Pharmacological evidences for the stimulation of calcium-sensing receptors by nifedipine in gingival fibroblasts.

OBJECTIVE

To investigate pharmacologically whether CaSRs are involved in the Ca(2+) antagonist-induced [Ca(2+)]i elevation in gingival fibroblasts.

MATERIALS AND METHODS

Gin-1 cells, normal human gingival fibroblasts, were used as the material. The [Ca(2+)] i was measured with fura-2/AM, a Ca(2+)-sensitive fluorescent dye.

RESULTS

At first, we confirmed the existence of CaSRs in these cells by showing that [Ca(2+)] i was elevated by high concentrations of extracellular Ca(2+) and by prototypic agonists of the CaSR such as gentamicin. The action of gentamicin was antagonized by inhibitors of phospholipase C (PLC), inositol trisphosphate (IP(3)) receptors, NSCCs, and, importantly, by the CaSR antagonist, NPS2390. Furthermore, the action of gentamicin was potentiated by activators of PLC and protein kinase C (PKC). This confirmed the pathway components mediating Ca(2+) responses to a known agonist of the CaSR. We then investigated whether nifedipine (an L-type Ca(2+) channel blocker) stimulates CaSRs to elevate [Ca(2+)] i via a similar mechanism. Nifedipine Ca(2+) responses were dose-dependently blocked by NPS2390 and by the same inhibitors of PLC, IP(3) receptors, and NSCCs that disrupted the action of gentamicin. Calphostin C (a PKC inhibitor) and TMB-8 (an inhibitor of Ca(2+) release from stores) also inhibited the nifedipine-induced [Ca(2+)] i elevation.

CONCLUSION

These findings suggest that CaSRs are involved in the nifedipine-induced [Ca(2+)] i elevation in gingival fibroblasts.