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Abstract
Phenotype screening has become an important tool for the discovery of active components in traditional Chinese medicine. Anshen Buxin Six Pills (ASBX) are a traditional Mongolian medicine used for the treatment of neurosis in clinical settings. However, its antidepressant components have not been explicitly identified and studied. Here, the antidepressant effect of ASBX was evaluated in adult zebrafish. High performance liquid chromatography-mass spectrometry (HPLC-Q-TOF-MS/MS) was combined with zebrafish behavior trajectory analysis to screen and identify the antidepressant-active extract fraction and active components of ASBX. Finally, the antidepressant effect of the active ingredients were verified by the behavior, pathology, biochemical indices and protein level of adult fish. The novel tank driving test (NTDT) showed that ASBX can effectively improve the depressive effect of reserpine on zebrafish. Petroleum ether and dichloromethane extracts of ASBX were screened as antidepressant active extracts. Costunolide (COS) and dehydrocostus lactone (DHE) were screened as the active components of ASBX. COS had been shown to significantly improve the depressive behavior, nerve injury and neurotransmitter levels (5-hydroxytryptamine (5-HT) and norepinephrine (NE)) of zebrafish by inhibiting the high expression of serotonin transporter and norepinephrine transporter induced by reserpine suggesting the antidepressant effect of COS may be related to its effect on 5-HT and NE pathways. This study provided a phenotype based screening method for antidepressant components of traditional Chinese medicines, so as to realize the separation, identification and activity screening of components at the same time.
Highlights
Traditional Chinese medicines (TCM) have been shown to have an important role in the treatment of central nervous system (CNS) diseases (Li et al, 2016)
Gradient extraction was carried out, each solvent was 300 ml each time, extracted for 3 times, and the solvent was removed by rotary distillation, petroleum ether extract (PEE), dichloromethane extract (DME), ethyl acetate extract (EAE), water-saturated nbutanol extract (BUE), and aqueous extracts (AQE) were obtained
According to the mass spectrum information, we identified that the components contained in fraction 26 are costunolide and costic acid, and the components contained in fraction 28 are dehydrocostus lactone (See for mass spectrum information in Supplementary Table S1)
Summary
Traditional Chinese medicines (TCM) have been shown to have an important role in the treatment of central nervous system (CNS) diseases (Li et al, 2016). High-performance liquid chromatography-mass spectrometry (HPLC-Q-TOF-MS/MS) technology combined with target-based (Hou et al, 2020) and cell-based activity detection methods (Zhang et al, 2021), has been widely used in screening for lead compounds. It is not suitable for screening active ingredients where the target is unknown. Due to the complexity of the mechanism of central nervous system diseses (Krahn et al, 2020), such as depression, the screening methods based on cells and targets can not show the complex physiological activities of the nervous system. A rapid screening method of neuroactive drugs needs to be established
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