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Abstract Abstract: Long-term blue light exposure beyond the risk limit is an important factor that causes retinal degenerative diseases, and damage to the retinal pigment epithelium is the key source of retinal light damage. In this study, we investigated the damaging effects of 448 nm blue light on human retinal pigment epithelial (ARPE-19) cells and the mode of apoptosis. ARPE-19 cells were irradiated with 25 mW/cm2 blue light for 3, 6, 9 and 12 hours, and cell activity was detected immediately by Calcein AM staining 24 h after irradiation. RT-PCR was used to examine the expression of necrotic apoptosis related genes 24 hours after irradiation, and the Western blotting was used to examine the expression of necroptosis-related and DNA-damage related proteins. The cell morphology changed significantly in the 9-hour and 12-hour groups, the cells swelled, the intercellular space was enlarged, and a large number of cell fragments and non-adhesive cells were observed. Compared with the normal group, the expression levels of the necroptosis-related genes RIPK1 and RIPK3 were significantly higher 3～9 hours after blue light irradiation, and reached the maximum value at 9 h. The expression levels of necroptosis-related proteins RIPK1, RIPK3, P-MLKL markers significantly increased with the increase of irradiation time after 6～12 h blue light irradiation, and reached the highest value at 12 h. The expression level of DNA damage-related γ-H2AX protein increased significantly after 3～12 h blue light irradiation, and reached the highest value at 12 h. The expression of necroptosis-related genes reached the maximum after 9 hours of blue light irradiation, while the expression of necroptosis-related proteins reached the maximum after 12 hours irradiation. It was speculated that 25 mW/cm2 blue light irradiation for 12 h may cause the necroptosis in ARPE-19 cells, and the degree of DNA damage may be related to the occurrence of necroptosis.
Key words: blue light; ARPE-19 cells; necroptosis; DNA damage; photochemical damage
（Acta Laser Biology Sinica, 2023, 32(1): 036-042）
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