OBJECTIVE: The research investigated the mechanisms of CZG\'s improvement in insomnia by regulating cAMP/CREB signaling pathway and metabolic profiles.
METHODS: The main components of CZG were characterized by liquid chromatography-mass spectrometry (LC-MS). Subsequently, these validated components were applied to network pharmacological analysis to predict signaling pathways associated with insomnia. We evaluated the effect of CZG on BV-2 cells in vitro. We also evaluated the behavioral indexes of CUMS combined with PCPA induced insomnia in rats. HE staining and Nissl staining were used to observe the pathological damage of hippocampus. ELISA was used to detect the levels of various neurotransmitters, orexins, HPA axis, and inflammatory factors in insomnia rats. Then we detected the expression of cAMP/CREB signaling pathway through ELISA, WB, and IHC. Finally, the metabolomics was further analyzed by using UHPLC-QTOF-MS/MS to investigate the changes in the hippocampus of insomnia rats and the possible metabolic pathways were also speculated.
RESULTS: The results of CZG in vitro experiments showed that CZG has protective and anti-inflammatory effects on LPS induced BV-2 cells. A total of 161 chemical components were identified in CZG. After conducting network pharmacology analysis through these confirmed components, we select the cAMP/CREB signaling pathway for further investigate. The behavioral research results on insomnia rats showed that CZG significantly prolonged sleep time, mitigated brain tissue pathological damage, and exhibited liver protective properties. CZG treats insomnia by regulating the content of various neurotransmitters, reducing levels of orexin, HPA axis, and inflammatory factors. It can also treat insomnia by upregulating the expression of the cAMP/CREB signaling pathway. Hippocampus metabolomics analysis identified 69 differential metabolites associated with insomnia. The metabolic pathways related to these differential metabolites have also been predicted.
CONCLUSIONS: These results indicate that CZG can significantly prolong sleep time. CZG is used to treat insomnia by regulating various neurotransmitters, HPA axis, inflammatory factors, cAMP/CREB signaling pathways, and metabolic disorders.
目的:本研究探讨CZG通过调节cAMP/CREB信号通路和代谢谱改善失眠的机制。
方法:通过液相色谱-质谱(LC-MS)表征CZG的主要成分。随后,这些经过验证的成分被应用于网络药理学分析,以预测与失眠相关的信号通路.我们在体外评价了CZG对BV-2细胞的影响。我们还评估了CUMS联合PCPA诱导的大鼠失眠的行为指标。采用HE染色和Nissl染色观察海马的病理损伤。用ELISA法检测各种神经递质的水平,orexins,HPA轴,失眠大鼠的炎症因子。然后通过ELISA检测cAMP/CREB信号通路的表达,WB,IHC。最后,通过使用UHPLC-QTOF-MS/MS进一步分析代谢组学,以研究失眠大鼠海马的变化,并推测可能的代谢途径。
结果:CZG的体外实验结果表明,CZG对LPS诱导的BV-2细胞具有保护和抗炎作用。在CZG中鉴定出总共161种化学成分。通过这些确认的成分进行网络药理学分析后,我们选择cAMP/CREB信号通路进行进一步研究。对失眠大鼠的行为学研究结果表明,CZG显著延长了睡眠时间,减轻脑组织病理损伤,并表现出肝脏保护特性。CZG通过调节各种神经递质的含量来治疗失眠,降低食欲素水平,HPA轴,和炎症因子。它还可以通过上调cAMP/CREB信号通路的表达来治疗失眠。海马代谢组学分析确定了69种与失眠相关的差异代谢物。还预测了与这些差异代谢物相关的代谢途径。
结论:这些结果表明CZG可以显着延长睡眠时间。CZG用于通过调节各种神经递质来治疗失眠,HPA轴,炎症因子,cAMP/CREB信号通路,和代谢紊乱。