Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing 211198, China.
J Pharm Biomed Anal. 2011 Apr 28;55(1):101-8. doi: 10.1016/j.jpba.2010.12.015. Epub 2010 Dec 21.
Sheng-Mai-San (SMS), a traditional Chinese multiherbal formula, is widely used in clinic for the treatment of myocardial ischemia (MI) in China. Recently, it has been shown that the change of neurotransmitters in central nervous system is closely related to the pathogenesis of MI, whether SMS might affect the neurotransmitters at central nervous system in MI patients has not been studied yet. In this study, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of two types of neurotransmitters (neuropeptides and monoaminergic neurotransmitters) in rat brain tissue was developed. The instrument was operated under the multiple reaction monitoring (MRM) mode using electrospray ionization (ESI) in the positive ion mode. A good linear relationship with coefficients ≥0.99 was achieved over the concentration ranges of 10-1000ngmL(-1) for 5-hydroxytryptamine (5-HT) and norepinephrine (NE); 2-250ngmL(-1) for methionine-enkephalin (M-ENK) and leucine-enkephalin (L-ENK). Quantification limit for 5-HT and NE was 4.0ngmL(-1); and 2.0ngmL(-1) for M-ENK and L-ENK. The intra- and inter-day precision was less than 15% and accuracy was within ±15%. The analysis revealed significant reductions at the levels of 5-HT (p<0.01), NE (p<0.01), M-ENK (p<0.05) and L-ENK (p<0.01) in the MI group compared to the control group. These findings demonstrate that myocardial ischemia reduces the concentrations of 5-HT, NE, M-ENK and L-ENK in rat brains, while SMS shows protective effects on MI associated with reversing these four neurotransmitters to sham levels.
生脉散（SMS）是一种传统的中药复方，在中国临床上广泛用于治疗心肌缺血（MI）。最近的研究表明，中枢神经系统中神经递质的变化与 MI 的发病机制密切相关，但是否 SMS 可能会影响 MI 患者中枢神经系统中的神经递质尚未进行研究。在这项研究中，开发了一种用于同时测定大鼠脑组织中两种类型神经递质（神经肽和单胺类神经递质）的液相色谱-串联质谱（LC-MS/MS）方法。仪器在正离子模式下采用电喷雾电离（ESI），以多反应监测（MRM）模式操作。5-羟色胺（5-HT）和去甲肾上腺素（NE）的浓度范围为 10-1000ngmL（-1）时，5-HT 和 NE 的线性关系良好，相关系数≥0.99；甲硫氨酸脑啡肽（M-ENK）和亮氨酸脑啡肽（L-ENK）的浓度范围为 2-250ngmL（-1）时，M-ENK 和 L-ENK 的线性关系良好，相关系数≥0.99。5-HT 和 NE 的定量限为 4.0ngmL（-1）；M-ENK 和 L-ENK 的定量限为 2.0ngmL（-1）。日内和日间精密度均小于 15％，准确度在±15％范围内。分析表明，与对照组相比，MI 组 5-HT（p<0.01），NE（p<0.01），M-ENK（p<0.05）和 L-ENK（p<0.01）的水平显着降低。这些发现表明，心肌缺血会降低大鼠大脑中 5-HT，NE，M-ENK 和 L-ENK 的浓度，而 SMS 对与 MI 相关的神经递质具有保护作用，可将这四种神经递质恢复至 sham 水平。
