Composition for promoting intestinal development and application thereof

Abstract

The invention relates to a composition for promoting intestinal development and application thereof. The composition comprises lactose-N-neotetraose and 3 '-sialic acid lactose. According to the composition for promoting intestinal development, the transcriptional level of genes such as has1, has2 and has3 related to intestinal glycocalyx development can be up-regulated, and the transcriptional level of ext2 can be up-regulated, so that the intestinal glycocalyx development is synergistically promoted.

Description

Technical Field

[0001] The present invention relates to the technical field of compositions containing human milk oligosaccharides, and particularly relates to a composition for promoting intestinal development and its application. 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 also plays a role in preventing pathogens from entering the human body. In addition, it also regulates the crosstalk between the 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 neonatal intestinal barrier function 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 children 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 the milk of other mammals. In infant formula, an important function of HMOs is to stimulate the colonization of the gastrointestinal tract by the microbiota and promote the development of intestinal barrier function. One possible way to achieve this is to stimulate the development of the epithelial glycocalyx on intestinal epithelial cells. The glycocalyx on neonatal intestinal epithelial cells provides binding sites for commensal microorganisms, and a well-developed glycocalyx can prevent pathogen adhesion and act as a barrier to luminal toxins and enzymes.

[0004] Currently, there are few clinical intervention studies on the direct regulatory effects of HMOs on intestinal cells and structures. However, preclinical research data indicate that HMOs have a positive effect on intestinal maturation. Intestinal cell line cultures such as Caco-2 are usually used in vitro to study the mechanisms by which nutrients and microbial metabolites affect epithelial integrity. It has been demonstrated that N-lacto-neotetraose (LNnT), 2'-fucosyllactose (2'-FL), and 3'-sialyllactose (3'-SL) can promote the expression of tight junction proteins and increase cell differentiation along the crypt-villus axis; 2'-fucosyllactose and 3-fucosyllactose have a positive effect on the expression of mucin glycoproteins. In in vivo animal experiments, it was found that supplementation with 2'-FL can significantly increase epithelial integrity and reduce the occurrence of colitis. However, there are few studies on the effect of HMOs on the glycocalyx, especially on the study of the function of the glycocalyx. Summary of the Invention

[0005] Technical Problem Therefore, the first object of the present invention is to provide an application of 3'-sialyllactose (3'-SL) and lacto-N-neotetraose (LNnT) in synergistically promoting the development of the glycocalyx.

[0006] The second object of the present invention is to provide an application of lacto-N-neotetraose (LNnT) and 3'-sialyllactose (3'-SL) in a specific ratio in synergistically promoting the development of the glycocalyx.

[0007] The third object of the present invention is to provide an application of lacto-N-neotetraose (LNnT) and 3'-sialyllactose (3'-SL) in a specific ratio in synergistically promoting the transcriptional level of genes related to the development of the glycocalyx, thereby promoting the development of the intestinal glycocalyx.

[0008] Technical solution This application explored the effects of LNnT and 3'-SL on the development and function of the glycocalyx of intestinal epithelial cells Caco-2 at the cellular level, laying a theoretical foundation for the application of LNnT and 3'-SL.

[0009] According to one aspect of the present invention, there is provided a composition comprising lacto-N-neotetraose and 3'-sialyllactose.

[0010] In one embodiment, the ratio of lacto-N-neotetraose to 3'-sialyllactose is: 1:1 to 3:1 by weight, preferably 1:1, 1:2, 2:1 and 3:1 by weight, more preferably 1:1 or 3:1 by weight, and most preferably 3:1 by weight.

[0011] In one embodiment, in the composition, the concentration of lacto-N-neotetraose is: 0.17 to 0.35 mg / mL, and the concentration of 3'-sialyllactose is: 0.05 to 0.3 mg / mL.

[0012] Preferably, in the composition, the concentration of lacto-N-neotetraose is: 0.2 to 0.3 mg / mL, preferably 0.2 to 0.27 mg / mL; the concentration of 3'-sialyllactose is: 0.06 to 0.2 mg / mL, preferably 0.08 to 0.18 mg / mL.

[0013] In a specific embodiment of the present invention, when the above ratio of lacto-N-neotetraose to 3'-sialyllactose is satisfied, the concentrations of lacto-N-neotetraose and 3'-sialyllactose can be any value within the above concentration ranges.

[0014] On the other hand, the present invention provides an application of the composition as described above in the preparation of foods or health products.

[0015] In still another aspect, the present invention provides an application of the composition in the preparation of a product for promoting the development of the intestinal glycocalyx.

[0016] In one embodiment, the composition is used to up-regulate the transcription level of genes related to intestinal glycocalyx development, thereby promoting intestinal glycocalyx development.

[0017] In a specific embodiment, the composition is used to up-regulate the transcription levels of has1, has2, and has3, promote the synthesis of hyaluronic acid synthase in the glycocalyx layer, and thereby promote the secretion of hyaluronic acid in the glycocalyx layer.

[0018] Preferably, the composition is used to up-regulate the transcription level of has2, promote the synthesis of hyaluronic acid synthase in the glycocalyx layer, and thereby promote the secretion of hyaluronic acid in the glycocalyx layer.

[0019] In another specific embodiment, the composition is used to up-regulate the transcription level of ext2, thereby promoting the synthesis of heparan sulfate in the glycocalyx layer.

[0020] Beneficial effects The composition provided by the present invention, which contains lacto-N-neotetraose and 3'-sialyllactose, after being combined in a specific ratio, can synergistically up-regulate the transcription levels of glycocalyx development-related genes has1, has2, and has3, as well as the transcription level of ext2, thereby synergistically promoting intestinal glycocalyx development. In particular, it significantly up-regulates the transcription levels of has2 and ext2, thereby synergistically promoting intestinal glycocalyx development. Brief description of the drawings

[0021] Figure 1 Shows the effect of LNnT on the transcription level of gpc1 in Caco-2 cells, Figure 2 Shows the effect of 3'-SL on the transcription level of gpc1 in Caco-2 cells; Figure 3 , Figure 4 , Figure 5 Respectively show the effects of LNnT on the transcription levels of has1, has2, and has3 in Caco-2 cells, Figure 6 , Figure 7 , Figure 8 Respectively show the effects of 3'-SL on the transcription levels of has1, has2, and has3 in Caco-2 cells; Figure 9 And Figure 10 Respectively show the effects of LNnT on the transcription levels of ext1 and ext2 in Caco-2 cells, Figure 11 And Figure 12 Respectively show the effects of 3'-SL on the transcription levels of ext1 and ext2 in Caco-2 cells; Figure 13 Shows the effect of LNnT and 3'-SL on the transcription level of gpc1 in Caco-2 cells; Figure 14 ,Figure 15 , Figure 16 Shows the effects of the compositions of LNnT and 3'-SL on the transcriptional levels of has1, has2, and has3 in Caco-2 cells respectively; Figure 17 , Figure 18 Shows the effects of the compositions of LNnT and 3'-SL on the transcriptional levels of ext1 and ext2 in Caco-2 cells respectively. Detailed implementation mode

[0022] In order to make the objectives, technical solutions, and advantages of the present invention clearer, the present invention will be described in detail below in conjunction with specific embodiments. The described embodiments are part of the embodiments of the present invention, rather than all of the embodiments.

[0023] Human milk oligosaccharides (HMOs) are diverse and mainly include neutral HMOs and acidic HMOs. Neutral HMOs such as 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), N-lactotetraose (LNT), and lacto-N-neotetraose (LNnT); acidic HMOs, such as 3'-sialyllactose (3'-SL), 6'-sialyllactose (6'-SL), and sialyllacto-N-tetraose (LST).

[0024] The glycocalyx is composed of glycans and proteins. Proteoglycans are generally considered the most important components of the glycocalyx and form the framework of the glycocalyx. In addition, the glycocalyx contains glycosaminoglycan chains linked to the core proteins of proteoglycans, among which heparan sulfate (HS) and hyaluronic acid (HA) are the main glycosaminoglycan components in the glycocalyx.

[0025] The genes related to glycocalyx development in Caco-2 cells include: the transcriptional levels of glypican 1 (gpc1), hyaluronan synthase 1 (has1, has2, has3), and exostosin glycosyltransferase 1 (ext1, ext2) genes were measured. Among them, gpc1 is the coding gene of glypican 1 (GPC1), and GPC1 is the protein backbone of the glycocalyx layer and an important carrier of glycosaminoglycan chains including hyaluronic acid (HA) and heparan sulfate (HS). has1, has2, and has3 are the coding genes of hyaluronan synthase (HAS), and HAS is crucial for the synthesis of HA, and HA is a highly viscous component of the intestinal mucus layer, which is responsible for tissue repair, stability, and anti-inflammatory effects. Compared with HAS1 and HAS2, HAS3 synthesizes low-molecular-weight HA, and low-molecular-weight HA is essential for the development of intestinal stem cells. In addition, ext1 and ext2 are involved in the elongation of heparan sulfate (HS) chains and are responsible for integrating HS chains with nucleotide sugars in the Golgi apparatus, and HS synthesis supports organogenesis, growth factor signaling, and bacterial adhesion.

[0026] Unless otherwise specified, the raw materials and equipment used in the present invention are common raw materials and equipment in the art, and the methods used in the present invention are conventional methods in the art.

[0027] Unless otherwise specified, the parts are parts by weight, % is % by weight, and the temperature is in degrees Celsius. In the following examples, the following raw materials and methods are used.

[0028] Example Example 1: Caco-2 cell culture Human colon adenocarcinoma cell line Caco-2 was cultured in MEM complete medium containing 20% high-quality fetal bovine serum (purchased from Nanjing Senbeijia Biotechnology Co., Ltd.), and grew in 25 cm 2 cell culture flasks, and were cultured in a carbon dioxide incubator at 37 °C, 5% CO2, and 95% relative humidity. The culture medium was changed every one or two days. When the cells grew to 80% - 90% confluence, they were digested with 0.25% trypsin digestion solution and subcultured at a ratio of 1:3.

[0029] Example 2: Effects of LNnT and 3'-SL on the development of the glycocalyx layer of Caco-2 cells Adjust the density of Caco-2 cells to 1×10 5 cells / cm 2 , inoculated into 24-well plates, and the corresponding intervention substances, namely a composition of LNnT and 3'-SL (both purchased from Suzhou Yixi Biotechnology Co., Ltd.) in a specific ratio, were added to the MEM complete medium according to the grouping, and co-incubated for 36 h. The MEM medium was used as a blank control and incubated for 36 h (see Table 2 below).

[0030] After the incubation, total cellular RNA was extracted and reverse transcribed, and the transcriptional levels of genes related to glycocalyx development in Caco-2 cells, including glypican 1 (gpc1), hyaluronan synthases 1 (has1, has2, has3), and exostosin glycosyltransferases 1 (ext1, ext2), were measured using qPCR (Nanjing Novoprotein Biotechnology Co., Ltd.).

[0031] Table 1 qPCR primer sequences

[0032] Table 2 Co-incubation methods of a composition of LNnT and 3'-SL in a specific ratio, and LNnT and 3'-SL alone

[0033] Examples 2-1 to 2-6 respectively verified the effects of different concentrations of individual LNnT and individual 3'-SL on the development of the glycocalyx layer of Caco-2 cells.

[0034] The specific steps are as follows: Adjust the density of Caco-2 cells to 1×10 5 cells / cm 2 , inoculate into a 24-well plate, and add specific concentrations of LNnT and 3'-SL to the MEM complete medium according to the grouping, and incubate for 36 h. The final concentrations of 3'-SL in the MEM complete medium are 0.05 mg / mL, 0.1 mg / mL, 0.2 mg / mL, and 0.4 mg / mL, respectively. The final concentrations of LNnT in the MEM complete medium are 0.05 mg / mL, 0.3 mg / mL, and 0.6 mg / mL, respectively. After incubation, extract the total cellular RNA, perform reverse transcription, and use qPCR to measure the transcriptional levels of genes related to glycocalyx development, phosphatidylinositol proteoglycan 1 (gpc1), hyaluronan synthase 1 (has1, has2, has3), and exostosin glycosyltransferase 1 (ext1, ext2) in Caco-2 cells. The results are shown in Figures 1 to 12 .

[0035] Effect of individual LNnT on the development of the protein backbone of the glycocalyx layer in Example 2-1 Figure 1 Shows the effect of LNnT on the transcriptional level of gpc1 in Caco-2 cells.

[0036] As Figure 1 can be seen, only 0.05 mg / mL of LNnT can up-regulate the transcriptional level of gpc1, but there is no significant difference compared with the control group. It shows that LNnT has a certain promoting effect on the development of the protein backbone of the glycocalyx layer, but the effect is not obvious.

[0037] Effect of individual 3'-SL on the development of the protein backbone of the glycocalyx layer in Example 2-2 Figure 2 Shows the effect of 3'-SL on the transcriptional level of gpc1 in Caco-2 cells.

[0038] As Figure 2 can be seen, all four intervention concentrations of 3'-SL selected can up-regulate the transcriptional level of gpc1. Among them, 0.05 mg / mL, 0.1 mg / mL, and 0.2 mg / mL of 3'-SL can significantly up-regulate the transcriptional level of gpc1 (p < 0.01, p < 0.001). It shows that 0.05 mg / mL, 0.1 mg / mL, and 0.2 mg / mL of 3'-SL have a certain promoting effect on the development of the protein backbone of the glycocalyx layer.

[0039] Example 2-3 Effect of LNnT alone on hyaluronan synthase in the glycocalyx Figure 3 、 Figure 4 、 Figure 5 Showed the effects of LNnT on the transcriptional levels of has1, has2, and has3 in Caco-2 cells, respectively.

[0040] From Figure 3 、 Figure 4 、 Figure 5 It can be seen that LNnT can up-regulate the transcriptional levels of has1, has2, and has3, but there is no significant difference compared with the control group. It shows that only 0.05 mg / mL of LNnT has a certain promoting effect on the secretion of hyaluronic acid in the glycocalyx, but there is no significant difference compared with the control group.

[0041] Example 2-4 Effect of 3'-SL alone on hyaluronan synthase in the glycocalyx Figure 6 、 Figure 7 、 Figure 8 Showed the effects of 3'-SL on the transcriptional levels of has1, has2, and has3 in Caco-2 cells, respectively.

[0042] As Figure 6 、 Figure 7 、 Figure 8 Shown, 3'-SL at the four selected intervention concentrations could up-regulate the transcriptional levels of has1, has2, and has3. Among them, 3'-SL at 0.05 mg / mL and 0.1 mg / mL could significantly up-regulate the transcriptional level of has1 (p < 0.01), 3'-SL at 0.1 mg / mL could significantly up-regulate the transcriptional level of has2 (p < 0.01), and 3'-SL at 0.05 mg / mL and 0.1 mg / mL could significantly up-regulate the transcriptional level of has3 (p < 0.005, p < 0.001). It shows that low-concentration 3'-SL has a certain promoting effect on the secretion of hyaluronic acid in the glycocalyx (Note: The significant difference here is compared with the control group and is significant. No significant difference marked means the difference is not significant. Even if some seem to have a large difference, it may be due to too large an error, resulting in no significant difference).

[0043] Example 2-5 Effect of LNnT alone on heparan sulfate in the glycocalyx Figure 9 and Figure 10 Showed the effects of LNnT on the transcriptional levels of ext1 and ext2 in Caco-2 cells, respectively.

[0044] As Figure 9 and Figure 10As shown, only LNnT at 0.05 mg / mL can up-regulate the transcription level of ext1, but not significantly (p > 0.05). LNnT at the three selected intervention concentrations has no effect on the transcription level of ext2. This indicates that the promoting effect of LNnT alone on the synthesis of heparan sulfate in the glycocalyx layer is very weak.

[0045] Example 2-6 Effect of 3’-SL Alone on Heparan Sulfate in the Glycocalyx Layer Figure 11 and Figure 12 respectively show the effects of 3’-SL on the transcription levels of ext1 and ext2 in Caco-2 cells.

[0046] As Figure 11 and Figure 12 shown, among the four selected intervention concentrations of 3’-SL, except that 0.4 mg / mL has no up-regulating effect on the transcription level of ext1, the other concentrations can up-regulate the transcription levels of ext1 and ext2. However, only 3’-SL at 0.1 mg / mL can significantly up-regulate the transcription level of ext1 (p < 0.01), and 3’-SL at 0.05 mg / mL, 0.1 mg / mL, and 0.2 mg / mL can all significantly up-regulate the transcription level of ext2 (p < 0.05, p < 0.01). This indicates that 3’-SL has a certain promoting effect on the synthesis of heparan sulfate in the glycocalyx layer.

[0047] Example 3 Effect of the Composition of LNnT and 3’-SL on the Development of the Glycocalyx Layer in Caco-2 Cells Examples 3-1 to 3-3 respectively verified the effect of the composition of LNnT and 3’-SL at a specific ratio on the development of the glycocalyx layer in Caco-2 cells.

[0048] The specific steps are as follows: Adjust the density of Caco-2 cells to 1×10 5 cells / cm 2 , inoculate them into 24-well plates, and add the composition of LNnT and 3’-SL at a specific ratio, as well as LNnT and 3’-SL alone, to the MEM complete medium according to the grouping, and incubate for 36 h. Use the MEM medium as the blank control and incubate for 36 h.

[0049] The ratios of LNnT to 3’-SL added and their final concentrations in the complete MEM medium were 1:1 (0.175 mg / mL LNnT + 0.175 mg / mL 3’-SL), 2:1 (0.235 mg / mL LNnT + 0.115 mg / mL 3’-SL), 1:2 (0.115 mg / mL LNnT + 0.235 mg / mL 3’-SL), and 3:1 (0.263 mg / mL LNnT + 0.088 mg / mL 3’-SL), respectively. The final concentration of either LNnT or 3’-SL alone in the complete MEM medium was 0.35 mg / mL. After co-incubation for 36 h, total cellular RNA was extracted at the end of the incubation, reverse transcribed, and the transcriptional levels of genes related to glycocalyx development, namely glypican 1 (gpc1), hyaluronan synthases 1, 2, and 3 (has1, has2, has3), and exostosin glycosyltransferases 1 and 2 (ext1, ext2) in Caco-2 cells were determined using qPCR. The results are shown in Figures 13 to 18 。

[0050] Example 3-1 Effect of the LNnT and 3’-SL composition on the protein backbone of the glycocalyx layer Figure 13 The effect of LNnT and 3’-SL on the transcriptional level of gpc1 in Caco-2 cells is shown.

[0051] From Figure 13 it can be seen that the compositions of LNnT and 3’-SL at each selected ratio and 0.35 mg / mL of LNnT and 3’-SL had no significant effect on the transcriptional level of gpc1, indicating that the compositions of LNnT and 3’-SL and high concentrations of LNnT and 3’-SL had no significant promoting effect on the development of the protein backbone of the glycocalyx layer (Note: The significance here refers to the difference between groups).

[0052] Example 3-2 Effect of the LNnT and 3’-SL composition on hyaluronan synthases in the glycocalyx layer Figure 14 、 Figure 15 、 Figure 16 The effect of the LNnT and 3’-SL composition on the transcriptional levels of has1, has2, and has3 in Caco-2 cells is shown, respectively.

[0053] From Figure 14 it can be seen that the compositions of LNnT and 3’-SL at a ratio of 1:1 and 3:1 significantly upregulated the transcriptional level of has1, and the upregulation effect on the transcriptional level of has1 was significantly higher than that of 0.35 mg / mL of LNnT and 3’-SL alone.

[0054] From Figure 15It can be seen that the compositions with the selected LNnT and 3'-SL ratios of 2:1 and 3:1 significantly up-regulated the transcriptional level of has2, and the composition with a ratio of 3:1 had a significantly higher up-regulating effect on the has2 transcriptional level than LNnT and 3'-SL alone at 0.35 mg / mL.

[0055] It can be seen from Figure 16 that the compositions with the selected LNnT and 3'-SL ratios of 1:2 and 3:1 significantly up-regulated the transcriptional level of has3, and the composition with a ratio of 3:1 had a significantly higher up-regulating effect on the has3 transcriptional level than LNnT alone at 0.35 mg / mL.

[0056] In summary, the LNnT and 3'-SL compositions at specific ratios have a significant promoting effect on the synthesis of hyaluronic acid in the glycocalyx layer. When the ratio of LNnT to 3'-SL is 3:1, the up-regulating effects on has1 and has2 are significantly higher than those of LNnT and 3'-SL at 0.35 mg / mL, and the up-regulating effect on has3 is significantly higher than that of LNnT at 0.35 mg / mL, indicating that when the ratio of LNnT to 3'-SL is 3:1, it has a significant synergistic promoting effect on the synthesis of hyaluronic acid in the glycocalyx layer.

[0057] Example 3-3 Effects of the LNnT and 3'-SL Composition on Heparan Sulfate in the Glycocalyx Layer Figure 17 and Figure 18 show the effects of the LNnT and 3'-SL composition on the transcriptional levels of ext1 and ext2 in Caco-2 cells, respectively.

[0058] As Figure 17 shown, the selected LNnT and 3'-SL compositions at various ratios and LNnT and 3'-SL at 0.35 mg / mL had no significant effect on the ext1 transcriptional level, indicating that the LNnT and 3'-SL composition and high-concentration LNnT and 3'-SL had no significant promoting effect on the development of the protein skeleton in the glycocalyx layer.

[0059] As Figure 18 shown, the composition with the selected LNnT and 3'-SL ratio of 3:1 significantly up-regulated the transcriptional level of ext2, and the up-regulating effect of the composition with a ratio of 3:1 on the ext2 transcriptional level was significantly higher than that of LNnT alone at 0.35 mg / mL. It indicates that the LNnT and 3'-SL composition at a ratio of 3:1 can promote the synthesis of heparan sulfate in the glycocalyx layer by synergistically promoting the transcriptional level of ext2.

[0060] From the above experimental results, it can be seen that only 0.05 mg / mL of LNnT upregulates the transcriptional levels of has1, has2, and has3, indicating that LNnT alone has a certain promoting effect on the secretion of hyaluronic acid in the glycocalyx layer. However, compared with the control group, the difference is not significant. Only 0.05 mg / mL of 3'-SL can significantly upregulate the transcriptional levels of has1 and has3, indicating that low-concentration 3'-SL has a certain promoting effect on the secretion of hyaluronic acid in the glycocalyx layer. However, after combining LNnT and 3'-SL in a certain ratio in the present invention, the LNnT and 3'-SL composition at a specific ratio has a significant promoting effect on the synthesis of hyaluronic acid in the glycocalyx layer. Among them, when the ratio of LNnT to 3'-SL is 3:1, the upregulating effect on has1 and has2 is significantly higher than that of 0.35 mg / mL of LNnT and 3'-SL, and the upregulating effect on has3 is significantly higher than that of 0.35 mg / mL of LNnT, indicating that when the ratio of LNnT to 3'-SL is 3:1, it has a significant synergistic promoting effect on the synthesis of hyaluronic acid in the glycocalyx layer.

[0061] The selected three intervention concentrations of LNnT (used alone) had no significant effect on the transcriptional levels of ext1 and ext2 (p > 0.05), indicating that LNnT alone has a weak promoting effect on the synthesis of heparan sulfate in the glycocalyx layer. Among the selected three intervention concentrations of 3'-SL (used alone), except that 0.4 mg / mL had no upregulating effect on the transcriptional level of ext1, the other concentrations could upregulate the transcriptional levels of ext1 and ext2, but none of them were significant (p > 0.05). This indicates that 3'-SL has a weak promoting effect on the synthesis of heparan sulfate in the glycocalyx layer. However, after combining LNnT and 3'-SL in a certain ratio in the present invention, the composition with the ratio of LNnT to 3'-SL of 3:1 significantly upregulated the transcriptional level of ext2, and the upregulating effect of the composition with the ratio of 3:1 on the transcriptional level of ext2 was significantly higher than that of using 0.35 mg / mL of LNnT alone. This indicates that the composition with the ratio of LNnT to 3'-SL of 3:1 can promote the synthesis of heparan sulfate in the glycocalyx layer by synergistically promoting the transcriptional level of ext2.

[0062] In summary, after combining LNnT and 3'-SL in a certain ratio in the present invention, it can synergistically upregulate the transcriptional levels of has1, has2, and has3, as well as the transcriptional level of ext2.

[0063] When 3’-SL is added to co-culture with Caco-2 cells before they adhere and grow, it can simulate the state during intestinal growth and maturation. In this state, 3’-SL can significantly promote the development of the glycocalyx layer of Caco-2 cells. On this basis, LNnT and 3’-SL are formulated and combined, and at a specific ratio, they have a significant synergistic promoting effect on the development of the glycocalyx layer. It shows that supplementing a composition of LNnT and 3’-SL at a specific ratio during the stage of intestinal development and maturation can significantly promote the development of the intestinal glycocalyx layer, thereby promoting the overall healthy development of the intestine.

Claims

1. A composition for promoting intestinal development, characterized in that, Containing lacto-N-neotetraose and 3'-sialyllactose.

2. The composition according to claim 1, wherein The ratio of the lacto-N-neotetraose to the 3'-sialyllactose is: 1:1 to 3:1 by weight.

3. The composition according to claim 1 or 2, characterized in that, In the composition, the concentration of the lacto-N-neotetraose is: 0.17 to 0.35 mg / mL, and the concentration of the 3'-sialyllactose is: 0.05 to 0.3 mg / mL.

4. The composition according to claim 1 or 2, characterized in that, In the composition, the concentration of the lacto-N-neotetraose is: 0.2 to 0.3 mg / mL, and the concentration of the 3'-sialyllactose is: 0.06 to 0.2 mg / mL.

5. Use of the composition according to any one of claims 1 to 4 in the preparation of a food or a health product.

6. Use of the composition according to any one of claims 1 to 4 in the preparation of a product for promoting the development of the intestinal glycocalyx.

7. The application according to claim 6, wherein The composition is used to up-regulate the transcriptional level of genes related to the development of the intestinal glycocalyx, thereby promoting the development of the intestinal glycocalyx.

8. The application according to claim 6 or 7, characterized in that, The composition is used to up-regulate the transcriptional levels of has1, has2, and has3, promote the synthesis of hyaluronic acid synthase in the glycocalyx layer, and thereby promote the secretion of hyaluronic acid in the glycocalyx layer.

9. The application according to claim 8, wherein The composition is used to up-regulate the transcriptional level of has2, promote the synthesis of hyaluronic acid synthase in the glycocalyx layer, and thereby promote the secretion of hyaluronic acid in the glycocalyx layer.

10. The application according to claim 6 or 7, characterized in that, The composition is used to up-regulate the transcriptional level of ext2, thereby promoting the synthesis of heparan sulfate in the glycocalyx layer.

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