抗抑郁新化合物SIPI-C和SIPI-F对PC12细胞内游离钙离子浓度的影响

Effect of new antidepressants SIPI-C and SIPI-F on cytosolic free Ca^(2+) concentration in PC12 cells

目的研究抗抑郁新化合物SIPI-C和SIPI-F对PC12细胞内游离钙离子浓度([Ca2+]i)的影响,初步探讨其神经毒性的机制。方法 接种PC12细胞于经胶原Ⅰ包被的培养皿,加入钙离子荧光探针Fluo-3/AM染色后,用激光共聚焦显微镜分别记录①有钙外液中,10μmo.lL-1的SIPI-A,B,C和F对PC12细胞[Ca2+]i的影响;②有钙外液中,1,10和100μmol.L-1的SIPI-C和SIPI-F对[Ca2+]i的影响;③有钙外液中,硝苯地平10μmo.lL-1对10μmo.lL-1的SIPI-C或SIPI-F作用的影响;④无钙细胞外液中,10μmo.lL-1SIPI-C和SIPI-F对[Ca2+]i的影响。结果 有钙外液中,SIPI-A 10μmol.L-1给药后[Ca2+]i下降,10μmol.L-1的SIPI-B,SIPI-C和SIPI-F分别使[Ca2+]i增加27%(P<0.05),84%(P<0.05)和87%(P<0.01);SIPI-C和SIPI-F明显升高[Ca2+]i;同时给予SIPI-C 10μmol.L-1和硝苯地平10μmo.lL-1或SIPI-F 10μmo.l L-1和硝苯地平10μmo.lL-1,给药后荧光强度立即上升达到峰值,随后下降,[Ca2+]i分别增加24%和15%(P<0.05)。在无钙外液中,SIPI-C和SIPI-F 10μmo.lL-1分别使[Ca2+]i增加16%和18%(P<0.01)。结论 抗抑郁化合物SIPI-C和SIPI-F可以引起PC12细胞中[Ca2+]i显著增加,此影响可能与其神经毒性有关。

OBJECTIVE To investigate the effect of SIPI-C and SIPI-F on cytosolic free Ca2＋ concentration （[Ca2＋l]） in PC12 cells. METHODS PC12 cells were cultured in culture dish coated with collagen I .After incubation at 37℃ for 40 -50 min in 10 pmol-L-1 Fluo-3/AM enriched media, the E Ca2 ＋ I i changes were continuously measured by the confocal laser scanning microscope： ① effect of SIPI-A, B, C and F 10 iJmol·L-1 on [Ca2＋] in normal extracellular fluid; ② effect of SIPI-C and SIPI-F 1, 10 and 100 iJmoI.L-1 on [Ca2＋]1 in normal extracellular fluid; ③ effect of nifedipine 10 iJmol. L-1 on effects of SIPI-C or SIPI-F 10 pmol. L-1 in normal extracellular fluid; ④ effect of 10 pmol. L-1SIPI-C or SIPI-F on [ Ca2 ＋] in non-calcium extracellular fluid. RESULTS SIPI-A 10 pmol· L-1 decreased [ Ca2.＋]1 i while SIPI-B, SIPI-C and SIPI-F 10 pmol. L-1 resulted in the elevation of intracellular calcium by 27%, 84% and 87% in normal extracellular fluid. In SIPI-C or SIPI-F 10 pmol· L-1 combined with nifedipine 10 iJmol·L-1 group, ECa2＋ 3i was elevated by 24% and 15% after application of compounds. At the same concentration, SIPI-C and SIPI-F resulted in the elevation of [ Ca2.＋]1by 16% and 18% in non-calcium extracellular fluid. CONCLUSION SIPI-C and SIPI-F increase[-Ca2＋]1 which could be related to SIPI-C and SIPI-F induced neurotoxicity.