Aromatherapy based on essential oil (EO) has been widely used for alleviating pain and intense where no compound EO reports its application on pharmacological effects. In order to explore the active pharmaceutical ingredients (API) and mechanism of a compound EO, a blend of Artemisia argyi, Boswellia carterii, Commiphora myrrha, Cinnamomum cassia, Zingiber oj-jicinale, and Ilex pubescens EO, in treating neck and shoulder pain (NSP). Network pharmacology hyphenated with mice model was employed to investigate. Gas chromatography-mass spectrometry (GC-MS) was applied for the identification of constituents in compound EO. Lastly, transdermal absorption of compound EO was studied before verifying analgesic and anti-inflammatory effects in mice. Totally, 75 compounds were tentatively identified through GC-MS, predicting 46 potential analgesic targets. Moreover, 11 core targets were obtained through network topology screening. Animal test resulted that the compound EO had significantly stronger anti-inflammatory and analgesic effects compared to single EO. Multiple API in compound EO affected on targets and exerted therapeutic effects on NSP through multiple pathways. Afterwards, eucalyptol, camphor, and borneol from compound EO exhibited a sustained-release effect, which provide scientific basis to illustrate the application of compound EO in clinical.
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