Abstract
Purpose :
Müller cell is the main retinal glial cell that plays an important role in maintaining retina normal physiological function. All of retinal trauma, glaucoma, and diabetic retinopathy can activate Müller cell, upregulate Glial fibers acidic protein (GFAP) and vimentin expression to aggravate inflammatory injury. This study aimed to investigate Müller cell activation in retinal damage caused by endotoxin-induced uveitis (EIU)
Methods :
EIU animal model was established. Western blot, qRT-PCR, and confocal immunofluorescence microscopy were applied to detect GFAP and vimentin expression at 12 h and 24 h after lipopolysaccharides (LPS) injection. Transwell assay and WST-1 cell proliferation assay test were used to determine the impact of RAW264.7 cells stimulated by lipopolysaccharide (LPS) on Müller cells migration and proliferation ability. Enzyme-linked immunosorbent assay (ELISA) was performed to test cytokines level in the supernatant.
Results :
GFAP and vimentin protein and mRNA expression increased significantly in EIU rat retina. Immunofluorescence locating GFAP and vimentin protein in internal limiting membrane (ILM), retinal ganglion cell layer (RGC) and inner plexiform layer (IPL), suggesting Müller cell was activated. LPS activated RAW264.7 cell promoted Müller cell migration and proliferation ability. TNF-α and IL-6 level elevated in the supernatant of LPS activated RAW264.7 cell.
Conclusions :
Müller cell was activated and upregulated GFAP and vimentin protein to induce retinal inflammatory injury in EIU rat model. LPS activated macrophage secreting TNF-α and IL-6 to activate Müller cell. This study contributed to discuss the molecular mechanism of EIU retinal injury and provided new thought for uveitis treatment.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.