Protective effects of CY-09 and astaxanthin on NaIO-induced photoreceptor inflammation the NLRP3/autophagy pathway.

Aim: To study the effect of the NLRP3/autophagy pathway on the photoreceptor inflammatory response and the protective mechanism of CY-09 and astaxanthin (AST).

Methods: ICR mice were intraperitoneally injected NaIO, CY-09, AST successively and divided into 5 groups, including the control, NaIO, NaIO+CY-09, NaIO+AST, and NaIO3+CY-09+AST groups. Spectral domain optical coherence tomography and flash electroretinogram were examined and the retina tissues were harvested for immunohistochemistry, enzyme linked immunosorbent assay (ELISA), and Western blotting. Retinal pigment epithelium cell line (ARPE-19 cells) and mouse photoreceptor cells line (661W cells) were also treated with NaIO, CY-09, and AST successively. Cell proliferation was assessed by cell counting kit-8 (CCK-8) assay. Apoptosis was analyzed by flow cytometry. Changes in autophagosome morphology were observed by transmission electron microscopy. Quantitative polymerase chain reaction (qPCR) was used to detect NLRP3 and caspase-1. NLRP3, caspase-1, cleaved caspase-1, p62, Beclin-1, and LC3 protein levels were measured by Western blotting. IL-1β and IL-18 were measured by ELISA.

Results: Compared with the control group, the activity of NaIO-treated 661W cells decreased within 24 and 48h, apoptosis increased, NLRP3, caspase-1, IL-1β and IL-18 levels increased, and autophagy-related protein levels increased (<0.05). Compared with NaIO group, CY-09 and AST inhibited apoptosis (<0.05), reduced NLRP3, caspase-1, IL-1β and IL-18 expression (<0.05), and inhibited autophagy. Compared with the other groups, CY-09 combined with AST significantly decreased NLRP3 expression and inhibited the expression of the autophagy-related proteins p62, Beclin-1, and LC3 and (<0.05).

Conclusion: CY-09 and AST inhibit NaIO-induced inflammatory damage through the NLRP3/autophagy pathway and . CY-09 and AST may protect retina from inflammatory injury.