A composition for improving asthenopia and use thereof

Abstract

The application discloses a composition for improving asthenopia and application, and the composition is compounded by longissimum bifidobacterium longum subsp i772, 2'-fucose lactose, lacto-N-neotetraose and lutein with a weight ratio of 1-100:460-1600:120-400:0.03-0.35. The composition for improving asthenopia has a synergistic effect in the function of relieving asthenopia, longissimum bifidobacterium longum subsp i772, 2'-fucose lactose, lacto-N-neotetraose and lutein can effectively relieve the apoptosis of eye cells caused by asthenopia, and the composition can be applied to infant formula milk powder, children's formula milk powder, fermented milk and health food.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of microbial engineering, and relates to a composition containing Bifidobacterium longum, in particular, a composition for improving visual fatigue and application. BACKGROUND

[0002] The eye is one of the most important organs in human senses, and eye health has always been the pursuit of the nation. The Global Eye Health Research Report released in 2020 shows that the concern and cognition of Chinese people for eye health are at the global leading level. At the same time, a study in 2021 shows that the incidence of visual fatigue among college students worldwide is 46-71%. Studies have shown that the main physiological mechanisms of visual fatigue are oxidative stress damage, inflammatory damage, cell aging, and excessive depletion of visual cells. Among them, excessive depletion of visual cells is the main cause of visual fatigue and visual decline in children and adolescents. When visual cells are excessively depleted, and the supply of nutrients required by them is not timely, the recovery time of the macula and retina will increase, thereby causing visual fatigue. Therefore, supplementing appropriate nutrients required by the eye can help alleviate the occurrence of visual fatigue. Existing technologies show that lutein is a good substance for visual cell nutrition supplementation. However, there are certain limitations in using lutein alone to alleviate visual fatigue, such as the need for long-term use, different effects on different people, and safety hazards caused by excessive use.

[0003] Human milk oligosaccharides (HMOs) are an important component of breast milk, with a variety of benefits, including promoting the growth of beneficial bacteria, enhancing the immune system, and preventing intestinal infections. In recent years, scientists have conducted more and more in-depth research on human milk oligosaccharides, and more and more products have begun to apply HMOs to infant formula milk powder and other foods.

[0004] Zebrafish (Danio rerio) is a highly developed model animal with a very similar retinal morphological structure to humans and other vertebrates. The eye of zebrafish accounts for a large proportion of the whole, and its transparent embryonic characteristics facilitate experimental operation and organ visualization. Therefore, the model organism zebrafish has advantages for eye experiments. SUMMARY

[0005] The present application aims to provide a composition for improving visual fatigue and application, which can effectively reduce the apoptosis of eye cells caused by visual fatigue.

[0006] To achieve the above-mentioned purpose, the technical solution adopted by the present application is:

[0007] A composition for improving asthenopia, the composition comprising Bifidobacterium longum subsp. Longum i772, 2'-fucosyllactose, lacto-N-neotetraose and lutein.

[0008] The Bifidobacterium longum subsp. Longum i772 has been deposited with the China General Microbiological Culture Collection Center on May 20, 2020, and the deposit number is CGMCC NO. 19856, and the bacteria have been disclosed in CN112458015B.

[0009] As a limitation, the weight ratio of the Bifidobacterium longum subsp. Longum i772, 2'-fucosyllactose, lacto-N-neotetraose and lutein is 1-100: 460-1600: 120-400: 0.03-0.35.

[0010] As another limitation, the viable bacterial count of the Bifidobacterium longum subsp. Longum i772 is 5x10 10 ~ 5x10 11 CFU / g.

[0011] The application also provides the use of the above-mentioned composition for improving asthenopia, the composition being used for preparing infant formula milk powder, child formula milk powder, fermented milk or health food with the efficacy of improving asthenopia.

[0012] As a limitation, the addition amount of the composition in the infant formula milk powder, child formula milk powder, fermented milk or health food is 0.5-4.7% by weight.

[0013] The application also provides an infant formula milk powder comprising the above-mentioned composition for improving asthenopia.

[0014] The application also provides a child formula milk powder comprising the above-mentioned composition for improving asthenopia.

[0015] The application also provides a health food comprising the above-mentioned composition for improving asthenopia.

[0016] Compared with the prior art, the application has the following technical progress:

[0017] The application provides a composition for improving visual fatigue, wherein Bifidobacterium longum subsp. longum i 772, 2'-fucosyllactose (2'-FL) and lacto-N-neotetraose (LNnt) have no effect on improving visual fatigue, and the composition is prepared by compounding the three with lutein, so that the effect of improving visual fatigue is significantly improved compared with lutein alone. The composition has a synergistic effect on relieving visual fatigue, and the effect is specifically shown in the following aspects: the fluorescence intensity of apoptotic cells in the eyes of zebrafish can be reduced, and the damage of cell tissues in the eyes can be improved. The composition can be applied to infant formula milk powder, children's formula milk powder, fermented milk and health foods. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 The fluorescence intensity data of apoptotic cells in the eyes of zebrafish after sample treatment in Example 5 of the application;

[0019] Figure 2 The fluorescence intensity graph of apoptotic cells in the eyes of zebrafish after sample treatment in Example 5 of the application, and the yellow dotted box is the analysis part;

[0020] Figure 3 The typical graph of the eye tissue structure of zebrafish after sample treatment in Example 5 of the application, and the red arrow is the analysis part. DETAILED DESCRIPTION

[0021] The application will be further described in detail through specific examples. It should be understood that the preferred examples described herein are only used to illustrate and understand the application, and are not used to limit the application.

[0022] The materials, reagents and the like used in the examples of the application can be obtained from commercial channels unless otherwise specified. The experimental methods not specified in the examples are usually carried out according to conventional conditions or according to the conditions recommended by the manufacturer.

[0023] Examples 1-3 improve the composition for visual fatigue and application

[0024] Examples 1-3 respectively provide a composition for improving visual fatigue, and the specific components and the amount of each component are shown in Table 1:

[0025] Table 1

[0026]

[0027]

[0028] Example 5 verifies the effect of the composition for improving visual fatigue

[0029] This example verifies the effect of the composition for improving visual fatigue, and the specific verification experiment is as follows:

[0030] I. Experimental animals:

[0031] Zebrafish were all raised in fish water at 28℃ (water quality: 200mg of instant sea salt was added to each 1L of reverse osmosis water, conductivity was 450-550μS / cm; pH was 6.5-8.5; hardness was 50-100mg / L CaCO3), provided by the fish breeding center of the company, the experimental animal use license number was: SYXK(Zhe)2022-0004, the feeding and management met the requirements of international AAALAC certification (certification number: 001458), IACUC ethical review number: IACUC-2023-6993-01;

[0032] Wild-type AB strain zebrafish were bred by natural pair mating, and the age was 1dpf.

[0033] II. Experimental grouping

[0034] Group 1: normal control group

[0035] Group 2: model control group

[0036] Group 3: glutathione

[0037] Group 4: Bifidobacterium longum i 772

[0038] Group 5: lutein

[0039] Group 6: i 772+2'-FL+LNnT

[0040] Group 7: i 772+2'-FL+LNnT+lutein

[0041] 30 zebrafish in each group, a total of 7 groups.

[0042] III. Experimental method

[0043] S1. Group 1 did not use intervention, and groups 2-7 were given mycophenolate mofetil to establish a zebrafish asthenopia model, and were all treated at 28℃ for 1 day;

[0044] S2. After S1 treatment, 10 zebrafish were randomly selected from each group for AO staining and placed under a fluorescence microscope for photography, and NIS-Elements D3.20 advanced image processing software was used for analysis and data collection. The fluorescence intensity of ocular apoptotic cells was analyzed, and the statistical analysis results of the index were used to evaluate the asthenopia improvement effect of the sample. The statistical processing results were expressed as mean±SE, and SPSS 26.0 software was used for statistical analysis. p<0.05 indicated that the difference had statistical significance. The experimental results are shown in Table 2, Figure 1 , Figure 2

[0045] ​Table 2 Intervention amount and visual fatigue improvement efficacy experimental results

[0046]

[0047] Note: *** means p < 0.001 compared with the model control group; &&& means p < 0.001 compared with the experimental group 4

[0048] As shown in Table 1, the fluorescence intensity of apoptotic cells in the eyes of group 2 (model control group) was significantly higher than that of group 1 (normal control group), and the fluorescence intensity of apoptotic cells in the eyes of group 3 (glutathione group) was restored and had no difference with the normal control group, indicating that the modeling was successful.

[0049] As shown in Table 1, Figure 1 and Figure 2 Under the conditions of this experiment, it was found that group 2 (model control group) had no obvious visual fatigue improvement efficacy compared with group 4 (long bifidobacterium i 772 alone) and group 6 (i 772+2'-FL+LNnT composition); group 3 (glutathione group), group 5 (lutein group) and group 7 (long bifidobacterium i 772+2'-FL+LNnT+lutein composition) all had visual fatigue improvement efficacy; Notably, comparing group 7 and group 5 alone, it was found that group 7 had a significant difference compared with group 5, indicating that the composition (i 772+2'-FL+LNnT+lutein) had more significant visual fatigue improvement efficacy than lutein alone.

[0050] S3. After S1 treatment, each group of zebrafish was fixed, dehydrated, embedded, sectioned, H&E stained and other steps, and eye histopathology observation and analysis was performed to evaluate the visual fatigue improvement efficacy of the sample; The experimental results are shown in Figure 3 .

[0051] As shown in Table 1, Figure 3 It was found that group 2 (model control group) zebrafish eye histopathology examination showed that retinal cells generally underwent apoptosis (red arrow), showing cell pyknosis and chromatin margination; The positive control group 3 (glutathione group) zebrafish eye histopathology examination showed that the apoptosis of retinal cells in the eyes was significantly improved, indicating that glutathione had visual fatigue improvement efficacy, specifically the improvement of eye cell apoptosis. Group 5 (lutein group) and group 7 (long bifidobacterium i 772+2'-FL+LNnT+lutein composition) zebrafish eye histopathology examination showed that the apoptosis of retinal cells was improved, indicating that the above-mentioned compositions had visual fatigue improvement efficacy, and the improvement effect of group 7 on eye cell apoptosis was closest to the positive drug (glutathione group) in histopathology; i 772 group and i 772+2'-FL+LNnT group retinal cells generally underwent apoptosis, indicating that the above-mentioned concentrations had no obvious visual fatigue improvement efficacy.

Claims

1. A composition for improving eye strain, characterized in that, The composition comprises Bifidobacterium longum subsp. i772, 2'-fucolactose, lactose-N-neotetrasaccharide and lutein; The Latin name of the *Bifidobacterium longum* subspecies i772 is... Bifidobacterium longum subsp. Longum The strain was deposited on May 20, 2020, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.19856.

2. The composition for improving eye strain according to claim 1, characterized in that, The weight ratio of Bifidobacterium longum subsp. i772, 2'-fucolactose, lactose-N-neotetrasaccharide, and lutein is 1–100: 460–1600: 120–400: 0.03–0.

35.

3. The composition for improving eye strain according to claim 1 or 2, characterized in that, The viable count of the *Bifidobacterium longum* subspecies i772 was 5 × 10⁻⁶. 10 ~5×10 11 CFU / g.

4. The application of the composition for improving eye strain according to any one of claims 1-3, characterized in that, The composition is used to prepare infant formula, children's formula, fermented milk, or health food with the effect of improving eye fatigue.

5. The application of the composition for improving eye strain according to claim 4, characterized in that, The composition is added in infant formula, children's formula, fermented milk or health food at a weight percentage of 0.5% to 4.7%.

6. An infant formula milk powder, characterized in that, A composition comprising any one of claims 1-3 for improving eye strain.

7. A type of infant formula milk powder, characterized in that, A composition comprising any one of claims 1-3 for improving eye strain.

8. A health food product, characterized in that, A composition comprising any one of claims 1-3 for improving eye strain.

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