Use of indole acetic acid in the preparation of medicaments for treating optic nerve damage

Abstract

The application relates to a compound indole acetic acid, and application of the compound indole acetic acid as an active ingredient in the preparation of a medicament or food for treating optic nerve damage. In vivo administration of the indole acetic acid is beneficial to maintaining the number of retinal ganglion cells and the thickness of the retinal nerve fiber layer, inhibiting the activation of retinal microglia cells, inhibiting the activation of a retinal inflammation channel and the expression of inflammatory mediators, and protecting visual function.

Description

Technical Field

[0001] This invention relates to a new use of a known product, and more particularly to a compound, which has biological effects in protecting retinal ganglion cells, protecting visual function, inhibiting microglia activation, inhibiting the activation of retinal inflammatory pathways and the expression of inflammatory mediators, and its application as an active ingredient in the manufacture of pharmaceuticals and medical devices. Background Technology

[0002] Glaucoma is a group of diseases characterized by optic nerve atrophy and visual field defects, with pathologically elevated intraocular pressure being the main risk factor. Statistics show that glaucoma is the leading cause of irreversible blindness worldwide, and the number of people suffering from glaucoma globally is projected to reach 111.8 million by 2040, imposing a severe medical and economic burden on patients. Therefore, breakthroughs in its treatment are urgently needed.

[0003] Numerous compounds, proteins, and nucleic acids have been successfully developed and applied to the treatment of glaucoma, and many medical devices are also used in the clinical diagnosis and treatment of glaucoma.

[0004] Indoleacetic acid (IAA), first discovered in plants by Frits Warmolt Went in 1928, is one of the most common auxins in plants. Current research indicates that IAA has protective effects in mouse models of liver cancer and chronic obstructive pulmonary disease, and can alleviate oxidative stress and inflammation in a mouse model of non-alcoholic fatty liver disease. However, the role of IAA in glaucoma and related ocular neurodegenerative diseases has not yet been reported. Summary of the Invention

[0005] One object of the present invention is to provide a compound, which, as an active ingredient, is used for the protection of visual function, protection of retinal nerve cells, regulation of microglia activation and regulation of inflammatory factor expression, thereby facilitating the treatment of optic nerve damage.

[0006] Another object of the present invention is to provide a compound indoleacetic acid, which, as an active ingredient, is used for the protection of optic nerve damage and is beneficial for the treatment of glaucoma.

[0007] Another object of the present invention is to provide a medicament comprising the compound indoleacetic acid, for the treatment of glaucoma.

[0008] Another object of the present invention is to provide a medical device comprising the compound indoleacetic acid for the treatment of glaucoma.

[0009] Indoleacetic acid was first discovered in plants by Frits Warmolt Went in 1928 and is the most common auxin in plants. However, more and more studies have also shown that there is also indoleacetic acid in mammals, the main sources including endogenous and exogenous. Among them, endogenous indoleacetic acid may be produced by decarboxylation of intracellular tryptophan or oxidative deamination of tryptophan, while exogenous indoleacetic acid is usually produced by microorganisms in the mammalian gut. Current studies have shown that indoleacetic acid has a protective effect on liver cancer model mice, chronic obstructive pulmonary disease model mice, etc., and can relieve oxidative stress and inflammatory response in the liver of non-alcoholic fatty liver mouse models.

[0010] Indoleacetic acid is administered to glaucoma model mice to maintain the number of retinal ganglion cells and the thickness of the retinal nerve fiber layer, inhibit retinal microglial cell activation, inhibit retinal inflammatory pathway activation and inflammatory mediator expression, and protect visual function.

[0011] A bioreactor having lipid membranes or vesicle-formed microparticles containing nucleic acids therein, from which indoleacetic acid is produced.

[0012] A microorganism that produces indoleacetic acid.

[0013] Another microorganism that is genetically edited to contain exogenous genetic material and thus produces indoleacetic acid.

[0014] Another microorganism that includes a plasmid that includes exogenous genetic material and is regulated to produce indoleacetic acid.

[0015] In a specific embodiment, the exogenous genetic material is a B. fragilis indoleacetic acid-producing gene.

[0016] In another specific embodiment, the exogenous genetic material is more than 97% homologous to a B. fragilis indoleacetic acid-producing gene.

[0017] In another specific embodiment, the exogenous genetic material is more than 95% homologous to a B. fragilis indoleacetic acid-producing gene.

[0018] In another specific embodiment, the exogenous genetic material is more than 93% homologous to a B. fragilis indoleacetic acid-producing gene.

[0019] In another specific embodiment, the exogenous genetic material is more than 90% homologous to a B. fragilis indoleacetic acid-producing gene.

[0020] In another specific embodiment, the exogenous genetic material is more than 85% homologous to a B. fragilis indoleacetic acid-producing gene.

[0021] In another specific embodiment, the exogenous genetic material is more than 80% homologous to the B. fragilis indoleacetate-producing gene.

[0022] In another specific embodiment, the exogenous genetic material is more than 75% homologous to the B. fragilis indoleacetate-producing gene.

[0023] In another specific embodiment, the exogenous genetic material is more than 70% homologous to the B. fragilis indoleacetate-producing gene.

[0024] B. fragilis, purchased from Microbiotek (Shanghai) Co., Ltd., accession number: ATCC25285.

[0025] A drug is prepared with indoleacetic acid as the active ingredient, which is delivered into the body to alleviate retinal damage and retinal ganglion cell loss, including but not limited to maintaining the number of retinal ganglion cells, maintaining the thickness of the retinal nerve fiber layer, improving visual function, reducing retinal microglial activation, reducing retinal inflammation pathway activation and inflammatory mediator expression.

[0026] A drug is prepared with an indoleacetate-producing bioreactor as the active ingredient, which is delivered into the body (e.g., the gastrointestinal tract) to alleviate retinal damage and retinal ganglion cell loss, including but not limited to maintaining the number of retinal ganglion cells, maintaining the thickness of the retinal nerve fiber layer, improving visual function, reducing retinal microglial activation, reducing retinal inflammation pathway activation and inflammatory mediator expression.

[0027] A drug is prepared with an indoleacetate-producing microorganism as the active ingredient, which is delivered into the gastrointestinal tract to reduce optic nerve damage, reduce retinal ganglion cell loss, inhibit retinal microglial activation, inhibit retinal nerve inflammation, and protect visual function.

[0028] The indoleacetic acid of the present application is used on in vitro cells or in vivo cells to promote optic nerve repair, and it should be understood that the indoleacetic acid is used as a drug for the purpose of the present application.

[0029] Indoleacetic acid is mixed with other excipients to prepare a drug (preparation) for ocular optic nerve damage diseases (e.g., glaucoma).

[0030] These pharmaceutical excipients can be the conventional ones used in various preparations, such as, but not limited to, isotonic agents, buffers, flavoring agents, excipients, fillers, binders, disintegrants, and lubricants, etc. They can also be selected for use in accordance with the substance, such as emulsifiers, solubilizers, bacteriostatic agents, analgesics, and antioxidants, etc. Such excipients can effectively improve the stability and solubility of the compounds contained in the composition or modify the release rate and absorption rate of the compounds, thereby improving the metabolism of various compounds in the living body and enhancing the administration effect of the composition.

[0031] In the aqueous solution injection, the excipients generally include isotonic agents and buffers, and necessary emulsifiers (such as Tween-80, Pluronic, and Poloxamer, etc.), solubilizers, and bacteriostatic agents, etc. In addition, it also includes other pharmaceutically acceptable pharmaceutical excipients, such as antioxidants, pH regulators, and analgesics, etc.

[0032] The excipients used for preparing oral liquid preparations generally include solvents, and necessary flavoring agents, bacteriostatic agents, emulsifiers, and coloring agents, etc.

[0033] The excipients used for preparing tablets generally include fillers (such as starch, powdered sugar, dextrin, lactose, compressible starch, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, and mannitol, etc.), binders (such as ethanol, starch paste, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, gelatin solution, sucrose solution, and aqueous or alcoholic solution of polyvinylpyrrolidone, etc.), disintegrants (such as dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, and cross-linked sodium carboxymethyl cellulose), and lubricants (such as magnesium stearate, microfine silica, talc, hydrogenated vegetable oil, polyethylene glycol 4,000, polyethylene glycol 6,000, and magnesium lauryl sulfate, etc.), etc.

[0034] The excipients used for preparing emulsions are generally water, oil (such as fatty acid), emulsifier, and necessary preservatives and flavoring agents, etc.

[0035] The excipients used for preparing granules are similar to those for tablets, but the granulation process is different. The prepared granules are mixed with a flow aid as needed and then filled into capsules to obtain capsule preparations.

[0036] Various excipients are combined with the compound to form a dosage form that is advantageous for drug delivery, such as, but not limited to, aqueous injection, powder injection, pill, powder, tablet, patch, suppository, emulsion, cream, gel, granule, capsule, aerosol, spray, powder spray, sustained-release preparation, and controlled-release preparation. In addition, excipients such as, but not limited to, gelatin, albumin, chitosan, polyether, and polyester macromolecular materials such as, but not limited to, polyethylene glycol, polyurethane, polycarbonate, and copolymers thereof can be used to achieve specific drug delivery purposes or methods, such as, but not limited to, sustained-release drug delivery, controlled-release drug delivery, and pulse drug delivery. The term "advantageous for drug delivery" mainly refers to, but is not limited to, improving therapeutic effect, improving bioavailability, reducing toxic side effects, and improving patient compliance.

[0037] The indole acetic acid of the present application is combined with other excipients, such as chemical coupling, to further improve the efficacy of the compound, the in vivo release period, and the extension of the drug delivery period. These excipients are usually polymers, such as polyesters, polyethers, and polyamides.

[0038] Indole acetic acid is used as an active ingredient loaded on a medical material to make a medical device for delivering indole acetic acid into the body to relieve retinal damage and retinal ganglion cell loss, including but not limited to maintaining the number of retinal ganglion cells, maintaining the thickness of the retinal nerve fiber layer, improving visual function, reducing retinal microglial cell activation, reducing retinal inflammation pathway activation, and inflammatory mediator expression.

[0039] Indole acetic acid is loaded or coated on a stent material for making a medical device for repairing optic nerve damage. Common stent materials include PLA, PLGA, GelMA, chitosan, and gamma PGA, etc. It is also mixed with biocompatible degradable materials to make microneedles and microneedle arrays, or loaded in metal microneedles to make microneedle chips. When the microneedle pierces the skin, indole acetic acid is released in the dermis layer to form microspheres or gels, which adhere to the mucosa, improving the ability of optic nerve repair. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 Figure 2 shows the retinal ganglion cell protection of mice on the 28th day after glaucoma modeling. a is a typical field of view of the mouse retinal smear under fluorescence confocal microscope (scale bar = 30 μm), β-III tubulin (red) marks retinal ganglion cells, b is a schematic diagram of the position of the field of view in a, c is a statistical diagram of the survival rate of retinal ganglion cells in each group of mice, * indicates a statistically significant difference between the two groups, p < 0.05, *** indicates a statistically significant difference between the two groups, p < 0.001.

[0041] Figure 2Figure for the protection of retinal nerve fiber layer thickness and retinal ganglion cell number in glaucoma model mice on the 28th day after modeling; wherein, a is a typical field of view of hematoxylin-eosin staining of retinal sections of mice in each group (scale = 50 microns), b is a statistical diagram of the number of cells in the retinal ganglion cell layer (GCL) of each group, c is a statistical diagram of the thickness of the retinal nerve fiber layer (RNFL), d is a statistical diagram of the thickness of the inner plexiform layer (IPL), * indicates that there is a statistically significant difference between the two groups, p<0.05, ** indicates that there is a statistically significant difference between the two groups, p<0.01, *** indicates that there is a statistically significant difference between the two groups, p<0.001;

[0042] Figure 3 Figure for the improvement of visual function of indole acetic acid on glaucoma model mice; wherein, a is a statistical diagram of the number of mice in each group that first stepped on the cliff side in the visual cliff experiment, b is a statistical diagram of the total duration of the mice in each group staying on the cliff side in the visual cliff experiment, * indicates that there is a statistically significant difference between the two groups, p<0.05, *** indicates that there is a statistically significant difference between the two groups, p<0.001;

[0043] Figure 4 Figure for the inhibition of indole acetic acid on the activation of retinal microglia cells in glaucoma model mice; wherein, a is a typical field of view of fluorescence confocal microscope of mouse retinal smear (scale = 50 microns), Iba1 (green) marks microglia cells, b is a statistical diagram of the relative number of Iba1 positive cells in each group, c is a statistical diagram of the relative amount of Iba1 cumulative optical density, *** indicates that there is a statistically significant difference between the two groups, p<0.001;

[0044] Figure 5Figure for verification results of inhibiting retinal neuroinflammatory pathway activation and inflammatory mediator expression in glaucoma model mice by indole acetic acid; wherein a is the Western blotting diagram of iNOS, p-NFκB and NFκB in each group, b is the quantitative statistical diagram of p-NFκB protein expression level normalized by NFκB in each group, c is the quantitative statistical diagram of iNOS protein expression level normalized by β-tubulin in each group, d is the Western blotting diagram of TNF-α, IL-1β and IL-6 in each group, e is the quantitative statistical diagram of TNF-α protein expression level normalized by β-tubulin in each group, f is the quantitative statistical diagram of IL-1β protein expression level normalized by β-tubulin in each group, g is the quantitative statistical diagram of IL-6 protein expression level normalized by β-tubulin in each group, h is the relative quantitative statistical diagram of mRNA expression of TNF-α, IL-1β, IL-6 and iNOS in each group, i is the immunofluorescence diagram of TNF-α, IL-1β, IL-6 and iNOS localization and relative expression in GCL (ganglion cell layer), IPL (inner plexiform layer) and INL (inner nuclear layer) of retinal sections of mice in each group (scale bar = 50 μm), * indicates that there is a statistical difference between two groups, p<0.05, ** indicates that there is a statistical difference between two groups, p<0.01, *** indicates that there is a statistical difference between two groups, p<0.001. DETAILED DESCRIPTION

[0045] The technical solutions of the present application are described in detail below with reference to the drawings. The embodiments of the present application are only used to illustrate the technical solutions of the present application and not to limit. Although the present application is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present application can be modified or replaced equivalently without departing from the spirit and scope of the technical solutions of the present application, which should be covered in the scope of claims of the present application.

[0046] The reagents used in the embodiments of the present application are purchased from Biyun Tian Company if not specified.

[0047] The mice used in the present embodiment are 6-8 week old wild type C57BL / 6J mice, which are raised under standard conditions.

[0048] 1) Establishing glaucoma mouse model

[0049] Select 6-8 week old male C57BL / 6J mice with a body weight of 18-25 g, which are purchased from Shanghai Jesuit Experimental Animal Co., Ltd. The mice are randomly divided into three groups, namely the control group (Control group), the glaucoma model group (MB group) and the glaucoma model combined with indole acetic acid intervention group (MB+IAA). The method for establishing the glaucoma model is as follows: after mixed anesthesia of the mice, 2 μl (concentration of about 5.0×10 6Microbeads (15 pm in diameter, Invitrogen) were injected into the anterior chamber. Intraocular pressure was measured using Icare TonoLab (Icare, Finland) at 1, 3, 7, 14, 21 and 28 days after surgery.

[0050] 2) Indoleacetic acid intervention

[0051] Indoleacetic acid powder was dissolved to 100 mg / mL using DMSO. On the day of establishing the mouse model of glaucoma, indoleacetic acid (20 mg / kg body weight) was injected intraperitoneally, followed by once daily administration.

[0052] 3) Histological and morphological analysis

[0053] After 14 or 28 days of glaucoma modeling, retinal smears or eyeball sections were prepared for histological and morphological analysis. When taking samples, first perform heart perfusion with normal saline and 4% paraformaldehyde, and after completion, remove the mouse eyeball and place it in FAS eyeball fixing solution (Servicebio) overnight. For retinas prepared for smears, the mouse retina was removed intact, flattened and fixed in 4% paraformaldehyde for 15 minutes, and then blocked with 5% goat serum and 0.3% Triton-X100 mixed in PBST for 1 hour. Incubate the primary antibody β-III tubulin (1:1000, Abeam) at 4°C overnight in a wet box, and then incubate the Alexa Fluor 594 labeled secondary antibody (1:200, Abbkine) for 2 hours. Use a fluorescence confocal microscope (Nikon) to take pictures. In the middle and periphery of the retina, 8 fields of view were selected and the number of β-III tubulin positive cells was counted using ImageJ. For eyeballs prepared for sections, after OCT embedding, use a freezing microtome to section (10 pm), and perform hematoxylin-eosin staining to analyze the retinal structure. Count the number of retinal ganglion cell layer (GCL) cells, retinal nerve fiber layer (RNFL) thickness and inner plexiform layer (IPL) thickness in 5 consecutive fields of view per section, average the values and normalize them with the control group.

[0054] 4) Mouse visual function detection

[0055] Visual cliff test was used to detect the visual function of mice. The experimental device was a transparent glass box (60 cm long, 60 cm wide, and 15 cm high), half of which was placed on the edge of the experimental table and formed the "cliff" side (90 cm from the ground); the other half was still placed on the experimental table and formed the "platform" side. A transparent platform (10 x 7 cm, 2 cm high) was placed in the center of the box. The experimental table and the ground were covered with alternating black and white patterns (pattern length 2 cm) to help the mice form depth vision. Light sources were placed 30 cm above the experimental area and 30 cm from the ground to illuminate the experimental area. At the beginning of the experiment, the mice were placed on the transparent platform. The first choice of direction (cliff side or platform side) and the number of mice first stepping towards the cliff side were recorded. In addition, the total time each mouse spent in the "cliff" area within the first two minutes was recorded. Each mouse was tested only once, and the platform and box were thoroughly cleaned after each experiment.

[0056] 5) Retinal immunofluorescence

[0057] The mouse retinal sections were prepared according to the above steps, and then incubated with 0.3% Triton X-100 for 15 minutes at room temperature, and then blocked with 5% goat serum (Boster) and 0.3% Triton X-100 in PBS for 1.5 hours. TNFα (1:200, Proteintech), IL-1β (1:200, Proteintech), IL-6 (1:200, Proteintech), iNOS (1:200, Abclonal), RAGE (1:200, Abmart), and Iba1 (1:100, Servicebio, Wuhan) primary antibodies were incubated overnight at 4°C in a humidified box. Alexa Fluor 488 anti-rabbit or Alexa Fluor 594 anti-mouse (1:200, Abbkine) secondary antibodies were incubated for 1 hour at room temperature. Nuclei were stained with DAPI (Invitrogen). Finally, a confocal microscope (Nikon) was used for imaging. Each group included at least 3 samples. To evaluate the degree of microglial activation, 5 fields were randomly selected from each retina, and the number of Iba1-positive cells and the integrated optical density (IOD) were calculated and averaged; each group included 6 samples. The results were analyzed using ImageJ software and standardized using the control group.

[0058] 6) RNA extraction and real-time quantitative PCR (qPCR)

[0059] Total RNA of retinal tissue was extracted using TRIzol reagent, and the concentration and purity of RNA were determined by Nanodrop (THERMOFISHER). Reverse transcription was completed using PrimeScript RT reagent kit (Takara). The primers used in the present application were synthesized by GenScript (Beijing) according to PrimerBank. In the gene expression analysis, β-actin was used as an internal reference, and the fold change of expression was calculated by the 2^−Δ(ΔCT) relative quantification method.

[0060] 7) Western blot

[0061] Total protein of retinal tissue was extracted using RIPA lysis buffer (Sangon Biotech). After electrophoretic separation of protein samples in 10% SDS-PAGE gel, the samples were transferred to a polyvinylidene fluoride (PVDF) membrane. Then, the membrane was blocked with 5% skim milk in Tris-buffered saline containing Tween-20. The membrane was incubated with primary antibodies at 4°C overnight, and the antibodies were as follows: TNFα (1:1000, Proteintech), IL-1β (1:1000, Proteintech), IL-6 (1:1000, Proteintech), iNOS (1:1000, Abclonal, Wuhan, China), NFκB (1:1000, Proteintech), p-NFκB (1:1000, Abeam). Then, the membrane was incubated with secondary antibodies (1:1000, Beyotime) at room temperature for 1 hour. The membrane was developed using a Tanon imaging system (Shanghai) and BeyoECL Moon developing solution (Beyotime), and the images were quantitatively analyzed by ImageJ software.

[0062] 8) Statistical method

[0063] The data were expressed as mean ± standard deviation, and statistical analysis was performed using GraphPad Prism 8.0 software. The t-test was used for statistical analysis between two groups, and when the number of groups was greater than 2, the one-way analysis of variance was used, followed by Turkey’s test. P<0.05 was considered to be statistically significant. Examples

[0064] This example verified the effect of indole acetic acid on relieving retinal damage and retinal ganglion cell loss in glaucoma model mice.

[0065] The retinal flat mounts of mice in each group were subjected to immunofluorescence staining and confocal microscopy, and a typical field of view is shown in FIG. 8a. The β-III tubulin positive cells in each field of view were counted and statistically analyzed (FIG. 8b). Figure 1 a. The retinal flat mounts of mice in each group were subjected to immunofluorescence staining and confocal microscopy, and a typical field of view is shown in FIG. 8a. The β-III tubulin positive cells in each field of view were counted and statistically analyzed (FIG. 8b). Figure 1b-c), it can be seen that the percentage of retinal ganglion cells of glaucoma model mice (MB, blue) is significantly reduced compared with the control group (Control, white), while the percentage of retinal ganglion cells is significantly increased after the administration of indole acetic acid (MB+IAA, red).

[0066] The eyeball sections of mice in each group were subjected to hematoxylin-eosin staining, and a typical field of view is shown in Figure 2 a. The cell density of the retinal ganglion cell layer (GCL), the thickness of the retinal nerve fiber layer (RNFL), and the thickness of the inner plexiform layer (IPL) of each group of mice were statistically analyzed. It can be seen that the GCL cell density and RNFL thickness of glaucoma model mice (MB, blue) are significantly reduced, while the GCL cell density and RNFL thickness are significantly increased after intraperitoneal injection of indole acetic acid (MB+IAA, red) Figure 2 b-d).

[0067] The above results show that indole acetic acid can maintain the number of retinal ganglion cells and the thickness of the retinal nerve fiber layer, proving that it can alleviate retinal damage and retinal ganglion cell loss. Example

[0068] This example verifies the effect of indole acetic acid on visual function indicators in the visual cliff experiment of glaucoma model mice.

[0069] The test shows that compared with the control group (Control), the number of mice first stepping on the cliff side (blue part) increases, and the total time of staying on the cliff side (blue part) significantly increases in glaucoma model mice (MB), while the visual function of glaucoma mice treated with indole acetic acid (MB+IAA) is protected, which is manifested as a decrease in the number of first stepping on the cliff side (MB+IAA, green part) Figure 3 a) and a decrease in the time of staying on the cliff side (MB+IAA, green part) Figure 3 b).

[0070] Therefore, indole acetic acid effectively improves the visual function of glaucoma model mice. Example

[0071] This example verifies the effect of indole acetic acid on inhibiting the activation of retinal microglial cells in glaucoma model mice.

[0072] In order to evaluate the effect of indole acetic acid on the activation state of retinal microglial cells in glaucoma model mice, we used Iba1 primary antibody (1:100, Servicebio) and Alexa Fluor 488 anti-rabbit secondary antibody to perform immunofluorescence staining on mouse retinal smears, and a typical field of view of confocal microscopy is shown in Figure 4a. Compared with the control group, the glaucoma model mice (MB, blue) showed higher number of Ibal positive cells and Ibal integrated optical density (IOD), while the indole acetic acid supplementation significantly reduced the number of Ibal positive cells and Ibal integrated optical density (IOD) Figure 4 b-c).

[0073] The above results show that indole acetic acid can inhibit the activation of retinal microglial cells in glaucoma model mice. Embodiment

[0074] This embodiment verifies the effect of indole acetic acid on inhibiting the activation of retinal inflammation pathway and the expression of inflammatory mediators in glaucoma model mice.

[0075] This embodiment uses real-time quantitative PCR, Western blot and immunofluorescence to detect the changes in the expression of key proteins and inflammatory mediators in the retinal inflammation pathway of glaucoma model mice. Compared with the control group, the expression of iNOS and the ratio of phosphorylated NFκB / NFκB in glaucoma model mice (MB) were significantly higher than those in the control group (Control), and the supplementation of indole acetic acid (MB+IAA) significantly alleviated the expression Figure 5 a-c). Compared with the control group, the expression of TNF-α, IL-1β and IL-6 in glaucoma model mice (MB) was up-regulated at the protein level Figure 5 d-g) and mRNA level Figure 5 h), and this was further verified by the results of immunofluorescence quantification Figure 5 i), while the supplementation of indole acetic acid significantly inhibited the expression of the above inflammatory factors Figure 5 d-i).

[0076] The above experimental results show that indole acetic acid can inhibit the activation of retinal inflammation pathway and the expression of inflammatory mediators in glaucoma model mice.

[0077] The above results show that indole acetic acid can be used as a drug for glaucoma optic nerve protection, and can be used for the treatment of glaucoma or eye optic nerve damage.

Claims

1. Application of the compound indoleacetic acid in the preparation of drugs or medical devices for treating glaucoma.

2. The application according to claim 1, characterized in that... The compound indoleacetic acid can reduce optic nerve damage caused by glaucoma and protect visual function.

3. The application according to claim 1, characterized in that... The compound indoleacetic acid can maintain the number of retinal ganglion cells and the thickness of the retinal nerve fiber layer.

4. The application according to claim 1, characterized in that... The compound indoleacetic acid can inhibit the activation of retinal microglia.

5. The application according to claim 1, characterized in that... Inhibits the activation of retinal inflammatory pathways and the expression of inflammatory mediators.

6. The application according to claim 1, characterized in that... The indoleacetic acid mentioned above is produced by a bioreactor.

7. The application according to claim 6, characterized in that... The bioreactor includes exogenous genetic material that is regulated to produce indoleacetic acid.

8. The application according to claim 7, characterized in that... The exogenous genetic material is the indoleacetic acid production gene of Bacteroides fragilis.

9. The application according to claim 6, characterized in that... The bioreactor is Bacteroides fragilis.

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