Clinical Chemistry of Newly-found Endogenous Bufalin Isomer in Saliva

内源性洋地黄样因子（EDLF）是水盐代谢的重要调节因子。近年来研究发现内源性洋地黄样因子主要由强心内酯和蟾蜍二烯内酯组成。蟾蜍灵是蟾蜍二烯内酯的一种，我们最近发现这种EDLF存在于哺乳动物体液中，尤其是唾液中。液相色谱-质谱（LC/MS）的串联质谱信号表明，蟾毒灵与蟾蜍中的蟾毒灵是一致的。这项研究是在国际动脉粥样硬化会议上公开的。在上述研究中，我们在人血浆中发现了一种新的蟾蜍二烯内酯，即海蟾毒精（MBG）的双氢加成形式，即端脂蟾毒精（TCB）。这是哺乳动物体液中TCB的首次报道。慢性肾功能衰竭患者血浆中TCB与MBG共存，且含量高于MBG。本研究提示蟾蜍类药物可能存在代谢和/或合成途径;在心血管领域，我们发现纳摩尔水平的哇巴因可使大鼠主动脉内皮细胞内钙离子增加，产生一氧化氮;洋地黄类药物的中毒也是临床领域的重要问题。113我们编辑了《药物毒理学分析与临床实验室中毒》系列，这些研究策略成为最近在小鼠肾上腺皮质细胞系Y-1中发现海蟾蜍毒素（MBT），以及连续少量给予MBT对大鼠产生高血压作用的重要基石。本研究首次证实MBT在哺乳动物体内的存在及其降压作用。MBT成为EDLF的新成员。

Endogenous digitalis-like factor (EDLF) act as important modulator for water and salt metabolism. From recent studies endogenous digitalis-like factor is consisted of cardenolides and bufadienolides. Bufalin is a member of bufadienolide, and we recently found that this EDLF was in mammalian body fluid, especially in saliva. The tandem mass spectrometric signals of liquid chromatography-mass spectrometry (LC/MS) indicated that saliva bufalin is the same as authentic bufalin from toad. This study was opened in the international meeting of atherosclerosis. But due to the slight recovery of saliva, structural study was incomplete such as NMR.During the above study, we found new bufadienolide, two-hydrogen added form of marinobufagin (MBG) that is telocinobufagin (TCB) in human plasma. This is the first report TCB in mammalian body fluid. Moreover, TCB in the plasma of chronic renal failure was co-existed with MBG and larger amount than MBG. This study indicated that metabolic and/or synthetic pathway may exist in mammalian bufadienolide.In the cardenolide field, we found that Nanomolar level of ouabain increases intracellular calcium to produce nitric oxide in rat aortic endothelial cells.The poisoning is also important problem in the clinical field of digitalis. We edited the series of toxicological analysis of drug and poisoning in the clinical laboratoryThese strategy of studies become the important foundation stone of recent impressive finding of marinobufotoxin (MBT) in mouse adrenocortical cell line, Y-1, and hypertensive effect on the rat by continuously and small amount giving of MBT. This is the first study, which proves the existence in mammal and hypertensive effect of MBT. MBT becomes is new member of EDLF.