ROLE OF NITRIC-OXIDE SYNTHASE INHIBITION IN LEUKOCYTE-ENDOTHELIUM INTERACTION IN THE RAT PIAL MICROVASCULATURE

We investigated the role of nitric oxide (NO) in leukocyte-endothelium interaction, blood-brain barrier (BBB) function and oxygen free-radic al production in the rat pial microcirculation. In a closed cranial wi ndow preparation (dura removed) over the parietal cortex of pentobarbi tal-anesthetized Wistar rats, NO synthase (NOS) was inhibited by syste mic and/or topical application of N-omega-nitro-L-arginine (L-NNA) und er physiological conditions and during leukotriene B-4 (LTB(4)) activa tion. Circulating leukocytes were labeled by intravenous injection of rhodamine 6G. We used a confocal laser scanning microscope (CLSM) and studied leukocyte rolling and sticking in pial veins and arteries befo re and after NOS inhibition. At the end of the experiments, sodium-flu orescein was injected intravenously to test BBB integrity. Brain corte x oxygen free-radical production was investigated in the cranial windo w preparation using lucigenin-enhanced chemiluminescence (CL). L-NNA a pplication did not lead to significant changes in leukocyte-endotheliu m interaction, BBB function, and oxygen free-radical production under physiological conditions [leukocyte-endothelium interaction: control ( n = 5), L-NNA systemically (n = 5), L-NNA topically (n = 5): at baseli ne rollers/100 mu m: 0.76 +/- 0.55, 0.64 +/- 0.94, 0.44 +/- 0.55 and s tickers/100 mu m: 0.90 +/- 0.28, 0.76 +/- 0.24, 0.84 +/- 0.42; at 60 m in rollers/100 mu m: 1.49 +/- 0.66, 1.21 +/- 0.99, 0.67 +/- 0.66 and s tickers/100 mu m: 1.04 +/- 0.20, 1.19 +/- 0.23, 1.21 +/- 0.54; oxygen free-radical production (n = 4): CL count before L-NNA application 35 +/- 17 cps, after 1 h of topical superfusion of L-NNA 38 +/- 14 cps; p < 0.05]. In contrast to the results achieved under physiological cond itions, a significant further increase of rolling leukocytes and BBB p ermeability occurred due to NOS inhibition under LTB(4)-activated cond itions [76 +/- 47% significant (p less than or equal to 0.01, n = 7) f urther increase of rollers/100 mu m due to 60 min L-NNA application fo llowing the activation period of 120 min LTB(4) superfusion]. Our resu lts support a modulatory role for NO in leukocyte-endothelium interact ion and BBB permeability in the pial microcirculation when this intera ction is increased.