通过生物测定法和鸟苷酸环化酶刺激同时测量内皮衍生舒张因子。
Simultaneous measurement of endothelium-derived relaxing factor by bioassay and guanylate cyclase stimulation.
作者信息
Kondo K, Mitchell J A, de Nucci G, Vane J R
机构信息
William Harvey Research Institute, St Bartholomew's Hospital Medical College, London.
出版信息
Br J Pharmacol. 1989 Oct;98(2):630-6. doi: 10.1111/j.1476-5381.1989.tb12637.x.
Abstract
- Endothelium-derived relaxing factor (EDRF) released by cultured endothelial cells (EC) from bovine aortae was measured by bioassay using pre-contracted strips of rabbit aorta and by radioimmunoassay of guanosine 3':5'-cyclic monophosphate (cyclic GMP) produced by stimulation of bovine lung soluble guanylate cyclase. 2. Bradykinin (Bk, 3 and 30 pmol) injected through a column of EC caused release of EDRF as detected by bioassay and increased cyclic GMP concentrations. Superoxide dismutase (SOD, 15 u ml-1) increased the amount of EDRF detected by the activation of soluble guanylate cyclase. 3. In the absence of endothelial cells, nitric oxide (NO, 1-2 microM), arachidonic acid (AA, 3-30 microM) or sodium nitroprusside (SNP, 1-100 microM) stimulated guanylate cyclase. Superoxide dismutase strongly increased the stimulation of guanylate cyclase induced by NO, but had little effect on the stimulation induced by SNP and no effect on the stimulation induced by AA. 4. Oxyhaemoglobin (10-300 microM) abolished the stimulation of guanylate cyclase by EDRF, NO or SNP but was much less effective as an inhibitor of AA-induced stimulation of guanylate cyclase. 5. These results demonstrate that measurement of guanylate cyclase stimulation by radioimmunoassay is a viable method for detecting EDRF release, especially useful when the drugs used interfere with bioassay tissues.
摘要
- 通过使用预先收缩的兔主动脉条进行生物测定以及对牛肺可溶性鸟苷酸环化酶受刺激产生的3':5'-环磷酸鸟苷（环鸟苷酸）进行放射免疫测定，来检测从牛主动脉培养的内皮细胞（EC）释放的内皮衍生舒张因子（EDRF）。2. 通过EC柱注射缓激肽（Bk，3和30 pmol），经生物测定检测到EDRF释放，并使环鸟苷酸浓度增加。超氧化物歧化酶（SOD，15 u/ml-1）通过激活可溶性鸟苷酸环化酶增加了检测到的EDRF量。3. 在没有内皮细胞的情况下，一氧化氮（NO，1-2 microM）、花生四烯酸（AA，3-30 microM）或硝普钠（SNP，1-100 microM）刺激鸟苷酸环化酶。超氧化物歧化酶强烈增强了由NO诱导的鸟苷酸环化酶刺激，但对SNP诱导的刺激影响很小，对AA诱导的刺激没有影响。4. 氧合血红蛋白（10-300 microM）消除了EDRF、NO或SNP对鸟苷酸环化酶的刺激，但作为AA诱导的鸟苷酸环化酶刺激的抑制剂效果要差得多。5. 这些结果表明，通过放射免疫测定法测量鸟苷酸环化酶刺激是检测EDRF释放的可行方法，当所用药物干扰生物测定组织时特别有用。
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