June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
PROTECTIVE EFFECT OF NITRO-OLEIC ACID ON OXIDATIVE STRESS, GLIOSIS AND PRO-ANGIOGENIC RESPONSE IN MÜLLER GLIAL CELLS
Author Affiliations & Notes
  • María Victoria Vaglienti
    Departamento de Bioquímica Clínica, Universidad Nacional de Cordoba Facultad de Ciencias Quimicas, Cordoba, Córdoba, Argentina
    Centro de Investigaciones en Bioquimica Clinica e Inmunologia, Cordoba, Córdoba, Argentina
  • Paula Virginia Subirada
    Instituto de Investigacion Medica Mercedes y Martin Ferreyra, Cordoba, Córdoba, Argentina
    Departamento de Bioquímica Clínica, Universidad Nacional de Cordoba Facultad de Ciencias Quimicas, Cordoba, Córdoba, Argentina
  • MARIANA BELEN JORAY
    CONICET, Instituto de Investigaciones en Recursos Naturales y Sustentabilidad José Sánchez Labrador J. S., Córdoba, Córdoba, Argentina
  • Pablo Federico Barcelona
    Departamento de Bioquímica Clínica, Universidad Nacional de Cordoba Facultad de Ciencias Quimicas, Cordoba, Córdoba, Argentina
    Centro de Investigaciones en Bioquimica Clinica e Inmunologia, Cordoba, Córdoba, Argentina
  • GUSTAVO BONACCI
    Departamento de Bioquímica Clínica, Universidad Nacional de Cordoba Facultad de Ciencias Quimicas, Cordoba, Córdoba, Argentina
    Centro de Investigaciones en Bioquimica Clinica e Inmunologia, Cordoba, Córdoba, Argentina
  • Maria Cecilia Sanchez
    Departamento de Bioquímica Clínica, Universidad Nacional de Cordoba Facultad de Ciencias Quimicas, Cordoba, Córdoba, Argentina
    Centro de Investigaciones en Bioquimica Clinica e Inmunologia, Cordoba, Córdoba, Argentina
  • Footnotes
    Commercial Relationships   María Vaglienti None; Paula Subirada None; MARIANA JORAY None; Pablo Barcelona None; GUSTAVO BONACCI None; Maria Sanchez None
  • Footnotes
    Support  Secretaría de Ciencia y Tecnología, Universidad Nacional de Córdoba (SECyT-UNC) Consolidar 2018–2021, Fondo para la Investigación Científica y Tecnológica (FONCyT), Proyecto de Investi-gación en Ciencia y Tecnología (PICT) 2015 No. 1314, and PIP (CONICET) 2021–2023
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 1604. doi:
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    • Get Citation

      María Victoria Vaglienti, Paula Virginia Subirada, MARIANA BELEN JORAY, Pablo Federico Barcelona, GUSTAVO BONACCI, Maria Cecilia Sanchez; PROTECTIVE EFFECT OF NITRO-OLEIC ACID ON OXIDATIVE STRESS, GLIOSIS AND PRO-ANGIOGENIC RESPONSE IN MÜLLER GLIAL CELLS. Invest. Ophthalmol. Vis. Sci. 2023;64(8):1604.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

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<0.05), as well as of HO-1 mRNA expression at 4h (p<0.05) which was abrogated by trigonelline (Nrf2 inhibitor). PMA and LPS increased ROS levels in MIO-M1 cells compared with control (p<0.001), whereas NO2-OA prevented the increase induced by both stimuli (p>0,05). α2M induced gliosis mediated by the increase of GFAP and vimentin expression (p<0.001). In addition to this effect α2M increased ROS levels compared to control (p<0.05). The treatment with NO2-OA before the 6h with α2M reduced the GFAP and ROS levels to the control level (p>0,05). When MGCs under hypoxia and inflammation IL-1β were incubated with NO2-OA significantly reduced VEGF levels (p<0.05). NO2-OA inhibited ECs tubulogenesis (p< 0.05) via Keap1/Nrf2 (p>0.05).

Conclusions : Our results highlight NO2-OA protective effect on oxidative damage, gliosis and the exacerbated pro-angiogenic response in MGCs.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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