Hydrogen peroxide-induced Ca2+ responses in CNS pericytes

Objective: The aims of the present study were to elucidate the interaction of reactive oxygen species (ROS) and Ca2+ response in central nervous system (CNS) pericytes. Methods: The intracellular Ca2+ concentration was measured using fluorescent Ca2+ indicator, fura-2, in cultured CNS pericytes. Results: Hydrogen peroxide evoked a dose-dependent increase in cytosolic Ca2+, which was completely inhibited by catalase. Removal of external Ca2+ or addition of nicardipine (1 μM) during application of hydrogen peroxide did not affect Ca2+ response. Incubation of the cells in Ca2+ free solution did not abolish but slightly reduced Ca2+ response by hydrogen peroxide. Ca2+ response to hydrogen peroxide was not altered by the depletion of intracellular Ca2+ by thapsigargin (1 μM). Pretreatment of the cells with tyrosine kinase inhibitor genistein (100 μM) or tyrphostin A47 (30 μM) significantly reduced Ca2+ increase by hydrogen peroxide. Conclusions: These results indicate that hydrogen peroxide evokes Ca2+ increase predominantly by release from intracellular Ca2+ store, which may be regulated by tyrosine kinases.