Pannexin-1 Inhibits Pathologic Neovascularization in Oxygen-Induced Retinopathy by Targeting the HIF-1α/VEGF Pathway...

Retinopathy of prematurity (ROP) is a leading cause of childhood blindness, characterized by hypoxia-driven pathological retinal neovascularization. Current treatments, such as laser therapy and anti-VEGF drugs, carry significant risks, highlighting the need for safer preventive strategies. This study investigates the role of Pannexin-1 (Panx1) in the pathogenesis of ROP and explores its potential as a therapeutic target by modulating the HIF-1α/VEGF pathway.

Human retinal microvascular endothelial cells (HRMECs) were obtained from fetal eyes at 26-32 weeks gestational age, with ethical approval from the Guangdong Women and Children Hospital (No: 202201258). Retinal tissue was digested using 25 g/L trypsin and cultured in DMEM/F12 medium supplemented with 100 g/L fetal bovine serum (FBS). Transcriptomic sequencing libraries were prepared using the TruSeq stranded total RNA kit, and sequencing was conducted on the Illumina 6000 platform. Differentially expressed genes (DEGs) were identified (p < 0.05, |log₂FC| > 1). Validation experiments included rt-qPCR, Western blotting, and CCK-8 assays. Statistical analysis was performed using SPSS 24.0 and R v4.1.2, with DEG analysis conducted via the limma package.

Transcriptomic sequencing revealed 567 differentially expressed mRNAs(345 upregulated, 222 downregulated) and indicated a significant upregulation of Panx1 expression in the ROP group compared to controls. Rt-qPCR and Western blotting confirmed hypoxia-induced overexpression of Panx1. Probenecid (PBC) treatment inhibited the hypoxia-driven upregulation of Panx1, HIF-1α, and VEGF at both mRNA and protein levels. CCK-8 assays showed that hypoxia significantly impaired HRMEC proliferation (24h: p < 0.05; 48h: p < 0.001), and PBC further suppressed this effect (p < 0.0001).

Panx1 inhibition (via PBC) suppresses the HIF-1α/VEGF signaling pathway and reduces hypoxia-induced endothelial cell proliferation, providing a novel preventive strategy for ROP.