Application of thrombospondin 1 (THBS-1) in preparation of medicine for preventing and/or treating age-related macular degeneration (AMD)

Abstract

The invention provides application of thrombospondin 1 (THBS-1) in preparation of a medicine for preventing and / or treating age-related macular degeneration (AMD), and belongs to the technical field of eye disease medicines. As the pathological process of the AMD comprises injury and death of retinal pigment epithelium (RPE), formation of choroidal neovascularization (CNV) and retinal inflammation, in-vitro cell experiments and in-vivo animal experiments are combined to verify the anti-inflammatory effect of the THBS-1 in experimental blue light induced retinal inflammation and verify the effect of the THBS-1 in inhibition of pathological neovascularization and the protection effect of retinal injury. Therefore, the invention provides the application of the THBS-1 in preparation of the medicine for preventing and / or treating the AMD.

Description

technical field

[0001] The invention belongs to the technical field of eye disease drugs, and in particular relates to the application of thrombospondin 1 in the preparation of drugs for preventing and / or treating age-related macular degeneration. Background technique

[0002] Retinal degenerative diseases mainly include age-related macular degeneration (AMD), diabetic retinopathy and retinitis pigmentosa, and are one of the leading causes of visual impairment worldwide. AMD is the most common form of retinal degenerative disease, which is mainly divided into two forms: non-neovascular AMD (dry AMD) and neovascular AMD (wet AMD). Dry AMD is associated with damage and death of the retinal pigment epithelium (RPE) and degeneration of photoreceptors, while wet AMD is usually attributed to the formation of choroidal neovascularization (CNV), which seriously affects the patient's vision.

[0003] Retinal inflammation is an important pathological process in the pathogenesis of AM...

Examples

Embodiment 1

[0038] Thrombospondin 1 (THBS-1) can effectively inhibit the retinal inflammatory response induced by blue light

[0039] 1. Cell culture

[0040] ARPE-19 cells (human retinal pigment epithelial cell line) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Cells were cultured in high-glucose DMEM medium containing 10% FBS and 1% penicillin-streptomycin double antibody at 37°C and 5% CO 2 Cultured in an incubator, and after about 2 to 3 days, the cells were subcultured at a ratio of 1:3.

[0041] 2. THBS-1 administration treatment

[0042] For in vitro experiments, 4 nM recombinant human thrombospondin-1 (THBS-1) was added to cells in a serum-starved state and incubated for at least 6 hours before blue light irradiation.

[0043] 3. Description of cell grouping

[0044] THBS-1 administration treatment + blue light irradiation group: 8×10 3 The density of cells / well was seeded in a 96-well plate, and after the cells were treated according to the ...

Embodiment 2

[0076] Thrombospondin 1 (THBS-1) can effectively inhibit pathological neovascularization

[0077] 1. hMEC-1 cell level experiment

[0078] 1. Cell Culture

[0079] hMEC-1 cells (human microvascular endothelial cell line) were purchased from ATCC cell bank in the United States. Cells were cultured in ECM (endothelial cell medium) containing 5% FBS, 1% penicillin-streptomycin double antibody and 1% endothelial cell growth supplement (ECGS) at 37°C, 5% CO 2 Culture in an incubator, change the medium halfway every day, and perform cell passage at 1:2 after about 2 to 3 days.

[0080] 2. THBS-1 administration treatment

[0081] When detecting the expression of phosphorylated protein in hMEC-1 cells, the cells were pretreated with 4 μM THBS-1 solution for 6 h under serum starvation state, and 20 ng / mL VEGF was added to the medium without ECGS for 30 min.

[0082] 3. Cell grouping

[0083] THBS-1 administration treatment+VEGF treatment group: at a density of 8×10 3 The cells / we...

Embodiment 3

[0118] Thrombospondin 1 (THBS-1) significantly protects the retina from blue light-induced retinal damage

[0119] 1. ARPE-19 cell level experiment

[0120] 1. Cell culture

[0121] ARPE-19 cells (human retinal pigment epithelial cell line) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Cells were cultured in high-glucose DMEM medium containing 10% FBS and 1% penicillin-streptomycin double antibody at 37°C and 5% CO 2 Cultured in an incubator, and after about 2 to 3 days, the cells were subcultured at a ratio of 1:3.

[0122] 2. THBS-1 administration treatment

[0123] For in vitro experiments, 4 nM recombinant human thrombospondin-1 (THBS-1) was added to cells in a serum-starved state and incubated for at least 6 hours before blue light irradiation.

[0124] 3. Cell grouping

[0125] THBS-1 administration treatment + blue light irradiation group: 8×10 3 The density of cells / well was seeded in a 96-well plate, and after the cells were trea...

technical field

[0001] The invention belongs to the technical field of eye disease drugs, and in particular relates to the application of thrombospondin 1 in the preparation of drugs for preventing and / or treating age-related macular degeneration. Background technique

[0002] Retinal degenerative diseases mainly include age-related macular degeneration (AMD), diabetic retinopathy and retinitis pigmentosa, and are one of the leading causes of visual impairment worldwide. AMD is the most common form of retinal degenerative disease, which is mainly divided into two forms: non-neovascular AMD (dry AMD) and neovascular AMD (wet AMD). Dry AMD is associated with damage and death of the retinal pigment epithelium (RPE) and degeneration of photoreceptors, while wet AMD is usually attributed to the formation of choroidal neovascularization (CNV), which seriously affects the patient's vision.

[0003] Retinal inflammation is an important pathological process in the pathogenesis of AM...

Examples

Embodiment 1

[0038] Thrombospondin 1 (THBS-1) can effectively inhibit the retinal inflammatory response induced by blue light

[0039] 1. Cell culture

[0040] ARPE-19 cells (human retinal pigment epithelial cell line) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Cells were cultured in high-glucose DMEM medium containing 10% FBS and 1% penicillin-streptomycin double antibody at 37°C and 5% CO 2 Cultured in an incubator, and after about 2 to 3 days, the cells were subcultured at a ratio of 1:3.

[0041] 2. THBS-1 administration treatment

[0042] For in vitro experiments, 4 nM recombinant human thrombospondin-1 (THBS-1) was added to cells in a serum-starved state and incubated for at least 6 hours before blue light irradiation.

[0043] 3. Description of cell grouping

[0044] THBS-1 administration treatment + blue light irradiation group: 8×10 3 The density of cells / well was seeded in a 96-well plate, and after the cells were treated according to the ...

Embodiment 2

[0076] Thrombospondin 1 (THBS-1) can effectively inhibit pathological neovascularization

[0077] 1. hMEC-1 cell level experiment

[0078] 1. Cell Culture

[0079] hMEC-1 cells (human microvascular endothelial cell line) were purchased from ATCC cell bank in the United States. Cells were cultured in ECM (endothelial cell medium) containing 5% FBS, 1% penicillin-streptomycin double antibody and 1% endothelial cell growth supplement (ECGS) at 37°C, 5% CO 2 Culture in an incubator, change the medium halfway every day, and perform cell passage at 1:2 after about 2 to 3 days.

[0080] 2. THBS-1 administration treatment

[0081] When detecting the expression of phosphorylated protein in hMEC-1 cells, the cells were pretreated with 4 μM THBS-1 solution for 6 h under serum starvation state, and 20 ng / mL VEGF was added to the medium without ECGS for 30 min.

[0082] 3. Cell grouping

[0083] THBS-1 administration treatment+VEGF treatment group: at a density of 8×10 3 The cells / we...

Embodiment 3

[0118] Thrombospondin 1 (THBS-1) significantly protects the retina from blue light-induced retinal damage

[0119] 1. ARPE-19 cell level experiment

[0120] 1. Cell culture

[0121] ARPE-19 cells (human retinal pigment epithelial cell line) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Cells were cultured in high-glucose DMEM medium containing 10% FBS and 1% penicillin-streptomycin double antibody at 37°C and 5% CO 2 Cultured in an incubator, and after about 2 to 3 days, the cells were subcultured at a ratio of 1:3.

[0122] 2. THBS-1 administration treatment

[0123] For in vitro experiments, 4 nM recombinant human thrombospondin-1 (THBS-1) was added to cells in a serum-starved state and incubated for at least 6 hours before blue light irradiation.

[0124] 3. Cell grouping

[0125] THBS-1 administration treatment + blue light irradiation group: 8×10 3 The density of cells / well was seeded in a 96-well plate, and after the cells were trea...

Images