Swelling in the isolated perfused cornea induced by 12(R)hydroxyeicosatetraenoic acid.
OBJECTIVE
To evaluate the effect of 12(R)hydroxyeicosatetraenoic acid (12(R)HETE) on corneal swelling when directly perfused to human and rabbit corneal endothelium.


METHODS
Excised rabbit and human corneas were mounted in the in vitro specular microscope and the endothelium was perfused with 12(R)HETE at 10(-5), 10(-6), and 10(-7) mol/l. Both 12(R)HETE and 12(S)HETE were compared at equal molar (10(-6) mol/l) concentrations. The reversal of 12(R)HETE and ouabain corneal swelling was also compared. Endothelial permeability to carboxyfluorescein was measured after 12(R)HETE perfusion. High-performance liquid chromatographic analysis confirmed that 12(R)HETE remained in the perfusion media.


RESULTS
12(R)HETE caused a dose-dependent corneal swelling of 25 +/- 2, 24 +/- 1, and 14 +/- 0.5 microns/hr at 10(-5), 10(-6), and 10(-7) mol/l, respectively. Equal molar concentrations (10(-6) mol/l) of 12(S)HETE did not cause corneal swelling. Removal of the 12(R)HETE from the perfusion media resulted in reversal of corneal swelling whereas corneal swelling induced by ouabain did not reverse after ouabain removal. 12(R)HETE (10(-6) mol/l) perfused to the human corneal endothelium inhibited temperature reversal corneal thinning when compared to the paired corneal endothelium perfused with BSS Plus (Alcon Laboratories, Inc., Fort Worth, TX). Na/K adenosine triphosphatase activity was inhibited by 10(-6) mol/l ouabain by 35%, 10(-6) mol/l 12(R)HETE by 54%, and 10(-6) mol/l 12(S)HETE by 0.5%. Endothelial permeability to carboxyfluorescein was unaffected by 12(R)HETE.


CONCLUSIONS
12(R)HETE causes corneal swelling by inhibiting endothelial pump function. This inhibition of transport appears to be at least partly mediated by inhibition of endothelial Na/K adenosine triphosphatase.
Edelhauser
H F
HF
Department of Ophthalmology, Emory University School of Medicine, Atlanta, Georgia.
Geroski
D H
DH
Woods
W D
WD
Holley
G P
GP
Laniado-Schwartzman
M
M
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D000368:Aged; D000818:Animals; D002463:Cell Membrane Permeability; D002851:Chromatography, High Pressure Liquid; D003316:Corneal Diseases / Q000139:chemically induced* / Q000201:enzymology / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D004487:Edema / Q000139:chemically induced* / Q000201:enzymology / Q000473:pathology; D004728:Endothelium, Corneal / Q000187:drug effects / Q000648:ultrastructure; D005452:Fluoresceins / Q000378:metabolism; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000633:toxicity*; D010042:Ouabain / Q000633:toxicity; D010477:Perfusion; D011817:Rabbits; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013237:Stereoisomerism
D005452:Fluoresceins; D006893:Hydroxyeicosatetraenoic Acids; C024098:6-carboxyfluorescein; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D010042:Ouabain; D000254:Sodium-Potassium-Exchanging ATPase