Use of 2'-fucosyllactose in promoting intestinal development and preventing intestinal damage

Abstract

The use of 2'-fucosyllactose in the preparation of a food / drug for promoting intestinal glycocalyx development and preventing intestinal glycocalyx damage caused by lipopolysaccharide (LPS). The use of 2'-fucosyllactose in the preparation of a food / drug for the adjuvant treatment and / or adjuvant prevention of gastrointestinal infections, inflammatory bowel diseases, obesity or allergy caused by glycocalyx damage. Before adherent growth of Caco-2 cells, adding 2'-FL for co-culture with the Caco-2 cells can simulate the intestinal growth and maturation state. In the state, 2'-FL can significantly promote the development of the glycocalyx layer of Caco-2 cells, and can further prevent the glycocalyx layer damage caused by LPS on this basis.

Description

Application of 2′-fucosyllactose in promoting intestinal development and preventing intestinal damage

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application number 202410220178.0 filed with the China Patent Office on February 28, 2024, entitled “Application of 2'-fucosyllactose in promoting intestinal development and preventing intestinal damage”, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present application relates to the technical field of human milk oligosaccharides, and specifically to the application of 2'-fucosyllactose in promoting intestinal development and preventing intestinal damage. Background Art

[0004] The intestinal barrier is considered the gatekeeper of human health. It is responsible for the passage and absorption of nutrients, but also serves to prevent pathogens from entering the body. Furthermore, it regulates crosstalk between the intestinal luminal microbiota and macromolecules, and maintains tolerance and immune responses. After birth, this gatekeeping function of the intestinal barrier is not fully developed, and any disruption of the intestinal barrier function in neonates can have long-term consequences and may play a role in the development of gastrointestinal infections and inflammatory bowel disease (IBD). Furthermore, it influences the development of other diseases in childhood and adulthood, such as obesity and allergies.

[0005] Breast milk is the gold standard for infant nutrition, and human milk oligosaccharides (HMOs) are the primary components that distinguish it from other mammalian milks. In infant formula, a key function of HMOs is to stimulate microbial colonization of the gastrointestinal tract and promote the development of intestinal barrier function.

[0006] The glycocalyx of the neonatal intestinal epithelium provides binding sites for commensal microorganisms. A well-developed glycocalyx prevents the adhesion of pathogens and acts as a barrier to luminal toxins and enzymes. The glycocalyx is composed of glycans and proteins. Proteoglycans are generally considered the most important components of the glycocalyx and form the backbone of the glycocalyx. Furthermore, the glycocalyx contains glycosaminoglycan chains linked to the proteoglycan core protein. Heparan sulfate (HS) and hyaluronic acid (HA) are the main glycosaminoglycan components of the glycocalyx.

[0007] Currently, there are few clinical intervention studies on the direct regulatory effects of HMOs on intestinal cells and structures. Further research is necessary to lay a theoretical foundation for the future application of HMOs. Summary of the Invention

[0008] In view of this, this article provides the application of 2'-fucosyllactose in promoting intestinal glycocalyx development and preventing intestinal glycocalyx damage. This article explores the effects of 2'-fucosyllactose (2'-FL) on the development and function of intestinal epithelial cell Caco-2 glycocalyx at the cellular level.

[0009] In order to solve the technical problems raised in the background technology, this article adopts the following technical solutions:

[0010] In a first aspect, the present invention provides the use of 2'-fucosyllactose in the preparation of a food for promoting intestinal glycocalyx development and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0011] In one embodiment, the food includes at least one of infant formula milk powder, children's milk powder, adult milk powder, food for special medical purposes, nutritional products, liquid milk, complementary food, and non-alcoholic beverages.

[0012] In a second aspect, the present invention provides the use of 2'-fucosyllactose in the preparation of a medicament for promoting intestinal glycocalyx development and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0013] In a third aspect, provided herein is a use of 2'-fucosyllactose in promoting glycocalyx development in the intestine and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0014] In one embodiment, the drug further includes other acceptable carriers and / or excipients.

[0015] In one embodiment, the symptom promoting glycocalyx development in the intestine is selected from at least one of the following:

[0016] Protect the glycocalyx protein skeleton, maintain the normal synthesis of glycocalyx hyaluronic acid and maintain the normal synthesis of glycocalyx heparan sulfate.

[0017] In one embodiment, the expression of promoting glycocalyx development in the intestine is selected from at least one of the following: upregulating the transcription level of the glycocalyx development-related gene glypican 1, upregulating the transcription level of the hyaluronan synthase encoding gene, and upregulating the transcription level of the exosome glycosyltransferase encoding gene.

[0018] In one embodiment, the expression of promoting glycocalyx development in the intestine is selected from at least one of the following: upregulating the level of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA, and ext2 mRNA.

[0019] In one embodiment, the prevention of manifestations of glycocalyx damage caused by lipopolysaccharide (LPS) is selected from at least one of the following:

[0020] Prevent the damage of glycocalyx protein skeleton caused by LPS, prevent the damage of glycocalyx hyaluronan synthase caused by LPS and prevent the damage of glycocalyx heparan sulfate caused by LPS.

[0021] In one embodiment, the prevention of manifestations of glycocalyx damage caused by lipopolysaccharide (LPS) is selected from at least one of the following:

[0022] Up-regulation of the transcription level of glypican 1, a gene related to glycocalyx development, caused by LPS, up-regulation of the transcription level of the gene encoding hyaluronan synthase, caused by LPS, and up-regulation of the transcription level of the gene encoding exosome glycosyltransferase, caused by LPS.

[0023] In one embodiment, the manifestation of preventing glycocalyx damage caused by lipopolysaccharide (LPS) is selected from the group consisting of upregulating the decrease in the level of at least one of the following mRNAs caused by LPS:

[0024] gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA.

[0025] In one embodiment, when 2'-fucosyllactose is used to prepare a food for promoting intestinal glycocalyx development, the concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Alternatively, the concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL. Alternatively, the concentration of 2'-fucosyllactose is 1.2-2.4 mg / mL.

[0026] In one embodiment, the food includes at least one of infant formula milk powder, children's milk powder, adult milk powder, food for special medical purposes, nutritional products, liquid milk, complementary food, and non-alcoholic beverages.

[0027] In one embodiment, in the application of 2'-fucosyllactose in the preparation of a drug for promoting intestinal glycocalyx development, the application concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the application concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0028] In one embodiment, in the application of 2'-fucosyllactose in the preparation of food for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the application concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the application concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0029] In one embodiment, in the application of 2'-fucosyllactose in the preparation of a medicament for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the application concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the application concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0030] In a fourth aspect, the present invention provides the use of 2'-fucosyllactose in the preparation of food for the auxiliary treatment and / or auxiliary prevention of gastrointestinal infection, inflammatory bowel disease, obesity or allergy caused by glycocalyx damage.

[0031] In a fifth aspect, the present invention provides the use of 2'-fucosyllactose in the preparation of a medicament for treating and / or preventing gastrointestinal infection, inflammatory bowel disease, obesity or allergy caused by glycocalyx damage.

[0032] In a sixth aspect, the present invention provides the use of 2'-fucosyllactose in the adjuvant treatment and / or adjuvant prevention of gastrointestinal infection, inflammatory bowel disease, obesity or allergy caused by glycocalyx damage.

[0033] In one embodiment, in the application of 2'-fucosyllactose in the preparation of foods for the auxiliary treatment and / or auxiliary prevention of gastrointestinal infections and inflammatory bowel diseases, obesity and allergies caused by glycocalyx damage, the application concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the application concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0034] In one embodiment, in the application of 2'-fucosyllactose in the preparation of health foods for the auxiliary treatment and / or auxiliary prevention of gastrointestinal infections and inflammatory bowel diseases, obesity and allergies caused by glycocalyx damage, the application concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the application concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0035] In one embodiment, in the use of 2'-fucosyllactose in the preparation of a medicament for treating and / or preventing gastrointestinal infections and inflammatory bowel disease, obesity and allergies caused by glycocalyx damage, the concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0036] The beneficial effects of the above technical solution of this article are as follows:

[0037] This article provides an application of 2'-fucosyllactose. In this study, 2'-FL was added to Caco-2 cells during non-adherent growth to simulate the maturation phase of the intestine. Under these conditions, 2'-FL significantly promoted the development of the Caco-2 cell glycocalyx layer, further protecting against LPS-induced glycocalyx damage. This suggests that supplementing with a specific dose of 2'-FL during the maturation phase of the intestine can significantly promote the development of the intestinal glycocalyx layer and overall healthy intestinal development, protecting against damage from external factors such as LPS. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 shows the effects of different concentrations of 2'-FL on gpc1 mRNA transcription levels;

[0039] Figure 2 shows the effects of different concentrations of 2'-FL on the transcription levels of has1 mRNA, has2 mRNA, and has3 mRNA, where A shows the effects of different concentrations of 2'-FL on the transcription level of has1 mRNA, B shows the effects of different concentrations of 2'-FL on the transcription level of has2 mRNA, and C shows the effects of different concentrations of 2'-FL on the transcription level of has3 mRNA;

[0040] Figure 3 shows the effects of different concentrations of 2'-FL on the transcription levels of ext1 mRNA and ext2 mRNA, wherein A shows the effects of different concentrations of 2'-FL on the transcription level of ext1 mRNA, and B shows the effects of different concentrations of 2'-FL on the transcription level of ext2 mRNA;

[0041] Figure 4 shows the effects of different concentrations of 2'-FL and LPS on gpc1 mRNA transcription levels;

[0042] Figure 5 Effects of different concentrations of 2'-FL and LPS on the transcription levels of has1 mRNA, has2 mRNA, and has3 mRNA, where A shows the effects of different concentrations of 2'-FL and LPS on the transcription level of has1 mRNA, B shows the effects of different concentrations of 2'-FL and LPS on the transcription level of has2 mRNA, and C shows the effects of different concentrations of 2'-FL and LPS on the transcription level of has3 mRNA;

[0043] Figure 6 shows the effects of different concentrations of 2'-FL and LPS on the transcription levels of ext1 mRNA and ext2 mRNA, where A shows the effects of different concentrations of 2'-FL and LPS on the transcription level of ext1 mRNA, and B shows the effects of different concentrations of 2'-FL and LPS on the transcription level of ext2 mRNA. DETAILED DESCRIPTION

[0044] In order to further understand the present invention, preferred embodiments of the present invention are described below in conjunction with examples. However, it should be understood that these descriptions are only for further illustrating the features and advantages of the present invention, rather than limiting the present invention.

[0045] In a first aspect, the present invention provides the use of 2'-fucosyllactose in the preparation of a food for promoting intestinal glycocalyx development and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0046] According to some embodiments of the present invention, the food includes at least one of infant formula, children's milk powder, adult milk powder, food for special medical purposes, nutritional products, liquid milk, complementary food, and non-alcoholic beverages.

[0047] In a second aspect, the present invention provides the use of 2'-fucosyllactose in the preparation of a medicament for promoting intestinal glycocalyx development and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0048] According to some embodiments herein, the drug further comprises other acceptable carriers and / or excipients.

[0049] In a third aspect, provided herein is a use of 2'-fucosyllactose in promoting glycocalyx development in the intestine and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0050] In one embodiment, the symptom promoting glycocalyx development in the intestine is selected from at least one of the following:

[0051] Protect the glycocalyx protein skeleton, maintain the normal synthesis of glycocalyx hyaluronic acid and maintain the normal synthesis of glycocalyx heparan sulfate.

[0052] In one embodiment, the expression of promoting glycocalyx development in the intestine is selected from at least one of the following: upregulating the transcription level of the glycocalyx development-related gene glypican 1, upregulating the transcription level of the hyaluronan synthase encoding gene, and upregulating the transcription level of the exosome glycosyltransferase encoding gene.

[0053] In one embodiment, the expression of promoting glycocalyx development in the intestine is selected from at least one of the following: upregulating the level of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA, and ext2 mRNA.

[0054] In one embodiment, the prevention of manifestations of glycocalyx damage caused by lipopolysaccharide (LPS) is selected from at least one of the following:

[0055] Preventing LPS-induced damage to the glycocalyx protein skeleton, preventing LPS-induced damage to the glycocalyx hyaluronan synthase, and preventing LPS-induced damage to the glycocalyx heparan sulfate. In one embodiment, preventing manifestations of lipopolysaccharide (LPS)-induced glycocalyx damage includes, but is not limited to, restoring and maintaining the glycocalyx.

[0056] In one embodiment, the prevention of manifestations of glycocalyx damage caused by lipopolysaccharide (LPS) is selected from at least one of the following:

[0057] Up-regulation of the transcription level of glypican 1, a gene related to glycocalyx development, caused by LPS, up-regulation of the transcription level of the gene encoding hyaluronan synthase, caused by LPS, and up-regulation of the transcription level of the gene encoding exosome glycosyltransferase, caused by LPS.

[0058] In one embodiment, the manifestation of preventing glycocalyx damage caused by lipopolysaccharide (LPS) is selected from the group consisting of upregulating the decrease in the level of at least one of the following mRNAs caused by LPS:

[0059] gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA.

[0060] According to some embodiments of the present invention, in the application of 2'-fucosyllactose in the preparation of a food that promotes intestinal glycocalyx development, the application concentration of the 2'-fucosyllactose is 0.6 to 2.4 mg / mL. Optionally, the application concentration of the 2'-fucosyllactose is 0.6 to 1.2 mg / mL. Optionally, the application concentration of 2'-fucosyllactose is 1.2 to 2.4 mg / mL. 2'-fucosyllactose can greatly upregulate has1, has2, and has3 transcription levels at 0.6 mg / mL; 2'-fucosyllactose can greatly upregulate gpc1 transcription levels at 0.6 to 1.2 mg / mL; 2'-fucosyllactose can greatly upregulate ext1 and ext2 transcription levels at 0.6 mg / mL. 2'-fucosyllactose at 0.6-1.2 mg / mL can greatly prevent the down-regulation of gpc1 transcription levels caused by LPS, thereby protecting the glycocalyx protein skeleton; 2'-fucosyllactose at 0.6-2.4 mg / mL can greatly prevent the down-regulation of has2 and has3 transcription levels caused by LPS, thereby maintaining the synthesis of hyaluronic acid in the glycocalyx; 2'-fucosyllactose at 0.6-2.4 mg / mL can greatly prevent the down-regulation of ext1 and ext2 transcription levels caused by LPS, thereby maintaining the synthesis of heparan sulfate in the glycocalyx.

[0061] According to some embodiments of the present invention, in the application of 2'-fucosyllactose in the preparation of health foods for promoting intestinal glycocalyx development, the application concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the application concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0062] According to some embodiments of the present invention, in the application of 2'-fucosyllactose in the preparation of a drug for promoting intestinal glycocalyx development, the application concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the application concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0063] According to some embodiments herein, in the application of 2'-fucosyllactose in the preparation of food for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the application concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL, optionally, the application concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0064] According to some embodiments herein, in the application of 2'-fucosyllactose in the preparation of a health food for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the application concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the application concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0065] According to some embodiments herein, in the use of 2'-fucosyllactose in the preparation of a medicament for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the concentration of 2'-fucosyllactose is 0.6-2.4 mg / mL. Optionally, the concentration of 2'-fucosyllactose is 0.6-1.2 mg / mL.

[0066] In a fourth aspect, the present invention provides the use of 2'-fucosyllactose in the preparation of food for the auxiliary treatment and / or auxiliary prevention of gastrointestinal infections and inflammatory bowel diseases, obesity and allergies caused by glycocalyx damage.

[0067] According to some embodiments of the present invention, the food includes at least one of infant formula, children's milk powder, adult milk powder, food for special medical purposes, nutritional products, liquid milk, complementary food, and non-alcoholic beverages.

[0068] In a fifth aspect, the present invention provides the use of 2'-fucosyllactose in the preparation of a medicament for treating and / or preventing gastrointestinal infections and inflammatory bowel diseases, obesity and allergies caused by glycocalyx damage.

[0069] In a sixth aspect, the present invention provides the use of 2'-fucosyllactose in the adjuvant treatment and / or adjuvant prevention of gastrointestinal infection, inflammatory bowel disease, obesity or allergy caused by glycocalyx damage.

[0070] The inflammatory bowel disease is selected from any one of acute colitis, ulcerative colitis, Crohn's disease, microscopic colitis, diversion colitis, Behçet's disease immuno-oncology colitis, chemotherapy or radiation colitis, graft-versus-host disease colitis, collagenous colitis, lymphocytic colitis and pouchitis.

[0071] According to some embodiments herein, in the use of 2'-fucosyllactose in preparing foods for the adjuvant treatment and / or adjuvant prevention of gastrointestinal infections and inflammatory bowel diseases, obesity or allergies caused by glycocalyx damage, the concentration of 2'-fucosyllactose is 0.6 to 2.4 mg / mL, optionally, the concentration of 2'-fucosyllactose is 0.6 to 1.2 mg / mL.

[0072] According to some embodiments herein, in the use of 2'-fucosyllactose in the preparation of health foods for the auxiliary treatment and / or auxiliary prevention of gastrointestinal infections and inflammatory bowel diseases, obesity or allergies caused by glycocalyx damage, the concentration of 2'-fucosyllactose is 0.6 to 2.4 mg / mL, optionally, the concentration of 2'-fucosyllactose is 0.6 to 1.2 mg / mL.

[0073] According to some embodiments herein, in the use of 2'-fucosyllactose in the preparation of a medicament for treating and / or preventing gastrointestinal infections, inflammatory bowel disease, obesity or allergies caused by glycocalyx damage, the concentration of 2'-fucosyllactose is 0.6 to 2.4 mg / mL. Optionally, the concentration of 2'-fucosyllactose is 0.6 to 1.2 mg / mL.

[0074] The present invention will be further described below through some specific examples.

[0075] The raw materials used in the following examples are as follows:

[0076] Human colon adenocarcinoma cell line Caco-2: purchased from Nanjing Senbeijia Biotechnology Co., Ltd.

[0077] MEM complete medium: purchased from Nanjing Senbeijia Biotechnology Co., Ltd.

[0078] Modified MRS medium: purchased from Haibo Biotechnology Co., Ltd.;

[0079] Lipopolysaccharide (LPS): purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0080] Caco-2 cell culture: Human colon adenocarcinoma cells Caco-2 were cultured in MEM complete medium containing 20% ​​high-quality fetal bovine serum and grown at 25 cm 2Culture the cells in a flask in a CO2 incubator at 37°C, 5% CO2, and 95% relative humidity. Change the culture medium every one or two days. When the cells reach 80%-90% confluency, digest them with 0.25% trypsin solution and subculture at a 1:3 ratio.

[0081] Example 1

[0082] Effects of 2'-FL on the development of intestinal epithelial cell glycocalyx

[0083] 1.1 Experiment: Adjust the Caco-2 cell density to 1×10 5 pieces / cm 2 Cells were seeded in 24-well plates and incubated for 3 days in complete MEM medium with 0.6 mg / mL, 1.2 mg / mL, and 2.4 mg / mL of 2'-FL. The cells were then incubated for 3 days using MEM medium as a blank control. Total RNA was extracted and reverse transcribed, and the transcript levels of genes involved in glycocalyx development, such as glypican 1 (gpc1), hyaluronan synthases (has1, has2, has3), and exosome glycosyltransferases (ext1, ext2), were measured by qPCR. qPCR primer sequences are shown in Table 1.

[0084] The qPCR reaction kit was manufactured by Nanjing Novozymes Biotechnology Co., Ltd.; model: AceQ Universal U+Probe Master Mix V2. The amplification protocol was as follows: Step 1: 37°C for 2 min, cycle 1; Step 2: 95°C for 5 min, cycle 1; Step 3: 95°C for 10 s, 60°C for 30 s, cycle 45.

[0085] Table 1

[0086] 1.2 Test Results

[0087] 1.2.1 Effects of different concentrations of 2'-FL on the protein backbone of the glycocalyx

[0088] gpc1 is the gene encoding the glypican 1 (GPC1) proteoglycan. GPC1 serves as the protein backbone of the glycocalyx and is an important carrier of glycosaminoglycan chains, including HA and heparan sulfate (HS). HA is a highly viscous component of the intestinal mucus layer, responsible for tissue repair, stability, and anti-inflammatory effects. Figure 1 shows the effects of different concentrations of 2'-FL on gpc1 mRNA transcription levels. As shown in Figure 1, all three selected intervention concentrations of 2'-FL significantly upregulated gpc1 transcription levels (p < 0.05). With increasing 2'-FL concentration, the upregulation of gpc1 transcription levels initially increased and then decreased, reaching a maximum at 1.2 mg / mL.

[0089] 1.2.2 Effects of different concentrations of 2'-FL on hyaluronan synthase in the glycocalyx

[0090] has1, has2, and has3 are genes encoding hyaluronic acid (HA) synthase. HAS is crucial for the synthesis of HA, a highly viscous component of the intestinal mucus layer responsible for tissue repair, stability, and anti-inflammatory effects. The main function of HAS1 and HAS2 is to catalyze the polymerization reaction between glucosamine and UDP-glucose to synthesize hyaluronic acid. Hyaluronic acid plays an important structural and biological function in the extracellular matrix, including intercellular signaling, cell migration, and tissue repair. Compared with HAS1 and HAS2, HAS3 synthesizes low-molecular-weight HA, which is essential for the development of intestinal stem cells. Figure 2 shows the effects of different concentrations of 2'-FL on the transcription levels of has 1 mRNA, has2 mRNA, and has3 mRNA, where A shows the effects of different concentrations of 2'-FL on the transcription levels of has1 mRNA, B shows the effects of different concentrations of 2'-FL on the transcription levels of has2 mRNA, and C shows the effects of different concentrations of 2'-FL on the transcription levels of has3 mRNA. As can be seen from Figure 2, the three selected intervention concentrations of 2'-FL can significantly upregulate the transcription levels of has1, has2 and has3 (p < 0.05). The upregulation effect of 2'-FL on the transcription levels of has1, has2 and has3 decreases with increasing concentration and reaches the maximum value at 0.6 mg / mL.

[0091] 1.2.3 Effects of different concentrations of 2'-FL on heparan sulfate

[0092] The ext1 and ext2 genes are involved in the elongation of heparan sulfate (HS) chains and are responsible for integrating HS chains into nucleotide sugars in the Golgi apparatus. Heparan sulfate (HS) and hyaluronic acid (HA) are the major glycosaminoglycan components of the glycocalyx. HS synthesis supports organogenesis, growth factor signaling, and bacterial adhesion. Figure 3 shows the effects of different concentrations of 2'-FL on the transcript levels of ext1 and ext2 mRNA. (A) Shows the effect of different concentrations of 2'-FL on the transcript levels of ext1 mRNA, and (B) Shows the effect of different concentrations of 2'-FL on the transcript levels of ext2 mRNA. As shown in Figure 3, all three selected intervention concentrations of 2'-FL significantly upregulated the transcript levels of ext1 and ext2 (p < 0.05). The upregulatory effect of 2'-FL decreased with increasing concentration, reaching a maximum at 0.6 mg / mL.

[0093] Example 2

[0094] Preventive effect of 2'-FL on LPS-induced glycocalyx damage

[0095] 2.1 Experiment: Adjust the Caco-2 cell density to 1×10 5 pieces / cm 2 Cells were seeded in 24-well plates and incubated in MEM complete medium at concentrations of 0.6 mg / mL, 1.2 mg / mL, and 2.4 mg / mL, respectively. After 3 days of incubation, the supernatant was removed and fresh MEM complete medium with LPS at a final concentration of 100 ng / mL was added for 24 hours. Cells supplemented with LPS but without 2'-FL served as the model group, while cells supplemented with neither LPS nor 2'-FL served as the blank group. Total RNA was extracted and reverse transcribed, and transcript levels of genes involved in glycocalyx development, glypican1 (gpc1), hyaluronan synthases (has1, has2, has3), and exosome glycosyltransferases (ext1, ext2), were measured in Caco-2 cells using qPCR. For qPCR primer sequences, see Table 1 above.

[0096] Lipopolysaccharide (LPS) is a molecule found in bacterial cell walls that can trigger an immune response and cause inflammation. When LPS enters the body, it may activate the immune system and lead to an inflammatory response. In certain cases, LPS may cause an inflammatory response that damages the glycocalyx (a complex of glycoproteins, sugars, and proteins).

[0097] 2.2 Test Results

[0098] 2.2.1 Preventive effect of different concentrations of 2'-FL on glycocalyx protein skeleton damage

[0099] Figure 4 shows the effects of different concentrations of 2'-FL and LPS on gpc1 mRNA transcription levels. As shown in Figure 4, after LPS intervention, gpc1 mRNA transcription levels in the model group were downregulated by 2-fold compared to the blank group, indicating that LPS may damage the glycocalyx protein skeleton by downregulating gpc1 transcription levels. After Caco-2 cells were co-cultured with different concentrations of 2'-FL for 3 days and then treated with LPS, gpc1 transcription levels were still upregulated compared to the blank group. 2'-FL concentrations of 0.6 mg / mL and 1.2 mg / mL significantly upregulated gpc1 transcription, indicating that 2'-FL can prevent the LPS-induced downregulation of gpc1 transcription levels and thereby protect the glycocalyx protein skeleton.

[0100] 2.2.2 Preventive effect of 2'-FL on LPS-induced damage of hyaluronan synthase in the glycocalyx of Caco-2 cells

[0101] Figure 5 Effects of different concentrations of 2'-FL and LPS on the transcription levels of has1 mRNA, has2 mRNA, and has3 mRNA, where A shows the effect of different concentrations of 2'-FL and LPS on the transcription level of has1 mRNA, B shows the effect of different concentrations of 2'-FL and LPS on the transcription level of has2 mRNA, and C shows the effect of different concentrations of 2'-FL and LPS on the transcription level of has3 mRNA.

[0102] As can be seen from Figure 5, after LPS intervention, the has1 transcription levels in the model group and different concentrations of 2'-FL prevention groups were significantly downregulated compared with the blank group (p < 0.05), indicating that LPS may damage the hyaluronan synthase 1 of Caco-2 cells by downregulating the transcription level of has1, and the has1 transcription level after 2'-FL prevention was still significantly different from that of normal cells (p < 0.05), but after 2.4 mg / mL 2'-FL prevention, the has1 transcription level was significantly upregulated compared with the model group, indicating that 2.4 mg / mL 2'-FL can prevent the decrease in has1 transcription level caused by LPS.

[0103] After LPS treatment, the transcription levels of has2 and has3 in the model group decreased by 2-fold compared to the blank group, indicating that LPS may damage hyaluronan synthases 2 and 3 in Caco-2 cells by downregulating their transcription levels. There were no significant differences in the transcription levels of has2 and has3 between the 2'-FL treatment groups at different concentrations and the blank group (p>0.05). Furthermore, treatment with 2.4 mg / mL 2'-FL significantly upregulated the transcription levels of has2 and has3 compared to the model group (p<0.05), indicating that 2'-FL can prevent the LPS-induced downregulation of has2 and has3 transcription levels, thereby maintaining glycocalyx hyaluronan synthesis.

[0104] 2.2.3 Preventive effect of 2'-FL on LPS-induced heparan sulfate damage in the glycocalyx of Caco-2 cells

[0105] Figure 6 shows the effects of different concentrations of 2'-FL and LPS on the transcription levels of ext1 mRNA and ext2 mRNA, where A shows the effects of different concentrations of 2'-FL and LPS on the transcription level of ext1 mRNA, and B shows the effects of different concentrations of 2'-FL and LPS on the transcription level of ext2 mRNA.

[0106] As shown in Figure 6, after LPS intervention, the transcription levels of ext1 and ext2 in the model group were downregulated by nearly 2-fold compared with the blank group, indicating that LPS may damage the glycocalyx heparan sulfate by downregulating the transcription levels of ext1 and ext2. After Caco-2 cells were co-cultured with different concentrations of 2'-FL for 3 days and then treated with LPS, the transcription levels of ext1 and ext2 were still upregulated compared with the blank group. Among them, the three concentrations of 2'-FL intervention groups significantly upregulated the transcription level of ext1 (p < 0.05), and the 1.2 mg / mL and 2.4 mg / mL 2'-FL intervention groups significantly upregulated the transcription level of ext2 (p < 0.05), indicating that 2'-FL can prevent the downregulation of ext1 and ext2 transcription levels caused by LPS, thereby maintaining the synthesis of glycocalyx heparan sulfate.

[0107] The above is the preferred embodiment of this invention. It should be noted that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles described in this invention. These improvements and modifications should also be considered as the scope of protection of this invention. Industrial Applicability

[0108] This article provides an application of 2'-fucosyllactose. In this study, 2'-FL was added to Caco-2 cells during non-adherent growth to simulate the maturation phase of the intestine. Under these conditions, 2'-FL significantly promoted the development of the Caco-2 cell glycocalyx layer, further protecting against LPS-induced glycocalyx damage. This suggests that supplementing with a specific dose of 2'-FL during the maturation phase of the intestine can significantly promote the development of the intestinal glycocalyx layer and overall healthy intestinal development, protecting against damage from external factors such as LPS.

Claims

1. Use of 2'-fucosyllactose in the preparation of food for promoting intestinal glycocalyx development and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

2. Use of 2'-fucosyllactose in the preparation of a medicament for promoting intestinal glycocalyx development and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

3. Use of 2'-fucosyllactose in promoting intestinal glycocalyx development and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

4. The use according to any one of claims 1 to 3, characterized in that The performance of promoting the development of glycocalyx in the intestine is selected from at least one of the following: Protect the glycocalyx protein skeleton, maintain the normal synthesis of glycocalyx hyaluronic acid and maintain the normal synthesis of glycocalyx heparan sulfate.

5. The use according to any one of claims 1 to 4, characterized in that The performance of promoting glycocalyx development in the intestine is selected from at least one of the following: upregulating the transcription level of the glycocalyx development-related gene glypican 1, upregulating the transcription level of the hyaluronan synthase encoding gene, and upregulating the transcription level of the exosome glycosyltransferase encoding gene.

6. The use according to any one of claims 1 to 5, characterized in that The performance of promoting glycocalyx development in the intestine is selected from at least one of the following: upregulating the level of gpc1 mRNA, the level of has1 mRNA, the level of has2 mRNA, the level of has3 mRNA, the level of ext1 mRNA and the level of ext2 mRNA.

7. The use according to any one of claims 1 to 3, characterized in that The manifestation of preventing glycocalyx damage caused by lipopolysaccharide (LPS) is selected from at least one of the following: Prevent the damage of glycocalyx protein skeleton caused by LPS, prevent the damage of glycocalyx hyaluronan synthase caused by LPS and prevent the damage of glycocalyx heparan sulfate caused by LPS.

8. The use according to any one of claims 1 to 3 or claim 7, characterized in that: The manifestation of preventing glycocalyx damage caused by lipopolysaccharide (LPS) is selected from at least one of the following: Up-regulation of the transcription level of glypican 1, a gene related to glycocalyx development, caused by LPS, up-regulation of the transcription level of the gene encoding hyaluronan synthase, caused by LPS, and up-regulation of the transcription level of the gene encoding exosome glycosyltransferase, caused by LPS.

9. The use according to any one of claims 1 to 6 or any one of claims 7 to 8, characterized in that: The manifestation of preventing glycocalyx damage caused by lipopolysaccharide (LPS) is selected from the group consisting of upregulating the decrease in the level of at least one of the following mRNAs caused by LPS: gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA.

10. The use according to any one of claims 1 to 9, characterized in that The application concentration of the 2'-fucosyllactose is 0.6-2.4 mg / mL.

11. The use according to any one of claims 1 to 9, characterized in that The application concentration of the 2'-fucosyllactose is 0.6-1.2 mg / mL.

12. The use according to any one of claims 1 to 9, characterized in that The application concentration of the 2'-fucosyllactose is 1.2-2.4 mg / mL.

13. Use of 2'-fucosyllactose in the preparation of food for the auxiliary treatment and / or auxiliary prevention of gastrointestinal infection, inflammatory bowel disease, obesity or allergy caused by glycocalyx damage.

14. The use according to claim 1 or 13, characterized in that The food is selected from at least one of infant formula milk powder, children's milk powder, adult milk powder, food for special medical purposes, nutritional products, liquid milk, complementary food and non-alcoholic beverages.

15. The use according to any one of claims 13-14, characterized in that The application concentration of the 2'-fucosyllactose is 0.6-2.4 mg / mL.

16. Use of 2'-fucosyllactose in the adjuvant treatment and / or adjuvant prevention of gastrointestinal infections, inflammatory bowel diseases, obesity or allergies caused by glycocalyx damage.

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