Application of reagent for inhibiting STAT3 gene expression in treatment of oxidative damage of trabecular meshwork cells

Abstract

The invention relates to application of a reagent for inhibiting STAT3 gene expression in treatment of oxidative damage of trabecular meshwork cells. According to the invention, a signal channel STAT3 to EIF4E to TGF-beta-SMAD < 2 / 3 > for regulating and controlling the oxidative damage of the trabecular meshwork cells is innovatively found, and the STAT3 activates the TGF-beta-SMAD < 2 / 3 > channel by combining with EIF4E3, so that the apoptosis, oxidative stress and fibrosis of the trabecular meshwork cells caused by the oxidative damage are promoted. After the expression of the STAT3 gene in trabecular meshwork cells is inhibited, the EIF4E3 level can be reduced, and the apoptosis, oxidative stress and fibrosis of the trabecular meshwork cells induced by EIF4E3 are improved. Therefore, a reagent for inhibiting STAT3 gene expression can be used for treating trabecular meshwork cell oxidative damage and eye diseases related to the trabecular meshwork cell oxidative damage, including glaucoma and the like.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to the treatment of eye diseases, specifically the application of a reagent that inhibits STAT3 gene expression in the treatment of oxidative damage to trabecular meshwork cells. Background Technology

[0002] The trabecular meshwork (TM) is composed of trabecular meshwork cells (TMC) within multiple layers of extracellular matrix (ECM). It plays a crucial role in the drainage of aqueous humor (AH) and controls intraocular pressure by regulating the outflow dynamics of aqueous humor.

[0003] Glaucoma is a group of eye diseases characterized by optic nerve atrophy and visual field defects, which can cause progressive optic nerve degeneration, decreased vision, and even irreversible blindness. Elevated intraocular pressure (IOP) is a major and common risk factor for glaucoma and visual field damage, and trabecular meshwork dysfunction is a major risk factor for both IOP elevation and glaucoma development. The main functions of trabecular meshwork cells include secreting efferent membrane condensation (ECM), producing specific enzymes, and phagocytizing aqueous humor (AH), helping to maintain aqueous humor outflow from the trabecular meshwork. Oxidative damage to trabecular meshwork cells can lead to cell death, impaired tissue integrity, and subsequent impaired aqueous humor outflow, resulting in elevated IOP. The integrity of trabecular meshwork cell function is crucial for resisting elevated IOP and further preventing glaucoma progression and visual field damage; increased oxidative damage is a significant pathogenic risk factor for glaucoma progression.

[0004] The protective and damaging mechanisms of immune responses to oxidative damage in the trabecular meshwork are not yet clear. Further research is expected to provide new targets and ideas for glaucoma treatment based on oxidative damage in the trabecular meshwork. Summary of the Invention

[0005] Therefore, the purpose of this invention is to provide the application of reagents that inhibit STAT3 gene expression in the treatment of oxidative damage in trabecular meshwork cells. The STAT3 gene can serve as an effective target for improving oxidative damage in trabecular meshwork cells and can be used in the treatment of related diseases, including glaucoma.

[0006] The first aspect of the present invention is to provide the use of an agent that inhibits STAT3 gene expression in the preparation of a medicament for treating oxidative damage to trabecular meshwork cells.

[0007] A second aspect of the present invention is the use of an agent that inhibits STAT3 gene expression in the preparation of a medicament for treating diseases related to oxidative damage in trabecular meshwork cells.

[0008] In some embodiments, the trabecular meshwork cell oxidative damage-related disease is glaucoma.

[0009] In some embodiments, the application includes suppressing the expression of the EIF4E3 gene after suppressing the expression of the STAT3 gene.

[0010] In some embodiments, the reagent is siRNA that inhibits STAT3 gene expression.

[0011] In some embodiments, the siRNA comprises a sense strand and a reaction strand, the nucleotide sequence of the sense strand being shown in SEQ ID NO: 1, and the nucleotide sequence of the antisense strand being shown in SEQ ID NO: 2.

[0012] A third aspect of the present invention is to provide a medicament for treating oxidative damage to trabecular meshwork cells, wherein the active ingredient of the medicament includes a reagent that inhibits the expression of the STAT3 gene;

[0013] The reagent is siRNA, which includes a sense strand and a reaction strand. The nucleotide sequence of the sense strand is shown in SEQ ID NO: 1, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO: 2.

[0014] A fourth aspect of the present invention is to provide a method for inhibiting oxidative damage to trabecular meshwork cells for non-therapeutic purposes, comprising the following steps: introducing a reagent for inhibiting STAT3 gene expression into a test sample, thereby inhibiting the expression of the STAT3 gene in the test sample.

[0015] In some embodiments, the reagent is siRNA that inhibits STAT3 gene expression.

[0016] In some embodiments, the siRNA comprises a sense strand and a reaction strand, the nucleotide sequence of the sense strand being shown in SEQ ID NO: 1, and the nucleotide sequence of the antisense strand being shown in SEQ ID NO: 2.

[0017] This invention has identified a novel target, STAT3, for treating oxidative damage in trabecular meshwork cells and provides an siRNA that effectively inhibits STAT3 gene expression in these cells. The invention innovatively discovers a signaling pathway regulating oxidative damage in trabecular meshwork cells: STAT3→EIF4E→TGF-β-SMAD2 / 3. STAT3 activates the TGF-β-SMAD2 / 3 pathway by binding to EIF4E3, thereby promoting oxidative damage-induced apoptosis, oxidative stress, and fibrosis in trabecular meshwork cells. Specific inhibition of STAT3 gene expression in trabecular meshwork cells using siRNA reduces EIF4E3 levels and improves EIF4E3-induced apoptosis, oxidative stress, and fibrosis. Therefore, agents that inhibit STAT3 gene expression can be used to treat oxidative damage in trabecular meshwork cells and related eye diseases, including glaucoma.

[0018] This invention also provides a method for inhibiting oxidative damage to trabecular meshwork cells for non-therapeutic purposes. This method is for scientific research use and can be used to study the pathogenesis and development mechanism of oxidative damage to trabecular meshwork cells and related diseases, and to develop potential therapeutic targets and drugs. Attached Figure Description

[0019] Figure 1 The results show the effects of H2O2 dose-dependent induction of apoptosis, oxidative damage, and fibrosis in HTM cells.

[0020] Figure 2 Experimental results showing that EIF4E3 knockdown can reverse H2O2-induced apoptosis, oxidative damage, and fibrosis in HTM cells.

[0021] Figure 3 The experimental results show that STAT3 promotes the expression of EIF4E3 by binding to the EIF4E3 promoter.

[0022] Figure 4 Knockdown of STAT3 can reverse H2O2-induced apoptosis, oxidative damage, and fibrosis in HTM cells by affecting EIF4E3 expression.

[0023] Figure 5 Experimental results for STAT3 binding to EIF4E3 to activate the TGF-β-SMAD2 / 3 pathway.

[0024] Figure 6 The results show that EIF4E3 regulates H2O2-induced apoptosis, oxidative damage, and fibrosis in HTM cells by activating the TGF-β-SMAD2 / 3 pathway. Detailed Implementation

[0025] To facilitate understanding of the present invention, a more complete description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0026] Unless otherwise specified, experimental methods in the following examples are generally performed under standard conditions or as recommended by the manufacturer. All commonly used chemical reagents used in the examples are commercially available products.

[0027] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used in this invention includes any and all combinations of one or more of the associated listed items.

[0028] The present invention will be further described in detail below with reference to specific embodiments.

[0029] Example 1

[0030] Exposure of human trabecular meshwork cells (HTM) to hydrogen peroxide (H2O2) is a widely used in vitro method for inducing oxidative stress damage, which can lead to HTM cell death, impaired tissue integrity, and subsequent aqueous humor outflow obstruction. This study utilizes H2O2-induced HTM cells to investigate the regulatory role and mechanism of H2O2-induced oxidative damage in HTM cells.

[0031] I. Experimental Methods

[0032] 1. Cell Culture and Processing

[0033] Immortalized HTM cells were purchased from Wuhan Shangen Biotechnology Co., Ltd., and were cultured in DMEM medium containing 10% fetal bovine serum (FBS), 2 mM L-glutamine, 110 mg / L sodium pyruvate and 1% penicillin-streptomycin (Beyotime, Beijing) at 37°C in a 5% CO2 incubator.

[0034] HTM cells in the logarithmic growth phase were exposed to 0 μM, 200 μM, 300 μM, or 400 μM H2O2 for 12 hours. 300 μM H2O2 was ultimately selected for further functional analysis.

[0035] 2. Cell transfection

[0036] (1) siRNA

[0037] siRNAs targeting EIF4E3 (si-EIF4E3) or STAT3 (si-STAT3) were synthesized and screened. The following siRNAs targeting STAT3 are provided as examples for comparison: si-STAT3#1 / 2 / 3, si-STAT3#1 / 2 / 3, si-EIF4E3, and the non-targeting siRNA (si-NC) were provided by Shanghai Gemma Gene Co., Ltd.

[0038] si-STAT3#1

[0039] Chain of Justice (SEQ ID NO: 1): 5'-UGAUUCUUCGUAGAUUGUGCU-3';

[0040] Antisense chain (SEQ ID NO: 2): 5'-CACAAUCUACGAAGAAUCAAG-3'.

[0041] si-STAT3#2

[0042] Chain of Justice (SEQ ID NO: 3): 5'-ACGUUAUCCAGUUUUCUAGCC-3';

[0043] Antisense chain (SEQ ID NO: 4): 5'-CUAGAAAACUGGAUAACGUCA-3'.

[0044] si-STAT3#3

[0045] Chain of Justice (SEQ ID NO: 5): 5'- UCAUUAAGUUUCUAAACAGCU-3';

[0046] Antisense chain (SEQ ID NO: 6): 5'- CUGUUUAGAAACUUAAUGAAA-3'.

[0047] si-EIF4E3

[0048] Chain of Justice (SEQ ID NO: 7): 5'-UGCUUUAAAAGUUAUGUGGGG-3'

[0049] Antisense strand (SEQ ID NO: 8): 5'-CCACAUAACUUUUAAAGCAGU-3'.

[0050] To inhibit the expression of EIF4E3 and STAT3 in HTM cells, HTM cells were transfected using si-STAT3#1 / 2 / 3, si-EIF4E3, and the control si-NC, respectively, as follows: HTM cells with 50%–60% confluence were placed in six-well plates. After 24 hours, the culture medium was aspirated and replaced with FBS-free DMEM medium. A sterile Eppendorf tube was taken, and 250 μL of Opti-MEM medium and 6.0 μL of Lipofectamine 2000 reagent were added, mixed thoroughly, and incubated at room temperature for 5 min. This tube was labeled as tube A. Another sterile Eppendorf tube was taken, and 250 μL of Opti-MEM medium and si-STAT3#1 / 2 / 3, si-EIF4E3, or the negative control si-NC at a final concentration of 100 nmol / L were added, mixed thoroughly, and incubated at room temperature for 5 min. This tube was labeled as tube B. The liquids from tubes A and B were then mixed and incubated at room temperature for 15 min. Take 100 μL of the above mixture and slowly add it to a 6-well cell culture plate. Mix well and incubate in an incubator for 12 h, then aspirate the culture medium. Subsequently, place the cells in DMEM culture medium containing 10% FBS and continue culturing for 48 h.

[0051] The relative expression levels of the STAT3 gene after transfection with si-STAT3#1, si-STAT3#2, and si-STAT3#3 were detected by RT-qPCR (three replicates). The results are shown in Table 1.

[0052] Table 1

[0053]

[0054] It is evident that si-STAT3#1 can better suppress the expression of the STAT3 gene, and further research will be conducted using si-STAT3#1.

[0055] (2) EIF4E3 overexpression

[0056] To overexpress EIF4E3 (Genbank accession number: NM_001134651.2) or STAT3 (Genbank accession number: NM_139276.3), their full-length forms were cloned into the pcDNA3.1 vector (Germage), with an empty vector (pcDNA) used as a control. The corresponding overexpression vectors and empty vectors were transformed into HTM cells using the same method as described above.

[0057] 3. MTT test

[0058] HTM cells were co-incubated with 20 μl MTT (5 mg / mL, Beyotime) in 96-well plates for 4 hours (5 × 10⁻⁶). 3Cells / well). Then discard the supernatant and add 200 μL DMSO (Sigma-Aldrich) to each well to dissolve the methylxanone. Record the OD value at 490 nm.

[0059] 4. EdU detection

[0060] 1 × 10 4 HTM cells were incubated for 3 hours in a medium containing 50 μM EdU solution (Raybro Biotech, Guangzhou, China) and then stained with Apollo. DAPI was used for staining the cell nuclei. Finally, images were taken to detect EdU-positive cells.

[0061] 5. Flow cytometry detection of apoptosis

[0062] HTM cells were resuspended in 500 µL of binding buffer (1 × 10⁻⁶). 6 The cells were stained in the solution (cells / mL), and then stained with 5 μL Annexin V-FITC and 5 μL PI (BET Biotech, Shanghai) for 15 min in the dark, and then analyzed by flow cytometry.

[0063] 6. Detection of SOD and MDA

[0064] Collect HTM cells, centrifuge (4000 g, 10 min) and collect the supernatant. Then, detect SOD and MDA levels according to the SOD detection kit or MDA detection kit (Nanjing Jiancheng).

[0065] 7. Western blot detection

[0066] Proteins were extracted from HTM cells by lysis on ice, then separated with a 15% separating gel and transferred to a PVDF membrane. Next, the membrane was incubated overnight at 4°C on a shaker with primary antibodies against fibronectin (#AF5335, 1:1000), α-SMA (#AF1032, 1:1000), collagen I (#AF7001, 1:1000), EIF4E3 (#DF8652, 1:2000), STAT3 (#AF6294, 1:800), TGF-β (#AF1027, 1:1000), SMAD3 (#AF6362, 1:800), SMAD2 (#AF6449, 1:800), and GAPDH (#AF7021, 1:5000) (Affinity, Brisbane, Australia). After incubation with the secondary antibody, the cells were developed and photographed using an ECL chemiluminescence detection kit (Beyotime).

[0067] 8. Chromatin Immunoprecipitation Method for Detection

[0068] The JASPAR database (https: / / jaspar.genereg.net / ) predicts that STAT3 has two binding sites in the promoter region of EIF4E3. Chromatin immunoprecipitation (ChIP) was used to verify the interaction between the STAT3 protein and the EIF4E3 promoter. HTM cells were fixed with 1% formaldehyde, and the cross-linked protein-DNA complex fragments were broken down into 200–500 bp fragments using sonication. Immunoprecipitation was performed using an anti-STAT antibody (ab178941, Abcam, Cambridge, England). Rabbit IgG antibody was used as a negative control. After elution, the immunoprecipitated chromatin was detected by qRT-PCR.

[0069] 9. Dual-luciferase reporter assay

[0070] The wild-type (WT) binding site of STAT3 in the EIF4E3 promoter region (site 1, nucleotide sequence as shown in SEQ ID NO: 9, SEQ ID NO: 9: TTTCCTAGAAG) and the mutant sequence at site 1 (MUT, nucleotide sequence as shown in SEQ ID NO: 10, SEQ ID NO: 10: TTTCCATGAAG) were inserted into the psiCHECK-2 vector to construct the luciferase vector (WT / MUT-EIF4E3) (Promega, Beijing). The recombinant plasmid was co-transfected with si-NC or si-STAT3 into HTM cells as follows: HTM cells in logarithmic growth phase were seeded in 96-well plates (2 × 10⁻⁶ cells / wells). 3 (Cells / well) were incubated at 37°C, 5% CO2, and 95% RH for 24 h. Transfection was performed when the cell density reached 70%. Solution A was prepared by thoroughly mixing 10 μL of fetal bovine serum-free DMEM medium with 0.3 μL of Lipofectamine™ 2000 Transfection Reagent and incubating at room temperature for 5 min. Simultaneously, 0.16 μg of WT-EIF4E3, MUT-EIF4E3, si-STAT3, and si-NC were diluted with 10 μL of fetal bovine serum-free DMEM medium and thoroughly mixed to prepare Solution B. Solution A and Solution B were thoroughly mixed and incubated at room temperature for 20 min. The transfection mixture was then pipetted into 96-well plates and incubated at 37°C, 5% CO2, and 95% RH for 6 h. The culture medium was then replaced with DMEM medium containing 10% fetal bovine serum and antibiotics. Luciferase activity was measured 48 hours later.

[0071] 10. Statistical Analysis

[0072] Data are expressed as mean ± standard deviation (SD). Student's t-test was used for comparisons between two groups, and ANOVA and Tukey's test were used for comparisons among multiple groups. P < 0.05 was considered statistically significant.

[0073] II. Experimental Results

[0074] 1. H2O2 dose-dependently induces apoptosis, oxidative damage, and fibrosis in HTM cells.

[0075] HTM cells were treated with 0 μM, 200 μM, 300 μM, and 400 μM H2O2 for 12 hours, respectively. MTT and EdU assays showed that H2O2 dose-dependently inhibited HTM cell proliferation. Figure 1 China A, Figure 1 (B). Simultaneously, H2O2 dose-dependently induced apoptosis in HTM cells ( Figure 1 In addition, H2O2 treatment decreased SOD activity in HTM cells, while MAD content increased in a dose-dependent manner. Figure 1 D, Figure 1 (E). Western blot results showed that H2O2 treatment dose-dependently increased the levels of fibronectin, α-SMA, and collagen I protein fibrosis in HTM cells ( Figure 1 (Middle F).

[0076] 2. EIF4E3 knockdown can reverse H2O2-induced apoptosis, oxidative damage, and fibrosis in HTM cells.

[0077] Studies have shown that EIF4E3 is involved in chronic oxidative stress in HTM cells. Therefore, we investigated the role of EIF4E3 in HTM cell damage to explore its potential function in glaucoma.

[0078] like Figure 2 As shown in Figure A, Western blot results indicated that H2O2 dose-dependently increased the EIF4E3 content in HTM cells. These results suggest that EIF4E3 may be involved in the occurrence of HTM cell damage and fibrosis. Transfection of HTM cells with si-NC or si-EIF4E3, followed by treatment with 300 μM H2O2 for 12 h, showed that compared with the si-NC group, si-EIF4E3 effectively reduced the increase in EIF4E3 in HTM cells induced by H2O2 treatment. Figure 2 (B). Functionally, EIF4E3 knockdown can eliminate H2O2-induced inhibition of HTM cell proliferation (B). Figure 2 C- Figure 2 E), enhanced apoptosis ( Figure 2 The content of F and SOD decreased (in the middle F) Figure 2 Elevated levels of G and MDA (in China) Figure 2 (H), and elevated levels of fibrosis in fibronectin, α-SMA, and collagen I proteins ( Figure 2 Middle I).

[0079] 3. STAT3 promotes EIF4E3 expression by binding to the EIF4E3 promoter.

[0080] According to predictions from the JASPAR database (https: / / jaspar.genereg.net / ), STAT3 has two binding sites in the promoter region of EIF4E3. Figure 3 (A). After H2O2 treatment, the expression of STAT3 in HTM cells increased in a dose-dependent manner. Figure 3 (B). Then, the interaction between STAT3 and EIF4E3 was verified. ChIP experiments showed that STAT3 binds to the EIF4E3 promoter at site 1 ( Figure 3 (C). Further, mutations were performed at site 1, and a dual-luciferase reporter assay was conducted. It was found that STAT3 silencing significantly reduced luciferase activity in the wild-type group, while there was no statistically significant difference in luciferase activity between the mutant and HTM cell groups. Figure 3 Next, Western blot was used to demonstrate the transfection efficacy of si-STAT3 and STAT3 overexpression plasmids in HTM cells. We found that in HTM cells, STAT3 silencing reduced EIF4E3 expression, while STAT3 overexpression increased EIF4E3 expression. Figure 3 Chinese E, Figure 3 (F). In summary, STAT3 promotes EIF4E3 expression by binding to the EIF4E3 promoter.

[0081] 4. STAT3 knockdown can reverse H2O2-induced apoptosis, oxidative damage, and fibrosis in HTM cells by affecting EIF4E3 expression.

[0082] Next, we assessed whether STAT3 affected H2O2-induced HTM cell damage using EIF4E3. HTM cells were transfected with si-STAT3 or si-STAT3 and EIF4E3 overexpression plasmids, respectively, and then treated with H2O2. Western blot results showed that si-STAT3 reduced the H2O2-induced increase in EIF4E3 expression, and transfection with EIF4E3 overexpression plasmids restored EIF4E3 expression. Figure 4 (A). In further functional analysis, we found that STAT3 silencing promoted the proliferation of HTM cells treated with H2O2 ( Figure 4 China B- Figure 4D), inhibited apoptosis ( Figure 4 E), increased SOD activity ( Figure 4 (in F), reduced MDA level ( Figure 4 (G), reduced fibronectin, α-SMA and collagen I ( Figure 4 The levels of profibrotic proteins in EIF4E3 were elevated; however, these effects mediated by STAT3 silencing were eliminated by EIF4E3 overexpression. Figure 4 China B- Figure 4 (H).

[0083] 5. STAT3 binds to EIF4E3 to activate the TGF-β-SMAD2 / 3 pathway.

[0084] Studies have found that the TGF-β signaling pathway is an important component of oxidative stress-induced fibrosis. Furthermore, TGF-β and its associated SMAD family are activated upon H2O2 stimulation, and TGF-β can mobilize Smad2 and Smad3 transcription factors to induce fibrosis. Therefore, we further investigated whether the TGF-β-related signaling pathway mediates the effect of EIF4E3 on H2O2-induced HTM cell damage.

[0085] We found that H2O2 increased the protein levels of TGF-β, SMAD2, and SMAD3 in HTM cells, while silencing these proteins by EIF4E3 decreased their levels in HTM cells. Figure 5 (A). Furthermore, knocking down STAT3 with si-STAT3 downregulated the protein levels of TGF-β, SMAD2, and SMAD3 in H2O2-treated HTM cells, while EIF4E3 overexpression reversed the effects induced by si-STAT3. Figure 5 (B). These results indicate that STAT3 regulates the activation of the TGF-β-SMAD2 / 3 pathway via EIF4E3.

[0086] 6. EIF4E3 regulates H2O2-induced apoptosis, oxidative damage, and fibrosis in HTM cells by activating the TGF-β-SMAD2 / 3 pathway.

[0087] Further investigation is needed to determine whether the TGF-β-SMAD2 / 3 pathway is involved in the effect of EIF4E3 on H2O2-induced HTM cell damage.

[0088] First, Western blot was used to verify the enhancement efficiency of the EIF4E3 plasmid. Figure 6 (A). Then it was found that in HTM cells treated with H2O2, proliferation inhibition induced by EIF4E3 overexpression ( Figure 6 China B- Figure 6 Increased D) and apoptosis ( Figure 6Treatment with SD-208 (a type I TGF-β receptor kinase inhibitor) attenuated the enhancement of oxidative stress induced by EIF4E3 overexpression. Furthermore, SD-208 treatment eliminated the enhancement induced by EIF4E3 overexpression. Figure 6 China F, Figure 6 In HTM cells treated with G and H2O2, the levels of fibrosis of fibronectin, α-SMA, and collagen I proteins increased. Figure 6 (H).

[0089] In summary, STAT3 binds to EIF4E3 and activates the TGF-β-SMAD2 / 3 pathway, thereby promoting H2O2-induced apoptosis, oxidative stress, and fibrosis in trabecular meshwork cells (HTM cells). Inhibiting STAT3 gene expression can effectively improve oxidative damage in HTM cells. Therefore, agents that inhibit STAT3 gene expression can be used to treat oxidative damage in trabecular meshwork cells and related eye diseases, including glaucoma.

[0090] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. Application of reagents that inhibit STAT3 gene expression in the preparation of drugs for treating oxidative damage to trabecular meshwork cells.

2. Application of reagents that inhibit STAT3 gene expression in the preparation of drugs for treating diseases related to oxidative damage in trabecular meshwork cells.

3. The application as described in claim 2, characterized in that, The disease associated with oxidative damage to trabecular meshwork cells is glaucoma.

4. The application as described in any one of claims 1 to 3, characterized in that, The application includes inhibiting the expression of the EIF4E3 gene after inhibiting the expression of the STAT3 gene.

5. The application as described in any one of claims 1 to 3, characterized in that, The reagent is siRNA that inhibits STAT3 gene expression.

6. The application as described in claim 5, characterized in that, The siRNA comprises a sense strand and a reaction strand, the nucleotide sequence of which is shown in SEQ ID NO: 1, and the nucleotide sequence of which is shown in SEQ ID NO:

2.

7. A drug for treating oxidative damage to trabecular meshwork cells, characterized in that, The active ingredient of the drug includes a reagent that inhibits STAT3 gene expression; The reagent is siRNA, which includes a sense strand and a reaction strand. The nucleotide sequence of the sense strand is shown in SEQ ID NO: 1, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO:

2.

8. A method for inhibiting oxidative damage to trabecular meshwork cells for non-therapeutic purposes, characterized in that, Includes the following steps: A reagent that inhibits STAT3 gene expression is introduced into the test sample, thereby inhibiting the expression of the STAT3 gene in the test sample.

9. The method as described in claim 8, characterized in that, The reagent is siRNA that inhibits STAT3 gene expression.

10. The application as described in claim 9, characterized in that, The siRNA comprises a sense strand and a reaction strand, the nucleotide sequence of which is shown in SEQ ID NO: 1, and the nucleotide sequence of which is shown in SEQ ID NO: 2.

Images