Bifidobacterium animalis subsp. Lactis composition for improving vision health and application thereof

Abstract

The invention belongs to the technical field of microbial engineering, discloses a bifidobacterium animalis subsp. Lactis composition for improving vision health and application thereof, and finds that the bifidobacterium animalis subsp. Lactis i797 has an outstanding effect on improving the vision health. The invention provides the animal bifidobacterium subsp. Lactis composition for improving the vision health, which contains the animal bifidobacterium subsp. Lactis i797, and can reduce the eyeball diopter of a myopia guinea pig model, remarkably improve the diopter of a low-myopia guinea pig model and relieve asthenopia, so that the aim of improving the vision health is fulfilled; the composition of the bifidobacterium animalis subsp. Lactis i797, the xanthophyll and the vitamin A in a specific proportion can remarkably improve vision health. The bifidobacterium animalis subsp. Lactis composition is suitable for preparing the bifidobacterium animalis subsp. Lactis composition for improving the vision health, and is used for preparing dairy products for improving the vision health or probiotics for improving the vision health.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial engineering, and relates to an application of animal bifidobacterium lactis subspecies, in particular to an animal bifidobacterium lactis subspecies composition for improving vision health and an application thereof. Background Art

[0002] With the popularity of electronic products such as smartphones and tablets, people spend more and more time in front of screens. Staring at electronic screens for a long time can lead to visual fatigue and accelerate the development of myopia or other eye problems. Vision health issues have become a public health issue that needs to be paid great attention to at all ages. Among them, according to surveys, the myopia rate among children and adolescents is continuing to rise. Among adults, as many as 93.67% of working people have symptoms of visual fatigue, and cataracts, presbyopia and other problems among middle-aged and elderly people should not be ignored.

[0003] Among the many vision health problems, myopia not only affects the quality of daily life, but may also lead to serious ophthalmic complications such as retinal detachment. Especially for adolescents, myopia can also affect educational achievements, employment opportunities, etc. At present, the research on myopia mainly focuses on preventing and relieving visual fatigue, and delaying the progression of myopia.

[0004] With the deepening of probiotic research, the "intestinal-eye" axis has become a new research direction that is expected to solve the problem of myopia. At present, there are many types and strains of known probiotics, and their specific mechanisms of action and efficacy are also different. Some strains have been reported, such as the Chinese invention patent with publication number CN116139235B and the name "A medicinal and edible dual-purpose composition with the effect of relieving visual fatigue in adolescents and its preparation method", which discloses that a Chinese herbal medicine composition containing Lactobacillus plantarum has the effect of relieving visual fatigue, but it only discloses that the composition is used to relieve visual fatigue caused by screen blue light, etc. Therefore, Lactobacillus plantarum can only be used to prevent and avoid the deepening of myopia, and cannot improve organic problems caused by myopia such as axial elongation and lens refractive power. Among other probiotics, the representative strain BB-12 is the most widely used animal Bifidobacterium lactis subspecies strain with the most published related papers. Its role in improving immunity, regulating intestinal and oral health, and relieving upper respiratory tract infections has been proven in both in vitro and human experiments. However, there is currently no research on the application of Bifidobacterium animalis subsp. lactis in vision health, especially in improving axial length of the eye. Summary of the invention

[0005] In order to solve the above problems existing in the prior art, the present invention screens strains with outstanding effects in improving vision health from dozens of probiotics. In particular, it is found that among the animal Bifidobacterium lactis subspecies that have been maturely used commercially, only the i797 strain has the effect of improving vision health. It is also unexpectedly found that when animal Bifidobacterium lactis subspecies i797 is used in combination with lutein and vitamin A in a specific ratio as a composition, it can significantly alleviate myopia-related problems such as axial elongation and lens refractive power, thereby achieving the purpose of improving vision health.

[0006] The invention provides a composition for improving vision health containing animal bifidobacterium lactis subspecies i797 and application thereof. The composition can achieve the purpose of improving vision health.

[0007] The present invention also provides the use of the animal Bifidobacterium lactis subspecies composition for improving vision health in the preparation of milk powder, yogurt or probiotics with the effect of improving vision health.

[0008] The present invention also provides milk powder, yogurt or probiotics containing the animal Bifidobacterium lactis subspecies composition for improving vision health.

[0009] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0010] A composition of Bifidobacterium animalis subspecies lactis for improving vision health, the composition comprising Bifidobacterium animalis subspecies lactis i797, lutein and vitamin A;

[0011] In the composition, the number of viable bacteria of Bifidobacterium animalis subspecies i797 is 2×10 6 -5×10 10 CFU / g or CFU / mL.

[0012] Among them, the animal Bifidobacterium lactis subsp. lactis strain i797 is disclosed in the Chinese invention patent with application number CN202010259210.8. It was deposited in the China General Microbiological Culture Collection Center of the China Microbiological Culture Collection Administration on August 20, 2019, with the deposit number CGMCC NO.18403, and its Latin name is Bifidobacterium animalissubsp.lactis.

[0013] The composition improves vision health by reducing axial elongation of the eye and increasing the refractive power of the lens.

[0014] As a further limitation of the present invention, the composition contains an amount ratio of 1×10 6 -1×10 9 CFU: 50-120μg: 45-70μg of Bifidobacterium animalis lactis subsp. i797, lutein and vitamin A.

[0015] The present invention also provides the application of the above-mentioned animal Bifidobacterium lactis subspecies composition for improving vision health, specifically for preparing milk powder, yogurt or probiotics.

[0016] As a limitation of the present invention, the Bifidobacterium lactis subsp. animalis composition for improving vision health is added in an amount of 0.6%-4.5% in milk powder, yogurt, fermented milk or probiotics, in percentage by weight.

[0017] The present invention also provides a milk powder comprising the Bifidobacterium lactis subsp. animalis composition for improving vision health as described in any one of the above technical solutions.

[0018] The present invention also provides a yogurt comprising the Bifidobacterium lactis subspecies composition for improving vision health described in any one of the above technical solutions.

[0019] The present invention also provides a probiotic agent, comprising the Bifidobacterium lactis subsp. animalis composition for improving vision health as described in any one of the above technical solutions.

[0020] Due to the adoption of the above technical solution, the present invention has the following beneficial effects compared with the prior art:

[0021] (1) The present invention discovered that animal Bifidobacterium lactis subsp. i797 has the effect of improving vision health, developed an animal Bifidobacterium lactis subsp. composition for improving vision health, and effectively utilized the mechanism of action of active animal Bifidobacterium lactis subsp. i797 to improve vision health by reducing axial elongation of the eye and increasing the refractive power of the lens.

[0022] (2) The components of the Bifidobacterium lactis subsp. i797 composition for improving vision health of the present invention synergize to enhance the efficacy, so that lutein and vitamin A, which have no obvious effect on reducing axial elongation and improving lens refractive power when used alone at a certain concentration, can be compounded with Bifidobacterium lactis subsp. i797 at a specific concentration and used in the form of a composition, thereby significantly improving the vision health effect of the composition product.

[0023] (3) The animal Bifidobacterium lactis subspecies composition for improving vision health in the present invention can be applied to products such as milk powder, yogurt, fermented milk and probiotics.

[0024] The invention is suitable for the preparation of an animal Bifidobacterium lactis subspecies composition capable of improving vision health, and is used for industrial production of milk powder, yogurt or probiotics capable of improving vision health. DETAILED DESCRIPTION

[0025] The present invention is further described in detail below through specific embodiments. It should be understood that the described embodiments are only used to explain the present invention, and are not intended to limit the present invention.

[0026] The instruments, materials, reagents, etc. used in the examples of the present invention can be obtained from commercial sources unless otherwise specified. The experimental methods without specific conditions in the examples are usually carried out under conventional conditions or under conditions recommended by the manufacturer. The dosage of reagents in the verification experiments of the examples of the present invention has been verified by the minimum toxic concentration detection experiment.

[0027] The instruments, materials and reagents are described as follows: disposable 5 ml syringe (Shanghai Kangdelai, China), electronic analytical balance (Precisa12A, Precisa, Switzerland), strip retinoscope (YZ24, Suzhou Liuliu Vision Technology Co., Ltd., China), guinea pig gavage needle (Zhongke Life Science, China), small animal keratometer (SN-001, STRIA.TECH, Germany), ophthalmic A / B type ultrasound diagnostic instrument (MD-2300S, Tianjin Mida Medical Technology Co., Ltd., China).

[0028] Compound tropicamide eye drops (Jinyao, Yongguang Pharmaceutical Co., Ltd., China), lidocaine hydrochloride eye drops (Sichuan Heyi Pharmaceutical, China), tobramycin dexamethasone eye ointment (saALCON-COUVREUR nv, USA), xylazine hydrochloride injection (Jilin Fangzheng Animal Pharmaceutical Co., Ltd., China), anesthetic: 5% chloral hydrate: 5 g of chloral hydrate was dissolved in about 60 ml of normal saline, and the volume was made up to 100 ml with normal saline, and stored at room temperature away from light.

[0029] Probiotics: Bifidobacterium animalis subsp. lactis i797 and Bifidobacterium animalis subsp. lactis JM-1038;

[0030] Among them, Bifidobacterium animalis subspecies lactis JM-1038 was isolated from the intestinal samples of healthy infants and preserved in the Junlebao strain resource bank.

[0031] Inactivation of probiotics: dilute the probiotics to the intragastric concentration with normal saline and boil at 100°C for 30 minutes.

[0032] In the verification experiment of the embodiment of the present invention: the experimental animals are calico guinea pigs, male, 5 months old, provided by Life Breeding Farm (China), Pukou District, Nanjing, and the certificate number is: SCXK (Su) 2024-0007.

[0033] Example 1 A Bifidobacterium lactis subsp. animalis composition for improving vision health

[0034] This embodiment is a composition containing animal Bifidobacterium lactis subspecies i797 for improving vision health, comprising 1×10 10CFU / g of animal Bifidobacterium lactis subsp. i797 100 μg (i.e., the total viable count of animal Bifidobacterium lactis subsp. i797 is 1×10 9 CFU), 112 μg of lutein and 59.5 μg of vitamin A were mixed to obtain an animal Bifidobacterium lactis subsp. composition for improving vision health by reducing axial elongation and increasing lens refractive power.

[0035] 1. Experimental design to verify the efficacy of improving vision health

[0036] (1.1) Setting and processing of each group:

[0037] Animal grouping: After 7 days of adaptive feeding, 110 guinea pigs were divided into 11 groups, namely: blank group, model group, positive group, inactivated group 797, inactivated group 1038, experimental group 797, experimental group 797 combination, experimental group 1038, experimental group 1038 combination, lutein group and vitamin A group, with 10 animals in each group.

[0038] Myopia modeling: The initial axial length, corneal curvature and refractive power of each group of guinea pigs were measured and recorded. Except for the blank group, all guinea pigs were modeled with myopia by form deprivation myopia: after anesthesia assisted by the muscle relaxant xylazine hydrochloride injection, a translucent balloon was used as an eye mask fixed on the right eye of the guinea pig's face, and the left eye was used as its own control eye. It was worn and raised for 2 weeks, and the eye mask was checked twice a day for any fall-off. If the eye mask was found to fall off, it was immediately re-fixed. After 2 weeks, the modeling of guinea pigs in each group was verified: the myopia modeling was successful when the guinea pig's refractive power decreased and the axial length increased, that is, when "relative myopia" was achieved. Among them, tobramycin dexamethasone eye ointment was applied around the eyes of the guinea pigs before and after form deprivation to prevent conjunctivitis infection.

[0039] Experimental treatment: The experimental period was 28 days. After the modeling verification of the 11 experimental groups, the guinea pigs in each group were gavaged every other day. The guinea pigs in the blank group, model group and positive group were given an equal volume of normal saline every day, and the guinea pigs in the positive group were additionally instilled with 1-2 drops of pirenzepine eye drops in the right eye every day.

[0040] Inactivated group 797, inactivated animal Bifidobacterium lactis subsp. i797 was administered orally daily, and the dose was equivalent to 5x10 9 CFU;

[0041] The inactivated group 1038 was given inactivated animal Bifidobacterium lactis subspecies JM-1038 by gavage every day, and the dose was equivalent to 5x10 9 CFU;

[0042] Experimental group 797 was given active probiotics animal Bifidobacterium lactis subsp. i797 by gavage daily, and the dose converted to human daily intake is 5x109 CFU;

[0043] The experimental group 797 composition was administered daily with the composition product of Example 1, wherein the doses of lutein, vitamin A and active animal Bifidobacterium lactis subsp. i797 were converted to 112 μg, 59.5 μg and 1×10 9 CFU;

[0044] The experimental group 1038 was given active probiotics Bifidobacterium animalis subspecies lactis JM-1038 by gavage every day, and the dose was equivalent to 5x10 9 CFU;

[0045] The experimental group 1038 composition was administered daily by gavage of a composition product prepared by mixing lutein, vitamin A and active animal Bifidobacterium lactis subspecies JM-1038, wherein the doses of lutein, vitamin A and active animal Bifidobacterium lactis subspecies JM-1038 were equivalent to 112 μg, 59.5 μg and 1x10 9 CFU.

[0046] The lutein group was given lutein by oral gavage daily, and the dose was equivalent to 112 μg of daily intake for humans.

[0047] The vitamin A group was given vitamin A by oral gavage daily, and the dose converted to human daily intake corresponds to 59.5 μg.

[0048] (1.2) Indicator measurement method:

[0049] One day after the last intragastric administration, the corneal curvature, axial length and refractive power of each group of guinea pigs were measured again, and the efficacy of each administered sample in improving vision health was evaluated by statistical analysis of the results of each index.

[0050] Corneal curvature measurement: The guinea pig corneal curvature radius was measured using a Keratometer small animal keratometer. After fixing the guinea pig, the measurement was performed in a dark room. The measurement was repeated more than 10 times for each eye, and the average value was taken after removing the obvious deviation values.

[0051] Axial length measurement: MEDA A / B ultrasound was used to measure the axial length of the right eye of the guinea pig. The propagation speed of ultrasound in different media in the eye was set as follows: 1534 m / s for the anterior chamber and vitreous body, and 1774 m / s for the lens. 0.4% lidocaine hydrochloride eye drops were used for corneal surface anesthesia before measurement. During measurement, the probe was aligned with the center of the cornea and perpendicular to the corneal plane. The measurement was repeated 5 times for each eye, and the average value was taken after removing the obvious deviation value.

[0052] Refractive power measurement: Before the measurement, the guinea pigs were dilated with compound tropicamide eye drops, once every 5 minutes for a total of three times. After the pupils were fully dilated, the retinoscope was used in a dark room to measure the refractive power of the left and right eyes of all guinea pigs, and the refractive power value was determined according to the principle of equivalent spherical lens. The measurement was repeated 3 times for each eye and the average value was taken.

[0053] The statistical results were expressed as sample mean ± standard deviation and were analyzed using SPSS10.0 software package. One-way ANOVA was used to compare different groups at different time points, paired sample T test was used for the same group at different time points, LSD was used for multiple comparisons between groups, and p < 0.05 was considered significant.

[0054] (II) The guinea pig myopia model was successfully constructed using the form deprivation method

[0055] After wearing the eye mask and feeding normally for 2 weeks, compared with the blank group, the axial length of the right eye (modeling eye) of the guinea pigs in the other groups increased significantly, p<0.01; and the refractive power decreased significantly, p<0.01. There were no significant differences in the axial length, refractive power and corneal curvature of the left eye (control eye) of the guinea pigs in each group. The results showed that the guinea pig myopia model was established. See Table 1 for details.

[0056] Table 1. Effects of form deprivation on guinea pig eyes

[0057]

[0058]

[0059] Note: Compared with the blank group, **p<0.01

[0060] (III) Results of the experiment to verify the efficacy of improving vision health

[0061] After 4 weeks of gavage intervention, compared with the model group, the axial length of the right eye (modeling eye) of the guinea pigs in the experimental group 797 composition group (lutein + vitamin A + active animal Bifidobacterium lactis subspecies i797 compound product group) was significantly reduced p<0.01, and the refractive power was significantly improved p<0.05; the axial length of the right eye (modeling eye) of the guinea pigs in the experimental group 797 group (active animal Bifidobacterium lactis subspecies i797) showed a downward trend, and the refractive power showed an upward trend. See Table 2 for details.

[0062] Table 2. Improvement of guinea pig myopia model by animal Bifidobacterium lactis subsp. i797 and its compound products

[0063]

[0064]

[0065] Note: Compared with the blank group, **p<0.01; compared with the model group, #p<0.05, ##p<0.01

[0066] The above results indicate that active animal Bifidobacterium lactis subspecies i797 has a tendency to improve visual health, specifically by reducing axial elongation and increasing the refractive power of the lens to achieve improved visual health. The combination product of active animal Bifidobacterium lactis subspecies i797 and lutein and vitamin A has the effect of improving visual health.

[0067] Example 2 A Bifidobacterium lactis subsp. animalis composition for improving vision health

[0068] The composition of this embodiment contains 5×10 11 CFU / g of animal Bifidobacterium lactis subsp. i797 100 μg (i.e., the total viable count of animal Bifidobacterium lactis subsp. i797 was 5×10 10 CFU), 50 μg of lutein and 45 μg of vitamin A, and mix them evenly to obtain an animal Bifidobacterium lactis subspecies composition for improving vision health.

[0069] Example 3 A Bifidobacterium lactis subsp. animalis composition for improving vision health

[0070] The composition of this embodiment contains 2×10 7 CFU / g of Bifidobacterium animalis subsp. lactis i797 100 μg (i.e., the total viable count of Bifidobacterium animalis subsp. lactis i797 was 2×10 6 CFU), lutein 120 μg and vitamin A 70 μg, and mix them evenly to obtain a composition of animal Bifidobacterium lactis subspecies for improving vision health.

[0071] Examples 4-7 are milk powders containing a composition of animal Bifidobacterium lactis subsp. for improving vision health.

[0072] The milk powder in Example 4 is added with 4.5% by weight of the Bifidobacterium lactis subsp. animalis composition for improving vision health in Example 3.

[0073] The milk powder in Example 5 is added with 2.4% by weight of the Bifidobacterium lactis subsp. animalis composition for improving vision health in Example 4.

[0074] The milk powder in Example 6 is added with 4.0% by weight of the Bifidobacterium lactis subsp. animalis composition for improving vision health in Example 2.

[0075] The milk powder in Example 7 is added with 0.6% by weight of the Bifidobacterium lactis subsp. animalis composition for improving vision health in Example 1.

[0076] Examples 8-12 are formula milk powders containing a composition of animal Bifidobacterium lactis subsp. for improving vision health.

[0077] The yogurt in Example 8 is added with 0.6% by weight of the Bifidobacterium animalis subsp. lactis composition for improving vision health in Example 1.

[0078] The yogurt in Example 9 is added with 1.6% by weight of the Bifidobacterium animalis subsp. lactis composition for improving vision health in Example 2.

[0079] The formula milk powder in Example 10 is added with 2.9% by weight of the Bifidobacterium lactis subsp. animalis composition for improving vision health in Example 4.

[0080] The formula milk powder in Example 11 is added with 4% by weight of the Bifidobacterium animalis subsp. lactis composition for improving vision health in Example 3.

[0081] The formula milk powder in Example 12 is added with 3.5% by weight of the Bifidobacterium animalis subsp. lactis composition for improving vision health in Example 4.

[0082] In another embodiment, the composition for improving vision health comprises 1×10 8 CFU: 120μg: 45μg or 1×10 7 CFU: 100 μg: 70 μg or 1 × 10 8 The composition of CFU: 120μg: 65μg of animal Bifidobacterium lactis subspecies i797, lutein and vitamin A has been proven to have a certain effect in improving vision health.

[0083] In other embodiments, the amount of the Bifidobacterium lactis subsp. animalis composition for improving vision health added to milk powder, yogurt or probiotics is a value between 0.6% and 4.5% by weight. The obtained products all have the effect of improving vision health.

[0084] It should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention is described in detail with reference to the above embodiments, those skilled in the art can still modify the technical solutions described in the above embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the scope of protection of the claims of the present invention.

Claims

1. An animal Bifidobacterium lactis subspecies composition for improving vision health, characterized in that: The composition comprises animal bifidobacterium lactis subspecies i797, lutein and vitamin A; In the composition, the number of viable bacteria of Bifidobacterium animalis subspecies i797 is 2×10 6 -5×10 10 CFU / g or CFU / mL.

2. The composition of Bifidobacterium lactis subsp. animalis for improving vision health according to claim 1, characterized in that: The composition contains a dosage ratio of 1×10 6 -1×10 9 CFU: 50-120μg: 45-70μg of Bifidobacterium animalis lactis subsp. i797, lutein and vitamin A.

3. The use of the animal Bifidobacterium lactis subsp. composition for improving vision health according to any one of claims 1 to 2, characterized in that: The animal Bifidobacterium lactis subspecies composition for improving vision health is used for preparing milk powder, yogurt or probiotics.

4. The use of the animal Bifidobacterium lactis subspecies composition for improving vision health according to claim 3, characterized in that: In terms of weight percentage, the Bifidobacterium lactis subspecies composition for improving vision health is added in an amount of 0.6%-4.5% in milk powder, yogurt or probiotics.

5. A milk powder, characterized in that: A composition for improving visual health comprising Bifidobacterium lactis subsp. animalis according to any one of claims 1 to 2.

6. A yogurt, characterized in that: A composition for improving visual health comprising Bifidobacterium lactis subsp. animalis according to any one of claims 1 to 2.

7. A probiotic, characterized in that: A composition for improving visual health comprising Bifidobacterium lactis subsp. animalis according to any one of claims 1 to 2.

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