Application of breast milk oligosaccharide in preparation of preparation for inhibiting proliferation of intestinal tract Ruminococcus

Abstract

The invention relates to an application of breast milk oligosaccharide in preparation of a preparation for inhibiting proliferation of intestinal tract Ruminococcus, the breast milk oligosaccharide is 2 '-FL and / or 6'-SL, the relative abundance of the intestinal tract Ruminococcus can be obviously reduced, the proliferation of the intestinal tract Ruminococcus can be inhibited, and the breast milk oligosaccharide can be applied to preparation of the preparation for inhibiting the proliferation of the intestinal tract Ruminococcus. The breast milk oligosaccharide is 2 '-FL or a composition of 2'-FL and 6 '-SL, compared with 2'-FL or a composition of 2 '-FL and 6'-SL, the breast milk oligosaccharide has a better inhibition effect on intestinal tract Ruminococcus, can be applied to preparation of a preparation for inhibiting abnormal proliferation of intestinal tract Ruminococcus of a patient suffering from the inflammatory bowel disease, and is particularly applied to preparation of a preparation for preventing enteritis and maintaining intestinal health.

Description

[0001] This application claims priority to the Chinese application number 202411451431X filed on October 17, 2024, with the invention name "Application of human milk oligosaccharides in the preparation of preparations for inhibiting the proliferation of intestinal Ruminococcus". Technical Field

[0002] The present invention relates to the field of biotechnology, and more specifically to an application of human milk oligosaccharide in preparing a preparation for inhibiting the proliferation of intestinal Ruminococcus. Background Art

[0003] Human Milk Oligosaccharides (HMOs) are a type of complex oligosaccharides with a high content in breast milk. They are composed of monosaccharides and their derivatives, sialic acid and other structural units connected by glycosidic bonds. At present, more than 150 HMOs have been confirmed, among which 2'-fucosyllactose (2'-FL) is mainly used in infant formula to enhance the baby's immune system, especially as a prebiotic to promote the growth of beneficial intestinal bacteria. 6'-sialyllactose (6'-SL) is mainly used as a prebiotic to promote the growth of beneficial intestinal bacteria.

[0004] Human microecology, especially intestinal microecology, has been reported to be directly or indirectly related to tumors, digestive system diseases (such as inflammatory bowel disease, chronic diarrhea), immune system diseases (such as asthma, allergies, eczema), metabolic diseases (such as obesity, diabetes), mental diseases (such as depression, autism), etc. Among them, intestinal microorganisms may be involved in the occurrence and development of intestinal diseases. Professor Zhang Hongjie published an article in the "Chinese Journal of Inflammatory Bowel Diseases" entitled "Progress in the Study of Intestinal Flora in Inflammatory Bowel Disease", which revealed the differential changes in the composition of intestinal flora in patients with inflammatory bowel diseases (such as Crohn's disease and ulcerative colitis). Among them, the intestinal Prevotella, Bifidobacterium, and Veillonella decreased in patients with Crohn's disease (CD), while Clostridium, Escherichia coli, Neisseria, and Ruminococcus increased; in patients with ulcerative colitis (UC), Coprococcus, Prevotella, and Roseburia decreased, while Escherichia coli, Ruminococcus, Gemmatimonas, Actinobacillus, and Enterococcus increased. It can be seen that there is a common abnormal increase in Ruminococcus in the intestines of patients with inflammatory bowel disease.

[0005] Although the cause of Crohn's disease is currently unknown, studies have found that there are great differences in the structure of intestinal flora between patients with the disease and healthy people. Among them, Ruminococcus is a flora associated with Crohn's disease, and its increased relative abundance is related to the severity of symptoms. In addition, many studies have shown that Ruminococcus is positively correlated with irritable bowel syndrome. For example, a research team from Hong Kong Baptist University in China found that active Ruminococcus is highly enriched in the intestines of patients with diarrhea-predominant irritable bowel syndrome (IBS-D). Human milk oligosaccharides, as prebiotics, are mainly used to promote the growth of beneficial intestinal bacteria. Therefore, further research on the effects of human milk oligosaccharides on the composition of intestinal flora will help to explore more applications of human milk oligosaccharides. Summary of the invention

[0006] In view of the above defects or improvement needs of the prior art, the present invention provides an application of human milk oligosaccharides in the preparation of a preparation for inhibiting the proliferation of Ruminococcus in the intestine. The purpose is to find that 2′-FL in human milk oligosaccharides and its combination with 6′-SL can significantly reduce the relative abundance of Ruminococcus in the intestine, and can be used to prepare a preparation for inhibiting the proliferation of Ruminococcus in the intestine, thereby solving the technical problem of the lack of application of 2′-FL and 6′-SL in intestinal flora inhibitors.

[0007] To achieve the above objectives, according to one aspect of the present invention, there is provided a use of human milk oligosaccharides in the preparation of a preparation for inhibiting the proliferation of intestinal Ruminococcus, wherein the human milk oligosaccharides are 2′-FL and / or 6′-SL, which are used in the preparation of a preparation for inhibiting the proliferation of intestinal Ruminococcus.

[0008] Preferably, in the use, the human milk oligosaccharide is 2′-FL, or a combination of 2′-FL and 6′-SL.

[0009] Preferably, the use is used to prepare a preparation for inhibiting abnormal proliferation of Ruminococcus in the intestine of patients with inflammatory bowel disease.

[0010] Preferably, the use is used for preparing a preparation for inhibiting abnormal proliferation of Ruminococcus in the intestine of patients with Crohn's disease or ulcerative colitis.

[0011] Preferably, the use is used to prepare a preparation for inhibiting abnormal proliferation of Ruminococcus in the intestine of patients with irritable bowel syndrome.

[0012] Preferably, the use is used for preparing a preparation for inhibiting abnormal proliferation of Ruminococcus in the intestine of patients with diarrhea-predominant irritable bowel syndrome.

[0013] Preferably, in the use, the human milk oligosaccharide is a composition of 2′-FL and 6′-SL.

[0014] Preferably, in the application, the mass ratio of 2′-FL to 6′-SL in the composition is 1.0-2.0:0.3-0.4.

[0015] Preferably, in the application, the mass ratio of 2′-FL to 6′-SL in the composition is 1.5-2.0:0.35-0.4.

[0016] Preferably, in the use, the preparation is in the form of powder, tablet, capsule, bead, elixir, solution or suspension.

[0017] Preferably, in the application, the human milk oligosaccharide is 2′-FL and / or 6′-SL, and is used for preparing a preparation for preventing enteritis and maintaining intestinal health.

[0018] Preferably, in the application, the human milk oligosaccharide is 2′-FL, or a combination of 2′-FL and 6′-SL, and is used to prepare a preparation for preventing enteritis caused by proliferation of Ruminococcus in the intestine.

[0019] In general, compared with the prior art, the above technical solution conceived by the present invention can achieve the following beneficial effects.

[0020] Since the present invention finds that 2′-FL and / or 6′-SL can significantly reduce the relative abundance of intestinal Ruminococcus, they can be used to prepare a preparation for inhibiting the proliferation of intestinal Ruminococcus. Compared with 2′-FL, the combined use of 2′-FL and 6′-SL has a better inhibitory effect on intestinal Ruminococcus. By inhibiting the proliferation of intestinal Ruminococcus, inflammatory bowel disease and / or irritable bowel syndrome caused by the proliferation of intestinal Ruminococcus can be prevented, or the progression of the disease can be slowed down. In particular, it can be used to prepare a preparation for preventing enteritis and maintaining intestinal health. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The effect of HMOs on the body weight of healthy mice;

[0022] Figure 2 The effect of HMOs on the organ / body weight ratio of mice;

[0023] Figure 3 To investigate the effects of HMOs treatment on intestinal flora;

[0024] Figure 4 The effects of HMOs treatment on different intestinal flora.

[0025] Figure 4 (A) is a histogram of relative abundance of species at the genus level, (B) is a heat map, and (C) is a comparative analysis of intestinal microorganisms with different relative abundances at the genus level (taking the top 20 in relative abundance). DETAILED DESCRIPTION

[0026] In order to further explain the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the specific implementation mode, structure, characteristics and effects of the present invention are described in detail below in combination with the accompanying drawings and preferred embodiments.

[0027] The present invention studies the effects of different doses of 2'-fucosyllactose (2'-FL) and 2'-fucosyllactose + 6'-sialyllactose (6'-SL) on intestinal flora, and the results show that the administration of 2'-FL and a combination of 2'-FL + 6'-SL can significantly reduce the relative abundance of Ruminococcus in the intestine, and can be used to prepare a preparation for inhibiting the proliferation of Ruminococcus in the intestine. In particular, compared with the use of 2'-FL alone, the combined use of 2'-FL and 6'-SL has a better inhibitory effect on Ruminococcus in the intestine.

[0028] Based on this, the present invention provides an application of human milk oligosaccharides in the preparation of a preparation for inhibiting the proliferation of intestinal Ruminococcus, wherein the human milk oligosaccharides are 2′-FL and / or 6′-SL, wherein the human milk oligosaccharides are the active ingredients of the preparation, which can be derived from nature, artificial synthesis or bacterial fermentation.

[0029] In some embodiments, the human milk oligosaccharide is 2′-FL, and its application amount is 30 to 60 mg / kg / d. It has a significant inhibitory effect on intestinal Ruminococcus. Within a certain application amount range, the higher the application amount, the better the inhibitory effect on intestinal Ruminococcus. Generally, the relative abundance of intestinal Ruminococcus is higher in patients with inflammatory bowel disease or irritable bowel syndrome. It can be used to prepare preparations for inhibiting the abnormal proliferation of intestinal Ruminococcus in patients with inflammatory bowel disease or irritable bowel syndrome.

[0030] In some embodiments, the human milk oligosaccharide is a composition of 2′-FL and 6′-SL, and the mass ratio of 2′-FL to 6′-SL in the composition is 1.0-2.0:0.3-0.4. In this ratio range, the added 6′-SL can enhance the inhibitory effect of 2′-FL on intestinal Ruminococcus, and can be particularly used to prevent inflammatory bowel disease (such as Crohn's disease and ulcerative colitis) or diarrhea-type irritable bowel syndrome, or slow down its progression; preferably, the mass ratio of the two is 1.5-2.0:0.35-0.4, and the composition has a better inhibitory effect on intestinal Ruminococcus. It can be used to prepare a preparation for inhibiting abnormal proliferation of intestinal Ruminococcus in patients with inflammatory bowel disease (such as Crohn's disease and ulcerative colitis) or diarrhea-type irritable bowel syndrome.

[0031] Furthermore, the application is used to prepare preparations for preventing enteritis and maintaining intestinal health.

[0032] Preferably, the human milk oligosaccharide is 2′-FL, or a combination of 2′-FL and 6′-SL, which is used to prepare a preparation for preventing enteritis caused by proliferation of Ruminococcus in the intestine.

[0033] In addition, the present invention also provides a preparation for inhibiting the abnormal proliferation of Ruminococcus in the intestine, wherein the active ingredients include 2′-FL and / or 6′-SL, and the preparation is used for inhibiting the abnormal proliferation of Ruminococcus in the intestine.

[0034] The forms of the preparations in the present invention include but are not limited to powders, tablets, capsules, beads, elixirs, solutions or suspensions; preferably, they also include the auxiliary ingredients required for different forms, such as one or more of excipients, disintegrants, granulating binders, and lubricants corresponding to tablet forms, enteric carriers, fillers, or stabilizers corresponding to capsule forms, and emulsifiers, diluents, etc. corresponding to suspensions or elixirs.

[0035] In some embodiments, the active ingredient of the preparation includes 2′-FL, which is used to inhibit the abnormal proliferation of Ruminococcus in the intestine, prevent Crohn's disease, ulcerative colitis or diarrhea-type irritable bowel syndrome or slow down the progression thereof; preferably, the preparation is applied to patients with inflammatory bowel disease and / or irritable bowel syndrome in an amount sufficient to inhibit the abnormal proliferation of Ruminococcus in the intestine of patients with inflammatory bowel disease and / or irritable bowel syndrome.

[0036] In some embodiments, the preparation further comprises 6′-SL, wherein the mass ratio of 2′-FL to 6′-SL is (1-2):(0.3-0.4), which is used to inhibit the abnormal proliferation of Ruminococcus in the intestine, prevent Crohn's disease, ulcerative colitis or diarrhea-predominant irritable bowel syndrome or slow down its progression; preferably, it is administered to patients with inflammatory bowel disease and / or irritable bowel syndrome in an amount sufficient to inhibit the abnormal proliferation of Ruminococcus in the intestine of patients with inflammatory bowel disease and / or irritable bowel syndrome.

[0037] More preferably, the mass ratio of 2′-FL to 6′-SL in the preparation is (1.5-2):(0.35-0.4).

[0038] In addition, the present invention also provides an application of human milk oligosaccharides in the preparation of a preparation for inhibiting the abnormal proliferation of uncultured rumenbacterium in the intestine, and the human milk oligosaccharides are 2′-FL and / or 6′-SL.

[0039] The following are examples

[0040] Human milk oligosaccharides mainly consist of three major categories: (1) fucosylated oligosaccharides, with 2'-fucosyllactose and 3'-fucosyllactose as representative substances; (2) sialylated oligosaccharides, with 3'-sialyllactose and 6'-sialyllactose as representative substances; (3) oligosaccharides formed by core sugar chain structures without fucosyl or sialyl groups, with lacto-N-tetraose and lacto-N-neotetraose as representative substances. The constituent monomers of 2'-fucosyllactose (2′-FL) include fucose, galactose and glucose, and its chemical name is α-L-fucopyranosyl-(1→2)-β-D-galactopyranosyl-(1→4)-D-glucopyranoside; while 6′-sialyllactose (6′-SL) is the most abundant sialylated human milk oligosaccharides (HMOs) in breast milk. In the following examples, both 2′-FL and 6′-SL are commercially available products and can be provided in the form of natural sources, synthetic sources, or bacterial fermentation sources.

[0041] Example 1 Safety evaluation of HMOs

[0042] Experimental animals: Healthy male BALB / c mice, 4 weeks old, weighing 14-17 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., with the production license number SCXK(E)2022-0030. The SPF-level animal laboratory of this center has a temperature of 20-26°C and a humidity of 40-70%, and the use license number is SYXK(E)2023-0045.

[0043] Conversion of gavage dose: The dose of mice was converted according to the dose ratio of mice to humans of 9.1:1.

[0044] Taking pure 2′-FL as an example, the concentration of 2′-FL is 1.15 g / L. In this example, based on the fact that a 15 kg infant drinks 450 mL of milk per day, the dosage ingested by the human body is calculated as follows:

[0045] 1.15g / L×450mL / 15kg=34.5mg / kg.BW / d;

[0046] Then, according to the mouse to human dose ratio of 9.1:1, the dose ingested by mice is 34.5×9.1=313.95 mg / kg.BW / d.

[0047] After one week of adaptive feeding observation of the experimental mice, they were randomly divided into 5 groups, with 8 mice in each group, namely blank control group (NC), low-dose 2′-FL group (LF), high-dose 2′-FL group (HF), low-dose 2′-FL+6′-SL group (LFS), and high-dose 2′-FL+6′-SL group (HFS). The LF, HF, LFS and HFS groups were gavaged with 1.15 g / L 2′-FL, 2.0 g / L 2′-FL, 1.15 g / L 2′-FL+0.30 g / L 6′-SL, and 2.0 g / L 2′-FL+0.37 g / L 6′-SL according to the human gavage concentration, and the daily gavage volume was 450 mL. The corresponding gavage volume of the mice was calculated according to the above conversion method. The NC group was gavaged with normal saline at the same dose according to the body weight of the mice. The test substance was administered by gavage once a day for 30 consecutive days. The test was performed 30 days after gavage, and the weight of mice in each group was recorded during the gavage process. The weight results of mice before and after the test are shown in Table 1.

[0048] Table 1 Effects of HMOs on the body weight of healthy mice (n=8)

[0049]

[0050]

[0051] From the test results in Table 1, we can see that the weight of mice in each group increased before and after oral administration, and there was no significant difference in the weight gain of mice in each group compared with the blank control group. In addition, all mice had no abnormal appearance during the experimental period, with clean ears, shiny hair, bright eyes and no odor, agile movements, and normal appetite and sleep. The weight of mice increased steadily during the experiment. The specific results are as follows: Figure 1 shown.

[0052] Determination of organ / body weight ratio: According to the experimental method determined in the "Catalogue of Health Functions Allowed to Be Claimed by Health Foods (2020 Edition)" issued by the State Administration for Market Regulation, the organ / body weight ratio of each group of mice was determined as follows:

[0053] After the administration was completed, the weights of all mice were measured. The mice were sacrificed by cervical dislocation, and the thymus and spleen were taken. The weights of the thymus and spleen were recorded. Finally, the ratios of the thymus and spleen to the body weight were calculated respectively as follows:

[0054] Thymus / body weight = thymus weight (mg) / mouse body weight (g);

[0055] Spleen / body weight = spleen weight (mg) / mouse body weight (g);

[0056] The measurement results are as Figure 2 shown, where Figure 2 the left in the figure is the thymus / body weight ratio, and the right is the spleen / body weight ratio.

[0057] The immune organs are the material basis for the animal body to realize immune functions, and their development status directly affects the strength of the body's immune function. Among them, the thymus and spleen are the two most important immune organs. The immune function of the animal body can be reflected by the thymus index and spleen index. From Figure 2 the measurement results, it can be seen that according to the gavage dose in this example, there were no significant differences in the measurement results of the thymus index and spleen index of each group of mice, indicating that HMOs had no significant effect on the organ / body weight ratio of mice.

[0058] Example 2 Inhibitory effect of HMOs on harmful bacteria in the intestinal flora of mice

[0059] Experimental animals: Healthy male BALB / c mice, 4 weeks old, weighing 14 - 17 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., with the production license number SCXK(E)2022 - 0030. The SPF - level animal laboratory of this center has a temperature of 20 - 26°C and a humidity of 40 - 70%, and the use license number is SYXK(E)2023 - 0045.

[0060] After one - week adaptive feeding and observation of the experimental mice, they were randomly divided into 5 groups, with 8 mice in each group, namely the blank control group (NC), low - dose 2′ - FL group (LF), high - dose 2′ - FL group (HF), low - dose 2′ - FL + 6′ - SL group (LFS), and high - dose 2′ - FL + 6′ - SL group (HFS). The daily gavage volume of the mice in the LF, HF, LFS, and HFS groups was the same as that in Example 1. The NC group was gavaged with the same dose of normal saline according to the body weight of the mice once a day. After continuously gavaging the test substances for 30 days, fecal samples were collected and stored in a - 80°C low - temperature refrigerator for later measurement, as follows:

[0061] According to the instructions of the bacterial DNA kit, bacterial microbial genomic DNA was extracted from the intestinal contents. Bacterial primer 27F (AGRGTTTGATYNTGGCTCAG) and reverse primer 1492R (TASGGHTACCTTGTTASGACTT) were used to amplify the bacterial genome V3-V4 variable region. The PCR amplification products were sent to Beijing Biomike Biotechnology Co., Ltd. for high-throughput sequencing. Chromatographic peak extraction and quantitative analysis were performed using Tracefinder software (Thermo Fisher Scientific, Waltham, MA, USA). The sequencing data were analyzed on the cloud platform (https: / / international.biocloud.net / zh / dashboard) of Beijing Biomike Biotechnology Co., Ltd. The spliced ​​raw data were filtered using lima v1.7.0, cutadapt 1.9.1, and UCHIME v4.2 software. Reads were clustered at a similarity level of 97.0% using Usearch software to obtain OTUs. The Alpha and Beta diversity indices of the samples were evaluated using QIIME2 software, and the results are as follows:

[0062] (1) Effects of different HMOs on the richness of intestinal flora in healthy mice

[0063] Alpha diversity analysis can reflect the species diversity within a single sample, including species richness (i.e., the number of different species in a community) and species evenness (i.e., how evenly these species are distributed in the community). The main indicators of alpha diversity include Chao1, ACE, Shannon, and Simpson indices. Chao1 and ACE can be used to measure the richness of species in a community, while Shannon and Simpson indices can be used to measure the diversity of communities in a sample.

[0064] Depend on Figure 3 A shows that there is no significant difference in ACE index and Chao1 index among the groups (P>0.05), indicating that there is no significant difference in community richness among the groups; Shannon and Simpson indexes also show that the community diversity in each group of samples is basically consistent (P>0.05). It can be seen that 2′-FL and 6′-SL have no significant effect on the community richness of each group of intestinal microbiota within a certain range of addition.

[0065] (2) Effects of different HMOs on the composition of intestinal flora in healthy mice

[0066] Beta diversity compares the differences in microbial communities between samples. It is used to measure the similarity or difference in species composition between different samples, and can reflect the changes in species composition of microbial communities as the environment changes. Among them, UniFrac distance is a metric based on evolutionary relationships, which can reflect the differences in microbial community structure between samples. Beta diversity is measured by comparing the distance values ​​between samples. The closer the distance between samples, the more similar the species composition. This embodiment uses PCA based on unweighted_unifrac to reduce the dimensionality of the sample distance matrix and then makes an intuitive judgment on the sample relationship on a two-dimensional plane. The results are as follows: Figure 3 As shown in B.

[0067] Depend on Figure 3 The results in Figure B show that the HF group was completely separated from the NC group, and the community composition of the two groups was quite different, while the LF group, LFS group, and HFS group all partially overlapped with the NC group, among which the LF group and the HFS group were close, and the fecal microbial community composition of the two groups was similar. It can be seen that different HMOs have different effects on the composition of intestinal microbiota, among which the supplementation of high doses (2.0 g / L) of 2′-FL has a more obvious effect on the species composition of the intestinal community.

[0068] (3) Effects of different HMOs on harmful bacteria in the intestines of healthy mice

[0069] This example compares the community composition and relative abundance of cecal microorganisms in mice in different treatment groups at the phylum level. Figure 3 In Figure C, it was found that the cecal flora of each treatment group at the phylum level were mainly Firmicutes, Bacteroidetes, Verrucomicrobia, Actinobacteria and Proteobacteria, which accounted for more than 90% of the total cecal bacteria, among which Firmicutes and Bacteroidetes were the dominant phyla in each group. The difference analysis of each treatment group at these phyla levels is as follows:

[0070] Compared with the NC group, all experimental groups had no significant effect on the overall relative abundance of Firmicutes and Bacteroidetes at the phylum level (P>0.05), among which the relative abundance of Firmicutes and Bacteroidetes in the NC group was 35.9% and 55.3%, respectively, in the LF group was 39.8% and 55.7%, respectively, in the HF group was 36.6% and 52.4%, respectively, in the LFS group was 31.5% and 49.5%, respectively, and in the HFS group was 38.0% and 55.4%. It can be seen that the intervention of 2′-FL or 2′-FL+6′-SL had almost no effect on the overall relative abundance of Firmicutes and Bacteroidetes at the phylum level.

[0071] Compared with the NC group, the relative abundance of Verrucomicrobia in the HF group was significantly increased (P<0.05), with the relative abundance of Verrucomicrobia in the NC group being 0.19% and the relative abundance of Verrucomicrobia in the HF group being 6.10%.

[0072] Compared with the NC group, the relative abundance of Actinobacteria in the LFS group was significantly increased (P<0.05), with the relative abundance of Actinobacteria in the NC group being 1.07% and the relative abundance of Actinobacteria in the LFS group being 3.05%.

[0073] In addition, compared with the NC group, the relative abundance of Cyanobacteria in the LF, HF and HFS groups was significantly decreased (P<0.05). The relative abundance of Cyanobacteria in the NC group was 2.15%, and the relative abundance of Cyanobacteria in the LF, HF and HFS groups were 0.21%, 0.42% and 0.61%, respectively.

[0074] Furthermore, at the genus level, we focused on evaluating the top 20 intestinal microorganisms in relative abundance. The relative abundance histogram of species at the genus level for each group is shown in Figure 2. Figure 4 As shown in A.

[0075] Depend on Figure 4As shown in Figure A, the relative abundance of the genus unclassified_Muribaculaceae was high in each group, with the largest number and dominance, and its relative abundance in the NC, LF, HF, LFS, and HFS groups was 23.0%, 20.3%, 20.9%, 19.4%, and 21.8%, respectively, with no significant difference among the groups. Unclassified_Muribaculaceae was reported as a producer of SCFAs and was associated with tryptophan metabolites, and its increased abundance played a vital role in improving the intestinal barrier and structure.

[0076] In addition, a heat map was used to compare the relative abundance of these 20 genus (genera) by horizontal clustering. The results are as follows: Figure 4 As shown in B, the small squares in the figure represent genes, and the color gradient from blue to red represents the relative abundance from low to high. ANOVA one-way analysis of variance and Tukey test were used to analyze the significance between groups. The results are shown in Figure 4 As shown in C, different letters in the figure indicate significant differences among different groups (P<0.05).

[0077] Depend on Figure 4 As shown in Figures B and C, compared with the NC group, the relative abundance of Ruminococcus in the intestines of mice in the LF, HF, LFS and HFS groups decreased, among which the relative abundance of Ruminococcus in the intestines of mice in the HF, LFS and HFS groups decreased significantly (P<0.05), and the relative abundance of the beneficial bacteria Akkermansia was significantly increased, and the relative abundance of Ruminococcus in the intestines of mice in the LFS and HFS groups was lower than that in the LF and HF groups, respectively. It can be seen that the combination of 6′-SL and 2′-FL can enhance the inhibitory effect of 2′-FL on intestinal Ruminococcus, and the combination of the two has a better effect in inhibiting the proliferation of intestinal Ruminococcus. It can be seen that 2′-FL and its combination with 6′-SL can be used to prepare preparations for inhibiting the abnormal proliferation of intestinal Ruminococcus.

[0078] Furthermore, the changes in the relative abundance of Ruminococcus gnavus, an important member of Ruminococcus, were compared in different groups. The results showed that compared with the NC group, the relative abundance of Ruminococcus gnavus in the feces of mice in the LF, HF, LFS and HFS groups was significantly reduced (P<0.05), especially the combination of 2′-FL and 6′-SL had a better inhibitory effect.

[0079] Existing studies have shown that Ruminococcus colonizing the human intestine is associated with the occurrence and development of a variety of diseases. For example, the study "Identification of colorectal cancer progression-associated intestinal microbiome and predictive signature construction" showed that the increase of Ruminococcus is associated with the progression of colorectal cancer (CRC). And the study "Ruminococcus gnavus, a member of the human gut microbiome associated with Crohn's disease, produces an infammatory polysaccharide" published by Matthew T. Henke et al. showed that R. gnavus colonizing the human intestine synthesizes and secretes complex glucorhamnose polysaccharides with rhamnose main chain and glucose side chain. This glucorhamnose effectively induces dendritic cells to secrete inflammatory cytokines, thereby causing inflammatory bowel disease. In addition, the research team led by Zhaoxiang Bian and Kailiang Wang from Hong Kong Baptist University published an article titled "Ruminococcus gnavus plays a pathogenic role in diarrhea-predominant irritable bowel syndrome by increasing serotonin biosynthesis" in Cell Host & Microbe, showing that Ruminococcus gnavus, which is highly enriched in the intestines of IBS-D patients, can stimulate the biosynthesis of 5-HT in the intestine through the decomposition of dietary essential amino acids, thereby promoting the development of IBS-D.

[0080] In summary, inhibiting the proliferation of Ruminococcus in the intestine is beneficial to prevent the occurrence or slow down the progression of inflammatory bowel disease or diarrhea-predominant irritable bowel syndrome.

[0081] In addition, Akkermansia is generally considered to be negatively correlated with inflammation levels, and its increased abundance is conducive to preventing the occurrence of inflammation; uncultured_rumen_bacterium is negatively correlated with propionic acid and butyric acid, while propionic acid helps maintain the stability of the intestinal environment, and butyric acid can reduce intestinal inflammation, which may have a certain therapeutic effect on diseases such as inflammatory bowel disease. The combination of 2′-FL or 2′-FL+6′-SL can not only significantly reduce the proliferation of intestinal harmful bacteria Ruminococcus, but also significantly promote the growth of beneficial bacteria Akkermansia and inhibit the proliferation of uncultured_rumen_bacterium. It can be seen that the preparation containing 2′-FL or 2′-FL+6′-SL is more conducive to preventing or slowing down inflammatory bowel disease caused by abnormal proliferation of Ruminococcus.

[0082] In summary, 2′-FL or 2′-FL+6′-SL intervention can inhibit the proliferation of harmful bacteria Ruminococcus in the intestine and reduce its relative abundance, and can be used to prepare preparations that inhibit the proliferation of intestinal Ruminococcus.

[0083] The human milk oligosaccharides described in the present invention have a certain preventive effect on inflammatory bowel disease or irritable bowel syndrome caused by the proliferation and abnormal proliferation of Ruminococcus, and can be used to prepare preparations for preventing enteritis and maintaining intestinal health. In particular, the human milk oligosaccharides are 2′-FL, or a combination of 2′-FL and 6′-SL, which can be used to prepare preparations for preventing enteritis caused by the proliferation of intestinal Ruminococcus.

[0084] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technical personnel in this field can make some changes or modify the technical contents disclosed above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. An application of human milk oligosaccharides, characterized in that: The human milk oligosaccharide is 2'-FL and / or 6'-SL, and is used for preparing a preparation for inhibiting the proliferation of intestinal Ruminococcus.

2. The use according to claim 1, characterized in that The human milk oligosaccharide is 2′-FL or a combination of 2′-FL and 6′-SL.

3. The use according to claim 2, characterized in that It is used to prepare a preparation for inhibiting the abnormal proliferation of Ruminococcus in the intestines of patients with inflammatory bowel disease.

4. The use according to claim 3, characterized in that The invention is used for preparing a preparation for inhibiting abnormal proliferation of Ruminococcus in the intestine of patients with Crohn's disease or ulcerative colitis.

5. The use according to claim 2, characterized in that It is used to prepare a preparation for inhibiting the abnormal proliferation of Ruminococcus in the intestines of patients with irritable bowel syndrome.

6. The use according to claim 5, characterized in that It is used to prepare a preparation for inhibiting the abnormal proliferation of Ruminococcus in the intestines of patients with diarrhea-type irritable bowel syndrome.

7. The use according to any one of claims 3 to 6, characterized in that: The human milk oligosaccharide is a composition of 2′-FL and 6′-SL.

8. The use according to claim 7, characterized in that The mass ratio of 2′-FL to 6′-SL in the composition is 1.0-2.0:0.3-0.

4.

9. The use according to claim 1, characterized in that The human milk oligosaccharide is 2′-FL and / or 6′-SL, and is used for preparing a preparation for preventing enteritis and maintaining intestinal health.

10. The use according to claim 9, characterized in that The human milk oligosaccharide is 2′-FL or a combination of 2′-FL and 6′-SL, and is used for preparing a preparation for preventing enteritis caused by the proliferation of Ruminococcus in the intestine.

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