CN119700739B

CN119700739B - Application of fenofibrate in treating Fuchs corneal endothelial dystrophy

Info

Publication number: CN119700739B
Application number: CN202510216096.3A
Authority: CN
Inventors: 王丽强; 黄一飞; 李晓琦; 曾湘纹; 肖国城
Original assignee: Third Medical Center of PLA General Hospital
Current assignee: Third Medical Center of PLA General Hospital
Priority date: 2025-02-26
Filing date: 2025-02-26
Publication date: 2025-06-20
Anticipated expiration: 2045-02-26
Also published as: CN119700739A

Abstract

The invention relates to the technical field of biological medicine, in particular to application of fenofibrate in treating Fuchs corneal endothelial malnutrition. The thickness of the elastic layer behind cornea in the FECD morbidity process can be effectively reduced by using fenofibrate drugs, the density reduction, cell damage and the like of corneal endothelial cells are reduced, and finally the decompensation of the corneal endothelial cells is improved.

Description

Application of fenofibrate in treating Fuchs corneal endothelial dystrophy

Technical Field

The invention relates to the technical field of biological medicine, in particular to application of fenofibrate in treating Fuchs corneal endothelial malnutrition.

Background

Fuchs corneal endothelial dystrophy (Fuchs' endothelial corneal dystrophy, FECD), also known as trichomonas cornea, is an age-related complex genetic disease that manifests as the formation of extracellular matrix deposits and apoptosis of corneal endothelial cells (Corneal Endothelial Cells, CECs).

FECD clinically divides into two categories, early and late. The early-onset FECD is rare and is caused by the point mutation of the pathogenic group Col8a2, the incidence rate of the late-onset FECD is higher, the early-onset FECD is frequently 50-60 years old, the early-onset FECD is a degenerative disease which is formed by the combined action of gene mutation and environmental factors, the incidence rate of female people is 3-4 times that of male people, and the disease condition of female people is more serious. Delayed FECD is characterized clinically by corneal drop warts (guttae), thickening of the elastic layer behind the cornea, and progressive loss of corneal endothelial cells.

The formation of corneal drop warts on the posterior elastic layer (DM) is the earliest and most typical pathological feature of FECD. CECs are attached to a posterior elastic layer consisting of two layers, the surface adjacent to the cornea stroma being formed for fetal time, called anterior banding (Anterior Banded Layer or FETAL LAYER, ABL), the posterior non-banding (Posterior Nonbanded Layer, PNBL) surface adjacent to the endothelial surface being formed by CECs secretion, which thickens year by year with age. The thickness of the back elastic layer of the FECD patient is about 4 times that of the normal person, the back non-banded layer is thinned or disappeared, the back elastic layer is replaced by the abnormally formed collagen banded layer (collagenous banded layer, CBL), a loose fibrous layer is formed under the cornea endothelium of part of the patient, and the cornea drop-shaped wart is attached to the collagen banded layer or the fibrous layer.

Human corneal endothelial cells are nonrenewable cells, and when the density of the corneal endothelial cells is less than 500 corneal endothelial cells per mm ², symptoms such as corneal edema, painful epithelial bulla, corneal haze and the like can occur, and serious cases even lead to blindness. Delayed type FECD is the most important disease causing decompensation of corneal endothelial function, the traditional means is corneal endothelial transplantation, and in recent years, it has been found that after the corneal endothelium in the central area of the cornea where corneal drop-like warts appear is torn off and removed, migration of the corneal endothelium can be promoted by locally applying rosudil (ripasudil) in combination, but damage to the corneal endothelium cannot be inhibited, which is the only effective drug for clinical use in treating FECD at present.

Thickening of the posterior elastic layer and formation of corneal drop-like warts are the primary links in the FECD pathogenesis and are key factors leading to subsequent corneal endothelial injury, apoptosis and decompensation. At present, no effective means for inhibiting abnormal proliferation of the post-elastic layer is reported, and no medicine for treating FECD for the incision point from the aspect of lipid metabolism exists.

Disclosure of Invention

The invention discovers that the abnormal energy metabolism of the cornea endothelium of the FECD patient has correlation with the remodelling of extracellular matrix, and can effectively inhibit the thickening of the elastic layer after cornea by regulating and controlling the lipid metabolism of the cornea endothelium so as to realize the function of protecting the cornea endothelial cells. Based on the method, the lipid metabolism of the cornea endothelium is regulated and controlled to serve as a target point for treating the cornea endothelium fibrosis and the thickening of the posterior elastic layer, so that the tendency of thickening of the posterior elastic layer is reduced, and the apoptosis and the shedding of the cornea endothelium are reversed or reduced, thereby treating the FECD from the source.

Thus, the present invention provides the use of fenofibrate for the treatment of Fuchs corneal endothelial dystrophy, in particular, for the treatment of corneal endothelial fibrosis and post-elastic layer thickening in Fuchs corneal endothelial dystrophy by modulating lipid metabolism of the corneal endothelium using fenofibrate to act on the PPARa target. Experiments show that the oral administration of fenofibrate can inhibit the thickening of the cornea posterior elastic layer after the decompensation of cornea endothelium, obviously relieve the damage of cornea endothelium and inhibit the apoptosis of cornea endothelium, thereby treating Fuchs cornea endothelial malnutrition at the source.

In order to achieve the above purpose, the technical scheme provided by the invention is as follows:

The invention provides an application of fenofibrate in treating Fuchs corneal endothelial dystrophy.

In the present invention, the Fuchs corneal endothelial dystrophy is delayed.

In the present invention, the mode of application is oral administration of fenofibrate.

In the present invention, the application mode is to prepare an eye drop containing fenofibrate.

In the invention, the fenofibrate can reduce the thickening of the elastic layer after cornea and/or reduce the apoptosis and shedding of cornea endothelial cells by regulating the lipid metabolism of the cornea endothelial cells.

In the invention, the fenofibrate has a PPARa as an action target for regulating lipid metabolism of corneal endothelial cells.

The invention also provides application of PPARa as a target in screening or developing treatment of Fuchs corneal endothelial dystrophy.

The invention further provides a medicament for the treatment of Fuchs corneal endothelial dystrophy, characterized in that the medicament comprises a therapeutically effective amount of fenofibrate.

According to the invention, the fenofibrate drug can inhibit thickening of the elastic layer after cornea and protect cornea endothelial cells by regulating lipid metabolism of cornea endothelium.

Specifically, by using fenofibrate drugs, the thickness of the elastic layer behind the cornea in the FECD morbidity process can be effectively reduced, the density reduction of the corneal endothelial cells and the cell damage are improved, and finally the corneal endothelial cell decompensation is improved. The method can be used for treating Fuchs corneal endothelial malnutrition fundamentally without surgery or immunosuppression.

Drawings

FIGS. 1A-1C are graphs of cornea morphology and transparency of mice at different time nodes of injury by slit lamp photographs;

FIG. 2 is a statistical plot of the corneal opacity of mice;

FIGS. 3A-3C are OCT diagrams of corneal states of different lesion time nodes of a mouse;

FIG. 4 is a graph showing statistics of OCT test mouse cornea thickness;

FIG. 5 is a morphology of corneal endothelial cells from different injury time nodes of mice observed by in vivo confocal microscopy;

FIGS. 6A-6C are HE staining and PSR staining to observe corneal stroma morphology and the post-corneal elastic layer;

FIG. 7 is a morphology of the elastic layer after observation of the cornea of the mouse by a transmission electron microscope experiment;

FIG. 8 is a statistical plot of the thickness of the elastic layer after transmission electron microscopy of the mouse cornea.

Detailed Description

Hereinafter, the present invention will be described in detail with reference to exemplary embodiments, but the present invention is not limited to these embodiments. The present invention is embodied in the various forms described below, but should not be construed as limited to the exemplary embodiments set forth herein. Accordingly, the detailed description and examples of the invention will convey the scope of the invention to those skilled in the art and are to be construed as being within the scope of the invention.

The invention discovers that fenofibrate medicine is used for regulating and controlling lipid metabolism of corneal endothelium for the first time, reduces the thickness of the elastic layer after cornea in the FECD pathogenesis process, further improves the reduction of the density of corneal endothelial cells, reduces cell damage and finally improves the decompensation of the corneal endothelial cells. Specifically, the fenofibrate drug has PPARa as an action target for regulating lipid metabolism of corneal endothelial cells.

The fenofibrate drug of the present invention is used in such a manner that fenofibrate is added to the feed of mice in a proportion of 0.2% (2 mg/kg), and the feeding amount of the mice is calculated as 4.5g per day. In some embodiments, fenofibrate may be used in an eye drop formulation with a single dose of 0.1g, three times daily.

EXAMPLE 1 Effect of oral administration of 0.2% fenofibrate on mouse corneal endothelial cells

1-1 Experimental methods

Feed containing 0.2% fenofibrate was prepared and experimental mice were divided into a normal feed group and a 0.2% fenofibrate feed group. The right eye of the experimental mouse is irradiated with 750J/cm ² UVA, the irradiation process shields the left eye, the left eye is a control group, and the right eye is a UVA damage group.

After the completion of the injury experiment, the mice were further divided into normal feed-fed groups and feed-fed groups containing 0.2% fenofibrate.

After chronic damage of the corneal endothelium is induced, continuous observation is carried out for 1 week, 2 weeks and 3 weeks, and a slit lamp is periodically used for photographing and observing the transparency of the cornea of the mouse, then the thickness of the cornea of the mouse is detected by OCT (optical coherence tomography) 1 month after the damage, the morphology of the corneal endothelium of the mouse is observed by confocal, the thickness of the corneal stroma and the morphology of the elastic layer after the cornea are observed by HE (HE) staining, and the structures such as the elastic layer after the cornea of the mouse are observed by a transmission electron microscope.

1-2 Experimental results

Figures 1A-1C show the damage of the cornea of mice in different experimental groups one week, two weeks and three weeks after UVA damage, and experimental results show that the cornea damage degree of mice in three weeks after UVA damage is more stable, and the corneal haze degree of the UVA irradiated group fed with common feed is more serious than that of the UVA irradiated group fed with 0.2% fenofibrate.

Figure 2 also shows the corneal haze of the different experimental groups one month after UVA damage, and the experimental results show that the UVA-irradiated group fed with normal feed had more severe corneal haze than the UVA-irradiated group fed with 0.2% fenofibrate.

Fig. 3A-3C show schematic diagrams of anterior segment OCT of mouse cornea one week, two weeks, and three weeks after UVA injury, respectively, of different experimental groups, and experimental results show that the degree of corneal edema of the group fed with 0.2% fenofibrate is significantly reduced compared with that of the group fed with ordinary feed.

Fig. 4 shows the cornea thickness of the different experimental groups one month after UVA injury, and the experimental results show that the UVA-irradiated group fed with normal feed had a greater cornea thickness than the UVA-irradiated group fed with 0.2% fenofibrate.

Fig. 5 shows schematic diagrams of confocal microscopy of cornea living bodies of mice three weeks after UVA injury in different experimental groups, in which the cells of the UVA-irradiated group fed with normal feed were enlarged, irregular in cell morphology and reduced in cell density in the same field of view, while the UVA-irradiated group fed with 0.2% fenofibrate showed no significant reduction in cell density and uniform cell size.

The experimental results show that 0.2% fenofibrate orally taken has the protection and treatment effects on corneal endothelial cells.

Example 2 protection of fenofibrate against corneal injury

2-1 Experimental method

At a time point of 1 month after UVA damage, paraffin sections were taken from the eyeballs of mice for immunohistochemical staining, HE staining was observed for cell damage, and sirius (PSR) staining was observed for collagen changes in the cornea.

At a time point of 1 month after UVA damage, a transmission electron microscopy experiment was performed to observe the change of the post-elastic layer.

2-2 Experimental results

Figure 6A shows the change in the morphology of the mouse cornea stroma and the change in the elastic layer after cornea in different experimental groups one month after UVA injury.

The experimental result of HE staining shows that after UVA injury, the corneal epithelial layer of the mouse becomes thinner (p < 0.001), the thickness of the corneal endothelial cells becomes thinner, the density is reduced, the cell nucleus staining is deepened, the matrix layer is obviously thickened, the thickness is increased from about 60 micrometers to about 90 micrometers (p < 0.001), and the thickness is far more than that of the cornea caused by technical factors such as slice manufacturing.

The experimental results of HE staining also showed that the corneal epithelial cells fed with 0.2% fenofibrate were thinner than the normal group, but the degree of corneal epithelial thinning was not different between before and after UVA irradiation in this group, which may be related to factors such as weight loss caused by feeding fenofibrate, but not UVA irradiation damage factors, compared to the UVA irradiation group fed with normal feed, the corneal endothelial thickness of the UVA irradiation group fed with 0.2% fenofibrate remained more (p < 0.001), and the corneal endothelial nuclei were not significantly condensed, the corneal endothelial cell layer thickness was variable, the corneal stroma thickness was greatly reduced (p < 0.001), indicating that the degree of corneal stroma edema was reduced. (see FIGS. 6B, 6C).

In addition, HE staining results also show that the UVA irradiation group fed with normal feed has a more obvious layer of post-elastic layer staining, which is most likely to be related to the thickening of the post-elastic layer thickness, and meanwhile, after sirius staining, we can observe that the UVA irradiation group fed with normal feed deepens and thickens the post-injury elastic layer staining, but is limited to microscope magnification, and the post-elastic layer thickness cannot be statistically analyzed.

In order to further observe the change of the post-elastic layer, a transmission electron microscope experiment was performed. As shown in the results of the experiment in FIG. 7 and FIG. 8, the thickness of the elastic layer after cornea of the mice in the UVA irradiation group fed with 0.2% fenofibrate is not significantly changed (about 2 μm) as compared with that of the mice in the common feed-fed UVA irradiation group, and the thickness of the elastic layer after cornea of the mice in the UVA irradiation group is thickened by about 1/2 (about 3.0 μm).

The experimental results show that the oral administration of fenofibrate can obviously inhibit the thickening of the elastic layer after cornea.

As determined in the above results, the present invention can effectively reduce the thickness of the elastic layer after cornea in FECD onset process, improve the decrease of the density of corneal endothelial cells, cell damage, and finally improve the decompensation of corneal endothelial cells by using fenofibrate drug. The method can be used for treating Fuchs corneal endothelial malnutrition fundamentally without surgery or immunosuppression.

It should be understood that the exemplary embodiments described herein are to be considered in descriptive sense only and not for purposes of limitation. Descriptions of features or aspects in various embodiments should generally be considered as being possible for similar features or aspects in other embodiments.

Claims

1. The application of fenofibrate in preparing medicine for treating Fuchs corneal endothelial malnutrition.

2. The use according to claim 1, wherein said Fuchs corneal endothelial dystrophy is delayed.

3. The use according to claim 2, characterized in that the mode of application is oral administration of fenofibrate.

4. The use according to claim 2, characterized in that the mode of application is the preparation of an ophthalmic formulation comprising fenofibrate.

5. The use according to claim 2, wherein fenofibrate reduces the thickening of the post-corneal elastic layer and/or reduces the apoptotic shed of the corneal endothelium by modulating the lipid metabolism of the corneal endothelial cells.

6. The use according to claim 5, wherein the fenofibrate has a PPARa as a target for regulating lipid metabolism of corneal endothelial cells.