Pharmacological evidences for the stimulation of calcium-sensing receptors by nifedipine in gingival fibroblasts.
Pharmacological evidences for the stimulation of calcium-sensing receptors by nifedipine in gingival fibroblasts.
复制标题
在牙龈成纤维细胞中，硝苯地平刺激钙感应受体的药理证据。
DOI：
10.4103/0976-500x.77111
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发表时间：
2011-01
影响因子：
0.2
通讯作者：
Fujinami Y
中科院分区：
文献类型：
--
作者：
Hattori T;Ara T;Fujinami Y
探讨CaSRs是否参与钙拮抗剂诱导的牙龈成纤维细胞[Ca 2 +]i升高。Gin-1细胞，正常人牙龈成纤维细胞，被用作材料。[Ca ~（2+）] i用钙敏感荧光染料fura-2/AM测定。首先，我们证实了CaSRs在这些细胞中的存在，通过显示高浓度的细胞外Ca 2+和CaSR的原型激动剂如庆大霉素升高[Ca 2 +] i。庆大霉素的作用被磷脂酶C（PLC）、三磷酸肌醇（IP 3）受体、NSCC的抑制剂拮抗，重要的是，被CaSR拮抗剂NPS 2390拮抗。此外，PLC和蛋白激酶C（PKC）的激活剂可增强庆大霉素的作用。这证实了介导对CaSR的已知激动剂的Ca 2+响应的途径组分。然后，我们研究了硝苯地平（L-型Ca 2+通道阻滞剂）是否通过类似的机制刺激CaSRs升高[Ca 2 +] i。硝苯地平Ca 2+反应被NPS 2390和PLC、IP 3受体和NSCC的相同抑制剂阻断，这些抑制剂破坏庆大霉素的作用。PKC抑制剂Calphostin C和钙库释放抑制剂TMB-8也能抑制硝苯地平引起的[Ca ~（2+）] i升高。这些结果表明，CaSRs参与硝苯地平诱导的牙龈成纤维细胞[Ca 2 +] i升高。
To investigate pharmacologically whether CaSRs are involved in the Ca2+ antagonist-induced [Ca2+]i elevation in gingival fibroblasts. Gin-1 cells, normal human gingival fibroblasts, were used as the material. The [Ca2+] i was measured with fura-2/AM, a Ca2+-sensitive fluorescent dye. At first, we confirmed the existence of CaSRs in these cells by showing that [Ca2+] i was elevated by high concentrations of extracellular Ca2+ and by prototypic agonists of the CaSR such as gentamicin. The action of gentamicin was antagonized by inhibitors of phospholipase C (PLC), inositol trisphosphate (IP3) 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 Ca2+ responses to a known agonist of the CaSR. We then investigated whether nifedipine (an L-type Ca2+ channel blocker) stimulates CaSRs to elevate [Ca2+] i via a similar mechanism. Nifedipine Ca2+ responses were dose-dependently blocked by NPS2390 and by the same inhibitors of PLC, IP3 receptors, and NSCCs that disrupted the action of gentamicin. Calphostin C (a PKC inhibitor) and TMB-8 (an inhibitor of Ca2+ release from stores) also inhibited the nifedipine-induced [Ca2+] i elevation. These findings suggest that CaSRs are involved in the nifedipine-induced [Ca2+] i elevation in gingival fibroblasts.