Suppression of Ca²⁺ oscillations by SERCA inhibition in human alveolar type 2 A549 cells: rescue by ochratoxin A but not CDN1163

Aims: Lung type 2 alveolar cells, by secreting surfactant to lower surface tension, contribute to enhance lung compliance. Stretching, as a result of lung expansion, triggers type 1 alveolar cell to release ATP, which in turn stimulates Ca²⁺-dependent surfactant secretion by neighboring type 2 cells. In this report, we studied ATP-triggered Ca²⁺ signaling in human alveolar type 2 A549 cells. Main methods: Ca²⁺ signaling was examined using microfluorimetric measurement with fura-2 as fluorescent dye. Key findings: Ca²⁺ oscillations triggered by ATP relied on inositol 1,4,5-trisphosphate-induced Ca²⁺ release and store-operated Ca²⁺ entry. Pathological conditions such as influenza virus infection and diabetes reportedly inhibit sarcoplasmic/endoplasmic reticulum Ca²⁺ ATPase (SERCA). We found that a very mild inhibition of SERCA by cyclopiazonic acid (CPA) sufficed to decrease Ca²⁺ oscillation frequency and the percentage of cells exhibiting Ca²⁺ oscillations. Ochratoxin A (OTA), an activator of SERCA, could prevent the suppressive effects by CPA. Inhibition of SERCA by hydrogen peroxide also suppressed Ca²⁺ oscillations. Interestingly, hydrogen peroxide-induced inhibition was prevented by OTA but aggravated by CDN1163, an allosteric activator of SERCA. CDN1163 also had an untoward effect of releasing intracellular Ca²⁺. Significance: Different modes of activation of SERCA may determine the outcome of rescue of Ca²⁺ oscillations in case of SERCA inhibition in alveolar type 2 cells.