Potassium ion fluxes in corneal epithelial cells exposed to UVB

作者：Ubels John L^*; Van Dyken Rachel E; Louters Julienne R; Schotanus Mark R; Haarsma Loren D

来源：Experimental Eye Research, 2011, 92(5): 425-431.

DOI：10.1016/j.exer.2011.02.019

• apoptosis
• cornea
• corneal epithelium
• potassium ions
• potassium channels
• ultraviolet

The goal of this study was to investigate the efflux of K(+) from human corneal limbal epithelial cells (HCLE) exposed to ambient levels of UVB, which is known to cause apoptosis, and to examine the effect of K(+) channel blockers on loss of potassium induced by UVB. HCLE cells were exposed to 100-200 mJ/cm(2) UVB, followed by incubation in culture media with 5.5-100 mM K(+), BDS-1, Ba(2+) or ouabain. To measure intracellular cations, cells were washed in 280 mM sucrose and lysed in DI water. K(+) and Na(+) levels in lysates were measured by ion chromatography. HCLE cells showed maximal loss of K(i)(+) 10 min after exposure to UVB and 5.5 mM K(+) media, with recovery of normal K(+) levels after 90 min. Treatment with 1 mu M BDS-1 following UVB exposure reduced the loss of K(+) by HCLE cells. Exposure to 0.1-5 mM Ba(2+) inhibited UVB-induced K(+) loss in a time and dose-dependent manner. These results confirm that blocking channels in HCLE cells exposed to UVB prevents efflux of K(+), confirming that UVB activates K(+) channels in these cells. Electrophysiology data show that K(+) channels remain highly active at least 90 min after UVB exposure. HCLE cells exposed to UVB and incubated in 0.01-1 mu M ouabain did not recover from UVB-induced K(+) loss. These data suggest that the Na/K pump may act to restore [K(+)](i) to control levels in HCLE cells following UVB exposure and that the pump is not damaged by exposure to UVB. Incubation of HCLE cells exposed to UVB in medium with 25-100 mM K(+) media prevented K(+) efflux at extracellular concentrations as low as 25 mM (the concentration in tear fluid), maintaining control levels of K(i)(+). In all experiments inward fluxes and intracellular Na+ levels mirrored K(+) changes, albeit at the expected lower concentrations. The prevention of UVB-induced K(i)(+) loss by 25 mM K(o)(+) is consistent with the possible contribution of the relatively high K(+) concentration in tears to protection of the corneal epithelium from ambient UVB.

• 出版日期2011-5