Real-time imaging of mechanically and chemically induced ATP release in human lung fibroblasts

Extracellular adenosine 5′-triphosphate (ATP) acts as an inflammatory mediator of pulmonary fibrosis. We investigated the effects of mechanical and chemical stimuli on ATP release from primary normal human lung fibroblasts. We visualized the ATP release from fibroblasts in real time using a luminescence imaging system while acquiring differential interference contrast cell images with infrared optics. Immediately following a single uniaxial stretch for 1 s, ATP was released from a certain population of cells and spread to surrounding spaces. Hypotonic stress, which causes plasma membrane stretching, also induced the ATP release. Compared with the effects of mechanical stretch, ATP-induced release sites were homogeneously distributed. In contrast to the effects of mechanical stimuli, application of platelet-derived growth factor caused ATP release from small numbers of the cells. Our real-time ATP imaging demonstrates that there is a heterogeneous nature of ATP release from lung fibroblasts in response to mechanical and chemical stimuli.