TY - JOUR
T1 - Activation of protein kinase c by tumor necrosis factor-α in human non- pigmented ciliary epithelium
AU - Prasanna, Ganesh
AU - Dibas, Adnan
AU - Brown, Kevin
AU - Yorio, Thomas
PY - 1998/10
Y1 - 1998/10
N2 - Previously, we have shown that tumor necrosis factor-α (TNF-α), a proinflammatory cytokine, increases the synthesis and release of endothelin- 1 (ET-1), a potent vasoactive peptide from human non-pigmented ciliary epithelial (HNPE) cells, in a protein kinase C (PKC)-dependent manner. Diacylglycerol (DAG) and intracellular calcium ([Ca2+](i)) are well known activators of PKC. Some cytokines induce PKC activation by stimulating phospholipase C that hydrolyzes phosphatidylinositol bisphosphate (PIP2) into IP3 (intracellular calcium mobilizer) and DAG. In this study, the existence of a similar pathway was evaluated in HNPE cells treated with TNF- α, using intracellular calcium ([Ca2+](i)) measurements, PKC translocation assays and thin-layer chromatography (TLC) for quantification of DAG. Incubation times for agonists and inhibitors ranged from 1-30 minutes. The increase in DAG levels with TNF-α treatment was consistent with the observed translocation of the calcium-dependent PKC α isoform from the cytosol to the plasma membrane. However, these observations were not accompanied by a concomitant increase in [Ca2+](i). Similar translocation responses were observed with phorbol ester (phorbol 12-myristate 13-acetate) treatment. Our results indicate that TNF-α-induced PKC activation in HNPE cells occurs as a result of elevated DAG levels and is not due to an increase in intracellular calcium. Activated PKC, could enhance the pro-inflammatory responses of TNF- α in part by increasing the production of endothelins in the eye.
AB - Previously, we have shown that tumor necrosis factor-α (TNF-α), a proinflammatory cytokine, increases the synthesis and release of endothelin- 1 (ET-1), a potent vasoactive peptide from human non-pigmented ciliary epithelial (HNPE) cells, in a protein kinase C (PKC)-dependent manner. Diacylglycerol (DAG) and intracellular calcium ([Ca2+](i)) are well known activators of PKC. Some cytokines induce PKC activation by stimulating phospholipase C that hydrolyzes phosphatidylinositol bisphosphate (PIP2) into IP3 (intracellular calcium mobilizer) and DAG. In this study, the existence of a similar pathway was evaluated in HNPE cells treated with TNF- α, using intracellular calcium ([Ca2+](i)) measurements, PKC translocation assays and thin-layer chromatography (TLC) for quantification of DAG. Incubation times for agonists and inhibitors ranged from 1-30 minutes. The increase in DAG levels with TNF-α treatment was consistent with the observed translocation of the calcium-dependent PKC α isoform from the cytosol to the plasma membrane. However, these observations were not accompanied by a concomitant increase in [Ca2+](i). Similar translocation responses were observed with phorbol ester (phorbol 12-myristate 13-acetate) treatment. Our results indicate that TNF-α-induced PKC activation in HNPE cells occurs as a result of elevated DAG levels and is not due to an increase in intracellular calcium. Activated PKC, could enhance the pro-inflammatory responses of TNF- α in part by increasing the production of endothelins in the eye.
UR - http://www.scopus.com/inward/record.url?scp=0031740398&partnerID=8YFLogxK
U2 - 10.1089/jop.1998.14.401
DO - 10.1089/jop.1998.14.401
M3 - Article
C2 - 9811229
AN - SCOPUS:0031740398
VL - 14
SP - 401
EP - 412
JO - Journal of Ocular Pharmacology and Therapeutics
JF - Journal of Ocular Pharmacology and Therapeutics
SN - 1080-7683
IS - 5
ER -