Is ouabain an authentic endogenous mammalian substance derived from the adrenal?
Is ouabain an authentic endogenous mammalian substance derived from the adrenal?
复制标题
哇巴因是一种源自肾上腺的真正的内源性哺乳动物物质吗？
DOI：
10.1161/01.hyp.23.5.632
复制
发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Stocco,DM
中科院分区：
文献类型：
--
作者：
Doris,PA;Jenkins,LA;Stocco,DM
最近有报道称瓦巴因是一种内源性哺乳动物物质，由肾上腺皮质释放，存在于正常血浆中。我们试图证实并扩展这一观察结果。使用本实验室开发的瓦阿因放射免疫分析法，我们用高效液相色谱法（HPLC）分离健康志愿者的正常人血浆，以确定瓦阿因的免疫反应性，并将这种免疫反应性与真正的瓦阿因进行比较。在大多数受试者中，未观察到用真正的瓦巴因配制的免疫反应性。一些受试者有低水平的瓦阿因免疫反应物质，但这主要是由于与多种非瓦阿因物质的交叉反应。在对进入医疗重症监护病房的10名患者收集的血浆进行分析后，获得了类似的结果。用牛肾上腺皮质细胞培养的无血清培养基显示出一定的瓦巴因免疫反应性。为了确定这种物质是否可能是胆固醇侧链裂解的类固醇产物，我们对类固醇生成进行了化学阻断，这有效地抑制了这些细胞产生黄体酮，但对这种培养基中的瓦巴因免疫反应性没有一致的影响。在条件培养基中，用22- r - oh -胆固醇刺激牛肾上腺皮质细胞的类固醇生成并不会增加瓦巴因的免疫反应性。随后对牛肾上腺皮质细胞条件培养基中瓦阿因免疫反应性的高效液相色谱研究表明，在无血清培养基中，正宗的瓦阿因不占瓦阿因免疫反应性的大部分。对牛肾上腺皮质细胞在最低盐和葡萄糖培养基中培养的研究表明，免疫反应性的小峰值可能与真正的瓦巴因相对应。（摘要删节250字）
Ouabain has recently been reported to be an endogenous mammalian substance released by the adrenal cortex and present in normal plasma. We have attempted to confirm and extend this observation. Using a ouabain radioimmunoassay developed in this laboratory, we fractionated by high-performance liquid chromatography (HPLC) normal human plasma from healthy volunteers to determine the presence of ouabain immunoreactivity and compare this immunoreactivity with authentic ouabain. In most subjects no ouabain immunoreactivity that coeluted with authentic ouabain was observed. Some subjects had ouabain-immunoreactive material present at low levels, but it was largely attributable to cross-reactivity with diverse substances found not to be ouabain. Similar results were obtained after analysis of plasma collected from 10 patients entering a medical intensive care unit. Studies of serum-free medium conditioned by bovine adrenocortical cells showed some ouabain immunoreactivity. To determine whether this material might be a steroid product of cholesterol side-chain cleavage, we performed chemical blockade of steroidogenesis, which effectively suppressed progesterone production by these cells but had no consistent effect on ouabain immunoreactivity in this medium. Stimulation of steroidogenesis with 22-R-OH-cholesterol in bovine adrenocortical cells did not produce any increase in the ouabain immunoreactivity present in conditioned medium. Subsequent HPLC studies of ouabain immunoreactivity in bovine adrenocortical cell-conditioned medium indicated that authentic ouabain did not account for most of the ouabain immunoreactivity in serum-free medium. Studies with bovine adrenocortical cells incubated in a minimal salt and glucose medium indicated a small peak of immunoreactivity that may correspond to authentic ouabain.(ABSTRACT TRUNCATED AT 250 WORDS)