Jagged1-Notch1 Signaling Pathway Induces M1 Microglia to Disrupt the Barrier Function of Retinal Microvascular Endothelial Cells

• Published in: Current eye research 2024-10, Vol.49 (10), p.1098-1106
• Main Authors: Wu, Xiyu, Zhu, Haoxian, Liu, Junbin, Ouyang, Shuyi, Lyu, Zheng, Jin, Yeanqi, Chen, Xinyu, Meng, Qianli
• Format: Article
• Language: English
• Subjects: Apoptosis; Blood-Retinal Barrier; Blotting, Western; Cells, Cultured; Endothelial Cells - metabolism; Endothelium, Vascular - metabolism; Enzyme-Linked Immunosorbent Assay; Humans; Jagged-1 Protein - genetics; Jagged-1 Protein - metabolism; Microglia - metabolism; Real-Time Polymerase Chain Reaction; Receptor, Notch1 - genetics; Receptor, Notch1 - metabolism; Retinal Vessels - cytology; Retinal Vessels - metabolism; Signal Transduction - physiology
• ISSN: 0271-3683; 1460-2202; 1460-2202
• DOI: 10.1080/02713683.2024.2357601

Microglia-related inflammation is closely linked to the pathogenesis of retinal diseases. The primary objective of this research was to investigate the impact and mechanism of M1 phenotype microglia on the barrier function of retina microvascular endothelial cells. Quantitative polymerase chain reactions and western blot techniques were utilized to analysis the mRNA and protein expressions of M1 and M2 markers of human microglial clone 3 cell line (HMC3), as well as the levels of Notch ligands and receptors under the intervention of lipopolysaccharide (LPS) or interleukin (IL)-4. ELISA was utilized to detect the pro-inflammatory and anti-inflammatory cytokines from HMC3 cells. The cellular tight junction and apoptosis of human retinal microvascular endothelial cells (HRMECs) were assessed by western blot and fluorescein isothiocyanate-dextran permeability assay. The inhibitors of Notch1 and RNA interference (RNAi) targeting Jagged1 were used to assess their contribution to the barrier function of vascular endothelial cells. Inducible nitric oxide synthase (iNOS) and IL-1β were considerably elevated in LPS-treated HMC3, while CD206 and Arg-1 markedly elevated under IL-4 stimulation. The conditioned medium derived from LPS-treated HMC3 cells promoted permeability, diminished the expression of zonula occludens-1 and Occludin, and elevated the expression of Cleaved caspase-3 in HRMECs. RNAi targeting Jagged1 or Notch1 inhibitor could block M1 HMC3 polarization and maintain barrier function of HRMECs. Our findings suggest that Jagged1-Notch1 signaling pathway induces M1 microglial cells to disrupt the barrier function of HRMECs, which may lead to retinal diseases.