Protective effect of Nitro-Oleic Acid on Oxidative Stress, Gliosis and Pro-Angiogenic Response in Müller Glial Cells

VAGLIENTI, MARÍA VICTORIA; SUBIRADA, PAULA VIRGINIA; JORAY, MARIANA B.; BARCELONA, PABLO F.; BONACCI, GUSTAVO; SÁNCHEZ, MARIA CECILIA

New Orleans

Congreso; Annual Meeting of The Association for Research in Vision and Ophthalmology ? ARVO; 2023

ARVO

Purpose : Inflammation and oxidative stress are involved in proliferative retinopathies (PR). PR occurs with increase of VEGF, alterations of retinal blood barrier, extravasation of α2-macroglobulin (α2M) and gliosis in Müller Glial cells (MGCs). Nitro-fatty acids are electrophilic lipid mediators with anti-inflammatory and cytoprotective properties. Our aim was to investigate whether nitro-oleic acid (NO2-OA) could be beneficial against oxidative stress, gliosis and pro-angiogenic response in MGCs.Methods : MIO-M1 cells were treated or not with NO2-OA, and HO-1 was measured by WB at 8 and 16h post-stimulus (N=4). MIO-M1 cells were incubated 1h with trigonelline (1 and 25μM) before adding NO2-OA (5μM) for 4h. The HO-1 mRNA expression was measured by qRT-PCR (N=3). 6h after NO2-OA treatment, MIO-M1 cells were challenged by PMA (1µM) or LPS (1ng/ml) for 30min and ROS levels were determined by DCF assay (N=3). Then, MIO-M1 cells were treated or not with NO2-OA for 30 min before adding α2M for 2 to 6h and the GFAP, Vimentin and HO-1 levels were measured by WB (N=3) whereas ROS levels were determined by DCF assay (N=3). MIO-M1 cells were incubated under hypoxic and proinflammatory (IL-1β) conditions with NO2-OA or vehicle, and the VEGF expression was determined by qRT-PCR (N=3). BAEC were seeded in Matrigel and treated with vehicle or NO2-OA (5µM) with the addition of VEGF (10ng/mL) (N=3). GraphPad Prism program was employed for statistical analysis.Results : NO2-OA increased HO-1 protein expression at 8 and 16h post-stimulus (p