Application of ferroptosis inhibitor in preparation of medicine for preventing and treating intraocular hypertension

Abstract

The invention relates to the technical field of biological medicine, in particular to application of a ferroptosis inhibitor in preparation of a medicine for preventing and treating intraocular hypertension. Research finds that the ferroptosis inhibitor can effectively slow down fibrosis of trabecular meshwork cells caused by SiO, and evidence and thought are provided for treating intraocular hypertension caused by SiO filling.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the application of a ferroptosis inhibitor in the preparation of a drug for preventing and treating high intraocular pressure. Background Art

[0002] Silicone oil (SiO) helps maintain adhesion between the retinal neuroepithelium and the retinal pigment epithelium. Since its introduction into ophthalmic surgery by Cibis et al. in 1962, SiO has become the main material for intraocular tamponade after pars plana vitrectomy (PPV). It has been shown to be beneficial in the treatment of proliferative vitreoretinopathy and extensive tractional retinal detachments associated with proliferative diabetic retinopathy. In addition, in the treatment of ocular trauma, SiO can effectively reduce the risk of postoperative bleeding and ensure retinal adhesion.

[0003] However, there are also reports that vitreous filling with SiO is closely associated with postoperative complications such as corneal lesions, cataracts, and glaucoma. Among them, increased intraocular pressure (IOP) is one of the most common complications after the use of silicone oil in the vitreous cavity. Possible causes include pupillary obstruction, postoperative inflammation, and migration of silicone oil into the anterior chamber. In clinical situations, if the patient is unable to maintain a prone position after surgery, the lower density of SiO may cause it to migrate to the upper part of the eyeball, close to the lens and possibly spread to the anterior chamber. This migration can lead to mechanical obstruction of aqueous humor filtration. In addition, the presence of SiO in the anterior chamber leading to extensive infiltration of the trabecular meshwork is another potential cause of increased IOP after vitrectomy. However, the specific mechanism of increased IOP due to the migration of silicone oil into the anterior chamber remains unclear.

[0004] The current way to solve this complication is still to use intraocular pressure-lowering drugs, but this cannot fundamentally solve the high intraocular pressure that occurs after SiO vitreous filling. It is urgent to develop drugs targeting the mechanism of this disease. Summary of the invention

[0005] In view of this, the present invention provides the use of a ferroptosis inhibitor in the preparation of a drug for preventing and treating ocular hypertension. The present invention shows that a ferroptosis inhibitor can effectively slow down the fibrosis of trabecular meshwork cells caused by SiO2, and has a significant preventive and therapeutic effect on ocular hypertension caused by SiO2 filling.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] Application of ferroptosis inhibitors in the preparation of drugs for preventing and treating ocular hypertension.

[0008] In the above application, the ocular hypertension is ocular hypertension caused by silicone oil.

[0009] When HTMC fibrosis occurs, the structure and function of the trabecular meshwork change, and the aqueous humor cannot flow out smoothly, which can easily cause increased intraocular pressure. The present invention found that ferroptosis inhibitors can significantly inhibit the high expression of HTMC fibrosis-related proteins (such as Fibronectin, Myocilin and α-SMA) caused by SiO, thereby improving or slowing down the fibrosis of trabecular meshwork cells caused by SiO, and thus playing a role in preventing and treating ocular hypertension.

[0010] In the above application, the ocular hypertension is also accompanied by glaucoma.

[0011] In the above application, the prevention and treatment includes: regulating the expression of HTMC fibrosis-related proteins and / or alleviating HTMC fibrosis caused by SiO.

[0012] Furthermore, the HTMC fibrosis-related protein includes at least one of ACSL4, COX2, GPX4, Fibronectin, Myocilin and α-SMA.

[0013] Furthermore, the regulation of the expression of HTMC fibrosis-related proteins includes:

[0014] Inhibit the expression of at least one of Fibronectin, Myocilin and α-SMA.

[0015] In the above application, the ferroptosis inhibitor includes Fer-1.

[0016] In the above application, the effective concentration of Fer-1 is 1-10 μM, specifically 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM, 7 μM, 8 μM, 9 μM, and 10 μM.

[0017] The present invention also provides a drug for preventing and treating ocular hypertension, comprising a ferroptosis inhibitor Fer-1 and a pharmaceutically acceptable excipient, or comprising a ferroptosis inhibitor Fer-1, SiO and a pharmaceutically acceptable excipient.

[0018] The above-mentioned drugs also include effective amounts of other drugs for preventing and treating ocular hypertension.

[0019] Among the above-mentioned drugs, the dosage forms include tablets, capsules, drops, pills and injections.

[0020] The present invention also provides a method for treating ocular hypertension caused by SiO, comprising: administering a ferroptosis inhibitor Fer-1 or administering a pharmaceutical composition containing the ferroptosis inhibitor Fer-1.

[0021] The present invention also provides a method for preventing ocular hypertension caused by SiO, comprising: simultaneously administering ferroptosis inhibitor Fer-1 and SiO.

[0022] In the application and method of the present invention, the ferroptosis inhibitor Fer-1 can be used to prevent ocular hypertension caused by SiO. The two can exist in the same preparation in the form of a mixture, or can exist in separate preparations.

[0023] The present invention provides the use of a ferroptosis inhibitor in the preparation of a drug for preventing and treating high intraocular pressure. Studies have shown that ferroptosis inhibitors can regulate the expression of HTMC fibrosis-related proteins and effectively slow down the fibrosis of trabecular meshwork cells caused by SiO2, providing evidence and ideas for the treatment of high intraocular pressure caused by SiO2 filling. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Figure 1 shows the changes in HTMCs proliferation and expression of Fibronectin, Myocilin, and α-SMA when co-cultured with SiO. A, Edu test detected the proliferation of trabecular meshwork cells. The results showed that the proliferation rate of trabecular meshwork cells gradually decreased with the increasing concentration of silicone oil (n = 6), magnification 100, scale bar = 25 μm; B, protein expression of Fibronectin, Myocilin, and α-SMA in HTMCs co-cultured with different concentrations of SiO (n = 6); C, labeling of Fibronectin in HTMCs co-cultured with SiO; representative images show immunohistochemistry of HTMCs labeled with DAPI (blue), phalloidin (red), and fibronectin (green). Magnification 200, scale bar = 25 μm. Compared with the control group, *P < 0.05, **P < .01, ***P < .001, ****P < .0001;

[0025] Figure 2 shows the protein expression of ACSL4, COX2, GPX4, and ferritin in HTMCs co-cultured with different concentrations of SiO (n = 6). Compared with the control group, *P < 0.05, **P < .01, ***P < .001, ****P < .0001;

[0026] Figure 3 shows the effects of ferroptosis inducer (erastin) or inhibitor (fer-1) on ferroptosis-related indicators. A, Protein expression of ACSL4, COX2, GPX4, and ferritin in HTMCs treated with SiO and erastin / fer-1 (n = 6); B, Labeling of ferritin in HTMCs treated with SiO and erastin / fer-1; Representative images depict immunohistochemistry of HTMCs labeled with DAPI (blue), phalloidin (red), and ferritin (green). Magnification 200, scale bar = 25 μm. *P < 0.05, **P < .01, ***P < .001, ****P < .0001 compared with the control group;

[0027] Fig. 4 Protein expression of fibronectin, myocilin, and α-SMA in HTMCs treated with SiO and erastin / fer-1 (n = 6). *P < 0.05, **P < .01, ***P < .001, ****P < .0001 compared with the control group. DETAILED DESCRIPTION

[0028] The present invention provides the use of ferroptosis inhibitors in the preparation of drugs for the prevention and treatment of ocular hypertension. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It is particularly important to point out that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0029] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0030] The present invention will be further described below in conjunction with embodiments:

[0031] Example 1

[0032] The cells were co-cultured with different concentrations of SiO (5%, 10%, 15%, 20%) for 48 hours. The EdU assay showed that the proliferation rate of HTMCs decreased in a concentration-dependent manner (Figure 1A). At the same time, we evaluated the expression of fibrosis-related proteins (Fibronectin, myocilin, and α-SMA) in HTMCs after co-culture with SiO. Western blot analysis showed that the levels of Fibronectin, myocilin, and α-SMA gradually increased with increasing SiO concentrations (Figure 1B). This indicates that SiO plays an important role in promoting HTMC fibrosis. In addition, immunofluorescence staining experiments provide a more direct visualization effect, showing that the fluorescence intensity of Fibronectin in HTMCs increased significantly after co-culture with SiO ( Figure 1 C). The results showed that SiO not only inhibited the proliferation of HTMCs but also enhanced the fibrotic response in HTMCs.

[0033] Furthermore, in this example, HTMCs were co-cultured with different concentrations of SiO and the expression of key ferroptosis markers was detected. Western blot analysis showed that the expression of pro-ferroptosis proteins ACSL4 and COX2 also increased with increasing SiO concentrations. In contrast, the expression of GPX4 and Ferritin decreased. These changes suggest that SiO may enhance the ferroptosis process in HTMCs, and this effect is dose-dependent (Figure 2). These data further support the hypothesis that the role of SiO in ocular complications may involve the induction of ferroptosis in HTMCs.

[0034] The above experimental results showed that SiO intervention affected the fibrosis of HTMCs and changed their ferroptosis level.

[0035] Example 2

[0036] In this example, HTMCs co-cultured with SiO were treated with ferroptosis inducer (erastin, 10 μM) or inhibitor (ferrostatin-1, Fer-1, 1 μM). The concentration of ferroptosis inducer in the culture medium was 10 μM, and the concentration of inhibitor was 1 μM. The inhibitory effect of ferroptosis inhibitor on HTMCs fibrosis was evaluated.

[0037] The efficacy of ferroptosis modulators was determined by Western blot analysis, and the results showed that ferroptosis markers in HTMCs were significantly changed after treatment with erastin or Fer-1 (Figure 3A). Immunofluorescence staining showed that erastin significantly reduced ferritin fluorescence, while Fer-1 increased ferritin fluorescence, indicating successful regulation of ferroptosis (Figure 3B). Further analysis showed that induction of ferroptosis led to a significant increase in the expression of fibrosis-related proteins, while inhibition of ferroptosis led to a decrease in their expression (Figure 4). These findings suggest that SiO2 causes HTMC fibrosis by regulating ferroptosis, resulting in ocular hypertension, and that ferroptosis inhibitors can reverse this process and inhibit SiO2-induced HTMC fibrosis, thereby improving and / or preventing and treating ocular hypertension caused by SiO2 filling after vitrectomy.

[0038] The above are only preferred embodiments of the present invention. It should be pointed out that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Application of ferroptosis inhibitors in the preparation of drugs for the prevention and treatment of ocular hypertension.

2. The use according to claim 1, characterized in that: The ocular hypertension is ocular hypertension caused by silicone oil.

3. The use according to claim 1 or 2, characterized in that: The ocular hypertension is also associated with glaucoma.

4. The use according to any one of claims 1 to 3, characterized in that: The prevention and treatment includes: regulating the expression of HTMC fibrosis-related proteins and / or alleviating HTMC fibrosis caused by SiO.

5. The use according to claim 4, characterized in that: The HTMC fibrosis-related proteins include at least one of ACSL4, COX2, GPX4, Fibronectin, Myocilin and α-SMA.

6. The use according to claim 4 or 5, characterized in that: The regulating the expression of HTMC fibrosis-related proteins comprises: Inhibit the expression of at least one of Fibronectin, Myocilin and α-SMA.

7. The use according to any one of claims 1 to 6, characterized in that: The ferroptosis inhibitors include Fer-1.

8. The use according to claim 7, characterized in that: The effective concentration of Fer-1 is 1-10 μM.

9. A drug for preventing and treating ocular hypertension, characterized in that: It includes the ferroptosis inhibitor Fer-1 and pharmaceutically acceptable excipients, or includes: the ferroptosis inhibitor Fer-1, SiO and pharmaceutically acceptable excipients.

10. The drug according to claim 9, characterized in that The dosage form of the drug includes tablets, capsules, drops, pills or injections.

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