Comparison Of Extraction Process Of Acorus Tatarinowii-Alpinia Oxyphylla Miq Essential Oil And Study On The Effect Of Inhalation Administration On AD

The Zhenxin Xingshui Yizhi Fang?Xingshui Fang?from the"Qian Jin Yi Fang",composed of Polygala tatarinowii,Acorus tatarinowii,Fructus Alpinae Oxyphyllae three herbs,has the effect of treating convulsions,lethargy,forgetfulness.The prescription is rich in volatile components,its material basis and whether Acorus tatarinowii-Alpinia oxyphylla Miq essential oils has anti-AD effect,remains to be studied.In this study,GC-MS was used to analyze the composition of essential oils extracted by steam distillation and supercritical CO₂.The network pharmacology of drug-target,drug-target-disease and protein pathway networks were used to guide process selection and predict the activity of essential oils in the treatment of AD,then verified by animal experiments.The main contents of this paper are as follows:1.Extraction process researchThe essential oils obtained by two different extraction methods were analyzed by GC-MS.A total of 35 volatile components were identified by steam distillation,with ethers?54.70%?,alkenes?25.82%?,ketones?2.88%?and alcohols?1.68%?,and 49species were identified by supercritical CO₂ extraction,Ethers?37.97%?,alkenes?20.26%?,ketones?16.81%?,alkanes?7.40%?and alcohols?0.92%?were identified.Through the component-target-Alzheimer's disease network,it is predicted that the compounds with strong interaction between the two methods are alcohol,ene,ether compounds,and the relative percentages in steam distillation?64.12%?are higher than supercritical extraction?45.35%?.However,supercritical extraction of alkanes?7.40%?and ketones?16.81%?did not reveal strong interactions,demonstrated that steam distillation is superior to supercritical extraction.2.Verification of the activity of essential oils in the treatment of ADFor the purpose of this study,the animal model of Alzheimer's Disease?AD?produced by administered unilaterally in lateral ventricle by injecting A?_1-42 and APP/PS1 double transgenic AD mouse model were used.The essential oils were inhalation administrated.Morris water maze test and a platform test were used to determine their cognitive effects.Nissl's staining was used to observe pathological changes in brain tissue.Meanwhile,Senile plaques?SP?were observed by employing Thioflavin-S staining.The expression of glucose transporter 1?GLUT1?and insulin receptor substrate 1?IRS-1?were analyzed using immunohistochemistry techniques.The levels of neurotransmitters such as acetylcholine?ACH?,glutamate?GLU?and?-aminobutyric acid?GABA?in the hippocampus were quantified by Elisa.The memory function was significantly reduced in model group,and severe brain injury and neuronal apoptosis were also observed in comparison to normal group?P<0.05,P<0.01?.Compared with the model group,Zhenxin Xingshui Yizhi Fang and Acorus tatarinowii-Alpinia oxyphylla Miq essential oil extract improved previous learning and memory impairments?P<0.05,P<0.01?,and the levels of SP,IRS-1,GLU,GABA?except essential oil low dose group?were significantly reduced?P<0.01?,meanwhile the levels of ACH and GLUT1 were significantly improved?P<0.05,P<0.01?.These results indicate that Acorus tatarinowii-Alpinia oxyphylla Miq essential oil extract could ameliorate cognitive deficits and GLUT1 and IRS-1could be a possible therapeutic target for AD.It may be an interesting approach to the treatment of Alzheimer's disease.