Fermented Lactobacillus mucilaginosus IOB802 and application of metagen thereof in improving eye health

Abstract

The invention provides application of fermented lactobacillus mucus IOB802 and a metagen thereof in improving eye health, and relates to the technical field of microbial fermentation. The strain improves the activity of retinal epithelial cells so as to enhance the antioxidant capacity, provide metabolic support, promote cell repair, maintain blood-retinal barrier, reduce apoptosis and other mechanisms, reduce the content of proinflammatory factors, reduce the content of vascular endothelial growth factors and reduce the expression level of inflammatory factors. The antioxidant enzyme activity and macular region edema can be improved, and the eye health can be improved.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fermentation, and particularly relates to application of fermented mucus lactobacillus IOB802 and postbiotics thereof in improving eye health. Background Art

[0002] The retina is a key tissue in the eye responsible for sensing light and transmitting visual signals. It is located at the back of the eyeball and is composed of multiple layers of cells, including photoreceptors (rods and cones) and ganglion cells. The health of the retina is directly related to the clarity and quality of vision. Once damaged, it may lead to decreased vision or even blindness. In recent years, with the popularization of electronic devices, people are increasingly exposed to blue light, which has a shorter wavelength and higher energy. It can penetrate the transparent medium of the eye and directly act on the retina. Studies have shown that long-term or high-intensity blue light exposure may cause damage to the retina, induce photochemical damage, oxidative stress response, and even lead to retinal cell death. This damage not only affects the visual quality, but also may increase the risk of fundus lesions such as macular degeneration. Therefore, the prevention and treatment of retinal damage is particularly important. It is not only related to the visual health of individuals, but also to the public health issues of the entire society.

[0003] Retinal damage often involves oxidative stress and inflammatory responses, which can impair retinal structure and function. Vascular endothelial growth factor (VEGF) is a key angiogenic factor. In pathological processes such as diabetic retinopathy, VEGF expression increases, promoting new blood vessel formation, leading to macular edema and preretinal neovascularization. VEGF levels are significantly increased in the aqueous humor and vitreous of patients with diabetic macular edema (DME), indicating that VEGF plays a key role in the progression of DME. Antioxidant enzymes such as glutathione S-transferase (GST) are expressed in retinal glial cells, exerting antioxidant functions and alleviating damage to retinal neurons.

[0004] Studies have shown that probiotics can limit intestinal inflammation and promote mucosal tissue repair by regulating VEGF signaling. For example, Saccharomyces boulardii can reduce basal VEGFR-2 phosphorylation, VEGFR-2 phosphorylation in response to VEGF, and activation of downstream kinases PLCγ and Erk1 / 2, thereby inhibiting angiogenesis. In addition, probiotics have been found to enhance antioxidant factors such as IL-10, total antioxidant status, total antioxidant capacity, and glutathione. They achieve their antioxidant effects by producing metabolites and antioxidant enzymes, increasing antioxidant capacity, and reducing host oxidant metabolites. The antioxidant activity of probiotics can not only reduce oxidative damage, but also exert its effects through mechanisms such as regulating host signaling pathways and reducing the activity of reactive oxygen species-producing enzymes. At the same time, Lactobacillus fermentans has strong antioxidant activity. Therefore, probiotics have potential health benefits in regulating VEGF content and antioxidant enzyme activity, providing a new perspective for the prevention and treatment of eye-related symptoms.

[0005] In summary, VEGF plays an important role in improving eye health. Therefore, it is urgent to find and develop a probiotic and its microecological preparation that can improve eye health. Summary of the invention

[0006] In view of the technical problems existing in the prior art, the present invention aims to provide an application of fermented Lactobacillus mucilaginosus IOB802 and its postbiotics in improving eye health.

[0007] Lactobacillus mucilaginosus fermentation Limosilactobacillusfermentum ) The IOB802 strain was deposited on August 5, 2021 in the General Microbiology Center of the China Microorganism Culture Collection Administration, referred to as CGMCC, with the address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and its deposit number is CGMCC No. 23120.

[0008] One of the purposes of the present invention is to provide a use of fermented Lactobacillus mucosus IOB802 in the preparation of a medicine for improving eye health. The fermented Lactobacillus mucosus IOB802 is classified as fermented Lactobacillus mucosus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

[0009] Preferably, the fermented Lactobacillus mucilaginosus IOB802 is used to prepare a medicine for improving eye health in the form of fermented Lactobacillus mucilaginosus IOB802 bacterial powder, and the preparation method of the fermented Lactobacillus mucilaginosus IOB802 bacterial powder comprises: The strain was placed in a closed culture at 30-37°C for 24-36 hours and subcultured twice to obtain a fermentation seed liquid; the fermentation seed liquid was inoculated into a fermentation liquid medium, the fermentation tank was stirred at a speed of 50-70 r / min, the temperature was 37°C, the pH was 6.2±0.2, and the closed culture was carried out for 12 hours; the fermentation liquid medium was centrifuged and the bacterial mud was obtained, and the fermented mucus Lactobacillus IOB802 powder was obtained after vacuum freeze-drying.

[0010] The second object of the present invention is to provide an application of fermented mucus lactobacillus IOB802 in preparing a health food for relieving visual fatigue. The fermented mucus lactobacillus IOB802 is classified as fermented mucus lactobacillus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

[0011] Preferably, the fermented Lactobacillus mucilaginosus IOB802 is used to prepare a health food for relieving visual fatigue in the form of fermented Lactobacillus mucilaginosus IOB802 bacterial powder, and the preparation method of the fermented Lactobacillus mucilaginosus IOB802 bacterial powder comprises: The strain was placed in a closed culture at 30-37°C for 24-36 hours and subcultured twice to obtain a fermentation seed liquid; the fermentation seed liquid was inoculated into a fermentation liquid medium, the fermentation tank was stirred at a speed of 50-70 r / min, the temperature was 37°C, the pH was 6.2±0.2, and the closed culture was carried out for 12 hours; the fermentation liquid medium was centrifuged and the bacterial mud was obtained, and the fermented mucus Lactobacillus IOB802 powder was obtained after vacuum freeze-drying.

[0012] The third object of the present invention is to provide a postbiotic application of fermented Lactobacillus mucilaginosus IOB802 in the preparation of a drug for improving eye health. The fermented Lactobacillus mucilaginosus IOB802 is classified as fermented Lactobacillus mucilaginosus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

[0013] Preferably, the preparation method of postbiotics by fermenting Lactobacillus mucilaginosus IOB802 comprises: The strain is placed in a closed culture at 30-37°C for 24-36 hours, subcultured twice, and fermentation seed liquid is obtained; the fermentation seed liquid is inoculated into the fermentation liquid culture medium, the fermentation tank is stirred at a speed of 50-70 r / min, the temperature is 37°C, the pH is 6.2±0.2, and the closed culture is carried out for 12 hours; after the fermentation liquid is inactivated, the materials are mixed evenly, and spray-dried to obtain fermentation mucus Lactobacillus IOB802 postbiotics.

[0014] The fourth object of the present invention is to provide an application of fermented mucus lactobacillus IOB802 postbiotics in the preparation of health food for relieving visual fatigue. The fermented mucus lactobacillus IOB802 is classified as fermented mucus lactobacillus ( Limosilactobacillusfermentum), and its deposit number is CGMCC No.23120.

[0015] Preferably, the preparation method of postbiotics by fermenting Lactobacillus mucilaginosus IOB802 comprises: The strain is placed in a closed culture at 30-37°C for 24-36 hours, subcultured twice, and fermentation seed liquid is obtained; the fermentation seed liquid is inoculated into the fermentation liquid culture medium, the fermentation tank is stirred at a speed of 50-70 r / min, the temperature is 37°C, the pH is 6.2±0.2, and the closed culture is carried out for 12 hours; after the fermentation liquid is inactivated, the materials are mixed evenly, and spray-dried to obtain fermentation mucus Lactobacillus IOB802 postbiotics.

[0016] Beneficial effects of the present invention: The invention provides an application of fermented mucinous lactobacillus IOB802 and postbiotics thereof in improving eye health.

[0017] The bacteria increased the vitality of retinal epithelial cells, thereby improving eye health through multiple mechanisms such as enhancing antioxidant capacity, providing metabolic support, promoting cell repair, maintaining the blood-retinal barrier and reducing cell apoptosis; fermented mucus Lactobacillus IOB802 reduced the levels of cellular proinflammatory factors IL-6, IL-1β, and TNF-α, alleviated inflammation, helped maintain the integrity of the blood-retinal barrier, prevented harmful substances from penetrating into the retina, reduced damage to visual cells, reduced the risk of tissue damage and cell apoptosis, enhanced the vitality and function of retinal cells, and improved their resistance to blue light. It also helped to restore the expression of antioxidant enzymes, enhanced the cells' ability to respond to oxidative stress caused by blue light, and reduced the accumulation of reactive oxygen species.

[0018] Fermented Lactobacillus mucinus IOB802 can effectively improve macular edema by reducing VEGF content, thereby improving eye health. Increased VEGF usually leads to abnormal proliferation and increased permeability of retinal blood vessels, causing macular edema, which in turn damages the function of visual cells. By inhibiting the production of VEGF, retinal edema can be reduced, the integrity of the blood-retinal barrier can be maintained, the inflammatory response can be reduced, and the nutrient supply of cells can be improved, thereby enhancing the vitality and resistance of retinal cells, enabling them to better cope with oxidative stress and damage caused by blue light, and promoting the overall health of the retina.

[0019] Fermented Lactobacillus mucilaginosus IOB802 can also increase the activity of catalase, superoxide dismutase and glutathione peroxidase; these enzymes play a key role in antioxidant defense, and can effectively remove reactive oxygen species (ROS) and hydrogen peroxide induced by blue light, reducing the damage of oxidative stress to visual cells. Specifically, SOD converts superoxide into oxygen and hydrogen peroxide, catalase decomposes hydrogen peroxide into water and oxygen, and glutathione peroxidase further removes hydrogen peroxide and lipid peroxides by utilizing glutathione. By increasing the activity of these antioxidant enzymes, the antioxidant capacity of retinal cells can be enhanced, cell damage and apoptosis can be reduced, thereby better protecting retinal health. At the same time, fermented Lactobacillus mucilaginosus IOB802 also reduces the content of proinflammatory factors IL-6, IL-1β, and TNF-α in mouse serum, improves the body's immune imbalance, and regulates the body's immune system. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 is the activity graph of retinal epithelial cells; Figure 2 is the level of IL-6, a pro-inflammatory factor in cells; Figure 3 is the level of IL-1β, a pro-inflammatory factor in cells; Figure 4 is the level of TNF-α, a pro-inflammatory factor in cells; Figure 5 is the vascular endothelial growth factor content in cells; Figure 6 is the activity of cell catalase; Figure 7 It is the activity of superoxide dismutase in cells; Figure 8 is the activity of cellular glutathione peroxidase; Fig. 9 is the level of pro-inflammatory factor IL-6 in serum; Fig.10 is the level of pro-inflammatory factor IL-1β in serum; Fig.11 is the level of pro-inflammatory factor TNF-α in serum; Fig.12 is the content of vascular endothelial growth factor in serum; Fig.13 is the activity of catalase in serum; Fig.14 It is the activity of superoxide dismutase in serum; Fig.15 is the activity of glutathione peroxidase in serum; Fig.16 is the malondialdehyde activity in serum; Fig.17Mouse eye tissue slices.

[0021] The fermented mucus lactobacillus provided by the present invention ( Limosilactobacillusfermentum ) The IOB802 strain was deposited on August 5, 2021 in the General Microbiology Center of the China Microorganism Culture Collection Administration, referred to as CGMCC, with the address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and its deposit number is CGMCC No. 23120. DETAILED DESCRIPTION

[0022] The invention provides an application of fermented mucinous lactobacillus IOB802 and postbiotics thereof in improving eye health.

[0023] According to a first aspect of the present invention, there is provided a use of fermented Lactobacillus mucilaginosus IOB802 in the preparation of a medicine for improving eye health, characterized in that the fermented Lactobacillus mucilaginosus IOB802 is classified and named as fermented Lactobacillus mucilaginosus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

[0024] In a preferred embodiment of the present invention, the fermented Lactobacillus mucosus IOB802 is used to prepare a medicine for improving eye health in the form of fermented Lactobacillus mucosus IOB802 powder, and the preparation method of the fermented Lactobacillus mucosus IOB802 powder comprises: The strain was placed in a closed culture at 30-37°C for 24-36 hours and subcultured twice to obtain a fermentation seed liquid; the fermentation seed liquid was inoculated into a fermentation liquid medium, the fermentation tank was stirred at a speed of 50-70 r / min, the temperature was 37°C, the pH was 6.2±0.2, and the closed culture was carried out for 12 hours; the fermentation liquid medium was centrifuged and the bacterial mud was obtained, and the fermented mucus Lactobacillus IOB802 powder was obtained after vacuum freeze-drying.

[0025] According to a second aspect of the present invention, there is provided an application of fermented mucus lactobacillus IOB802 in preparing a health food for relieving visual fatigue, wherein the fermented mucus lactobacillus IOB802 is classified as fermented mucus lactobacillus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

[0026] In a preferred embodiment of the present invention, the fermented Lactobacillus mucilaginosus IOB802 is used to prepare a health food for relieving visual fatigue in the form of fermented Lactobacillus mucilaginosus IOB802 bacterial powder, and the preparation method of the fermented Lactobacillus mucilaginosus IOB802 bacterial powder comprises: The strain was placed in a closed culture at 30-37°C for 24-36 hours and subcultured twice to obtain a fermentation seed liquid; the fermentation seed liquid was inoculated into a fermentation liquid medium, the fermentation tank was stirred at a speed of 50-70 r / min, the temperature was 37°C, the pH was 6.2±0.2, and the closed culture was carried out for 12 hours; the fermentation liquid medium was centrifuged and the bacterial mud was obtained, and the fermented mucus Lactobacillus IOB802 powder was obtained after vacuum freeze-drying.

[0027] According to a third aspect of the present invention, there is provided a use of a fermented Lactobacillus mucoides IOB802 postbiotic in the preparation of a drug for improving eye health, wherein the fermented Lactobacillus mucoides IOB802 is classified as fermented Lactobacillus mucoides ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

[0028] In a preferred embodiment of the present invention, the preparation method of postbiotics by fermenting Lactobacillus mucilaginosus IOB802 comprises: The strain is placed in a closed culture at 30-37°C for 24-36 hours, subcultured twice, and fermentation seed liquid is obtained; the fermentation seed liquid is inoculated into the fermentation liquid culture medium, the fermentation tank is stirred at a speed of 50-70 r / min, the temperature is 37°C, the pH is 6.2±0.2, and the closed culture is carried out for 12 hours; after the fermentation liquid is inactivated, the materials are mixed evenly, and spray-dried to obtain fermentation mucus Lactobacillus IOB802 postbiotics.

[0029] According to a fourth aspect of the present invention, there is provided an application of fermented mucus lactobacillus IOB802 postbiotics in the preparation of a health food for relieving visual fatigue, wherein the fermented mucus lactobacillus IOB802 is classified as fermented mucus lactobacillus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

[0030] In a preferred embodiment of the present invention, the preparation method of postbiotics by fermenting Lactobacillus mucilaginosus IOB802 comprises: The strain is placed in a closed culture at 30-37°C for 24-36 hours, subcultured twice, and fermentation seed liquid is obtained; the fermentation seed liquid is inoculated into the fermentation liquid culture medium, the fermentation tank is stirred at a speed of 50-70 r / min, the temperature is 37°C, the pH is 6.2±0.2, and the closed culture is carried out for 12 hours; after the fermentation liquid is inactivated, the materials are mixed evenly, and spray-dried to obtain fermentation mucus Lactobacillus IOB802 postbiotics.

[0031] In a preferred embodiment of the present invention, improving eye health includes reducing the content of vascular endothelial growth factor, reducing the expression level of inflammatory factors, and improving antioxidant enzyme activity and macular edema.

[0032] In a preferred embodiment of the present invention, alleviating visual fatigue includes reducing the content of vascular endothelial growth factor, reducing the expression level of inflammatory factors, and improving antioxidant enzyme activity and macular edema.

[0033] Example 1 Preparation of fermented Lactobacillus mucilaginosus IOB802 powder / postbiotic powder Activation of strains: Inoculate the strain into liquid culture medium at 1% inoculation volume, and culture at 37±2℃, and let it stand in a closed state. Observe the colony morphology and bacterial characteristics at different culture times. The culture requires that the seeds are uniform, consistent in morphology, and strong. The seed age of 24h in culture time is more in line with the conditions, so the best culture time for seeds is determined to be 24h. The activated seed liquid is obtained. The activated seed liquid is inoculated into the activation culture medium at 3% inoculation volume, and the culture conditions are 37±2℃, and let it stand in a closed state for 24h to obtain the fermented seed liquid.

[0034] Fermentation: According to the inoculation amount of 3%, the cultured secondary seed liquid was inoculated into the liquid culture medium. The fermentation tank was stirred at a speed of 50-70 r / min, 37°C, and cultured in a closed manner for 12 hours. During the fermentation process, food-grade sodium hydroxide was added for alkali supplementation to ensure that the pH of the fermentation liquid was maintained at 6.2±0.2.

[0035] Preparation of bacterial powder: After centrifugation of the fermentation medium, bacterial sludge was obtained, and the fermented mucus Lactobacillus IOB802 bacterial powder was obtained after vacuum freeze drying. The freeze drying conditions were pre-freezing at -40°C for 6 hours, freeze drying for 36±2 hours, and freeze drying was completed when the vacuum degree dropped to 8.0Pa.

[0036] Preparation of postbiotic powder: The fermentation liquid was inactivated at 85°C for 15 min. After inactivation, the materials were evenly mixed and spray-dried at an inlet air temperature of 175°C and an outlet air temperature of 70°C.

[0037] The liquid culture medium contains: peptone 10.0 g / L; yeast powder 9.0 g / L; glucose 10 g / L; stachyose 10 g / L; Tween 80 1.0 mL / L; dipotassium hydrogen phosphate 2.0 g / L; ammonium citrate 2.0 g / L; magnesium sulfate 0.2 g / L; manganese sulfate 0.05 g / L; calcium chloride 0.05 g / L. The culture medium is sterilized at 121°C for 20 min.

[0038] The activation medium composition is: peptone 5.0g / L; yeast powder 8.0g / L; glucose 10g / L; stachyose 10g / L; Tween 80 1.0mL / L; dipotassium hydrogen phosphate 2.0g / L; ammonium citrate 2.0g / L; magnesium sulfate 0.2g / L; manganese sulfate 0.05g / L; calcium chloride 0.05g / L.

[0039] Example 2 Cell experiments: 1. Establishment of a cell model of retinal oxidative stress damage induced by blue light The experiment was divided into three groups: blank group, model group and IOB802 live bacteria group.

[0040] Blank group: cells were cultured in cell maintenance medium, covered with black tape to prevent light, and the culture temperature was adjusted to 37°C. ARPE-19 cells with good growth status were cultured at 5×10 5 Cells of 100 cells / mL were inoculated into a culture plate containing cell growth medium and cultured until the cell density reached about 65-75%; Model group: A blue LED lamp with a wavelength of 430 nm was installed in the incubator and adjusted to a suitable height so that the light intensity reaching the surface of the cell culture plate was about 2500 ± 200 Lux; the culture temperature was adjusted to 37°C. ARPE-19 cells with good growth status were cultured at a rate of 5×10 5 Cells were inoculated into a culture plate containing cell growth medium at a concentration of 100 cells / mL. When the cell density reached about 65-75%, the cells were cultured in a cell maintenance medium for 6 hours and then exposed to blue light for 12 hours. IOB802 live bacteria group: Install a blue LED lamp with a wavelength of 430 nm in the incubator and adjust it to a suitable height so that the light intensity reaching the surface of the cell culture plate is about 2500 ± 200 Lux. Adjust the culture temperature to 37°C. Incubate ARPE-19 cells in good growth state at a rate of 5×10 5 The cells were inoculated into a culture plate containing cell growth medium at a concentration of 100 cells / mL. When the cell density reached about 65-75%, the cells were pretreated with culture medium containing different concentrations of bacterial solution for 6 h, and then the cells were exposed to blue light for 12 h.

[0041] 2. Fermentation of Lactobacillus mucilaginosus IOB802 improves the vitality of retinal epithelial cells Human retinal epithelial cells ARPE-19, Pronocell Biotech Co., Ltd. After APRE-19 cells with a growth density that can be passed on are blown and evenly distributed, the cell suspension is added to a 96-well cell culture plate at a volume of 100 μL per well, gently shaken, and placed in an incubator for 24 h. The supernatant is discarded, and cell maintenance fluid containing different concentrations of fermented mucus lactobacillus IOB802 is added in sequence. After continuing to culture for 24 h, 20 μL of MTT reagent (5.0 mg / mL) is added to each well in the dark, and placed in an incubator for incubation for 3-4 h. The supernatant is carefully discarded, 150 μL of DMSO is added to each well, and the formazan purple crystals are fully shaken until they are completely dissolved. The absorbance at 490 nm is measured and expressed as the percentage of the absorbance of the measured group cells / the absorbance of the normal cells.

[0042] MTT test results Figure 1 As shown in the figure, the survival rate of APRE-19 cells decreased after blue light pretreatment. Compared with the model group, the survival rate of cells was significantly improved after intervention with cell maintenance solution containing fermented Lactobacillus mucilaginosus IOB802 bacterial solution.

[0043] 3. Inflammatory factor content determination experiment: After the cell model was established, the supernatant was subsequently used to detect the expression of inflammatory factors. The specific experimental operations were completed according to the IL-6, IL-1β, and TNF-α ELISA kits.

[0044] Experimental results: Fundus neovascularization not only contains high levels of VEGF, but also is rich in a large number of inflammatory factors such as IL-6, IL-1β, and TNF, and they have a significant positive correlation with each other, indicating that angiogenesis and inflammation have a close interaction during the development of fundus neovascularization. Figure 2 , Figure 3 and Figure 4 As shown in the figure, compared with the blank group, the levels of IL-6, IL-1β, and TNF-α in the model group were significantly increased, while compared with the model group, after pretreatment with IOB802, the levels of IL-6, IL-1β, and TNF-α decreased. This is consistent with the effect of VEGF, which shows that VEGF is not only involved in angiogenesis, but also related to inflammatory response. IOB802 can not only reduce VEGF content, but also reduce the levels of other inflammatory factors at the same time, alleviating inflammatory damage to the eye.

[0045] 4. Vascular endothelial growth factor (VEGF) content determination experiment: After the cell model was established, the supernatant was subsequently used to detect the expression of inflammatory factors. The specific experimental operation was completed according to the VEGF ELISA kit.

[0046] Vascular endothelial growth factor (VEGF) is a signaling protein that promotes angiogenesis. It plays a key role in angiogenesis and vascular permeability. VEGF is a key cytokine that induces angiogenesis and is prone to cause a variety of ophthalmic diseases, especially in diseases such as age-related macular degeneration (ARMD), diabetic macular edema (DME) and retinal vein occlusion (RVO). Abnormal expression of VEGF can lead to the formation of new blood vessels and macular edema, thereby affecting vision. Figure 5 As shown in the figure, compared with the blank group, the VEGF level in the blue light exposure group was significantly increased, while compared with the blue light exposure group, the VEGF level was significantly decreased after IOB802 pretreatment. And it showed a positive correlation with the bacterial activity of IOB802.

[0047] 5. Antioxidant enzyme content determination experiment: The separated mouse retinal tissue was added to PBS at a mass volume ratio of 1:9, and the tissue was ground into a 10% retinal tissue homogenate under ice bath conditions. Centrifuged at 3000 rpm at 4°C for 15 min. The supernatant was collected and the protein concentration was determined, and then the experimental determination was completed according to the specific experimental operation methods of the CAT, SOD and GSH-Px kits.

[0048] Experimental results: The human body's antioxidant system mainly includes enzyme antioxidant system and non-enzymatic antioxidant system, among which the enzyme antioxidant system includes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px), etc. These antioxidant enzymes work together to exert antioxidant effects to protect tissues and eyes from damage by reactive oxygen species (ROS).

[0049] SOD is regarded as the first line of defense for the human body's anti-oxidation. It has the function of disproportionating superoxide anions, reducing the excessive accumulation of hydrogen peroxide, and alleviating oxidative damage in cells. In the structure of the eyeball, SOD decomposes peroxide into hydrogen peroxide, which is then converted into water and oxygen by CAT and GSH-Px to protect cells from damage caused by excessive concentrations of ROS. GSH-Px plays a role in protecting the transparency of the cornea and lens, and in the regeneration and repair of tissues. It reduces the accumulation of peroxide by converting hydrogen peroxide into water and oxygen. As part of the antioxidant system, CAT works together with SOD and GSH-Px to convert hydrogen peroxide into water and oxygen, reducing the accumulation of ROS.

[0050] like Figure 6 , Figure 7 and Figure 8As shown in the figure, compared with the blank group, the activities of CAT, SOD, and GSH-Px in the model group were significantly reduced, indicating that blue light irradiation would destroy the endogenous antioxidant system of cells and cause cellular oxidative stress; compared with the model group, the activities of antioxidant enzymes in ARPE-19 cells were significantly increased after pretreatment with IOB802. This indicates that there is an interaction between VEGF and antioxidant enzymes, and antioxidant enzymes may affect the expression and function of VEGF by regulating oxidative stress.

[0051] Example 3 Animal experiments Female BALB / c mice weighing about 18-20 g and free of specific pathogens (SPF) were selected and housed in an animal laboratory with a 12-h light-dark cycle and standard temperature and humidity (22 ± 3℃ and 30-60%). During the experiment, the mice were able to move freely and eat and drink as they pleased, and the bedding and drinking water were changed every day.

[0052] Mouse modeling and grouping: BALB / c mice were randomly divided into 5 groups according to the experimental groups, with 8 mice in each group, and randomly divided into blank group, model group, positive group, IOB802 live bacteria group and IOB802 postbiotic group.

[0053] All animals were given the corresponding test substances by gavage for 4 consecutive weeks. Mice in the blank group and model group were gavaged with 200 μL of normal saline at the same time every day, mice in the positive group were gavaged with 10 mg / kg lutein solution at the same time every day, and mice in the probiotic group were gavaged with 200 μL of 3*10 8 CFU / ml of fermented Lactobacillus mucilaginosus IOB802 powder, the postbiotic group mice were gavaged with 200 μL of 3*10 8 CFU / ml of postbiotics from fermentation of Lactobacillus mucilaginosus IOB802.

[0054] Light exposure was performed on groups other than the blank group. These mice were dark-adapted for 12 h before light exposure. After that, the mice were exposed to 5000 ± 500 Lux blue light for 12 h in an exposure box maintained at 23 ± 2°C for 7 consecutive days. During the light exposure period, the mice ate and drank water normally, and the corresponding test substances were continuously gavaged. After light exposure, the mice were kept under dim circulating light. During the experiment, the weight changes of the mice were recorded once a week. After the light exposure ended, blood was collected and the eyeballs were separated. One part of the eyeballs was stored in eyeball fixative for pathological sectioning, and the other part, the retina was quickly separated on ice and transferred to liquid nitrogen for storage for subsequent experimental determination.

[0055] 1. Determination of inflammatory factor content After the separated plasma was placed at 4°C for 12 h, it was centrifuged at 12,000 rpm for 15 min, and the upper serum was carefully collected. The specific experimental operation was completed according to the IL-6, IL-1β, and TNF-α ELISA kits.

[0056] like Fig. 9 , Fig.10 and Fig.11 As shown, compared with the blank group, the levels of IL-6, IL-1β, and TNF-α in the serum of mice in the model group were significantly increased; compared with the model group, the levels of IL-6, IL-1β, and TNF-α in the IOB802 live bacteria group and the IOB802 postbiotic group were reduced to a certain extent, and the improvement effect of postbiotics was more obvious.

[0057] 2. Determination of vascular endothelial growth factor (VEGF) content After the separated plasma was allowed to stand at 4°C for 12 h, it was centrifuged at 12,000 rpm for 15 min, and the upper serum was carefully collected. The specific experimental operation was completed according to the VEGF ELISA kit.

[0058] like Fig.12 As shown, compared with the blank group, the VEGF content in the serum of mice in the model group was significantly increased; compared with the model group, the VEGF content in the IOB802 live bacteria group and the IOB802 postbiotic group was reduced to a certain extent, and the improvement effect of postbiotics was more obvious.

[0059] 3. Determination of Antioxidant Enzyme Content The separated mouse retinal tissue was added to PBS at a mass-volume ratio of 1:9, and the tissue was ground into a 10% retinal tissue homogenate under ice bath conditions. Centrifuged at 4°C, 3000 rpm for 15 min. The supernatant was collected and the protein concentration was determined, and then the experimental determination was completed according to the specific experimental operation methods of the CAT, SOD, GSH-Px and MDA kits.

[0060] like Fig.13 , Fig.14 , Fig.15 and Fig.16 As shown, compared with the blank group, the activities of CAT, SOD, and GSH-Px in the retina of mice in the model group were significantly decreased, and the activity of MDA was significantly increased, which indicated that the antioxidant status of the damaged mice deteriorated; compared with the model group, the activities of CAT and GSH-Px in the IOB802 live bacteria and postbiotic group were increased to a certain extent, and the activities of SOD and MDA were decreased to a certain extent, and the improvement effect of the postbiotic group was more obvious, which indicated that both the IOB802 live bacteria and the postbiotic group could improve the antioxidant capacity of the mouse retina.

[0061] 4. HE Staining The eyeball tissue was fully immersed in the eyeball fixative for 24 hours, and then embedded in paraffin and sliced. The thickness of the paraffin slices was 4 μm. After drying at room temperature, they were stained with hematoxylin and eosin (H&E). The changes in the mouse retinal structure were observed under a microscope and photographs were recorded. Fig.17 shown.

[0062] As a photosensitive layer, the retina mainly transmits visual information from the outside world to the visual center of the brain through the optic nerve. The neural retina consists of nine layers of structures, including the ganglion cell layer, inner nuclear layer, outer nuclear layer, and photoreceptor layer. Retinal damage was evaluated by image analysis of H&E-stained retinal tissue. Representative images of retinal sections of each group are shown in the figure. In the blank group, the results of H&E staining showed that the retinal layer was intact, the cells were tightly arranged, the morphology was normal, the number of pigment granules in the retinal pigment epithelial cells was large, and the chromatin density in the cell nucleus was high. However, the histological changes in the retina of mice in the model group were obvious, the cells were scattered, the intercellular gaps became larger, the chromatin in the cell nucleus agglomerated, and the number of cell layers decreased. The administration of IOB802 live bacteria and postbiotics improved the overall morphology of the retina to a certain extent, thereby protecting the retina from blue light damage.

[0063] In summary, the fermented Lactobacillus muciniphila IOB802 postbiotic powder of the present invention has a positive effect on eye health, effectively improves retinal damage caused by continuous blue light exposure, and provides ideas for developing new products of fermented Lactobacillus muciniphila IOB802 postbiotic powder.

Claims

1. An application of fermented Lactobacillus mucilaginosus IOB802 in the preparation of a medicine for improving eye health, characterized in that: The fermented mucus lactobacillus IOB802 is classified and named as fermented mucus lactobacillus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

2. The use of a fermented Lactobacillus mucilaginosus IOB802 as claimed in claim 1 in preparing a medicine for improving eye health, characterized in that: The fermented Lactobacillus mucilaginosus IOB802 is used to prepare the medicine for improving eye health in the form of fermented Lactobacillus mucilaginosus IOB802 bacterial powder. The preparation method of the fermented Lactobacillus mucilaginosus IOB802 bacterial powder comprises: The strain was placed in a closed culture at 30-37°C for 24-36 hours and subcultured twice to obtain a fermentation seed liquid; the fermentation seed liquid was inoculated into a fermentation liquid medium, the fermentation tank was stirred at a speed of 50-70 r / min, the temperature was 37°C, the pH was 6.2±0.2, and the closed culture was carried out for 12 hours; the fermentation liquid medium was centrifuged and the bacterial mud was obtained, and the fermented mucus Lactobacillus IOB802 powder was obtained after vacuum freeze-drying.

3. An application of fermented mucus lactobacillus IOB802 in preparing a health food for relieving visual fatigue, characterized in that: The fermented mucus lactobacillus IOB802 is classified and named as fermented mucus lactobacillus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

4. The use of a fermented mucus lactobacillus IOB802 as claimed in claim 3 in preparing a health food for relieving visual fatigue, characterized in that: The fermented Lactobacillus mucilaginosus IOB802 is used to prepare the health food for relieving visual fatigue in the form of fermented Lactobacillus mucilaginosus IOB802 bacterial powder. The preparation method of the fermented Lactobacillus mucilaginosus IOB802 bacterial powder comprises: The strain was placed in a closed culture at 30-37°C for 24-36 hours and subcultured twice to obtain a fermentation seed liquid; the fermentation seed liquid was inoculated into a fermentation liquid medium, the fermentation tank was stirred at a speed of 50-70 r / min, the temperature was 37°C, the pH was 6.2±0.2, and the closed culture was carried out for 12 hours; the fermentation liquid medium was centrifuged and the bacterial mud was obtained, and the fermented mucus Lactobacillus IOB802 powder was obtained after vacuum freeze-drying.

5. An application of fermented Lactobacillus mucilaginosus IOB802 postbiotics in the preparation of a medicine for improving eye health, characterized in that: The fermented mucus lactobacillus IOB802 is classified and named as fermented mucus lactobacillus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

6. The use of a fermented Lactobacillus mucilaginosus IOB802 postbiotic in the preparation of a medicine for improving eye health as claimed in claim 5, characterized in that: The preparation method of the fermented Lactobacillus mucilaginosus IOB802 postbiotics comprises: The strain is placed in a closed culture at 30-37°C for 24-36 hours, subcultured twice, and fermentation seed liquid is obtained; the fermentation seed liquid is inoculated into the fermentation liquid culture medium, the fermentation tank is stirred at a speed of 50-70 r / min, the temperature is 37°C, the pH is 6.2±0.2, and the closed culture is carried out for 12 hours; after the fermentation liquid is inactivated, the materials are mixed evenly, and spray-dried to obtain fermentation mucus Lactobacillus IOB802 postbiotics.

7. Application of fermented Lactobacillus mucilaginosus IOB802 postbiotics in the preparation of health food for relieving visual fatigue, characterized in that: The fermented mucus lactobacillus IOB802 is classified and named as fermented mucus lactobacillus ( Limosilactobacillusfermentum ), and its deposit number is CGMCC No.23120.

8. The use of a fermented Lactobacillus mucilaginosus IOB802 postbiotic in the preparation of a health food for relieving visual fatigue as claimed in claim 7, characterized in that: The preparation method of the fermented Lactobacillus mucilaginosus IOB802 postbiotics comprises: The strain is placed in a closed culture at 30-37°C for 24-36 hours, subcultured twice, and fermentation seed liquid is obtained; the fermentation seed liquid is inoculated into the fermentation liquid culture medium, the fermentation tank is stirred at a speed of 50-70 r / min, the temperature is 37°C, the pH is 6.2±0.2, and the closed culture is carried out for 12 hours; after the fermentation liquid is inactivated, the materials are mixed evenly, and spray-dried to obtain fermentation mucus Lactobacillus IOB802 postbiotics.

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