Slit2 Promotes H₂O₂-Induced Lens Epithelial Cells Oxidative Damage and Age-Related Cataract

To analyze the role of Slit2 in lens epithelial cell oxidative damage and its underlying mechanism.

Human lens epithelial cells (SRA01/04 cells) and rat transparent lens were cultured with H₂O₂ to establish cell oxidative stress models and rat cataract models. Immunohistochemistry, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot assays were employed to detect Slit2 levels within age-related cataracts(ARC) lens anterior capsule samples, rat cataract models, and cell oxidative stress models. In this study, qRT-PCR and Western blot assays were performed to derermine E-cadherin, N-cadherin, occludens1(ZO-1), α-SMA(α‑smooth muscle actin), Bcl-2, Bax, p-AKT, and AKT levels. In addition, Flow cytometry were performed to examine reactive oxygen species (ROS) and cell apoptosis. Cell viability, invasion, and migration were detected by CCK8, Transwell, and Wound healing.

Increased expression of Slit2 was found in ARC lens anterior capsule samples, H₂O₂-induced rat cataract models, and Human lens epithelial cells (HLECs) oxidative stress models. H₂O₂ significantly increased cell apoptosis and ROS generation, also accelerating cell migration, invasion, and epithelial-mesenchymal transition (EMT). In addition, H₂O₂ treatment repressed AKT phosphorylation and cell viability. Knock-down of Slit2 promoted cell viability and AKT phosphorylation levels, as well as repressed cell invasion, migration, apoptosis, ROS production and EMT.

Slit2 promoted lens epithelial cells oxidative stress damage via the AKT signalling pathways, providing a novel insight in ARC treatment.