Gukovskaya A, Pandol S
Department of Medicine, Veterans Affairs Medical Center, San Diego, California.
Am J Physiol. 1994 Mar;266(3 Pt 1):G350-6. doi: 10.1152/ajpgi.1994.266.3.G350.
Guanosine 3',5'-cyclic monophosphate (cGMP) rise is one of the early events in neurotransmitter or hormone-induced cascade of reactions in pancreatic acinar cells. The mechanism of agonist-stimulated guanylyl cyclase activation in these cells remains, however, unknown. In the present work, mechanisms of cGMP rise, as well as of Ca2+ influx, induced by carbachol were studied on acinar cells isolated from rat and guinea pig pancreas. In both types of acinar cells, blocking nitric oxide (NO) production by inhibitors of NO synthase, NG-monomethyl-L-arginine (L-NMMA) or NG-nitro-L-arginine, abolished carbachol-induced cGMP rise in a dose-dependent manner. The inhibition was reversed by addition of excess L-arginine. L-NMMA also caused inhibition of the basal cGMP level, suggesting a role for NO in cGMP homeostasis in resting cells. Carbachol was found to increase [3H]arginine conversion to [3H]citrulline. This conversion was inhibited by L-NMMA. By contrast, inhibition of carbon monoxide production by Zn-protoporphyrin did not affect carbachol-stimulated cellular cGMP levels. There was no increase in cellular cGMP levels in response to exogenous arachidonic acid (AA). Blocking of lipoxygenase oxidation of AA by nordihydroguaiaretic acid did not produce any changes in carbachol-induced cGMP rise. Indomethacin, a cyclooxygenase inhibitor, increased basal cGMP level through L-NMMA-sensitive mechanism. Blockade of NO production inhibited carbachol-induced increase in 45Ca2+ uptake in both guinea pig and rat acinar cells. The concentration-response curves for inhibition by L-NMMA of 45Ca2+ uptake and cGMP formation were superimposable. L-NMMA also suppressed stimulation of Mn2+ quenching by carbachol in fura 2-loaded acini.(ABSTRACT TRUNCATED AT 250 WORDS)
3',5'-环磷酸鸟苷(cGMP)升高是胰腺腺泡细胞中神经递质或激素诱导的级联反应的早期事件之一。然而,这些细胞中激动剂刺激的鸟苷酸环化酶激活机制尚不清楚。在本研究中,对从大鼠和豚鼠胰腺分离的腺泡细胞研究了卡巴胆碱诱导的cGMP升高以及Ca2+内流的机制。在这两种类型的腺泡细胞中,一氧化氮合酶抑制剂Nω-甲基-L-精氨酸(L-NMMA)或Nω-硝基-L-精氨酸阻断一氧化氮(NO)生成,以剂量依赖方式消除了卡巴胆碱诱导的cGMP升高。加入过量L-精氨酸可逆转这种抑制作用。L-NMMA还导致基础cGMP水平降低,提示NO在静息细胞的cGMP稳态中起作用。发现卡巴胆碱可增加[3H]精氨酸向[3H]瓜氨酸的转化。这种转化受到L-NMMA的抑制。相比之下,锌原卟啉抑制一氧化碳生成并不影响卡巴胆碱刺激的细胞cGMP水平。对外源性花生四烯酸(AA)无细胞cGMP水平升高反应。去甲二氢愈创木酸阻断AA的脂氧合酶氧化对卡巴胆碱诱导的cGMP升高无任何影响。环氧化酶抑制剂吲哚美辛通过L-NMMA敏感机制增加基础cGMP水平。阻断NO生成抑制了豚鼠和大鼠腺泡细胞中卡巴胆碱诱导的45Ca2+摄取增加。L-NMMA抑制45Ca2+摄取和cGMP形成的浓度-反应曲线可叠加。L-NMMA还抑制了卡巴胆碱对fura 2负载腺泡中Mn2+淬灭的刺激作用。(摘要截断于250字)