期刊
出版社
ELSEVIER
DOI: 10.1016/j.jchromb.2021.122856
关键词
Neurotransmitters; LC-MS; Hypertension; Uncaria; Spontaneously hypertensive rats
资金
- National Natural Science Foundation of China [81673591]
- National Key R&D Program of China [2018YFC1707900, 2018YFC1707001]
This study established a sensitive, reproducible, and rapid liquid chromatography method with tandem mass spectrometry detection for simultaneous quantitative analysis of 16 neurotransmitters and their metabolites in rats. The intervention with Uncaria, a traditional Chinese medicine, showed significant effects on regulating the levels of neurotransmitters in hypertensive rats, indicating its potential in treating symptoms related to the nervous system. The results suggest a potential relationship between changes in plasma neurotransmitters and hypertension pathogenesis, providing valuable insights into the mechanisms of Uncaria in lowering blood pressure.
This study aimed to establish a sensitive, reproducible, and rapid liquid chromatography method with tandem mass spectrometry detection to perform simultaneous quantitative analysis of 16 neurotransmitters and their metabolites in rat plasma, including levodopa, dopamine, norepinephrine, epinephrine, L-tryptophan, kynurenic acid, serotonin, melatonin, choline, acetylcholine, histamine, phenylethylamine, as well as excitatory (L-glutamic acid and L-aspartic acid) and inhibitory (gamma-aminobutyric acid and L-glycine) neurotransmitters. These analytes were measured by ultra-high performance chromatography coupled with triple quadrupole mass spectrometry using a hydrophilic interaction chromatographic column (ethylene-bridged hybrid amide column). The internal standards of stable isotope labeling were used to improve the reliability of the results. Our method provided high linearity for all neurotransmitters (for all coefficients measured > 0.99), with inter- and intra-day accuracy from -14.82% to 17.49% and precision was between 0.89% and 17.70%. The method was subsequently verified in an animal study, where the intervention of five different Uncarias, the traditional Chinese medicine with hypotensive effects, was applied to the spontaneously hypertensive rats (SHRs). SHRs showed dysregulated plasma kynurenic acid, acetylcholine, and norepinephrine levels, and these neuroactive analytes were significantly restored by Uncaria treatment compared with the model group (SHR group). Compared with captopril, included as a positive control for its hypotensive effect, Uncaria had more effects on perturbing the levels of plasma neurotransmitters, which might indicate Uncaria's potential in treating symptoms related to the nervous system. These results suggested that the changes in the neurotransmitters and their metabolites in plasma may be related to the pathogenesis of hypertension. It also provided valuable information about the action mechanisms of Uncaria on its hypotensive effects.
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