INVESTIGADORES
SALVADOR Gabriela Alejandra
congresos y reuniones científicas
Título:
Role of phospholipase D pathway in the inflammatory response of the retinal pigment epithelium cells
Autor/es:
MATEOS, MELINA; KAMERBEEK, CONSTANZA; GIUSTO, NORMA; SALVADOR, GABRIELA ALEJANDRA
Lugar:
Puerto Varas
Reunión:
Congreso; XII PABMB Congress- XLIX Annual Meeting Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2013
Resumen:
The
retinal pigment epithelium (RPE) plays an important immunological role in the
retina and it is involved in many ocular inflammatory diseases that may end in
loss of vision and blindness. In this work the role of the phospholipase D
(PLD) pathway in the inflammatory response of the human RPE cells (ARPE-19) exposed
to bacterial lipopolysaccharide (LPS) was studied. ARPE-19 were exposed to LPS
(10 µg/ml) for 24 and 48 h. LPS treatment reduced cell viability by 15% after
48 h and increased NO production by 1 and 1.8 fold after 24 and 48 h,
respectively. Furthermore, 24 h LPS treatment strongly induced ciclooxygenase-2
(COX-2) expression and the activation of protein kinase C (PKCα/βII) and extracellular signal-regulated kinase
(ERK1/2). Transfected cells with EGFP-PLD plasmids
showed the typical subcellular localization of PLD1 (perinuclear) and PLD2 (plasma
membrane). Untransfected cells expressed both PLD isoforms. LPS treatment
increased PLD activity (measured as phosphatidylethanol formation) by 80% with
respect to the control. The presence of the PLD1 inhibitor (EVJ 0.15 µM) or the
PLD2 inhibitor (APV 0.5 µM) reduced LPS-induced PKCα/βII activation and COX-2 induction but only
PLD2 inhibition reduced ERK1/2 activation. Moreover, the inhibition of PLD2 and
ERK1/2 restored cell viability to control levels. Our results demonstrate for
the first time the participation of the PLD pathway in the inflammatory
response of RPE cells exposed to LPS.