Effects of Shear Stress on Intracellular Calcium Change and Histamine Release in Rat Basophilic Leukemia (RBL-2H3) Cells

Massage, one form of physical therapy, is widely used for a large number of musculoskeletal disorders, but its exact mechanism still remains to be elucidated. One hypothesis is that the shear stress caused by massage may induce cutaneous mast cells to release histamine, thereby improving the local tissue microcirculation of blood. In the present work, a mast cell line (rat basophilic leukemia cells, RBL-2H3) was used in vitro to study cellular responses to the stimulus of shear stress generated by a rotating rotor in a cell dish. The intraccllular calcium ([Ca2+](c)) was studied by confocal fluorescence microscopy with Fluo-3/AM staining and the released histamine was measured with a fluorescence spectrometer using o-phthalaldehyde (OPA) staining. An elevation of [Ca2+](c) occurred immediately after the shear stress, followed by histamine release. However, both [Ca2+](c) increase and histamine release disappeared when a Ca2+-free saline was used, indicating that the rise in the [Ca2+](c) is due to a Ca2+ influx from the extracellular buffer. Furthermore, Ruthenium red, a transient receptor potential vanilloid (TRPV) inhibitor, could effectively block the shear stress-induced histamine release, suggesting that TRPV membrane proteins are the likely targets of die shear stress. Because histamine is a well-known mediator of microvascular tissue dilation, these results may have an important impact on understanding the mechanism involved in massage therapy.