TY - JOUR
T1 - Tissue protection and endothelial cell signaling by 20-HETE analogs in intact ex vivo lung slices
AU - Jacobs, Elizabeth R.
AU - Bodiga, Sreedhar
AU - Ali, Irshad
AU - Falck, Aaron M.
AU - Falck, John R.
AU - Medhora, Meetha
AU - Dhanasekaran, Anuradha
N1 - Funding Information:
The authors acknowledge contributions of Stephanie Gruenloh, Ying Gao and Brian Weinberg and other members of the Jacobs and Medhora labs. Confocal microscopy was facilitated by Drs. Paula North, Suresh Kumar, and Children's Research Institute Imaging Core Laboratory. Financial support was provided by the Robert A. Welch Foundation ( GL 625910 ) and NIH ( DK38226; JRF )( HL49294; ERJ ). This material is the result of work supported with resources and the use of facilities at the Clement J. Zablocki VA Medical Center.
PY - 2012/10/1
Y1 - 2012/10/1
N2 - The capacity to follow cell type-specific signaling in intact lung remains limited. 20-hydroxyeicosatetraenoic acid (20-HETE) is an endogenous fatty acid that mediates signaling for a number of key physiologic endpoints in the pulmonary vasculature, including cell survival and altered vascular tone. We used confocal microscopy to identify enhanced reactive oxygen species (ROS) production in endothelial cell (EC)s in intact lung evoked by two stable analogs of 20-HETE, 20-5,14-HEDE (20- hydroxy. eicosa-5(Z),14(Z)- di. enoic acid) and 20-5,14-HEDGE (N-[20- hydroxy. eicosa-5(Z),14(Z)- dienoyl]. glycin. e). These analogs generated increased ROS in cultured pulmonary artery endothelial cells as well. 20-HETE analog treatment decreased apoptosis of pulmonary tissue exposed to hypoxia-reoxygenation (HR) ex vivo. Enhanced ROS production and apoptosis were confirmed by biochemical assays. Our studies identify physiologically critical, graded ROS from ECs in live lung tissue ex vivo treated with 20-HETE analogs and protection from HR-induced apoptosis. These methodologies create exciting possibilities for studying signaling by stable 20-HETE analogs and other factors in pulmonary endothelial and other lung cell types in their native milieu.
AB - The capacity to follow cell type-specific signaling in intact lung remains limited. 20-hydroxyeicosatetraenoic acid (20-HETE) is an endogenous fatty acid that mediates signaling for a number of key physiologic endpoints in the pulmonary vasculature, including cell survival and altered vascular tone. We used confocal microscopy to identify enhanced reactive oxygen species (ROS) production in endothelial cell (EC)s in intact lung evoked by two stable analogs of 20-HETE, 20-5,14-HEDE (20- hydroxy. eicosa-5(Z),14(Z)- di. enoic acid) and 20-5,14-HEDGE (N-[20- hydroxy. eicosa-5(Z),14(Z)- dienoyl]. glycin. e). These analogs generated increased ROS in cultured pulmonary artery endothelial cells as well. 20-HETE analog treatment decreased apoptosis of pulmonary tissue exposed to hypoxia-reoxygenation (HR) ex vivo. Enhanced ROS production and apoptosis were confirmed by biochemical assays. Our studies identify physiologically critical, graded ROS from ECs in live lung tissue ex vivo treated with 20-HETE analogs and protection from HR-induced apoptosis. These methodologies create exciting possibilities for studying signaling by stable 20-HETE analogs and other factors in pulmonary endothelial and other lung cell types in their native milieu.
KW - Confocal
KW - Dihydroethidium
KW - Eicosanoid
KW - Reactive oxygen species
KW - TUNEL
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U2 - 10.1016/j.yexcr.2012.06.005
DO - 10.1016/j.yexcr.2012.06.005
M3 - Article
C2 - 22687879
AN - SCOPUS:84864296910
SN - 0014-4827
VL - 318
SP - 2143
EP - 2152
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 16
ER -