HIV protease inhibitors (HIV PI) reduce morbidity and mortality of
HIV infection but cause multiple untoward effects. Because certain HIV PI evoke production of
reactive oxygen species (ROS) and volume-sensitive Cl(-) current (I(Cl,swell)) is activated by ROS, we tested whether HIV PI stimulate I(Cl,swell) in ventricular myocytes.
Ritonavir and
lopinavir elicited outwardly rectifying Cl(-) currents under isosmotic conditions that were abolished by the selective I(Cl,swell)-blocker DCPIB. In contrast,
amprenavir,
nelfinavir, and
raltegravir, an
integrase inhibitor, did not modulate I(Cl,swell) acutely.
Ritonavir also reduced action potential duration, but
amprenavir did not. I(Cl,swell) activation was attributed to ROS because
ebselen, an H(2)O(2) scavenger, suppressed
ritonavir- and
lopinavir-induced I(Cl,swell). Major ROS sources in cardiomyocytes are sarcolemmal
NADPH oxidase and mitochondria. The specific
NADPH oxidase inhibitor
apocynin failed to block
ritonavir- or
lopinavir-induced currents, although it blocks I(Cl,swell) elicited by osmotic swelling or stretch. In contrast,
rotenone, a mitochondrial e(-) transport inhibitor, suppressed both
ritonavir- and
lopinavir-induced I(Cl,swell). ROS production was measured in HL-1 cardiomyocytes with C-H(2)DCFDA-AM and mitochondrial membrane potential (ΔΨ(m)) with
JC-1. Flow cytometry confirmed that
ritonavir and
lopinavir but not
amprenavir,
nelfinavir, or
raltegravir augmented ROS production, and HIV PI-induced ROS production was suppressed by
rotenone but not
NADPH oxidase blockade. Moreover,
ritonavir, but not
amprenavir, depolarized ΔΨ(m). These data suggest
ritonavir and
lopinavir activated I(Cl,swell) via mitochondrial ROS production that was independent of
NADPH oxidase. ROS-dependent modulation of I(Cl,swell) and other
ion channels by HIV PI may contribute to some of their actions in heart and perhaps other tissues.