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. 2020 Oct 15:11:01320.
doi: 10.3389/fphar.2020.01320. eCollection 2020.

Regulation of GABAA and 5-HT Receptors Involved in Anxiolytic Mechanisms of Jujube Seed: A System Biology Study Assisted by UPLC-Q-TOF/MS and RT-qPCR Method

Liang Chen  1 , Xue Zhang  1 , Chun Hu  2 , Yi Zhang  1 , Lu Zhang  1 , Juntao Kan  1 , Bo Li  1 , Jun Du  1
Affiliations

Regulation of GABAA and 5-HT Receptors Involved in Anxiolytic Mechanisms of Jujube Seed: A System Biology Study Assisted by UPLC-Q-TOF/MS and RT-qPCR Method

Liang Chen et al. Front Pharmacol. .

Abstract

The increase of the prevalence of anxiety greatly impacts the quality of life in China and globally. As the most popular traditional Chinese medicinal ingredient for nourishing health and tranquilizing mind, Jujube seed (Ziziphus jujuba Mill., Rhamnaceae) (SZJ) has been proved to exert anxiolytic effects in previous reports. In this study, a system biology method assisted by UPLC-Q-TOF/MS and RT-qPCR was developed to systematically demonstrate the anxiolytic mechanisms of SZJ. A total of 35 phytochemicals were identified from SZJ extract (Ziziphus jujuba Mill. var. spinosa [Bunge] Hu ex H.F. Chow), which interact with 71 anxiolytic targets. Protein-protein interaction, genes cluster, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis were subsequently conducted, and results demonstrated that regulation of serotonergic and GABAergic synapse pathways were dominantly involved in the anxiolytic mechanisms of SZJ extract. The effects of SZJ extract on mRNA expressions of multiple GABAA (gamma-aminobutyric acid type A) and 5-HT (serotonin) receptors subtypes were further validated in human neuroblastoma SH-SY5Y cells using RT-qPCR. Results showed that SZJ extract (250 μg/mL) significantly up-regulated the mRNA level of GABRA1 and GABRA3 as well as HTR1A, HTR2A, and HTR2B in non-H2O2 treated SH-SY5Y cells. However, it exerted an inhibitive effect on the overexpressed mRNA of GABRA1, GABRA2, HTR1A, and HTR2A in H2O2 treated SH-SY5Y cells. Taken together, our findings suggest that anxiolytic mechanisms of SZJ mostly involve the regulation of GABAergic and serotonergic synapse pathways, especially a two-way modulation of GABRA1, HTR1A, and HTR2A. Our current results provide potential direction for future investigation of SZJ as an anxiolytic agent.

Keywords: 5-HT receptors; GABAA receptors; anxiety; anxiolytic mechanism; jujube seed; system biology.

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Figures

Figure 1
Figure 1
Chromatographic profile of SZJ extract using UPLC-Q-TOF/MS in negative ion mode.
Figure 2
Figure 2
Protein-protein interaction (PPI) network of anxiolytic effects-related targets of SZJ. Cytoscape (Version 3.6.1) was applied to construct the interactions downloaded from the STRING (interaction score set as high confidence >0.7). All the targets are represented by nodes, whereas the interactions between the targets are represented by edges. MCODE plug-in was applied to conduct cluster analysis. Different clusters are noted with different colors. The node size is proportional to its located cluster MCODE score.
Figure 3
Figure 3
Top 10 significantly enriched GO terms in biologic process (red), cellular components (green), and molecular function (blue) categories. The bubble diagram was made using JMP software 14.2.0 (SAS Institute Inc., USA). The bubble size is proportional to its involved targets percentage in the term.
Figure 4
Figure 4
The interaction networks of enriched biological processes. ClueGO was applied to analysis procedure, and multiple color circles indicate that they are involved in multiple biological processes.
Figure 5
Figure 5
Targets-pathway network associated with anxiolytic effects of SZJ extract. A cytoscape ClueGo plug-in was applied to enrich the pathways and construct the network.
Figure 6
Figure 6
Effects of SZJ extract on mRNA gene expressions of different subtypes of GABAA and 5-HT receptors. GraphPad Prism 7.0 software was applied for statistical analysis and graphing. All data were expressed as mean ± SD, and a two-way ANOVA followed by Tukey’s test was applied. Two-way ANOVA analysis results presented that, GABRA1, F (2, 12) =57.27: p < 0.0001; GABRA2, F (2, 12) =112.50: p < 0.0001; GABRA3, F (2, 12) =40.10: p < 0.0001; HTR1A, F (2, 12) =677.80: p < 0.0001; HTR2A, F (2, 12) =13.07: p = 0.0010; HTR1B, F (2, 12) =42.99: p < 0.0001; HTR2B, F (2, 12) =151.50: p < 0.0001. Post hoc Tukey’s test results were represented by comparing with non-H2O2 treated control group, *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001; compared with H2O2 treated control group, ###p < 0.001 and ####p < 0.0001.
Figure 7
Figure 7
Abstracted phytochemicals-targets-pathway sub-networks of serotoninergic synapse (A) and GABAergic pathways (B). Cytoscape (Version 3.6.1) was applied to construct the sub-networks.
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References

    1. Abdoul-Azize S. (2016). Potential Benefits of Jujube (Zizyphus Lotus L.) Bioactive Compounds for Nutrition and Health. J. Nutr. Metab. 2016, 2867470–2867470. 10.1155/2016/2867470 - DOI - PMC - PubMed
    1. Akirav I. (2011). The role of cannabinoids in modulating emotional and non-emotional memory processes in the hippocampus. Front. Behav. Neurosci. 5, 34–34. 10.3389/fnbeh.2011.00034 - DOI - PMC - PubMed
    1. Benke D., Barberis A., Kopp S., Altmann K.-H., Schubiger M., Vogt K. E., et al. (2009). GABAA receptors as in vivo substrate for the anxiolytic action of valerenic acid, a major constituent of valerian root extracts. Neuropharmacology 56, 174–181. 10.1016/j.neuropharm.2008.06.013 - DOI - PubMed
    1. Bindea G., Mlecnik B., Hackl H., Charoentong P., Tosolini M., Kirilovsky A., et al. (2009). ClueGO: a Cytoscape plug-in to decipher functionally grouped gene ontology and pathway annotation networks. Bioinf. (Oxford Engl.) 25, 1091–1093. 10.1093/bioinformatics/btp101 - DOI - PMC - PubMed
    1. Bouayed J., Rammal H., Soulimani R. (2009). Oxidative stress and anxiety: relationship and cellular pathways. Oxid. Med. Cell Longev. 2, 63–67. 10.4161/oxim.2.2.7944 - DOI - PMC - PubMed