of phosphoglobulin-1, it can also exist in the glycocalyx layer by binding with other proteoglycans. Its highly sulfated structure allows it to specifically recognize and bind to adhesion proteins of intestinal pathogens (such as E. coli and rotavirus), preventing direct contact between pathogens and intestinal epithelial cells. Simultaneously, it can regulate the recruitment and activation of intestinal immune cells (such as macrophages and dendritic cells), making it a crucial executor of the glycocalyx layer's immune defense function. Hyaluronic acid (HA) is another important glycosaminoglycan. Unlike HS, it usually exists in the glycocalyx layer in free form or bound to hyaluronic acid-binding proteins. It is highly hydrophilic and can increase the thickness and elasticity of the glycocalyx layer by absorbing water and swelling, enhancing its buffering capacity against mechanical damage (such as intestinal peristalsis friction). HA can also regulate cell proliferation and differentiation by binding to CD44 receptors on the surface of intestinal epithelial cells, promoting the repair and renewal of the glycocalyx layer.

[0055] Based on the above research, in order to develop a product that can effectively promote the development of the intestinal glycocalyx layer, this invention, through long-term research and numerous experiments, discovered that SN-2 palmitate triglyceride 1,3-dioleoyl-2-palmitoylglyceride (OPO structural lipid, hereinafter referred to as OPO) can upregulate the transcription level of the gene encoding phospholipoglobulin gpc1 in the body, thereby promoting the development of the protein backbone of the glycocalyx layer; SN-2 palmitate triglyceride OPO can upregulate the transcription levels of the genes encoding hyaluronic acid synthase 1 (has1), hyaluronic acid synthase 2 (has2), and hyaluronic acid synthase 3 (has3) in the body, thereby promoting the synthesis of hyaluronic acid in the glycocalyx layer; SN-2 palmitate triglyceride OPO can also upregulate the transcription levels of the genes encoding exochrome glycosyltransferase 1 (ext1) and exochrome glycosyltransferase 2 (ext2), thereby promoting the synthesis of heparan sulfate in the glycocalyx layer. In short, SN-2 palmitate triglyceride OPO can promote the development of the glycocalyx layer by promoting the development of important components in the intestinal glycocalyx layer (glycocalyx protein backbone, hyaluronic acid, and heparan sulfate), and further improve the intestinal barrier, regulate the intestinal flora, and improve intestinal health.

[0056] Therefore, the first aspect of the present invention provides the application of SN-2 palmitic acid triglyceride in promoting the development of the intestinal glycocalyx layer.

[0057] The novel applications of SN-2 palmitic acid triglycerides provided by this invention not only fill existing market gaps and overcome the limitations of existing products, but also offer new approaches to regulating gut microbiota, improving gut health, preventing or treating gastrointestinal infections caused by glycocalyx damage, preventing or treating inflammatory bowel disease caused by glycocalyx damage, preventing or treating intestinal barrier damage caused by glycocalyx damage, and preventing or treating gut microbiota dysbiosis caused by glycocalyx damage. This invention has significant necessity and broad market prospects, and is of great importance for improving gut health maintenance.

[0058] Further, SN-2 palmitate triglyceride includes at least one of OPO, 1-oleic-2-palmitoyl-3-linoleic acid triglyceride (OPL), and lipoprotein lipase (LPL); and in a preferred embodiment, the application concentration of OPO is 3-15 mg / mL, which can better promote the development of the glycocalyx layer. Exemplarily, the application concentration of OPO can be 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, and any value between any two of the above ranges.

[0059] In the above technical solution, promoting the development of the intestinal glycocalyx layer includes at least one of the following: upregulating the transcription level of the gene gpc1 encoding phosphoglobulin glycan, upregulating the transcription level of the gene has1 encoding hyaluronic acid synthase 1, upregulating the transcription level of the gene has2 encoding hyaluronic acid synthase 2, upregulating the transcription level of the gene has3 encoding hyaluronic acid synthase 3, upregulating the transcription level of the gene ext1 encoding exoprotein glycosyltransferase 1, and upregulating the transcription level of the gene ext2 encoding exoprotein glycosyltransferase 2.

[0060] Experimental results showed that intervention with SN-2 palmitate triglyceride OPO could upregulate the transcriptional levels of the following genes in human colon adenocarcinoma cells Caco-2: gpc1 (encoding phosphoglobulin glycan), has1 (encoding hyaluronic acid synthase 1), has2 (encoding hyaluronic acid synthase 2), has3 (encoding hyaluronic acid synthase 3), ext1 (encoding exochrome glycosyltransferase 1), and ext2 (encoding exochrome glycosyltransferase 2), thereby promoting the development of the intestinal glycocalyx.

[0061] Understandably, SN-2 palmitate triglyceride OPO can be used to promote the development of the intestinal glycocalyx layer, and can also be used to prepare agonists that promote the transcription of the gene encoding phosphoglobulin gpc1, the gene encoding hyaluronic acid synthase 1 has1, the gene encoding hyaluronic acid synthase 2 has2, the gene encoding hyaluronic acid synthase 3 has3, the gene encoding exochrome glycosyltransferase 1 ext1, and the gene encoding exochrome glycosyltransferase 2 ext2.

[0062] In the above technical solutions, promoting the development of the intestinal glycocalyx layer also includes at least one of the following: promoting the development of the protein backbone of the intestinal glycocalyx layer, promoting the synthesis of hyaluronic acid in the intestinal glycocalyx layer, and promoting the synthesis of heparan sulfate in the intestinal glycocalyx layer.

[0063] Experiments have verified that intervention with SN-2 palmitate triglyceride OPO can upregulate the transcriptional level of the gene encoding phospholipoglobulin g1 in human colon adenocarcinoma cells Caco-2, thereby promoting the development of the intestinal glycocalyx protein backbone and thus the development of the intestinal glycocalyx. SN-2 palmitate triglyceride OPO intervention can also upregulate the transcriptional levels of the genes encoding hyaluronic acid synthase 1 (has1), hyaluronic acid synthase 2 (has2), and hyaluronic acid synthase 3 (has3) in human colon adenocarcinoma cells Caco-2, thereby promoting the synthesis of hyaluronic acid in the intestinal glycocalyx and thus promoting the development of the intestinal glycocalyx. Furthermore, SN-2 palmitate triglyceride OPO intervention can upregulate the transcriptional levels of the genes encoding exokinin glycosyltransferase 1 (ext1) and exokinin glycosyltransferase 2 (ext2) in human colon adenocarcinoma cells Caco-2, thereby promoting the synthesis of heparan sulfate in the intestinal glycocalyx and thus promoting the development of the intestinal glycocalyx.

[0064] Understandably, SN-2 palmitate triglyceride OPO can be used to promote the development of the intestinal glycocalyx layer, and can also be used to prepare agonists that promote the development of the protein backbone of the intestinal glycocalyx layer, agonists that promote the synthesis of hyaluronic acid in the intestinal glycocalyx layer, and agonists that promote the synthesis of heparan sulfate in the intestinal glycocalyx layer.

[0065] A second aspect of the present invention provides the use of a composition of 1,3-dioleoyl-2-palmitoylglycerol and casein phosphopeptide in promoting the development of the intestinal glycocalyx.

[0066] Experiments have shown that OPO and casein phosphopeptide (CPP) have a synergistic effect. When used simultaneously, both significantly upregulate the transcription level of the gene encoding phosphoglobulin glycan (gpc1), thereby promoting the development of the glycocalyx protein backbone. Furthermore, they upregulate the transcription levels of the genes encoding hyaluronic acid synthase 1 (has1), hyaluronic acid synthase 2 (has2), and hyaluronic acid synthase 3 (has3), thus promoting hyaluronic acid synthesis in the glycocalyx. Simultaneous use also significantly upregulates the transcription levels of the genes encoding exochrome glycosyltransferase 1 (ext1) and exochrome glycosyltransferase 2 (ext2), thereby promoting heparan sulfate synthesis in the glycocalyx. The combined use of OPO and CPP can further promote the development of the glycocalyx by promoting the production of key components (glycocalyx protein backbone, hyaluronic acid, and heparan sulfate), effectively strengthening the intestinal barrier, regulating gut microbiota, and improving gut health.

[0067] Furthermore, when the application concentration of 1,3-dioleoyl-2-palmitoylglycerol is 3-15 mg / mL and the application concentration of casein phosphopeptide is 0.05-1.5 mg / mL, OPO and CPP have a good matching effect and synergistically promote the development of the glycocalyx layer. For example, the application concentration of CPP can be 0.05 mg / mL, 0.15 mg / mL, 0.25 mg / mL, 0.35 mg / mL, 0.45 mg / mL, 0.55 mg / mL, 0.65 mg / mL, 0.75 mg / mL, 0.85 mg / mL, 0.95 mg / mL, 1.00 mg / mL, 1.10 mg / mL, 1.20 mg / mL, 1.30 mg / mL, 1.40 mg / mL, 1.50 mg / mL, or any value between any two of the above ranges.

[0068] Furthermore, a concentration ratio of 1,3-dioleoyl-2-palmitoylglycerol to casein phosphopeptide of (10-70):1 can better leverage the synergistic effect of OPO and CPP, thereby further promoting the development of the glycocalyx layer. For example, the concentration ratio of OPO to CPP can be 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 60:1, 70:1, etc.

[0069] In the above technical solution, promoting the development of the intestinal glycocalyx layer includes at least one of the following: upregulating the transcription level of the gene gpc1 encoding phosphoglobulin glycan, upregulating the transcription level of the gene has1 encoding hyaluronic acid synthase 1, upregulating the transcription level of the gene has2 encoding hyaluronic acid synthase 2, upregulating the transcription level of the gene has3 encoding hyaluronic acid synthase 3, upregulating the transcription level of the gene ext1 encoding exoprotein glycosyltransferase 1, and upregulating the transcription level of the gene ext2 encoding exoprotein glycosyltransferase 2.

[0070] Experimental results showed that the combined intervention of OPO and CPP can upregulate the transcriptional levels of the following genes in human colon adenocarcinoma cells Caco-2: gpc1 (encoding phosphoglobulin glycan), has1 (encoding hyaluronic acid synthase 1), has2 (encoding hyaluronic acid synthase 2), has3 (encoding hyaluronic acid synthase 3), ext1 (encoding exochrome glycosyltransferase 1), and ext2 (encoding exochrome glycosyltransferase 2), thereby promoting the development of the intestinal glycocalyx.

[0071] It is understandable that the combination of OPO and CPP can be used to promote the development of the intestinal glycocalyx layer, and can also be used to prepare agonists that promote the transcription of the gene encoding phosphoglobulin gpc1, the gene encoding hyaluronic acid synthase 1 has1, the gene encoding hyaluronic acid synthase 2 has2, the gene encoding hyaluronic acid synthase 3 has3, the gene encoding exochrome glycosyltransferase 1 ext1, and the gene encoding exochrome glycosyltransferase 2 ext2.

[0072] In the above technical solutions, promoting the development of the intestinal glycocalyx layer also includes at least one of the following: promoting the development of the protein backbone of the intestinal glycocalyx layer, promoting the synthesis of hyaluronic acid in the intestinal glycocalyx layer, and promoting the synthesis of heparan sulfate in the intestinal glycocalyx layer.

[0073] Experiments have verified that combined intervention with OPO and CPP upregulates the transcriptional level of gpc1, the gene encoding phospholipoglobulin glycan, in human colon adenocarcinoma cells Caco-2, thereby promoting the development of the intestinal glycocalyx protein backbone and thus the development of the intestinal glycocalyx. Similarly, combined intervention with OPO and CPP upregulates the transcriptional levels of has1, has2, and has3, the genes encoding hyaluronic acid synthase 1, hyaluronic acid synthase 2, and hyaluronic acid synthase 3, in human colon adenocarcinoma cells Caco-2, thereby promoting the synthesis of hyaluronic acid in the intestinal glycocalyx and thus promoting the development of the intestinal glycocalyx. Furthermore, combined intervention with OPO and CPP upregulates the transcriptional levels of ext1 and ext2, the genes encoding exochrome glycosyltransferase 1 and exochrome glycosyltransferase 2, in human colon adenocarcinoma cells Caco-2, thereby promoting the synthesis of heparan sulfate in the intestinal glycocalyx and thus promoting the development of the intestinal glycocalyx.

[0074] It is understandable that the combination of OPO and CPP can be used to promote the development of the intestinal glycocalyx layer, and can also be used to prepare agonists that promote the development of the protein backbone of the intestinal glycocalyx layer, agonists that promote the synthesis of hyaluronic acid in the intestinal glycocalyx layer, and agonists that promote the synthesis of heparan sulfate in the intestinal glycocalyx layer.

[0075] A third aspect of the present invention provides the use of a composition in the preparation of products related to promoting the development of the intestinal glycocalyx, wherein the composition comprises glyceryl 1,3-dioleoyl-2-palmitoylglycerol and optionally casein phosphopeptide.

[0076] Furthermore, the products include at least one of food, health products, and medicines.

[0077] In a preferred embodiment, the food includes at least one of the following: infant formula, children's formula, adolescent formula, adult formula, dietary supplements, snacks, complementary foods, nutritional products, liquid milk, non-alcoholic beverages, and foods for special medical purposes.

[0078] For example, an infant formula, children's formula, adolescent formula, adult formula, dietary supplement, snack, complementary food, nutritional product, liquid milk, non-alcoholic beverage, or food for special medical purposes for promoting the development of the intestinal glycocalyx layer includes 1,3-dioleoyl-2-palmitoylglycerol and optionally casein phosphopeptide.

[0079] In a preferred embodiment, the health product includes at least one of the following: health foods, functional foods, functional beverages, functional powders, functional capsules, and functional powders, which enhance the intestinal barrier, regulate intestinal flora, or improve intestinal health.

[0080] In a preferred embodiment, the drug includes at least one of the following: prevention or treatment of gastrointestinal infection caused by glycocalyx damage, prevention or treatment of inflammatory bowel disease caused by glycocalyx damage, prevention or treatment of intestinal barrier impairment caused by glycocalyx damage, and prevention or treatment of intestinal flora imbalance caused by glycocalyx damage.

[0081] The aforementioned products also include carriers and / or physiologically acceptable excipients.

[0082] The carrier includes at least one of microcapsules, microspheres, nanoparticles, and liposomes.

[0083] Physiologically acceptable excipients include at least one of the following: fillers, flavoring agents, diluents, wetting agents, dispersants, binders, disintegrants, lubricants, color, flavor and aroma modifiers, solvents, solubilizers, cosolvents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, local analgesics, pH adjusters, and isotonic or isotropic modifiers.

[0084] In actual use, the above products can be administered via at least one of the following methods: oral administration, intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, sublingual administration, nasal administration, and transdermal administration.

[0085] The technical solution of this application will be further explained below with reference to specific embodiments. Experimental methods without specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise specified, all reagents used are commercially available or obtained through public channels.

[0086] In the following examples, the culture conditions for human colon adenocarcinoma cells Caco-2 were as follows: Caco-2 cells were cultured using MEM complete medium (containing 20% ​​fetal bovine serum) produced by Nanjing Senbeijia Biotechnology Co., Ltd., and seeded in 25cm...2 The cell culture flasks were placed in a carbon dioxide incubator at 37°C, 5% carbon dioxide, and 95% relative humidity. The culture medium was changed every 1-2 days. When the cell confluence reached 80%-90%, the cells were digested with 0.25% trypsin digestion solution and passaged at a volume ratio of 1:3.

[0087] Example 1: Experimental Grouping

[0088] The density of Caco-2 cells was adjusted to 1×10⁻⁶. 5 pcs / cm 2 Caco-2 cells were seeded in 24-well plates and grouped and treated according to the groups and intervention substances in Table 1. Specifically, the blank control group received no intervention and was cultured in MEM complete medium for 36 h; the low-dose 1,3-dioleoyl-2-palmitoylglycerol (OPO) group (3.36 mg / mL) was treated by adding OPO to MEM complete medium at a final concentration of 3.36 mg / mL and culturing for 36 h; the medium-dose OPO group (6.72 mg / mL) was treated by adding OPO to MEM complete medium at a final concentration of 6.72 mg / mL and culturing for 36 h; the high-dose OPO group (13.44 mg / mL) was treated by adding OPO to MEM complete medium at a final concentration of 13.44 mg / mL and culturing for 36 h; and the OPO and casein phosphopeptide (CPP) group was treated by adding OPO and CPP to MEM complete medium at final concentrations of 6.72 mg / mL and 0.28 mg / mL respectively and culturing for 36 h.

[0089] Table 1

[0090]

[0091] Example 2: Real-time quantitative PCR (qPCR) experiment

[0092] Total RNA was extracted and reverse transcribed from cells cultured for 36 h in Example 1. The relative transcription levels of genes related to glycocalyx development in Caco-2 cells were detected by qPCR. The qPCR primers are shown in Table 2. Genes related to glycocalyx development include: glypican 1 (gpc1), hyaluronan synthase 1 (has1), hyaluronan synthase 2 (has2), hyaluronan synthase 3 (has3), exostosin glycosyltransferase 1 (ext1), and exostosin glycosyltransferase 2 (ext2).

[0093] Table 2

[0094]

[0095] Example 3: Effect of OPO on the protein backbone of the glycocalyx layer

[0096] The glypican 1 (gpc1) gene is the gene encoding the proteoglycan GPC1. GPC1 is the protein backbone of the glycocalyx and an important carrier of glycosaminoglycan chains, including heparan sulfate (HS) and hyaluronic acid (HA).

[0097] The relative transcription level of the gpc1 gene obtained by qPCR in Example 2 is shown. Figure 1 . Figure 1 The results showed that the gpc1 gene transcription levels in the medium-dose OPO group (6.72 mg / mL) and the high-dose OPO group (13.44 mg / mL) were significantly higher than those in the control group (p<0.01). These results indicate that OPO can upregulate gpc1 transcription levels, thereby promoting the development of the glycocalyx protein backbone and thus promoting the development of the glycocalyx layer.

[0098] Example 4: Effect of OPO on hyaluronic acid synthase in the glycocalyx layer

[0099] The genes encoding hyaluronan synthase 1 (HAS1), hyaluronan synthase 2 (HAS2), and hyaluronan synthase 3 (HAS3) are the coding genes for hyaluronan synthase (HAS). HAS is crucial for the synthesis of hyaluronan (HA), a major glycosaminoglycan component of the glycocalyx and a highly viscous component of the intestinal mucus layer, responsible for tissue repair, stability, and anti-inflammatory effects. Compared to HAS1 and HAS2, HAS3 can synthesize low molecular weight HA, which is essential for the development of intestinal stem cells; therefore, the presence of HAS3 is particularly important.

[0100] The relative transcription level of the has1 gene obtained by qPCR in Example 2 is shown. Figure 2 The relative transcription level of the has2 gene is visible. Figure 3 The relative transcription level of the has3 gene is visible. Figure 4 . Figure 2 The results showed that the transcriptional level of the has1 gene in the medium-dose OPO group (6.72 mg / mL) and the high-dose OPO group (13.44 mg / mL) was significantly higher than that in the blank control group (p<0.001). Figure 3 The results showed that the transcriptional level of the has2 gene in the low-dose OPO group (3.36 mg / mL) and the medium-dose OPO group (6.72 mg / mL) was significantly higher than that in the blank control group (p<0.0001). Figure 4 The results showed that the transcriptional levels of the has3 gene in the low-dose OPO group (3.36 mg / mL) and the high-dose OPO group (13.44 mg / mL) were significantly higher than those in the control group (p<0.01); the transcriptional level of the has3 gene in the medium-dose OPO group (6.72 mg / mL) was significantly higher than that in the control group (p<0.0001). These results indicate that OPO can upregulate the transcriptional levels of has1, has2, and has3, thereby promoting the synthesis of HA in the glycocalyx layer and thus promoting the development of the glycocalyx layer.

[0101] Example 5: Effect of OPO on heparan sulfate in the glycocalyx layer

[0102] The exostosin glycosyltransferase 1 (ext1) and exostosin glycosyltransferase 2 (ext2) genes are involved in the elongation of the heparan sulfate (HS) chain and are responsible for integrating the HS chain with nucleotide sugars in the Golgi apparatus. HS is the main glycosaminoglycan component in the glycocalyx, which supports organogenesis, growth factor signaling, and bacterial adhesion.

[0103] The relative transcription level of the ext1 gene obtained by qPCR in Example 2 is shown. Figure 5 The relative transcription level of the ext2 gene is visible. Figure 6 . Figure 5 The results showed that the ext1 gene transcription level in the low-dose OPO group (3.36 mg / mL) was significantly higher than that in the blank control group (p<0.05); the ext1 gene transcription level in the medium-dose OPO group (6.72 mg / mL) was significantly higher than that in the blank control group (p<0.0001). Figure 6 The results showed that the transcriptional level of the ext2 gene in the medium-dose OPO group (6.72 mg / mL) was significantly higher than that in the blank control group (p<0.001). These results indicate that OPO can upregulate the transcriptional levels of ext1 and ext2, thereby promoting the synthesis of HS in the glycocalyx layer and thus promoting the development of the glycocalyx layer.

[0104] Example 6: Effects of OPO and CPP on the protein backbone of the glycocalyx layer

[0105] The relative transcription levels of the gpc1 gene in the OPO and CPP groups (OPO+CPP) obtained by qPCR in Example 2 are shown. Figure 7 .

[0106] Figure 7 The results showed that the gpc1 gene transcription level in the OPO and CPP group (OPO+CPP) was significantly higher than that in the blank control group (blank group) (p<0.0001), and also significantly higher than that in the medium-dose OPO group (6.72 mg / mL). These results indicate that the combination of OPO and CPP can significantly upregulate gpc1 transcription levels, further promoting the development of the glycocalyx protein backbone, thereby promoting glycocalyx development; and that OPO and CPP have a synergistic effect in promoting glycocalyx development.

[0107] Example 7: Effects of OPO and CPP on hyaluronic acid synthase in the glycocalyx layer

[0108] The relative transcription levels of the has1 gene in the OPO and CPP groups (OPO+CPP) obtained by qPCR in Example 2 are shown. Figure 8 The relative transcription level of the has2 gene is visible. Figure 9 The relative transcription level of the has3 gene is visible. Figure 10 .

[0109] Figure 8 The results showed that the has1 gene transcription level in the OPO and CPP groups (OPO+CPP) was significantly higher than that in the blank control group (blank group) (p<0.001), and also significantly higher than that in the OPO medium-dose group (6.72 mg / mL). Figure 9 The results showed that the has2 gene transcription level in the OPO and CPP groups (OPO+CPP) was significantly higher than that in the blank control group (blank group) (p<0.0001), and also significantly higher than that in the OPO medium-dose group (6.72 mg / mL). Figure 10 The results showed that the transcriptional level of the has3 gene in the OPO and CPP group (OPO+CPP) was significantly higher than that in the blank control group (blank group) (p<0.001), and also significantly higher than that in the OPO medium-dose group (6.72 mg / mL). These results indicate that the combination of OPO and CPP can upregulate the transcriptional levels of has1, has2, and has3, thereby promoting HA synthesis in the glycocalyx layer and thus promoting its development; moreover, OPO and CPP have a synergistic effect on promoting glycocalyx layer development, and the effect is superior to that of OPO alone (6.72 mg / mL).

[0110] Example 8: Effects of OPO and CPP on heparan sulfate in the glycocalyx layer

[0111] The relative transcription levels of the ext1 gene in the OPO and CPP groups (OPO+CPP) obtained by qPCR in Example 2 are shown. Figure 11 The relative transcription level of the ext2 gene is visible. Figure 12 .

[0112] Figure 11 The results showed that the ext1 gene transcription level in the OPO and CPP groups (OPO+CPP) was significantly higher than that in the blank control group (blank group) (p<0.01), and also significantly higher than that in the OPO medium-dose group (6.72 mg / mL). Figure 12 The results showed that the transcriptional levels of the ext2 gene in the OPO and CPP group (OPO+CPP) and the medium-dose OPO group (6.72 mg / mL) were significantly higher than those in the blank control group (blank group) (p<0.01). These results indicate that the combination of OPO and CPP can upregulate the transcriptional level of ext1, thereby promoting the synthesis of HS in the glycocalyx layer and thus promoting the development of the glycocalyx layer.

[0113] In summary, co-culturing Caco-2 cells with OPO before they adhere to the culture vessel can simulate the state of intestinal growth and maturation. Under this cellular state, OPO significantly promotes the development of the glycocalyx layer in Caco-2 cells. Furthermore, OPO can be used in combination with CPP to synergistically promote the development of the glycocalyx layer skeleton and the synthesis of heparan sulfate. These results indicate that supplementing with a specific dose of OPO or a combination of OPO and CPP in a specific ratio during the intestinal maturation stage can significantly promote the development of the intestinal glycocalyx layer, thereby promoting overall healthy intestinal development.

[0114] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Application of SN-2 palmitic acid triglyceride in promoting the development of intestinal glycocalyx.

2. The application according to claim 1, characterized in that, The SN-2 palmitate triglyceride includes at least one of 1,3-dioleo-2-palmitoyl glyceride, 1-oleo-2-palmito-3-linoleic acid triglyceride, and lipoprotein lipase. Preferably, the SN-2 palmitic acid triglyceride is 1,3-dioleoyl-2-palmitoyl glyceride; Preferably, the application concentration of the 1,3-dioleoyl-2-palmitoylglycerol is 3-15 mg / mL.

3. The application according to claim 1 or 2, characterized in that, The promotion of intestinal glycocalyx development includes: Upregulate the transcriptional level of gpc1, the gene encoding phosphoglobulin; and / or, Upregulate the transcriptional level of the hyaluronic acid synthase 1 encoding gene has1; and / or, Upregulate the transcriptional level of the hyaluronic acid synthase 2 encoding gene has2; and / or, Upregulate the transcriptional level of the hyaluronic acid synthase 3 encoding gene has3; and / or, Upregulate the transcriptional level of ext1, the gene encoding exochrome glycosyltransferase 1; and / or, Upregulate the transcriptional level of ext2, the gene encoding exochrome glycosyltransferase 2; and / or, Promotes the development of the protein backbone of the intestinal glycocalyx; and / or, Promotes hyaluronic acid synthesis in the glycocalyx layer of the intestine; and / or, Promotes the synthesis of heparan sulfate in the glycocalyx layer of the intestine. 4.1,3-Dioleoyl-2-palmitoylglycerol and casein phosphopeptide composition in promoting the development of the intestinal glycocalyx.

5. The application according to claim 4, characterized in that, The application concentration of the 1,3-dioleoyl-2-palmitoylglycerol is 3-15 mg / mL; and / or, The application concentration of the casein phosphopeptide is 0.05-1.5 mg / mL.

6. The application according to claim 4 or 5, characterized in that, The concentration ratio of 1,3-dioleoyl-2-palmitoylglycerol to casein phosphopeptide is (10-70):

1.

7. The application according to claim 4 or 5, characterized in that, The promotion of intestinal glycocalyx development includes: Upregulate the transcriptional level of gpc1, the gene encoding phosphoglobulin; and / or, Upregulate the transcriptional level of the hyaluronic acid synthase 1 encoding gene has1; and / or, Upregulate the transcriptional level of the hyaluronic acid synthase 2 encoding gene has2; and / or, Upregulate the transcriptional level of the hyaluronic acid synthase 3 encoding gene has3; and / or, Upregulate the transcriptional level of ext1, the gene encoding exochrome glycosyltransferase 1; and / or, Upregulate the transcriptional level of ext2, the gene encoding exochrome glycosyltransferase 2; and / or, Promotes the development of the protein backbone of the intestinal glycocalyx; and / or, Promotes hyaluronic acid synthesis in the glycocalyx layer of the intestine; and / or, Promotes the synthesis of heparan sulfate in the glycocalyx layer of the intestine.

8. The use of a composition in the preparation of products related to promoting the development of the intestinal glycocalyx layer, characterized in that, The composition comprises 1,3-dioleoyl-2-palmitoylglycerol and optionally casein phosphopeptide.

9. The application according to claim 8, characterized in that, The product includes at least one of food, health products, and medicine; Preferably, the food includes at least one of the following: infant formula, children's formula, adolescent formula, adult formula, dietary supplements, snacks, complementary foods, nutritional products, liquid milk, non-alcoholic beverages, and foods for special medical purposes. Preferably, the health products include at least one of the following: health foods, functional foods, functional beverages, functional powders, functional capsules, and functional powders, which improve the intestinal barrier, regulate intestinal flora, or improve intestinal health. Preferably, the drug includes at least one of the following: prevention or treatment of gastrointestinal infection caused by glycocalyx damage, prevention or treatment of inflammatory bowel disease caused by glycocalyx damage, prevention or treatment of intestinal barrier impairment caused by glycocalyx damage, and prevention or treatment of intestinal flora imbalance caused by glycocalyx damage.

10. The application according to claim 8 or 9, characterized in that, The product also includes a carrier and / or physiologically acceptable excipients; The carrier includes at least one of microcapsules, microspheres, nanoparticles, and liposomes; Physiologically acceptable excipients include at least one of the following: fillers, flavoring agents, diluents, wetting agents, dispersants, binders, disintegrants, lubricants, color, flavor and aroma modifiers, solvents, solubilizers, co-solvents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, local analgesics, pH adjusters, and isotonic or isotropic modifiers.

Images