Application of bifidobacterium longum subsp. Infantis CCFM1426 strain and vitamin A in preparation of medicine for improving physiological activity of vitamin A and / or relieving symptoms of colitis and / or xerophthalmia

Abstract

The invention discloses application of a bifidobacterium longum subsp. Infantis CCFM1426 strain and vitamin A in preparation of a medicine for improving the physiological activity of the vitamin A and / or relieving colitis and / or xerophthalmia symptoms. The probiotic agent provided by the invention comprises a bifidobacterium longum subsp. Infantis CCFM1426 strain with the preservation number of GDMCC No.65152 and vitamin A, and can be used for improving the retinoic acid level in the colon of a mouse with ulcerative colitis; the diamine oxidase level, the intestinal fatty acid binding protein level and the lipopolysaccharide binding level in serum of mice with ulcerative colitis are reduced; the corneal injury, basal vacuolation and conjunctival goblet cell injury of the xerophthalmia mice are relieved; the level of retinoic acid in serum of the dry-eye mouse is improved; and the levels of IL-6, IL-17 and TNF-alpha in serum of dry eye mice are reduced. Therefore, the probiotic agent disclosed by the invention has a relatively great application prospect in preparation of products for supplementing vitamin A and preventing and / or treating ulcerative colitis and xerophthalmia.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, and in particular relates to the application of Bifidobacterium longum subsp. infantis CCFM1426 strain and vitamin A in the preparation of drugs that enhance the physiological activity of vitamin A and / or relieve symptoms of colitis and / or dry eye syndrome. Background Technology

[0002] Vitamin A is a fat-soluble vitamin that the human body cannot synthesize on its own and must obtain through diet or supplements. It has various physiological functions, including maintaining vision, participating in tissue growth and cell differentiation, maintaining epithelial integrity, and regulating immune responses. Retinoic acid is the physiologically active form of vitamin A and plays an important role in maintaining the integrity of the mucosal barrier and regulating mucosal immunity.

[0003] Patients with intestinal diseases often experience severe disruption of the intestinal barrier, intestinal immunity, and gut microbiota, affecting the absorption of nutrients and easily inducing vitamin deficiencies. Consequently, these patients frequently face vitamin A deficiency and impaired retinoic acid signal transduction, preventing the effective absorption and conversion of vitamin A into retinoic acid, thus triggering a vicious cycle. In recent years, the concept of the gut-eye axis has become increasingly well-established. A regulatory network exists between the gut and the ocular surface, and intestinal damage in patients with intestinal diseases can affect ocular surface health through the gut-eye axis. Dry eye syndrome is a common ocular surface disease, primarily affecting the cornea and conjunctiva. Dry eye patients have a reduced number of conjunctival goblet cells, leading to decreased retinoic acid secretion and increased susceptibility to ocular epithelial damage. Furthermore, serum vitamin A deficiency is often closely linked to dry eye syndrome. Therefore, the need for vitamin A supplementation and enhancement of its physiological activity in the treatment of intestinal and ocular surface diseases is becoming increasingly urgent.

[0004] Currently, the methods of vitamin A supplementation for patients with enterophthalmopathy mainly focus on the dosage of vitamin A and its combination with other nutrients, neglecting consideration of vitamin A's physiological activity in the body and the level of its conversion to retinoic acid. This lack of focus may lead to drug dependence, adverse reactions, poor efficacy of vitamin A supplementation, and adverse effects from excessive or prolonged supplementation. Furthermore, current methods of synthesizing vitamin A using retinol or retinal through oxidation and isomerization are complex, prone to decomposition, costly, and generate byproducts, causing environmental pollution.

[0005] In conclusion, there is an urgent need to screen probiotics that can enhance the physiological activity of vitamin A and increase retinoic acid levels, in order to regulate the body's immune response, alleviate intestinal barrier damage and ocular surface damage, and provide a more effective and lasting treatment for the prevention and relief of dry eye syndrome and intestinal barrier damage that may lead to ocular surface diseases through the intestinal-ocular axis. Summary of the Invention

[0006] To address the shortcomings of the existing technologies, this invention provides an application of a probiotic agent that enhances the physiological activity of vitamin A. The aim is to solve the current problems where vitamin A supplementation focuses primarily on dosage and supplementation methods, neglecting consideration of enhancing vitamin A's physiological activity. Furthermore, the chemical synthesis of all-trans retinoic acid is complex, easily decomposed, resulting in high costs, environmental pollution, and requiring further safety confirmation.

[0007] The technical solution provided by this invention is as follows:

[0008] The use of probiotic agents in the preparation of products that enhance the physiological activity of vitamin A and / or relieve symptoms of colitis and / or dry eye syndrome, wherein the probiotic agents comprise Bifidobacterium longum subsp. infantis strain CCFM1426 with accession number GDMCC No. 65152 and vitamin A.

[0009] The described *Bifidobacterium longum* subspecies *CCFM1426* was deposited on September 20, 2024, at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC No:65152, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0010] Preferably, the amount of Bifidobacterium longum infantis subsp. CCFM1426 added to the probiotic agent is not less than 1×10⁻⁶. 9 CFU / mL or 1×10 9 CFU / g.

[0011] Preferably, the amount of vitamin A added to the probiotic agent is not less than 100 IU.

[0012] Preferably, the probiotic agent further includes food ingredients and / or food- or medically acceptable excipients or additives.

[0013] Preferably, the dosage form of the probiotic agent includes lyophilized powder, capsules, tablets, or granules.

[0014] Preferably, the product is one or more of a drug, food, or health product.

[0015] The present invention has the following beneficial effects:

[0016] This invention provides a probiotic preparation containing Bifidobacterium longum subsp. infantis CCFM1426 and vitamin A. This probiotic preparation has the effect of improving the physiological activity of vitamin A and preventing and / or alleviating ulcerative colitis and dry eye syndrome, specifically manifested in:

[0017] (1) Reduce colonic damage in individuals with ulcerative colitis;

[0018] (2) Increase colonic retinoic acid levels in individuals with ulcerative colitis;

[0019] (3) Downregulate serum diamine oxidase levels in individuals with ulcerative colitis;

[0020] (4) Downregulate serum levels of intestinal fatty acid-binding protein in individuals with ulcerative colitis;

[0021] (5) Downregulates serum lipopolysaccharide-binding protein levels in individuals with ulcerative colitis;

[0022] (6) Reduce corneal damage in individuals with dry eye syndrome;

[0023] (7) Reduce conjunctival goblet cell damage in individuals with dry eye syndrome;

[0024] (8) Increase serum retinoic acid levels in individuals with dry eye;

[0025] (9) Downregulate serum IL-6 levels in individuals with dry eye

[0026] (10) Downregulate serum IL-17 levels in individuals with dry eye

[0027] (11) Downregulate serum TNF-α levels in individuals with dry eye

[0028] Therefore, this probiotic agent enhances the physiological activity of vitamin A and has great application potential in the preparation of products (such as food, medicine or health food) for the prevention of vitamin A deficiency and the prevention / alleviation of ulcerative colitis and dry eye syndrome.

[0029] Preservation Instructions

[0030] Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou

[0031] Deposit date: September 20, 2024

[0032] Strain name: Bifidobacterium longum subsp. infantis

[0033] Latin name: *Bifidobacterium longum* subsp. *infantis*

[0034] Strain number: CCFM1426

[0035] Preservation Institution: Guangdong Provincial Center for Microbial Culture Collection

[0036] Abbreviation for depository institution: GDMCC

[0037] Registration number at the Depository Center: GDMCC No:65152 Attached Figure Description

[0038] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0039] Figure 1 Comparison of histopathological sections of colon tissue from different groups of mice with ulcerative colitis.

[0040] Figure 2 The graph shows the results of retinoic acid levels in the colon of mice in different experimental groups.

[0041] Figure 3 The graph shows the results of diamine oxidase levels in the serum of mice in different experimental groups.

[0042] Figure 4 The graph shows the serum levels of intestinal fatty acid-binding protein in mice from different experimental groups.

[0043] Figure 5 The graph shows the results of lipopolysaccharide-binding protein levels in the serum of mice in different experimental groups.

[0044] Figure 6 A comparison of the results of sodium fluorescein staining of the corneas of mice in different groups.

[0045] Figure 7 Comparison of corneal tissue pathological sections from different groups of experimental mice.

[0046] Figure 8 Comparison of conjunctival tissue pathological sections from different groups of experimental mice.

[0047] Figure 9 The graph shows the results of serum retinoic acid levels in mice from different experimental groups.

[0048] Figure 10 The graph shows the results of IL-6 levels in the serum of mice in different experimental groups.

[0049] Figure 11 The graph shows the results of IL-17 levels in the serum of mice in different experimental groups.

[0050] Figure 12 The graph shows the serum TNF-α levels in mice from different experimental groups. Detailed Implementation

[0051] To better understand the present invention, the present invention will be further described in detail below with reference to the embodiments and accompanying drawings. However, those skilled in the art will understand that the following embodiments are not intended to limit the scope of protection of the present invention, and any changes and variations made on the basis of the present invention are within the scope of protection of the present invention.

[0052] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0053] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0054] Test method:

[0055] 1. Method for detecting viable bacteria count: The national standard GB 4789.35-2016, "National Food Safety Standard - Microbiological Testing of Food - Lactic Acid Bacteria Detection", was adopted.

[0056] 2. Acidity testing method: GB 431334-2010 is adopted.

[0057] 3. Preparation of pathological sections from mouse colon: Colon tissue was extracted and immediately fixed in 10% neutral buffered formaldehyde solution for 24-48 hours. Next, the tissue was dehydrated with a gradient of alcohols, cleared with xylene, and then embedded in paraffin. The embedded tissue was cut into 4-5 micrometer thick sections and stained with hematoxylin and eosin (H&E) under a microscope. Finally, the tissue structure and pathological changes of the sections were observed and analyzed under a microscope.

[0058] The experimental results were statistically analyzed using Graphpad Prism. Unless otherwise specified, the results were compared with the model group. **** represents P < 0.0001, *** represents P < 0.001, ** represents P < 0.01, and * represents P < 0.05.

[0059] Raw materials used in the examples:

[0060] The SPF-grade C57BL / 6J male mice used in the following examples were purchased from Vital River Pharmaceuticals in Beijing.

[0061] The culture media involved in the following examples are as follows:

[0062] Each liter of MRS medium contains: 10g peptone, 10g beef extract, 20g glucose, 2g sodium acetate, 5g yeast extract, 2g diammonium hydrogen citrate, 2.6g K₂PO₄·3H₂O, 0.1g MgSO₄·7H₂O, 0.05g MnSO₄, 1mL Tween 80, and 0.5g cysteine. All reagents used to prepare the medium were purchased from Sinopharm Chemical Reagent Co., Ltd.

[0063] MRS solid culture medium formula (1L): peptone 10g, beef extract 10g, glucose 20g, sodium acetate 2g, yeast extract 5g, diammonium hydrogen citrate 2g, K₂PO₄·3H₂O 2.6g, MgSO₄·7H₂O 0.1g, MnSO₄ 0.05g, Tween 80 1mL, cysteine ​​phosphate 0.5g, agar 20g. All reagents used to prepare the culture medium were purchased from Sinopharm Chemical Reagent Co., Ltd.

[0064] Example 1: Identification and Culture of Strains

[0065] (1) Identification of strains

[0066] The target strain was inoculated into MRS solid medium and cultured anaerobically at 37°C for 24 hours. Single colonies were then picked and transferred to MRS liquid medium, and cultured anaerobically at 37°C for 24 hours. The resulting purified culture was mixed, centrifuged, and the supernatant was discarded to obtain bacterial cells. The bacterial cells were sent to the company for genome sequencing. The 16S rDNA sequence obtained from the sequencing was analyzed using GenBank, and the results showed it to be *Bifidobacterium longum* subsp. *infantitidis*.

[0067] (2) Cultivation of bacterial strains and preparation of bacterial suspension

[0068] After inoculating the strain into MRS solid medium and culturing at 37°C for 48 hours, its colonies were observed and the bacterial cells were observed under a microscope. It was found that the colonies were milky white, round and raised, with a smooth surface. The bacterial cells were straight rods with neat edges and round ends, usually existing singly, in pairs, or in a V-shape.

[0069] Bifidobacterium longum subsp. infantis CCFM1426 was inoculated into MRS liquid medium and cultured at 37°C for 24 h. Then, it was transferred to fresh MRS liquid medium at an inoculum volume of 4% and cultured under the same conditions for 24 h. After centrifugation at 8000×g for 15 min, the bacterial cells were collected. The bacterial cells were washed with 0.9% physiological saline and centrifuged again at 8000×g for 15 min. The supernatant was discarded and the bacterial cells were collected. The cells were resuspended in 30% glycerol solution (containing 0.05% L-cysteine) to prepare a resuspended solution, which was then frozen at -80°C for later use.

[0070] When using Bifidobacterium longum subsp. infantis CCFM1426 for gavage in mice, the sample was removed from -80℃, centrifuged at 8000×g for 15 min, the supernatant was discarded, and the sample was resuspended in sterile physiological saline to obtain a gavage concentration of 1×10⁻⁶. 9 CFU / mL bacterial suspension.

[0071] Example 2: The alleviating effect of probiotics on colonic inflammation in mice with ulcerative colitis

[0072] Six-week-old SPF-grade male C57BL / 6J mice were divided into four groups: normal group, model group, vitamin A treatment group, and probiotic group, with six mice in each group. The animals were housed at the Experimental Animal Center of Jiangnan University, fed with standard feed, at a constant temperature of 20-26℃, humidity of 40-70%, and maintained with a 12-hour light / dark cycle.

[0073] The experiment lasted 21 days. From day 8 to day 14, mice were anesthetized and ulcerative colitis was induced daily by gavage with 0.2 mL of a 3% sodium dextran sulfate solution. The vitamin A group was gavaged with 200 μL of a vegetable oil solution containing 0.5 mg vitamin A; the probiotic group was gavaged with 200 μL of a solution containing 0.5 mg vitamin A and 1 × 10⁻⁶ live bacteria. 9 CFU / mL CCFM1426 bacterial suspension; the normal group and model group were administered an equal volume of sterile saline via gavage as a control; all groups had free access to water and food. Mice were weighed after gavage each day until sacrifice on day 21.

[0074] The grouping and treatment methods for experimental animals are shown in Table 2:

[0075] Table 2 Grouping of experimental animals

[0076]

[0077] Mice were sacrificed on day 21, and their colons were collected to prepare pathological sections for histopathological analysis.

[0078] Depend on Figure 1 It was found that, compared with the control group mice, some crypts in the colon of the model group mice were destroyed, the crypts shifted upwards, and the crypt structure was twisted and branched; inflammatory cell infiltration was observed, basal plasma cells increased, lymphocytes aggregated, and hemorrhage was present. After probiotic supplementation, the inflammation was significantly relieved, the infiltration of inflammatory cells in the colon tissue of the mice was significantly reduced, the crypt damage was restored, the crypt morphology was more intact, and there was no obvious hemorrhage. Treatment with vitamin A alone did not significantly reduce lymphocyte aggregation.

[0079] The above results indicate that probiotics are more effective than vitamin A in relieving intestinal inflammation in mice with intestinal damage.

[0080] Example 3: Effect of probiotics on colonic retinoic acid levels in mice with ulcerative colitis

[0081] The mouse grouping and modeling methods were the same as in Example 2.

[0082] Mice were sacrificed on day 21 to obtain colon tissue. The colon tissue was homogenized, and the supernatant was collected at 12000 rpm for 15 min. The retinoic acid level in the supernatant was detected using an ELISA kit (Shanghai Enzyme-Link Biotechnology Co., Ltd.). The results are as follows: Figure 2 As shown.

[0083] Depend on Figure 2 The results showed that the retinoic acid level in the intestinal tissue of the model group mice (97.30±7.66 pg / mgprot) was significantly lower than that of normal mice (125.93±5.14 pg / mgprot). Compared with the model group mice, the retinoic acid level in the intestinal tissue of mice with intestinal damage significantly increased after intervention with probiotics (P<0.05). Notably, after probiotic intervention, the retinoic acid level in the intestine of mice with intestinal damage (119.21±6.58 pg / mgprot) returned to normal levels, and was 22.52% higher than that in the colon of the model group mice. Furthermore, the retinoic acid level in the colonic tissue of mice treated with probiotics was 6.83% higher than that in the vitamin A group (111.59±4.29 pg / mgprot).

[0084] The results showed that probiotics increased retinoic acid levels in the colonic tissue of mice with ulcerative colitis, and their therapeutic effect was slightly better than that of vitamin A.

[0085] Example 4: Effect of probiotics on serum diamine oxidase levels in mice with ulcerative colitis

[0086] The mouse grouping and modeling methods were the same as in Example 2.

[0087] Mice were sacrificed on day 21, and blood was collected by removing the eyeballs. The blood was centrifuged at 3000 rpm for 15 min to obtain the supernatant. Serum diamine oxidase levels were detected using an ELISA kit (Shanghai Enzyme-Link Biotechnology Co., Ltd.). Results are shown below. Figure 3 .

[0088] Depend on Figure 3 The results showed that the diamine oxidase level in the model group mice (3887.81±110.823 pg / mL) was significantly higher than that in the normal group mice (2775.00±142.821 pg / mL) (P<0.001). The serum diamine oxidase level in the probiotic group mice (3023.02±208.449 pg / mL) was significantly lower than that in the model group mice by 22.24% (P<0.01).

[0089] The above results indicate that probiotics significantly reduced the level of diamine oxidase in the serum of mice with ulcerative colitis.

[0090] Example 5: Effect of probiotics on serum intestinal fatty acid-binding protein levels in mice with ulcerative colitis. The mouse grouping and modeling methods were the same as in Example 2.

[0091] Mice were sacrificed on day 21, and blood was collected by removing the eyeballs. The blood was centrifuged at 3000 rpm for 15 min to obtain the supernatant, which was then used to detect the serum intestinal fatty acid binding protein level using an ELISA kit (Shanghai Enzyme-Link Biotechnology Co., Ltd.).

[0092] Depend on Figure 4 The results showed that the serum intestinal fatty acid-binding protein level in the model group (603.76±22.15 pg / mL) was significantly increased compared to the normal group (338.63±29.71 pg / mL); while the serum intestinal fatty acid-binding protein level in mice administered probiotics was significantly decreased by 29.91% (P<0.001) to 423.15±22.14 pg / mL compared to the model group. The serum intestinal fatty acid-binding protein level in mice treated with vitamin A via gavage was 20.18% lower than that in the model group.

[0093] The above experimental results indicate that probiotics can effectively reduce the level of intestinal fatty acid-binding protein, a protein related to intestinal barrier damage, in the serum of mice with ulcerative colitis, with a slightly better therapeutic effect than vitamin A alone. This indirectly suggests that probiotics can alleviate / reduce intestinal barrier damage in mice with ulcerative colitis.

[0094] Example 6: Effect of probiotics on serum lipopolysaccharide-binding protein levels in mice with ulcerative colitis

[0095] The mouse grouping and modeling methods were the same as in Example 2.

[0096] Mice were sacrificed on day 21, and blood was collected by removing the eyeballs. The blood was centrifuged at 3000 rpm for 15 min to obtain the supernatant, which was then used to detect the level of lipopolysaccharide-binding protein in the serum using an ELISA kit.

[0097] Depend on Figure 5 The results showed that the serum lipopolysaccharide-binding protein (LPG-BB) level in the model group (126.61±11.22 nmol / L) was significantly higher than that in the normal group (79.68±4.94 nmol / L). However, compared to the model group, the LPG-BB level in mice administered the probiotic formula via gavage was significantly lower by 27.86% (P<0.05), decreasing to 91.34±32.67 nmol / L. The serum intestinal fatty acid-binding protein (IFB) level in mice treated with vitamin A via gavage (101.06±9.23 nmol / L) was 20.18% lower than that in the model group.

[0098] The above experimental results indicate that probiotics can effectively reduce the level of lipopolysaccharide-binding protein, a protein associated with intestinal barrier damage, in the serum of mice with ulcerative colitis, and its effect is superior to vitamin A alone.

[0099] Example 7: The alleviating effect of probiotics on corneal damage in mice with dry eye syndrome

[0100] Six-week-old SPF-grade male C57BL / 6J mice were randomly divided into four groups: a normal control group, a model group, a vitamin A treatment group, and a CCFM1426 experimental group, with six mice in each group. The animals were housed at the Experimental Animal Center of Jiangnan University, fed standard feed, maintained at a constant temperature of 20-26℃ and humidity of 40-70%, and kept under a 12-hour light / dark cycle. All procedures in this animal experiment were performed in accordance with the "Jiangnan University Experimental Animal Management Regulations," and animal ethics were approved by the Experimental Animal Welfare and Ethics Committee of Jiangnan University.

[0101] The experiment lasted 21 days. From day 8 to day 14, mice were anesthetized and induced with dry eye by twice-daily eye drops of 5 μL of 0.2% benzalkonium chloride solution. From day 15 to day 21, the experimental group was administered 0.2 mL of a live bacteria solution (1 × 10⁻⁶) via gavage daily. 9 The mice were given a bacterial suspension at CFU / mL. The normal and model groups were administered an equal volume of sterile saline via gavage as a control. All groups had free access to water and food until sacrifice on day 21. Before sacrifice on day 21, the mice's corneas were observed using a slit lamp with a cobalt blue lens. The grouping and treatment methods for the experimental animals are shown in Table 3.

[0102] Table 3 Grouping of experimental animals

[0103]

[0104] Mice were sacrificed on day 21, and their eyeballs were collected to prepare pathological sections for histopathological analysis.

[0105] Depend on Figure 6 It was observed that, compared to the corneal tissue of the blank control group mice, the cornea of ​​the model group mice showed obvious patchy sodium fluorescein staining. After intervention with vitamin A or probiotics, the dry eye symptoms of the mice were significantly relieved, but obvious punctate and slight patchy staining still existed. Figure 7 It was found that, compared with the control group mice, the corneal layer of the model group mice was significantly thinner, and sparse gaps and vacuoles appeared in the corneal stroma. After vitamin A intervention, the corneal thickness was restored, and the symptoms of sparse gaps and vacuoles in the corneal stroma were alleviated. Gavage administration of probiotics alleviated the significant thinning of the corneal layer, but the symptoms of sparse gaps and vacuoles in the corneal stroma still existed.

[0106] The above results indicate that probiotics can alleviate corneal damage in mice with dry eye syndrome.

[0107] Example 8: The alleviating effect of probiotics on conjunctival goblet cell damage in mice with dry eye syndrome

[0108] The mouse grouping and modeling methods were the same as in Example 7.

[0109] Mice were sacrificed on day 21, and conjunctiva was obtained to prepare pathological sections for histopathological analysis.

[0110] Depend on Figure 8 It was found that, compared with the conjunctival tissue of the blank control group mice, the conjunctival goblet cells of the model group mice were atrophied and reduced. After intervention with vitamin A or probiotics, the number of conjunctival goblet cells in the mice recovered to some extent.

[0111] The above results indicate that probiotics can alleviate conjunctival goblet cell damage in mice with dry eye syndrome.

[0112] Example 9: Effect of probiotic preparation on serum retinoic acid levels in mice with dry eye syndrome

[0113] The mouse grouping and modeling methods were the same as in Example 7.

[0114] Mice were sacrificed on day 21, and blood was collected by removing the eyeballs. The blood was centrifuged at 3000 rpm for 15 min to obtain the supernatant, and the serum retinoic acid level was detected using an ELISA kit (Tianjin Kevino Biotechnology Co., Ltd.).

[0115] Depend on Figure 9 It was found that the serum retinoic acid level in the model group (295.43±8.58 pg / mL) was significantly lower than that in the normal group (360.92±16.86 g / mL) (P<0.01); while the serum retinoic acid level in mice administered probiotics by gavage was significantly increased by 20.68% (P<0.05) compared with the model group, reaching 356.52±5.80 pg / mL.

[0116] The results showed that probiotics could increase serum retinoic acid levels in mice with dry eye syndrome.

[0117] Example 10: Effect of probiotic preparation on serum IL-6 levels in mice with dry eye syndrome

[0118] The mouse grouping and modeling methods were the same as in Example 7.

[0119] Mice were sacrificed on day 21, and blood was collected by removing the eyeballs. The blood was centrifuged at 3000 rpm for 15 min to obtain the supernatant, and the serum IL-6 level was detected using an ELISA kit (Shanghai Enzyme-Link Biotechnology Co., Ltd.).

[0120] Depend on Figure 10 It was found that the serum IL-6 level in the model group (69.14±4.50 pg / mL) was significantly increased compared with that in the normal group (45.59±3.50 pg / mL) (P<0.01); while the serum IL-6 level in mice administered probiotics by gavage was significantly decreased by 40.18% (P<0.01) compared with the model group, decreasing to 41.36±3.69 g / mL.

[0121] The above experimental results indicate that probiotics can downregulate the level of the inflammatory factor IL-6 in the serum of mice with dry eye syndrome.

[0122] Example 11 Effect of probiotics on serum IL-17 levels in mice with dry eye

[0123] The mouse grouping and modeling methods were the same as in Example 7.

[0124] Mice were sacrificed on day 21, and blood was collected by removing the eyeballs. The blood was centrifuged at 3000 rpm for 15 min to obtain the supernatant, and the serum IL-17 level was detected using an ELISA kit (Shanghai Enzyme-Link Biotechnology Co., Ltd.).

[0125] Depend on Figure 11 It was found that the serum IL-17 level in the model group (139.25±5.40 pg / mL) was higher than that in the normal group (124.83±4.02 pg / mL); while the serum IL-17 level in mice administered probiotics by gavage was significantly lower than that in the model group by 20.05% (P<0.01), decreasing to 111.33±2.85 pg / mL.

[0126] The above experimental results indicate that probiotics can downregulate the level of the inflammatory factor IL-17 in the serum of mice with dry eye syndrome.

[0127] Example 12 Effect of probiotics on serum TNF-α levels in mice with dry eye

[0128] The mouse grouping and modeling methods were the same as in Example 7.

[0129] Mice were sacrificed on day 21, and blood was collected by removing the eyeballs. The blood was centrifuged at 3000 rpm for 15 min to obtain the supernatant, and the serum TNF-α level was detected using an ELISA kit (Shanghai Enzyme-Link Biotechnology Co., Ltd.).

[0130] Depend on Figure 12It was found that the serum TNF-α level in the model group (697.43±24.04 pg / mL) was higher than that in the normal group (641.20±14.01 pg / mL); while the serum TNF-α level in mice administered probiotics by gavage was significantly lower than that in the model group by 16.51% (P<0.01), decreasing to 582.31±6.97 pg / mL.

[0131] The above experimental results indicate that probiotics can downregulate the level of the inflammatory factor IL-17 in the serum of mice with dry eye syndrome.

[0132] While specific embodiments of the present invention have been described above, those skilled in the art should understand that the specific embodiments described are merely illustrative and not intended to limit the scope of the present invention. Equivalent modifications and variations made by those skilled in the art in accordance with the spirit of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. Use of a probiotic agent for the preparation of a product for increasing the physiological activity of vitamin A and / or for alleviating the symptoms of colitis and / or dry eye, characterized in that, The probiotic agent comprises Bifidobacterium longum subsp. infantis CCFM1426 strain with a preservation number of GDMCC No. 65152 and vitamin A.

2. Use according to claim 1, characterized in that, The amount of *Bifidobacterium longum* subsp. infantis CCFM1426 added to the probiotic is not less than 1×10⁻⁶. 9 CFU / mL or 1×10 9 CFU / g.

3. Use according to claim 1, characterized in that, The added amount of the vitamin A in the probiotic agent is not less than 100 IU.

4. Use according to claim 1, characterized in that, The probiotic agent further comprises food raw materials, and / or food or medically acceptable adjuvants or additives.

5. The use according to claim 1, characterized in that, The dosage form of the probiotic agent comprises lyophilized powder, capsule, tablet or granule.

6. Use according to claim 1, characterized in that, The product is one or more of a drug, food or health product.

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