| Anxiety disorder is a common neurological disease in clinical practice.Being in an anxious state for a long time can significantly reduce the quality of life,increase the risk of cancer,and even induce suicidal tendencies and even death in severe cases.Currently,antianxiety and depression drugs commonly used in clinical practice have varying degrees of toxic and side effects.Inhaling aromatic plant essential oil can alleviate anxiety and depression,promote mental balance,improve mental state,and have fewer adverse reactions.However,the instability,poor water solubility,and low bioavailability of volatile oils severely limit their application.Microemulsions have the advantages of thermodynamic stability,low viscosity,good drug dispersion,and easy absorption.They can improve the water solubility of plant essential oils,while improving their stability.At the same time,the combination of several aromatic plant essential oils with similar efficacy can enhance each other’s efficacy.Therefore,the study of using herbal compound aromatic plant essential oil to alleviate anxiety has certain significance.In this experiment,four traditional Chinese medicine aromatic plants,such as Acorus calamus,sandalwood,mint,and orange peel,were used as experimental materials to formulate their formulas,and then the extraction process of essential oil from the traditional Chinese medicine compound aromatic plants was optimized.The components were detected and analyzed by gas chromatography mass spectrometry.Then,the traditional Chinese medicine compound aromatic plant essential oil was prepared into a microemulsion,and its structural type identification,quality evaluation,and stability test were conducted.Finally,its anti-anxiety efficacy and mechanism were evaluated in cell and animal models,providing a new theoretical basis for the prevention and treatment of anxiety disorders using Chinese herbal compound aromatic preparations.The specific results of this study are as follows:(1)Using the extraction rate of volatile components from compound aromatic plants as an evaluation index,a single factor experiment was conducted to find that the material liquid ratio,ultrasonic time,extraction time,and sodium chloride concentration had significant effects on the extraction rate of volatile components from compound aromatic plants.The response surface optimization experiment design was carried out using DesignExpert software,and the optimal extraction process parameters were determined as follows:material liquid ratio 1:12,ultrasonic time 55 min,extraction time 5.5 h,and sodium chloride concentration 4%.Under these extraction process parameters,the extraction rate of compound aromatic plant essential oil is 1.88 ± 0.02%.The volatile components extracted according to the optimal process parameters were analyzed and identified by gas chromatography mass spectrometry,β-Asarone is the main component and accounts for55.75% of the total essential oil.(2)Using compound aromatic plant essential oil as the oil phase and the area of the microemulsion region in the pseudo ternary phase diagram as an indicator,suitable surfactants,cosurfactants,and Km values were compared and selected.Finally,the compound aromatic plant essential oil micro lotion with compound aromatic plant essential oil as oil phase,EL-35 as surfactant and glycerol as cosurfactant(Km=4)was prepared.It was judged to be oil in water micro lotion by staining method.The physicochemical properties of compound aromatic plant essential oil microemulsion were analyzed.The results showed that the particle size,potential,and PDI of the microemulsion were(18.84± 2.99)nm,(-1.111±0.248)mv,and 0.194±0.014,respectively.After centrifugation at different rotational speeds,the particle size,PDI,and potential of the microemulsion before and after centrifugation were basically unchanged,and the microemulsion was stable within 60 days of storage at room temperature.(3)Cytotoxicity experiments showed that the concentration of compound aromatic plant essential oil below 200 μg/ml had no significant impact on the proliferation of PC12 cells.In the 500 μM corticosterone injured PC12 cell model,the cell survival rate in the model group was significantly reduced,which was the final concentration of the corticosterone injured cell model.The compound aromatic plant essential oil had a certain protective effect on PC12 cells damaged by corticosterone.(4)Under the autonomous activity apparatus model and the elevated cross maze model,the compound aromatic plant essential oil could increase the total movement distance of rats in the autonomous behavior box,but there was no significant difference compared to the control group.Compound aromatic plant essential oil can increase the number of movements,residence time,and percentage of distance in the open arm of rats on an elevated cross maze model.By observing the pathological sections of rats in each group,it was concluded that the compound aromatic plant essential oil had no damage to the main organs and hippocampus of rats.Compared with the control group,compound aromatic plants can increase the content of GABA and 5-HT in serum and 5-HT in hippocampus;Reduce DA content in serum and GLU content in hippocampus(5)In a chronic unpredictable mild stimulation model,compound aromatic plant essential oils can reverse the slow weight gain,reduced sugar preference,reduced total distance of exercise in an autonomous activity box,and reduced anxiety and depression symptoms caused by CUMS modeling,indicating that they have good efficacy in alleviating anxiety and depression.Nessler’s staining results showed that the compound aromatic plant can improve the hippocampal tissue damage caused by CUMS modeling.The Elisa test results showed that the improvement of anxiety and depression induced by CUMS modeling by compound aromatic plant essential oil may be related to the increase of 5-HT and GABA in serum and hippocampus and the decrease of GLU levels in serum and hippocampus. |
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