Ando M, Tatematsu T, Kunii S, Nagata Y
Department of Physiology, Fujita Health University School of Medicine, Aichi-ken, Japan.
Brain Res. 1994 Jul 11;650(2):283-8. doi: 10.1016/0006-8993(94)91793-0.
Cyclic GMP (cGMP) production in rat superior cervical sympathetic ganglia (SCG) was markedly increased (ca. 7-9-fold) by the addition of either acetylcholine (ACh; 0.1 mM) or a muscarinic agonist, carbachol (Carb; 0.1 mM), in the presence of an inhibitor (3-isobutyl-1-methylxanthine) for cGMP hydrolytic enzyme during in vitro aerobic incubation at 37 degrees C for 5 min. The ACh-induced accumulation of cGMP in SCG was effectively blocked (-73%) by the further addition of atropine (10 microM), a muscarinic antagonist, whereas a nicotinic blocker, hexamethonium (10 microM) partially antagonized (-41%) this ACh stimulation. The inhibitory effect of hexamethonium on ACh-evoked ganglionic cGMP production was effectively augmented (-83%) by addition of NG-monomethyl-L-arginine (L-NMMA, 50 microM), a compound that inhibits nitric oxide (NO) synthesis from L-arginine. Comparable inhibition of cGMP formation was observed following application of L-NMMA to the SCG upon stimulation of Carb. In contrast, L-NMMA had no effect on the decreased level of ACh-evoked cGMP production caused by the muscarinic antagonist. The Carb-induced elevation of ganglionic cGMP synthesis was significantly reduced within 1 min of incubation in the medium containing hemoglobin (Hb; 20 microM), an agent that scavenges only the extracellular fraction of NO. Thereafter, the tissue cGMP formation attenuated to the control level by subsequent incubation for several minutes. Addition of protein kinase C (PKC) activator, 12-O-tetradecanoylphorbol 13-acetate (TPA; 1 microM) to the medium significantly decreased Carb-evoked cGMP synthesis (-61%) in SCG, whereas superoxide dismutase (SOD; 30 U/ml) only slightly suppressed the Carb stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)
在37℃有氧体外孵育5分钟的过程中,加入乙酰胆碱(ACh;0.1 mM)或毒蕈碱激动剂卡巴胆碱(Carb;0.1 mM),同时存在环鸟苷酸(cGMP)水解酶抑制剂(3 - 异丁基 - 1 - 甲基黄嘌呤),大鼠颈上神经节(SCG)中的cGMP生成显著增加(约7 - 9倍)。在SCG中,ACh诱导的cGMP积累被进一步加入的毒蕈碱拮抗剂阿托品(10 μM)有效阻断(-73%),而烟碱拮抗剂六甲铵(10 μM)则部分拮抗(-41%)这种ACh刺激。通过加入抑制L - 精氨酸合成一氧化氮(NO)的化合物NG - 单甲基 - L - 精氨酸(L - NMMA,50 μM),六甲铵对ACh诱发的神经节cGMP生成的抑制作用有效增强(-83%)。在刺激Carb后,将L - NMMA应用于SCG时,观察到对cGMP形成的类似抑制。相反,L - NMMA对毒蕈碱拮抗剂引起的ACh诱发的cGMP生成水平降低没有影响。在含有血红蛋白(Hb;20 μM)的培养基中孵育1分钟内,Carb诱导的神经节cGMP合成显著降低,血红蛋白仅清除细胞外部分的NO。此后,通过随后几分钟的孵育,组织cGMP形成减弱至对照水平。向培养基中加入蛋白激酶C(PKC)激活剂12 - O - 十四烷酰佛波醇13 - 乙酸酯(TPA;1 μM)可显著降低SCG中Carb诱发的cGMP合成(-61%),而超氧化物歧化酶(SOD;30 U/ml)仅轻微抑制Carb刺激。(摘要截断于250字)