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Novel effect of N-palmitoyl-L-serine phosphoric acid on cytosolic Ca2+ levels in human osteoblasts
Authors:Jan Chung-Ren  Lu Yih-Chau  Jiann Bang-Ping  Chang Hong-Tai  Wang Jue-Long  Chen Wei-Chung  Huang Jong-Khing
Institution:Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan 813.
Abstract:The effect of N-palmitoyl-L-serine phosphoric acid (L-NASPA), which has been used as an inhibitor of lysophosphatidic acid receptors, on intracellular Ca2+ concentration (Ca2+]i) in human osteosarcoma MG63 cells was measured by using fura-2. L-NASPA (0.1-10 microM) caused a rapid and transient plateau Ca2+]i rise in a concentration-dependent manner (EC50=0.5 microM). The L-NASPA-induced Ca2+]i rise was partly reduced by removal of extracellular Ca2+ but was not altered by L-type voltage-gated Ca2+ channel blockers. In Ca2+-free medium, thapsigargin, an inhibitor of the endoplasmic reticulum Ca2+-ATPase, induced a Ca2+]i rise, after which the increasing effect of L-NASPA on Ca2+]i was completely inhibited; also, pretreatment with L-NASPA partly reduced thapsigargin-induced Ca2+]i rise. U73122, an inhibitor of phospholipase C, abolished histamine (but not L-NASPA)-induced Ca2+]i rise. Overnight incubation with 1 microM L-NASPA did not affect cell proliferation, but 10-20 microM L-NASPA exerted 4% and 15% inhibition, respectively. Collectively, L-NASPA rapidly increased Ca2+]i in MG63 cells by evoking both extracellular Ca2+ influx and intracellular Ca2+ release, and is cytotoxic at higher concentrations.
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