Application of casein phosphopeptide in promoting development of intestinal glycocalyx layer and preparation of related products

Abstract

The application provides application of casein phosphopeptide in promoting development of a sugar layer of an intestinal tract and preparation of related products. The casein phosphopeptide can up-regulate transcription levels of a coding gene gpc1 of phospholipid globulin polysaccharide, 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 exosin glycosyltransferase 1 and a coding gene ext2 of exosin glycosyltransferase 2, thereby promoting development of a protein skeleton of the sugar layer, synthesis of hyaluronic acid and synthesis of heparan sulfate, further promoting development of the sugar layer, and achieving the effects of perfecting an intestinal tract barrier, regulating intestinal flora and improving intestinal health.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to the application of a casein phosphopeptide in promoting the development of the intestinal glycocalyx and the preparation of related products. Background Technology

[0002] In the physiological structure of the intestine, the glycocalyx is a crucial and intricate structure covering the surface of intestinal epithelial cells, primarily composed of glycans and proteins. A well-developed intestinal glycocalyx plays an irreplaceable core role in maintaining intestinal health: on the one hand, it effectively prevents pathogen adhesion and invasion, significantly reducing the risk of intestinal infection; on the other hand, it provides specific binding sites for symbiotic microorganisms in the intestine. Therefore, a well-developed intestinal glycocalyx is an important foundation for maintaining the balance and stability of the intestinal microbial community, which is crucial for ensuring the normal functioning of the intestinal barrier and maintaining the homeostasis of the overall intestinal ecological environment. For example, during neonatal intestinal development, it can competitively inhibit pathogen adhesion by providing specific binding sites for symbiotic microorganisms and act as a natural barrier against luminal toxins and enzymes.

[0003] Currently, although gut health has received widespread attention and various 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 remain scarce. The mechanisms of action of existing gut health products mostly focus on regulating the quantity and types of gut microbiota, influencing gut health indirectly. Their direct effects on the core structure of the intestinal epithelial glycocalyx lack clear evidence. Even the few products involving intestinal barrier repair primarily focus on enhancing tight junctions between intestinal epithelial cells, without targeting the development of the intestinal epithelial glycocalyx as a core objective, resulting in significant limitations in maintaining gut health.

[0004] Therefore, developing a technology or product that can effectively promote the development of the intestinal glycocalyx layer is not only of great technical necessity, but also fills the functional gap in existing intestinal health products. It has important clinical value and application prospects for improving the stability of intestinal barrier function and optimizing the balance of intestinal microecology, and is of irreplaceable significance for breaking through the bottleneck of existing intestinal health maintenance technology and improving the level of intestinal health management. Summary of the Invention

[0005] This invention provides the application of casein phosphopeptide in promoting the development of the intestinal glycocalyx.

[0006] In the applications described above, the concentration of casein phosphopeptide is 0.01-0.5 mg / mL.

[0007] In the application described above, the concentration of casein phosphopeptide is 0.10-0.20 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 also provides the application of casein phosphopeptide in the preparation of products related to promoting the development of the intestinal glycocalyx.

[0019] The applications described above include at least one of health supplements and pharmaceuticals.

[0020] As described above, in which,

[0021] As described above, the health supplements include at least one of the following: functional beverages, functional powders, functional capsules, and functional powders, which enhance the intestinal barrier, regulate gut microbiota, or improve gut health.

[0022] As described above, 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.

[0023] In the applications described above, the product also includes a carrier and / or physiologically acceptable excipients;

[0024] The carrier includes at least one of microcapsules, microspheres, nanoparticles, and liposomes;

[0025] 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.

[0026] This invention, through long-term research and extensive experiments, has discovered that casein phosphopeptides can upregulate the transcriptional level of the gene encoding phosphoglobulin glucan (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, casein phosphopeptides 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 casein phosphopeptides provided by this invention not only fill existing market gaps and overcome the limitations of existing products, but also offer new approaches to improving the intestinal barrier, regulating gut microbiota, enhancing intestinal 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 impairment 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 intestinal health maintenance. Attached Figure Description

[0027] Figure 1 This is a diagram showing the relative transcriptional level of the gpc1 gene in one embodiment of the present invention;

[0028] Figure 2 This is a diagram showing the relative transcriptional level of the has1 gene in one embodiment of the present invention;

[0029] Figure 3 This is a diagram showing the relative transcriptional level of the has2 gene in one embodiment of the present invention;

[0030] Figure 4 This is a diagram showing the relative transcriptional level of the has3 gene in one embodiment of the present invention;

[0031] Figure 5This is a diagram showing the relative transcriptional level of the ext1 gene in one embodiment of the present invention;

[0032] Figure 6 This is a diagram showing the relative transcriptional level of the ext2 gene in one embodiment of the present invention. Detailed Implementation

[0033] 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.

[0034] 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.

[0035] Based on the above research, in order to develop a product that can effectively promote the development of the intestinal glycocalyx, this invention, through long-term research and extensive experiments, has discovered that casein phosphopeptides (CPP) can upregulate the transcriptional level of the gene encoding phosphoglobulin glucan, gpc1, thereby promoting the development of the glycocalyx protein backbone. Casein phosphopeptides 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), thereby promoting the synthesis of hyaluronic acid in the glycocalyx. Furthermore, casein phosphopeptides can 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. In short, casein phosphopeptides can promote the development of the intestinal glycocalyx by promoting the development of important components (glycocalyx protein backbone, hyaluronic acid, and heparan sulfate), and further achieve the effects of improving the intestinal barrier, regulating the intestinal flora, and improving intestinal health.

[0036] Therefore, this invention provides an application of casein phosphopeptide in promoting the development of the intestinal glycocalyx. Casein phosphopeptide is a bioactive polypeptide obtained from casein using biotechnology, and it possesses highly phosphorylated properties.

[0037] The novel applications of casein phosphopeptides provided by this invention not only fill existing market gaps and overcome the limitations of existing products, but also offer new approaches to improving the gut microbiota barrier, regulating gut flora, 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.

[0038] In the above technical solution, when the application concentration of casein phosphopeptide is 0.01-0.5 mg / mL, it can better promote the development of the glycocalyx layer. For example, the application concentration of casein phosphopeptide can be 0.01 mg / mL, 0.05 mg / mL, 0.10 mg / mL, 0.15 mg / mL, 0.20 mg / mL, 0.25 mg / mL, 0.30 mg / mL, 0.35 mg / mL, 0.40 mg / mL, 0.45 mg / mL, 0.50 mg / mL, or any value between any two of the above ranges.

[0039] In a preferred embodiment, when the application concentration of casein phosphopeptide is 0.10-0.20 mg / mL, it can further promote the development of the protein backbone of the intestinal glycocalyx layer, as well as the synthesis of hyaluronic acid and heparan sulfate. For example, the application concentration of casein phosphopeptide can be 0.10 mg / mL, 0.11 mg / mL, 0.12 mg / mL, 0.13 mg / mL, 0.14 mg / mL, 0.15 mg / mL, 0.16 mg / mL, 0.17 mg / mL, 0.18 mg / mL, 0.19 mg / mL, 0.20 mg / mL, or any value between any two of the above ranges.

[0040] 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.

[0041] Experiments have shown that intervention with casein phosphopeptide 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.

[0042] It is understandable that casein phosphopeptides 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 glycan 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.

[0043] 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.

[0044] Experiments have verified that intervention with casein phosphopeptides 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. Intervention with casein phosphopeptides 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, intervention with casein phosphopeptides 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.

[0045] It is understandable that casein phosphopeptides 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.

[0046] Based on the above research, the second aspect of the present invention provides the application of casein phosphopeptide in the preparation of products related to promoting the development of the intestinal glycocalyx.

[0047] Furthermore, the products include at least one of the following: health supplements and medicines.

[0048] Health supplements may include at least one of the following: functional beverages, functional powders, functional capsules, and functional powders, which can improve the intestinal barrier, regulate the intestinal flora, or improve intestinal health.

[0049] The medication may include 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.

[0050] The aforementioned products also include carriers and / or physiologically acceptable excipients.

[0051] The carrier includes at least one of microcapsules, microspheres, nanoparticles, and liposomes.

[0052] 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.

[0053] 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.

[0054] 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.

[0055] 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.

[0056] Example 1: Experimental Grouping

[0057] 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 intervened according to the groups and intervention substances listed in Table 1. Specifically, the blank control group received no intervention and was cultured in MEM complete medium for 36 h; the low-dose casein phosphopeptide group (0.028 mg / mL) was treated by adding casein phosphopeptide to MEM complete medium to a final concentration of 0.028 mg / mL and culturing for 36 h; the medium-dose casein phosphopeptide group (0.112 mg / mL) was treated by adding casein phosphopeptide to MEM complete medium to a final concentration of 0.112 mg / mL and culturing for 36 h; and the high-dose casein phosphopeptide group (0.448 mg / mL) was treated by adding casein phosphopeptide to MEM complete medium to a final concentration of 0.448 mg / mL and culturing for 36 h.

[0058] Table 1

[0059]

[0060] Example 2: Real-time quantitative PCR (qPCR) experiment

[0061] 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).

[0062] Table 2

[0063]

[0064] Example 3: Effect of casein phosphopeptides on the protein backbone of the glycocalyx layer

[0065] 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).

[0066] 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 low-dose casein phosphopeptide group (0.028 mg / mL) and the medium-dose casein phosphopeptide group (0.112 mg / mL) were significantly higher than those in the control group (p<0.0001). These results indicate that casein phosphopeptide can upregulate gpc1 transcription levels, thereby promoting the development of the glycocalyx protein backbone and thus promoting the development of the glycocalyx layer.

[0067] Example 4: Effect of casein phosphopeptides on hyaluronic acid synthase in the glycocalyx layer

[0068] 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.

[0069] 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 group (0.112 mg / mL) and the high-dose group (0.448 mg / mL) of casein phosphopeptide was significantly higher than that in the blank control group (p<0.001). Figure 3 The results showed that the transcription level of the has2 gene in the medium-dose casein phosphopeptide group (0.112 mg / mL) was significantly higher than that in the blank control group (p<0.001), and the transcription level of the has2 gene in the high-dose casein phosphopeptide group (0.448 mg / mL) was significantly higher than that in the blank control group (p<0.01). Figure 4 The results showed that the transcriptional level of the has3 gene in the low-dose casein phosphopeptide group (0.028 mg / mL) was significantly higher than that in the control group (p<0.01), the transcriptional level in the medium-dose casein phosphopeptide group (0.112 mg / mL) was significantly higher than that in the control group (p<0.0001), and the transcriptional level in the high-dose casein phosphopeptide group (0.448 mg / mL) was significantly higher than that in the control group (p<0.001). These results indicate that casein phosphopeptide 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.

[0070] Example 5: Effect of casein phosphopeptide on heparan sulfate in glycocalyx layer

[0071] 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.

[0072] 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 transcriptional levels of the ext1 gene in the low-dose casein phosphopeptide group (0.028 mg / mL), the medium-dose casein phosphopeptide group (0.112 mg / mL), and the high-dose casein phosphopeptide group (0.448 mg / mL) were significantly higher than those 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 casein phosphopeptide group (0.112 mg / mL) was higher than that in the blank control group (p<0.05). These results indicate that casein phosphopeptide 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.

[0073] In summary, co-culturing Caco-2 cells with casein phosphopeptide during non-adherent growth can simulate the state of intestinal growth and maturation. Under this cellular state, casein phosphopeptide significantly promotes the development of the glycocalyx layer in Caco-2 cells, indicating that supplementing with casein phosphopeptide during the intestinal maturation stage can significantly promote the development of the intestinal glycocalyx layer, thereby promoting overall healthy intestinal development.

[0074] 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. The application of a casein phosphopeptide in the preparation of health products related to promoting the development of the intestinal glycocalyx layer.

2. The application according to claim 1, characterized in that, The application concentration of the casein phosphopeptide is 0.01-0.5 mg / mL.

3. The application according to claim 1 or 2, characterized in that, The application concentration of the casein phosphopeptide is 0.10-0.20 mg / mL.

4. 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.

5. The application according to claim 1, characterized in that, The health products include at least one of the following: functional beverages, functional powders, functional capsules, and functional powders, which improve the intestinal barrier, regulate intestinal flora, or improve intestinal health.

6. The application according to claim 5, characterized in that, The health products also include carriers 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, pH adjusters, and isotonic or isotropic modifiers.

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