Application of 2 '-fucosyllactose in promoting intestinal development and preventing intestinal injury

Abstract

The invention provides the application of the 2 '-fucosyllactose in promoting intestinal development and preventing intestinal injury. According to the invention, when Caco-2 cells do not grow in an adherent manner, 2 '-FL is added and co-cultured with the Caco-2 cells, so that the state of mature intestinal growth can be simulated, the 2'-FL can significantly promote the development of the glycocalyx layer of the Caco-2 cells in the state, and on this basis, the damage to the glycocalyx layer caused by LPS can be further prevented. It is shown that by supplementing a specific dosage of 2 '-FL in the intestinal mature stage, development of the intestinal glycocalyx layer can be remarkably promoted, overall healthy development of the intestinal tract can be promoted, and invasion of external factors such as LPS can be prevented.

Description

Technical Field

[0001] The invention relates to the technical field of human milk oligosaccharides, and in particular to the application of 2'-fucosyllactose in promoting intestinal development and preventing intestinal damage. Background Art

[0002] The intestinal barrier is considered the gatekeeper of human health. The intestinal barrier is responsible for the passage and absorption of nutrients, but it also plays a role in preventing pathogens from entering the body. In addition, it has the function of regulating the crosstalk between the intestinal luminal microbiota and macromolecules, and maintaining tolerance and immune response. After birth, this gatekeeping function of the intestinal barrier is not fully developed, and any disruption of the intestinal barrier function in the newborn will have long-term effects and may play a role in the development of gastrointestinal infections and inflammatory bowel disease (IBD). In addition, it also affects the development of other diseases in childhood and adulthood, such as obesity and allergies.

[0003] Breast milk is the gold standard for infant nutrition, and human milk oligosaccharides (HMOs) are the main components that distinguish breast milk from other mammalian milk. In infant formula, an important function of HMOs is to stimulate the colonization of the gastrointestinal tract by microbiota and promote the development of intestinal barrier function.

[0004] The glycocalyx on the neonatal intestinal epithelium provides binding sites for commensal microorganisms, and a well-developed glycocalyx can prevent the adhesion of pathogens and serve as a barrier to luminal toxins and enzymes. The glycocalyx is composed of glycans and proteins. Proteoglycans are generally considered to be the most important components of the glycocalyx and form the skeleton of the glycocalyx. In addition, the glycocalyx contains glycosaminoglycan chains connected to the core protein of the proteoglycan, among which heparan sulfate (HS) and hyaluronic acid (HA) are the main glycosaminoglycan components in the glycocalyx.

[0005] 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

[0006] In view of this, the present invention provides the use of 2′-fucosyllactose in promoting intestinal glycocalyx development and preventing intestinal glycocalyx damage. The present invention explores the effect of 2′-fucosyllactose (2'-FL) on the development and function of intestinal epithelial cell Caco-2 glycocalyx at the cellular level.

[0007] In order to solve the technical problems raised in the background technology, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides the use of 2′-fucosyllactose in the preparation of food for promoting intestinal glycocalyx development and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0009] Furthermore, 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.

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

[0011] In a third aspect, the present invention provides the use of 2′-fucosyllactose in the preparation of a medicine for promoting the development of intestinal glycocalyx and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0012] Furthermore, the medicine also includes other acceptable carriers and / or excipients.

[0013] Furthermore, the manifestation of promoting intestinal glycocalyx development includes: upregulating the transcription level of at least one of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA.

[0014] Furthermore, the prevention of glycocalyx damage caused by lipopolysaccharide (LPS) includes: upregulating the decrease in transcription level of at least one of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA caused by LPS.

[0015] Furthermore, in the application of 2′-fucosyllactose in the preparation of food for promoting intestinal glycocalyx development, the application concentration of the 2′-fucosyllactose is 0.6-2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6-1.2 mg / mL.

[0016] Furthermore, 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.

[0017] Furthermore, in the application of 2′-fucosyllactose in the preparation of health food for promoting intestinal glycocalyx development, the application concentration of the 2′-fucosyllactose is 0.6-2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6-1.2 mg / mL.

[0018] Furthermore, in the application of 2′-fucosyllactose in the preparation of a medicine for promoting intestinal glycocalyx development, the application concentration of the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0019] Furthermore, in the application of 2′-fucosyllactose in the preparation of food for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the application concentration of the 2′-fucosyllactose is 0.6-2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6-1.2 mg / mL.

[0020] Furthermore, in the application of 2′-fucosyllactose in the preparation of health food for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the application concentration of 2′-fucosyllactose is 0.6-2.4 mg / mL, preferably, the application concentration of 2′-fucosyllactose is 0.6-1.2 mg / mL.

[0021] Furthermore, in the application of 2′-fucosyllactose in the preparation of a medicine for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the application concentration of the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0022] 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 and inflammatory bowel disease, obesity and allergy caused by glycocalyx damage.

[0023] In a fifth aspect, the present invention provides the use of 2′-fucosyllactose in the preparation of health food for the auxiliary treatment and / or auxiliary prevention of gastrointestinal infection and inflammatory bowel disease, obesity and allergy caused by glycocalyx damage.

[0024] In a sixth aspect, the present invention provides the use of 2′-fucosyllactose in the preparation of a medicine for treating and / or preventing gastrointestinal infections and inflammatory bowel diseases, obesity and allergies caused by glycocalyx damage.

[0025] Furthermore, in the application of the 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 the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0026] Furthermore, in the application of the 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 the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0027] Furthermore, in the application of the 2′-fucosyllactose in the preparation of medicines for treating and / or preventing gastrointestinal infections and inflammatory bowel diseases, obesity and allergies caused by glycocalyx damage, the application concentration of the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0028] The beneficial effects of the above technical solution of the present invention are as follows:

[0029] The present invention provides an application of 2'-fucosyllactose. In the present invention, 2'-FL is added to Caco-2 cells when they are not attached to the wall for co-culture, which can simulate the state of intestinal growth and maturation. In this state, 2'-FL can significantly promote the development of the glycocalyx layer of Caco-2 cells, and on this basis, it can further prevent the damage of the glycocalyx layer caused by LPS. It shows that supplementing a specific dose of 2'-FL in the intestinal maturation stage can significantly promote the development of the intestinal glycocalyx layer and promote the overall healthy development of the intestine, and prevent the damage of external factors such as LPS. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The effect of different concentrations of 2'-FL on the transcription level of gpc1 mRNA;

[0031] Figure 2 The effects of different concentrations of 2'-FL on the transcription levels of has1 mRNA, has2 mRNA and has3 mRNA, where A is the effect of different concentrations of 2'-FL on the transcription level of has1 mRNA, B is the effect of different concentrations of 2'-FL on the transcription level of has2 mRNA, and C is the effect of different concentrations of 2'-FL on the transcription level of has3 mRNA;

[0032] Figure 3 The effects of different concentrations of 2'-FL on the transcription levels of ext1 mRNA and ext2 mRNA, where A is the effect of different concentrations of 2'-FL on the transcription level of ext1 mRNA, and B is the effect of different concentrations of 2'-FL on the transcription level of ext2 mRNA;

[0033] Figure 4The effect of different concentrations of 2'-FL and LPS on the transcription level of gpc1 mRNA;

[0034] 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 is the effect of different concentrations of 2'-FL and LPS on the transcription level of has1 mRNA, B is the effect of different concentrations of 2'-FL and LPS on the transcription level of has2 mRNA, and C is the effect of different concentrations of 2'-FL and LPS on the transcription level of has3 mRNA;

[0035] Figure 6 The effects of different concentrations of 2'-FL and LPS on the transcription levels of ext1 mRNA and ext2 mRNA, where A is the effect of different concentrations of 2'-FL and LPS on the transcription level of ext1 mRNA, and B is the effect of different concentrations of 2'-FL and LPS on the transcription level of ext2 mRNA; DETAILED DESCRIPTION

[0036] 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 for limiting the present invention.

[0037] In a first aspect, the present invention provides the use of 2′-fucosyllactose in the preparation of food for promoting intestinal glycocalyx development and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0038] According to some embodiments of the present invention, 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.

[0039] In a second aspect, the present invention provides the use of 2′-fucosyllactose in the preparation of a health food for promoting the development of intestinal glycocalyx and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0040] In a third aspect, the present invention provides the use of 2′-fucosyllactose in the preparation of a medicine for promoting the development of intestinal glycocalyx and / or preventing glycocalyx damage caused by lipopolysaccharide (LPS).

[0041] According to some embodiments of the present invention, the medicine further includes other acceptable carriers and / or excipients.

[0042] According to some embodiments of the present invention, the performance of promoting intestinal glycocalyx development includes: upregulating the transcription level of at least one of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA. Preferably, the 2′-fucosyllactose can upregulate the transcription level of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA.

[0043] According to some embodiments of the present invention, the manifestation of preventing glycocalyx damage caused by lipopolysaccharide (LPS) includes: upregulating the decrease in the transcription level of at least one of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA caused by LPS. Preferably, the 2′-fucosyllactose can upregulate the decrease in the transcription level of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA caused by LPS.

[0044] According to some embodiments of the present invention, in the application of 2′-fucosyllactose in the preparation of food for promoting intestinal glycocalyx development, the application concentration of the 2′-fucosyllactose is 0.6-2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6-1.2 mg / mL.

[0045] According to some embodiments of the present invention, in the application of 2′-fucosyllactose in the preparation of a health food for promoting intestinal glycocalyx development, the application concentration of the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0046] According to some embodiments of the present invention, in the application of 2′-fucosyllactose in the preparation of a medicine for promoting intestinal glycocalyx development, the application concentration of the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0047] According to some embodiments of the present invention, in the application of 2′-fucosyllactose in the preparation of food for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the application concentration of the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0048] According to some embodiments of the present invention, 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 the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0049] According to some embodiments of the present invention, in the application of 2′-fucosyllactose in the preparation of a medicine for preventing glycocalyx damage caused by lipopolysaccharide (LPS), the application concentration of the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0050] 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 and inflammatory bowel disease, obesity and allergy caused by glycocalyx damage.

[0051] According to some embodiments of the present invention, 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.

[0052] In a fifth aspect, the present invention provides the use of 2′-fucosyllactose in the preparation of health food for the auxiliary treatment and / or auxiliary prevention of gastrointestinal infection and inflammatory bowel disease, obesity and allergy caused by glycocalyx damage.

[0053] In a sixth aspect, the present invention provides the use of 2′-fucosyllactose in the preparation of a medicine for treating and / or preventing gastrointestinal infections and inflammatory bowel diseases, obesity and allergies caused by glycocalyx damage.

[0054] According to some embodiments of the present invention, in the application of the 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 the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0055] According to some embodiments of the present invention, in the application of the 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 the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0056] According to some embodiments of the present invention, in the use of the 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 application concentration of the 2′-fucosyllactose is 0.6 to 2.4 mg / mL, preferably, the application concentration of the 2′-fucosyllactose is 0.6 to 1.2 mg / mL.

[0057] The present invention will be further described below through some specific embodiments.

[0058] The raw materials used in the following examples are as follows:

[0059] Human colon adenocarcinoma cell Caco-2: purchased from Nanjing Senbeijia Biotechnology Co., Ltd.

[0060] MEM complete medium: purchased from Nanjing Senbeijia Biotechnology Co., Ltd.;

[0061] Modified MRS medium: purchased from Haibo Biotechnology Co., Ltd.;

[0062] Lipopolysaccharide (LPS): purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0063] 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 2 The cell culture flask was placed in a carbon dioxide incubator at 37°C, 5% CO2, and 95% relative humidity. The culture medium was changed every other day or two. When the cells grew to 80% to 90% confluence, they were digested with 0.25% trypsin digestion solution and subcultured at a ratio of 1:3.

[0064] Example 1

[0065] Effects of 2'-FL on the development of intestinal epithelial cell glycocalyx

[0066] 1.1 Experiment: Adjust the Caco-2 cell density to 1×10 5 Pieces / cm 2 , inoculated in 24-well plates, and 2'-FL was added to the MEM complete medium at concentrations of 0.6 mg / mL, 1.2 mg / mL, and 2.4 mg / mL according to the grouping. After incubation for 3 days, MEM medium was used as a blank control. Total cell RNA was extracted and reverse transcribed, and qPCR was used to measure the transcription levels of glycocalyx development-related genes glypican 1 (gpc1), hyaluronan synthase 1 (has1, has2, has3), and exosome glycosyltransferase 1 (ext1, ext2) in Caco-2 cells. The qPCR primer sequences are shown in Table 1.

[0067] Table 1

[0068]

[0069]

[0070] 1.2 Test Results

[0071] 1.2.1 Effects of different concentrations of 2'-FL on the protein backbone of the glycocalyx

[0072] GPC1 is the gene encoding the Glypican 1 (GPC1) proteoglycan, which is the protein backbone of the glycocalyx and 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 The effect of different concentrations of 2'-FL on the transcription level of gpc1 mRNA is shown in Figure 1 It can be seen that the three selected intervention concentrations of 2'-FL can significantly upregulate the transcription level of gpc1 (p < 0.05). As the concentration of 2'-FL increases, the upregulation of the transcription level of gpc1 shows a trend of first increasing and then decreasing, reaching a maximum value at 1.2 mg / mL.

[0073] 1.2.2 Effects of different concentrations of 2'-FL on hyaluronan synthase in glycocalyx

[0074] HAS1, HAS2, and HAS3 are genes encoding hyaluronic acid (HA) synthase. HAS is essential for the synthesis of HA, a highly viscous component of the intestinal mucus layer that is 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 necessary for the development of intestinal stem cells. Figure 2 The effects of different concentrations of 2'-FL on the transcription levels of has1mRNA, has2mRNA and has3mRNA, where A is the effect of different concentrations of 2'-FL on the transcription level of has1mRNA, B is the effect of different concentrations of 2'-FL on the transcription level of has2mRNA, and C is the effect of different concentrations of 2'-FL on the transcription level of has3mRNA. Figure 2It can be seen that 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.

[0075] 1.2.3 Effects of different concentrations of 2'-FL on heparan sulfate

[0076] The ext1 and ext2 genes are involved in the elongation of heparan sulfate (HS) chains and are responsible for the integration of 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 The effects of different concentrations of 2'-FL on the transcription levels of ext1 mRNA and ext2 mRNA, where A is the effect of different concentrations of 2'-FL on the transcription level of ext1 mRNA, and B is the effect of different concentrations of 2'-FL on the transcription level of ext2 mRNA. Figure 3 It can be seen that the three selected intervention concentrations of 2'-FL can significantly upregulate the transcription levels of ext1 and ext2 (p < 0.05), and the upregulation effect of 2'-FL decreases with increasing concentration and reaches the maximum value at 0.6 mg / mL.

[0077] Example 2

[0078] Preventive effect of 2'-FL on glycocalyx damage induced by LPS

[0079] 2.1 Experiment: Adjust the Caco-2 cell density to 1×10 5 Pieces / cm 2 , seeded in 24-well plates, and 2'-FL was added to the MEM complete medium at concentrations of 0.6 mg / mL, 1.2 mg / mL, and 2.4 mg / mL according to the grouping. After 3 days of co-incubation, the culture supernatant was removed, and fresh MEM complete medium with a final concentration of 100 ng / mL LPS was added for 24 hours. The cells with LPS but no 2'-FL were used as the model group, and the cells without LPS and 2'-FL were used as the blank group. Total cell RNA was extracted and reverse transcribed, and the transcription levels of glycocalyx development-related genes glypican 1 (gpc1), hyaluronan synthase 1 (has1, has2, has3), and exosome glycosyltransferase 1 (ext1, ext2) in Caco-2 cells were measured by qPCR. The qPCR primer sequences are shown in Table 1 above.

[0080] LPS (lipopolysaccharide) is a molecule found in bacterial cell walls that can cause an immune response and lead to inflammation. When LPS enters the body, it may activate the immune system, leading to an inflammatory response. In some cases, LPS may cause an inflammatory response that causes damage to the glycocalyx (a complex of glycoproteins, sugars and proteins).

[0081] 2.2 Test Results

[0082] 2.2.1 Effect of different concentrations of 2'-FL on the prevention of glycocalyx protein skeleton damage

[0083] Figure 4 The effect of different concentrations of 2'-FL and LPS on the transcription level of gpc1 mRNA is shown in Figure 4 It can be seen that after LPS intervention, the transcription level of gpc1 mRNA in the model group was downregulated by 2 times compared with the blank group, indicating that LPS may damage the glycocalyx protein skeleton by downregulating the transcription level of gpc1. After Caco-2 cells were co-cultured with different concentrations of 2'-FL for 3 days and then intervened with LPS, it was found that the transcription level of gpc1 was still upregulated compared with the blank group, among which 2'-FL at concentrations of 0.6 mg / mL and 1.2 mg / mL was significantly upregulated, indicating that 2'-FL can prevent the downregulation of gpc1 transcription level caused by LPS, thereby protecting the glycocalyx protein skeleton.

[0084] 2.2.2 Preventive effect of 2'-FL on LPS-induced damage of hyaluronan synthase in the glycocalyx of Caco-2 cells

[0085] 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.

[0086] Depend on Figure 5 It can be seen that 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 transcription level of has1 after 2'-FL prevention was still significantly different from that of normal cells (p < 0.05), but after 2.4 mg / mL2'-FL prevention, the has1 transcription level was significantly upregulated compared with the model group, indicating that 2.4 mg / mL2'-FL can prevent the decrease in has1 transcription level caused by LPS.

[0087] After LPS intervention, the transcription levels of has2 and has3 in the model group decreased by 2 times compared with the blank group, indicating that LPS may damage the hyaluronan synthase 2 and 3 of Caco-2 cells by downregulating the transcription levels of has2 and has3. There was no significant difference in the transcription levels of has2 and has3 between the prevention groups with different concentrations of 2'-FL and the blank group (p>0.05), and the transcription levels of has2 and has3 after prevention with 2.4 mg / mL 2'-FL were significantly upregulated compared with the model group (p<0.05), indicating that 2'-FL can prevent the downregulation of has2 and has3 transcription levels caused by LPS, thereby maintaining the synthesis of glycocalyx hyaluronic acid.

[0088] 2.2.3 Preventive effect of 2'-FL on LPS-induced heparan sulfate damage in Caco-2 cell glycocalyx

[0089] Figure 6 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.

[0090] Depend on Figure 6 It can be seen that after LPS intervention, the transcription levels of ext1 and ext2 in the model group were nearly 2 times lower than those in 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 intervened with LPS, it was found that 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 could significantly upregulate the transcription level of ext1 (p < 0.05), and the 1.2 mg / mL and 2.4 mg / mL concentrations of 2'-FL intervention groups could significantly upregulate 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.

[0091] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

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

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

3. Use of '-fucosyllactose in the preparation of a medicine for promoting the development of glycocalyx in the intestine 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 manifestation of promoting the development of glycocalyx in the intestine includes: upregulating the transcription level of at least one of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA.

5. The use according to any one of claims 1 to 3, characterized in that: The prevention of glycocalyx damage caused by lipopolysaccharide (LPS) includes upregulating the decrease in the transcription level of at least one of gpc1 mRNA, has1 mRNA, has2 mRNA, has3 mRNA, ext1 mRNA and ext2 mRNA caused by LPS.

6. The use according to any one of claims 1 to 3, characterized in that: The application concentration of the 2'-fucosyllactose is 0.6-2.4 mg / mL.

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

8. The food according to claim 1 or 7, characterized in that 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.

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

10. The use according to any one of claims 7 to 9, characterized in that: The application concentration of the 2'-fucosyllactose is 0.6-2.4 mg / mL.

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