Ohkuma S, Katsura M, Higo A, Shirotani K, Hara A, Tarumi C, Ohgi T
Department of Pharmacology, Kawasaki Medical School, Matsushima, Kurashiki, Japan.
J Neurochem. 2001 Jan;76(2):341-50. doi: 10.1046/j.1471-4159.2001.00045.x.
The effect of peroxynitrite (OONO-) on voltage-dependent Ca2+ channels (VDCCs) was examined by measuring [45Ca2+] influx into mouse cerebral cortical neurones. OONO- time- and dose-dependently increased [45Ca2+] influx and this increase was abolished by manganese (III) tetrakis (4-benzoic acid) porphyrin, a scavenger for OONO-. Inhibition of cyclic GMP (cGMP) formation did not alter the OONO(-)-induced [45Ca2+] influx. OONO-, as well as 30 mm KCl, significantly increased fluorescence intensity of cell-associated bis-(1,3-dibutylbarbituric acid) trimethine oxonol (bis-oxonol). Tetrodotoxin and membrane stabilizers such as lidocaine dose-dependently suppressed OONO(-)-induced [45Ca2+] influx. Although each of 1 microM nifedipine and 1 microM omega-agatoxin VIA (omega-ATX) significantly inhibited the OONO(-)-induced [45Ca2+] influx and the concomitant presence of these agents completely abolished the influx, 1 microM omega-conotoxin GVIA (omega-CTX) showed no effect on the influx. On the other hand, OONO- itself reduced 30 mM KCl-induced [45Ca2+] influx to the level of [45Ca2+] influx induced by OONO- alone, and the magnitude of this reduction was as same as that of KCl-induced [45Ca2+] influx by omega-CTX. These results indicate that OONO- increases [45Ca2+] influx into the neurones through opening P/Q- and L-type VDCCs subsequent to depolarization, and inhibits the influx through N-type VDCCs.
通过测量[45Ca2+]流入小鼠大脑皮质神经元的情况,研究了过氧亚硝酸盐(OONO-)对电压依赖性Ca2+通道(VDCCs)的影响。OONO-呈时间和剂量依赖性地增加[45Ca2+]流入,且这种增加被OONO-清除剂四(4-苯甲酸)锰卟啉消除。抑制环鸟苷酸(cGMP)形成并未改变OONO-诱导的[45Ca2+]流入。OONO-以及30 mM KCl显著增加了细胞相关的双(1,3-二丁基巴比妥酸)三甲川草酚(双草酚)的荧光强度。河豚毒素和膜稳定剂如利多卡因呈剂量依赖性地抑制OONO-诱导的[45Ca2+]流入。尽管1 microM硝苯地平和1 microMω-阿加毒素VIA(ω-ATX)均显著抑制OONO-诱导的[45Ca2+]流入,且这些药物同时存在时完全消除了该流入,但1 microMω-芋螺毒素GVIA(ω-CTX)对该流入无影响。另一方面,OONO-本身将30 mM KCl诱导的[45Ca2+]流入降低至单独OONO-诱导的[45Ca2+]流入水平,且这种降低的幅度与ω-CTX对KCl诱导的[45Ca2+]流入的降低幅度相同。这些结果表明,OONO-通过在去极化后打开P/Q型和L型VDCCs增加[45Ca2+]流入神经元,并通过N型VDCCs抑制该流入。