Integrated Screening of Effective Anti-Insomnia Fractions of Zhi-Zi-Hou-Po Decoction via Drosophila melanogaster and Network Pharmacology Analysis of the Underlying Pharmacodynamic Material and Mechanism

Abstract

Insomnia is an anabatic epidemiology, while the mechanism is extremely complicated; it remains one of the major scientific challenges in life sciences. Because of the advantage of having a similar genetic background and circadian rhythm as those of humans, the Drosophila melanogaster model organism is hugely popular in sleep-related drug screening studies. Seven-day-old virgin D. melanogaster was used to establish the sleep deprivation model by repeated light stimulation at night. Using PySolo activity monitoring system and Drosophila activity as indices, the effective fractions of Zhi-Zi-Hou-Po decoction (ZZHPD) for insomnia were screened; the content of monoamine neurotransmitters dopamine (DA), 5-hydroxyindole-3-acetic acid (5-HIAA), Homovanillic acid (HVA), and 5-hydroxytryptamine (5-HT) in the brain of D. melanogaster were determined by high-performance liquid chromatography-electro-chemical detection. The herb-compound-target-disease target network were further constructed through network pharmacology to identify the potential targets and pathways of ZZHPD in the intervention of insomnia. Finally, the molecular docking method was used for evaluating the binding characteristics of important compounds from ZZHPD with related targets. The results showed that a certain dose of ZZHPD and its petroleum ether, dichloromethane, ethyl acetate, and n -butanol fractions could improve sleep. The dichloromethane fraction from ZZHPD extracts showed the best anti-insomnia effect among all extracts. It can also reduce the content of DA and HVA in the brain of D. melanogaster and increase 5-HT and 5-HIAA levels. The network pharmacology showed that the main active ingredients in ZZHPD included magnolol, honokiol, hesperidin, and so forth. According to the screening conditions, there were 71 targets and the result of KEGG enrichment analysis revealed that 73 pathways were associated with insomnia, which were primarily involved in inflammatory response, central neurotransmitter regulation, and apoptosis to relieve insomnia. The molecular docking results clarified that naringenin and apigenin have an intimate relationship with GABA A receptor, histamine H1, orexin receptor type 2, and interleukin-6. The mechanism of relieving insomnia is the result of the interaction of multi-components, multi-targets, and multi-pathways, which provides a certain theoretical basis for the treatment of insomnia and related diseases as well as clinical research.

Discovery of the Potential Novel Pharmacodynamic Substances From Zhi-Zi-Hou-Po Decoction Based on the Concept of Co-Decoction Reaction and Analysis Strategy

“…( Ma et al, 2021 ). As a classic Chinese prescription, ZZHPD has been proven to have a good effect on depression-like symptoms with few side effects ( Sun et al, 2021 ). However, due to the complex components and multi-target nature of ZZHPD, the mechanism in treating insomnia is still unclear.…”

Integrated Network Pharmacology Analysis and Experimental Validation to Investigate the Mechanism of Zhi-Zi-Hou-Po Decoction in Depression

“…These issues pose a big challenge for the evaluation of the clinical efficacy of TCM and for understanding of the material basis and molecular mechanisms responsible for its therapeutic efficacy. Fortunately, network pharmacology has evolved from systems biology and multidirectional pharmacology, thus revolutionizing the practice of TCM by shifting the paradigm of "one drug, one target, one disease" towards the model of "multicomponent, multitarget, multidisease," therefore making it possible to uncover the complex relationship between active components and their targets Yu et al, 2018;Sun et al, 2021).…”

Improvement of neurological recovery in the insomnia rats by Warming Yang Strategy through targeting SIRT4 by inhibiting inflammation and apoptosis

“…Meanwhile the promotion of Bax, downregulation of Bcl-2, and increase of apoptosis rate were observed after insomnia. 10 Previous studies indicated that silencing of SIRT4 exacerbated the expression of pro-inflammatory cytokines such as IL-1β, IL-6, IL-8, cyclooxygenase-2 (COX2), matrix metalloproteinase-9 (MMP-9), and intercellular adhesion molecule-1 (ICAM-1). 15 The upregulation of these genes is related to inflammation and vascular remodeling.…”

“…Continuous neuroinflammatory reaction can induce neuronal apoptosis and cause many central injuries. 10 Some neurobiological studies have shown that there is a correlation between neuronal apoptosis and insomnia. 11 Some antiinsomnia drugs present neuroprotective and antiapoptotic effects on neurons.…”