Application of narirutin in improving intestinal flora homeostasis of food allergy organism

Abstract

The invention belongs to the technical field of food functional ingredient nutriology research, and discloses application of narirutin in improving the intestinal flora homeostasis of food allergy organisms. The narirutin can adjust the relative abundance value of certain intestinal flora by adjusting the diversity of the intestinal flora of a food allergy body, improve the relative abundance of beneficial bacteria and harmful bacteria, and can influence the metabolism of short-chain fatty acid of the intestinal flora of the food allergy body, so that the food allergy body is improved. Further, the application of the narirutin in improving the intestinal flora homeostasis of food allergy organisms is realized. The application of narirutin in the research fields of nutriology, biology and pharmacology is expanded.

Description

Technical Field

[0001] The present invention belongs to the technical field of nutritional research on food functional components, and specifically relates to the application of naringin in improving the intestinal flora homeostasis of food-allergic organisms. Background Art

[0002] With the acceleration of the industrialization process, the number of people with food allergies has gradually increased. The imbalance of intestinal homeostasis in food-allergic people is the core link in the occurrence and development of allergic diseases, and maintaining the homeostasis of intestinal flora is an important way to prevent and treat food allergic diseases. At present, the main way to maintain the intestinal flora homeostasis in food-allergic people is to reasonably supplement probiotics. However, the existing probiotics have unstable curative effects on maintaining the intestinal flora homeostasis in food-allergic people and may cause digestive function disorders. With the acceleration of the promotion of the national health concept, flavonoids have received increasing attention due to their special functional activities and multi-dimensional health effects. Naringin is a flavonoid substance of natural origin and is widely present in the fruits of Rutaceae plants (grapefruit, pomelo, orange). It has biological activities such as antioxidant, anti-inflammatory, antiviral and immunomodulatory effects, and can be used as a functional food in the food and health industries.

[0003] At present, there has been no research on the application of naringin in improving the intestinal flora homeostasis of food-allergic organisms. The present invention helps to expand the application of naringin in the fields of nutrition, biology and pharmacology research. Summary of the Invention

[0004] The purpose of the present invention is to provide an application of naringin in improving the intestinal flora homeostasis of food-allergic organisms. The naringin provided by the present invention has the effect of improving the intestinal flora homeostasis of food-allergic organisms, can effectively regulate the abundance of intestinal flora, achieve the improvement of flora homeostasis, and prevent and treat the occurrence of food allergic reactions.

[0005] In order to achieve the above purpose, the technical solution of the present invention is as follows:

[0006] In the first aspect of the present invention, a flavonoid small molecule - naringin for maintaining the intestinal flora homeostasis of food-allergic organisms is provided, and the structure of the naringin is as Figure 1 shown;

[0007] In the second aspect of the present invention, a method for evaluating the intestinal flora homeostasis of food-allergic organisms is provided;

[0008] In the third aspect of the present invention, an application of the above naringin in the preparation of a product for maintaining the intestinal flora homeostasis of food-allergic organisms is provided.

[0009] In some examples of the present invention, the product has any one or more of the following functions:

[0010] (1) Naringin increases the diversity of gut microbiota in food allergy-prone organisms;

[0011] (2) Naringin increases the relative abundance of probiotics in the gut microbiota of food allergy-prone organisms;

[0012] (3) Naringin increases the relative abundances of probiotics g.Lachnospiraceae_NK4A136_group, g.norank_f_Lachnospiraceae, and g.unclassified_f_Lachnospiraceae, and g.Streptococcus;

[0013] (4) Naringin decreases the relative abundance of harmful bacteria in the gut microbiota of food allergy-prone organisms that are likely to cause food allergies;

[0014] (5) Naringin decreases the relative abundances of harmful bacteria g.Clostridium sensu stricto_1, g.Romboutsi, g.Turicibaccter, and g.Faecalibaculum;

[0015] (6) Naringin can maintain the gut microbiota homeostasis in food allergy-prone organisms by affecting the metabolism of short-chain fatty acids Description of the Drawings

[0016] Figure 1 The structure of naringin

[0017] Figure 2 Naringin affects the diversity of gut microbiota in food allergy-prone organisms

[0018] Figure 3 Naringin affects the relative abundance of gut microbiota in food allergy-prone organisms

[0019] Figure 4 Naringin affects the metabolism of short-chain fatty acids Detailed Implementation Modes

[0020] According to the following embodiments, the present invention can be better understood. However, those skilled in the art can easily understand that the content described in the embodiments is only used to illustrate the present invention and should not limit the present invention detailedly described in the claims.

[0021] A method for evaluating the gut microbiota homeostasis in food allergy-prone organisms using an animal model of the present invention is achieved through different groupings and different animal treatment methods for different groups.

[0022] (1) Experimental grouping

[0023] Healthy female BALB / c mice aged 4 - 6 weeks were randomly divided into 4 groups: blank control group (CON), sensitized model group (FA), 30 mg / kg naringin group (A3), and 50 mg / kg naringin group (A5).

[0024] (2) Establishment of the evaluation model

[0025] D0 - D4: CON and FA groups: gavaged with a certain amount of PBS; A3 and A5 groups: gavaged with a certain amount of OVA, the A3 group was gavaged with 30 mg / kg of naringin, and the A5 group was gavaged with 50 mg / kg of naringin;

[0026] D11 - D18: CON: injected intraperitoneally with PBS; FA group, A3 and A5 groups: injected intraperitoneally with OVA sensitization solution

[0027] D25 - D39: CON: gavaged with PBS 4 times a week for a total of 8 times; FA group, A3 and A5 groups: gavaged with OVA challenge solution 4 times a week for a total of 8 times

[0028] (3) Obtaining intestinal contents: Immediately after the mice were sacrificed, the cecum was dissected out, the surface of the cecum was rinsed with PBS, the cecal contents of each group of mice were collected aseptically, the samples were quickly placed in a centrifuge tube, the tube mouth was wrapped with sealing film, and stored in a -80 °C refrigerator for later use.

[0029] (4) High-throughput sequencing of intestinal flora 16S rDNA and detection of short-chain fatty acids

[0030] (5) Data analysis: All data analysis was performed on the Majorbio Cloud Platform (https: / / cloud.majorbio.com) as follows: The Alpha diversity indices Chao 1, Ace, Simpson, and Shannon indices were calculated using Mothur (version 1.30.2) software, dilution curves were constructed, and the Kruskal-Wallis rank sum test was used for inter-group difference analysis of Alpha diversity; PCoA analysis (principal coordinate analysis) based on the bray-curtis distance algorithm and hierarchical clustering analysis were used to test the similarity of the microbial community structure between samples, and the Kruskal-Wallis rank sum test was also used for multi-group difference comparison of single species; LEfSe analysis (LDA > 4, P < 0.05) was used to determine the flora with significant differences in abundance at the phylum to genus level between different groups.

[0031] Example 1 Naringin increases the diversity of intestinal flora in food-allergic organisms;

[0032] The structural formula of the naringin is as shown in the appendix Figure 1As shown, the naringin of the present invention can increase the diversity of the intestinal flora of food-allergic organisms, specifically as shown in Figure 2 A, B, C, D, and E in the appendix. Alpha diversity is a comprehensive index describing the diversity of the flora within a specific ecological environment, and the overall microbial community structure is evaluated by analyzing the richness and diversity of microorganisms in individual samples. Alpha diversity is commonly evaluated using the Chao 1, Ace, Shannon, and Simpson indices. The Chao 1 index and Ace index reflect species richness. The Simpson index and Shannon index reflect species diversity. The results of Alpha diversity are as shown in Figure 2 shown. The Chao 1, Ace, and Shannon indices of the FA group were all lower than those of the CON group, and the difference was not statistically significant. The Chao 1 and Ace indices of the A3 and A5 groups were significantly higher than those of the FA group, indicating that the microbial richness of the A3 and A5 groups was significantly higher than that of the FA group. The higher the Shannon index and the lower the Simpson index, the higher the species diversity. The Shannon index of the A5 group was significantly higher than that of the FA group, and the Simpson index was significantly lower than that of the FA group, indicating that the microbial diversity of the A5 group was significantly higher than that of the FA group. Beta diversity is used to reflect the difference in the composition of the microbial community between samples by analyzing the content of various taxa in the samples. Principal Coordinate Analysis (PCoA) is used to visualize the Beta diversity data. The closer the two sample points on the PCoA are, the more similar the species composition of the two samples. The scatter plot based on the PCoA scores shows that the community compositions of FA and CON, FA and A3, and FA and A5 are significantly separated.

[0033] Example 2 Naringin increases the relative abundance of probiotics in the intestinal flora of food-allergic organisms;

[0034] To further analyze the changes in the composition and structure of the intestinal flora, the relative abundances of the intestinal flora were analyzed at the phylum and genus levels, respectively.

[0035] The relative abundances of different species at the phylum level in different groups are as shown in the appendix Figure 2As shown in F and G. At the phylum level, the gut microbiota was mainly composed of p.Firmicutes, p.Actinobacteriota, and p.Bacteroidota. p.Firmicutes was the most dominant in each group of mice. The CON and FA groups were 84.16±12.89% and 90.55±4.76% respectively, and the A3 and A5 groups were 93.02±4.74% and 87.42±3.65% respectively. The relative abundance of FA was higher than that of CON, and the A5 group was the lowest. The changing trend of p.Actinobacteriota was similar, and the A3 group was the lowest. The relative abundances of p.Bacteroidota in the CON and FA groups were 7.08±5.48% and 1.16±0.83% respectively, and those in the A3 and A5 groups were 2.48±1.72% and 2.34±1.36% respectively. The relative abundance of FA was lower than that of CON, and the abundances in the A3 and A5 groups increased compared with those in the FA and OT groups. At the genus level, the top 10 genera were shown, including g.Lactobacillus, g.Clostridium sensu stricto 1, g.norank f norank o clostridia UCG-014, g.Turicibacter, g.Enterorhabdus, g.Faecalibaculum, g.Lachnosoiraceae NK4A136, etc., with relatively high abundances.

[0036] Analysis of the differences in the microbiota, as shown in the appendix Figure 2 As shown in H and I, the LEfSe (LDA Effect Size) linear discriminant method was used to analyze the data (screening conditions: LDA>4.0, p<0.05) to clarify the community differences in the gut microbiota of each group of mice. LEfSe is an analysis method used to find differentially abundant species between groups. The vertical axis represents the taxonomic units with significant differences between groups, and the horizontal axis intuitively shows the LDA analysis logarithmic score values of each taxonomic unit with a bar chart. The longer the length, the more significant the difference of the taxonomic unit. At the genus level, g.Lachnosoiraceae NK4A136 was the significantly different genus in the CON group; g.Adlercreutzia and g.Bifidobacterium were the significantly different genera in the FA group; g.Faecalibaculum in the low-dose naringin group had significant differences compared with other genera; g.norank f norank o clostridia UCG-014 was the dominant genus in the high-dose naringin group.

[0037] Example 3 Naringin increases the relative abundances of beneficial bacteria g.Lachnospiraceae_NK4A136_group, g.norank_f_Lachnospiraceae, g.unclassified_f_Lachnospiraceae, and g.Streptococcus;

[0038] As shown in A, B, C, and D in the appendix Figure 3 Due to the occurrence of food allergy reactions in the body, the relative abundance values of the probiotics g.Lachnospiraceae_NK4A136_group, g.norank_f_Lachnospiraceae, g.unclassified_f_Lachnospiraceae, and g.Streptococcus in the FA group were significantly lower than those in the CON group. With the action of naringin, the A3 and A5 groups were able to significantly increase the relative abundances of these four probiotics to varying degrees. This shows that naringin can maintain the intestinal homeostasis of food-allergic organisms by increasing the relative abundances of four typical probiotics in the intestinal flora.

[0039] Example 4 Naringin reduces the relative abundances of harmful bacteria in the intestinal flora of food-allergic organisms that are likely to cause food allergies, which is achieved by reducing the relative abundances of harmful bacteria g.Clostridium sensu stricto_1, g.Romboutsis, g.Turicibaccter, and g.Faecalibaculum.

[0040] As shown in E, F, G, and H in the appendix Figure 3 Due to the occurrence of food allergy reactions in the body, the relative abundance values of the harmful bacteria g.Clostridium sensu stricto_1, g.Romboutsis, g.Turicibaccter, and g.Faecalibaculum in the FA group were significantly higher than those in the CON group. With the action of naringin, the A3 and A5 groups were able to significantly reduce the relative abundances of these four harmful bacteria to varying degrees. This shows that naringin can maintain the intestinal homeostasis of food-allergic organisms by increasing the relative abundances of four typical harmful bacteria in the intestinal flora.

[0041] Example 5 Naringin can maintain the homeostasis of the intestinal flora of food-allergic organisms by affecting the metabolism of short-chain fatty acids

[0042] The content changes of SCFAs in the cecal contents of mice in each group were detected by GC-MS / MS to study whether naringin can also regulate the SCFAs metabolism of allergic mice. Figure 4The results showed that high and low doses of naringin intervention had different degrees of influence on the contents of acetic acid, propionic acid, isobutyric acid, valeric acid and isovaleric acid in the cecal contents of OVA-allergic mice, while there was no significant difference in the content of butyric acid and caproic acid after the intake of OVA and naringin. The acetic acid content in the FA group was significantly higher than that in the CON group, and it decreased significantly after intervention with different doses of naringin. The propionic acid content in the high dose naringin group was significantly lower than that in the FA group. The contents of isobutyric acid, valeric acid and isovaleric acid in the FA group were significantly higher than those in the CON group, and high and low doses of naringin could significantly reduce their contents to different degrees, restoring them to levels close to those of the normal control group. The above results showed that naringin could significantly regulate the content of SCFAs in OVA-induced allergic mice and make it tend to normal levels.

[0043] The correlation between intestinal flora and SCFAs mainly exists between differential bacterial genera and isobutyric acid, isovaleric acid, and propionic acid. g.Lachnospiraceae NK4A136 was extremely significantly negatively correlated with the contents of valeric acid, isobutyric acid, and isovaleric acid, and was significantly negatively correlated with the contents of acetic acid, propionic acid, and caproic acid. g.Alloprevotella was extremely significantly negatively correlated with the contents of valeric acid, isobutyric acid, and isovaleric acid, and was significantly negatively correlated with the content of acetic acid. g.Streptococcus was negatively correlated with the content of caproic acid. g.NorankfLachnospiraceae was negatively correlated with the content of valeric acid. g.Clostridium sensu stricto 1 and gBifidobacterium were extremely significantly positively correlated with the contents of valeric acid, isobutyric acid, and isovaleric acid, and gTuricibacter was significantly positively correlated with the contents of valeric acid, isobutyric acid, and isovaleric acid. OVA treatment significantly reduced the relative abundance of g Lachnospiraceae_NK4A136_group and increased the relative abundance of Clostridium sensu stricto 1 and g Turicibacter. Correlation analysis showed that g Lachnospiraceae NK4A136 was significantly negatively correlated with the contents of various short-chain fatty acids, while g.Clostridiumsensu stricto 1 and g.Turicibacter were significantly positively correlated with the contents of various short-chain fatty acids. This suggests that OVA increases the content of short-chain fatty acids by regulating the relative abundance of the above-mentioned genera, and naringin regulates the levels of these genera and adjusts the SCFAs content to normal.

Claims

1. An application of a flavonoid small molecule naringin, characterized in that: The use of naringin in improving the homeostasis of intestinal flora in food allergic organisms; The naringin can effectively regulate the diversity of intestinal flora and improve the homeostasis of intestinal flora in food allergic organisms. The naringin can effectively regulate the relative abundance of intestinal flora, thereby improving the homeostasis of intestinal flora in food allergic organisms. The naringin can effectively increase the relative abundance of beneficial intestinal flora, thereby improving the homeostasis of intestinal flora in food allergic organisms. The naringin can effectively reduce the relative abundance of harmful intestinal flora, thereby improving the homeostasis of intestinal flora in food allergic organisms.

2. The use according to claim 1, characterized in that: The naringin can effectively regulate the metabolism of short-chain fatty acids in the intestine, thereby improving the homeostasis of intestinal flora in food allergic organisms.

3. The use according to claim 1, characterized in that: The naringin can effectively regulate the Alpha diversity and Beta diversity of the intestinal flora of the body, thereby improving intestinal homeostasis.

4. The use according to claim 1, characterized in that: The naringin can effectively regulate the relative abundance of g. Lactobacillus, g. Clostridium sensu stricto 1, g. norankf noranko clostridia UCG-014, g. Turicibacter, g. Enterorhabdus, g. Faecalibaculum, and g. Lachnosoiraceae NK4A136 in the intestinal flora of the body, thereby improving intestinal homeostasis.

5. The use according to claim 1, characterized in that: The naringin can effectively regulate the intestinal flora of the body, increase the relative abundance of beneficial bacteria g.Lachnospiraceae_NK4A136_group, g.norank_f_Lachnospiraceae, g.unclassified_f_Lachnospiraceae, and g.Streptococcus, and achieve improvement of intestinal homeostasis.

6. The use according to claim 1, characterized in that: The naringin can effectively regulate the intestinal flora of the body, reduce the relative abundance of harmful bacteria g. Clostridium sensu stricto_1g. Romboutsis g. Turicibaccter and g. Faecalibaculum, and achieve improvement of intestinal homeostasis.

7. The use according to claim 2, characterized in that: Naringin regulates the metabolism of acetic acid, propionic acid, isobutyric acid, valeric acid and isovaleric acid by the intestinal flora of food allergic organisms, thereby improving intestinal homeostasis.

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