Available online 23 November 2022

Aconitine could induce neurotoxicity through excitatory toxicity caused by extracellular EAAs levels increasing.

The cross-talk among excitatory neurotoxicity, intracellular Ca2+ overload and oxidative stress was proved a mechanism of aconitine neurotoxicity.

Benzoylaconine and aconine were identified as two metabolites after aconitine stimulating HT22 cells.

The change of cell metabolites was real-time online investigated based on microfluidic chip-mass spectrometry.

Aconitine is a common and main toxic component of Aconitum, having toxicity of central nervous system. However, the mechanism of aconitine neurotoxicity is not yet clear. In this work, we made the perspective that excitatory amino acids could trigger excitotoxicity as a pointcut to explore the mechanism of neurotoxicity induced by aconitine. HT22 cells was simulated by aconitine and the changes of target cell metabolites were real-time online investigated based on a microfluidic chip-mass spectrometry system. Meanwhile, for confirming the metabolic mechanism of aconitine toxicity on HT22 cells, the levels of lactate dehydrogenase, intracellular Ca2+, reactive oxygen species, glutathione, superoxide dismutase and ratio of Bax/Bcl-2 protein were detected by molecular biotechnology. Integration of the detected results revealed that neurotoxicity induced by aconitine was associated with the process of excitotoxicity caused by glutamic acid and aspartic acid, which followed with accumulation of lactic acid and reduction of glucose. The surge of extracellular glutamic acid could further lead to a series of cascade reactions including intracellular Ca2+ overload and oxidative stress, and eventually result in cell apoptosis. In general, we illustrated a new mechanism of aconitine neurotoxicity and presented a novel analysis strategy that real-time online monitoring of cell metabolites could provide a new approach to mechanism analysis.

aconitine

neurotoxicity mechanism

HT22 cells

excitatory amino acids

microfluidic chip-mass spectrometry

© 2022 The Author(s). Published by Elsevier B.V. on behalf of Xi’an Jiaotong University.