Anti-alzheimer's Disease Effect Of Essential Oil From Aerial Part Of Salvia Miltiorrhiza Bunge.

Objective To optimize the extraction process of the essential oil from the aerial parts of Salvia miltiorrhiza Bunge..The composition was analyzed and the total terpenoids content was measured.The antioxidant capacity of the essential oil was tested through antioxidant tests in vitro,then the anti-AD effect of essential oil from aerial part of Salvia miltiorrhiza Bunge.was identified through pharmacological experiments in vivo of the AD mice model which was induced by D-gal plus AlCl₃.This research was aimed to provide evidences for finding new potential anti-AD drugs from traditional Chinese medicine and to further expand the comprehensive utilization of Salvia miltiorrhiza Bunge.resources.Methods To optimize the extraction process of steam distillation to obtain the essential oil from aerial parts of Salvia miltiorrhiza Bunge.through single factor experimental and orthogonal experimental design.The chemical compositions were analyzed through GC/GC-MS,and the method of measuring the total terpenoids of essential oil from aerial parts of Salvia miltiorrhiza Bunge.was established by using the colorimetry of 5% vanillin-glacial acetic acid solution combined with perchloric acid and?-Caryophyllene as reference substance.Then the content of essential oil was measured.The antioxidant capacity of the essential oil was tested by using DPPH free radical scavenging and trichloroacetic acid reduction method.The AD mice model was induced by D-gal plus AlCl₃,and then the mice in the experimental group were treated with essential oil from aerial parts of Salvia miltiorrhiza Bunge.The protective effects of essential oil on the memory impairment of mice were determined by Morris water maze test.The contents of amyliod-??A??and glial fibrillary acidic protein?GFAP?were measured by immunohistochemistry?IHC?and western blot?WB?,and the contents or activity of SOD,GSH,ROS,MDA,Ach and CAT in mice brain homogenate were analyzed through Biochemical Determinations?SOD,GSH,ROS,MDA,Ach and CAT kits were used?.Finally,the Realtime-PCR was used to measure the expression changes of inflammatory factors and A?-related protein in the mice brain.Results Orthogonal experiment was used to optimize the extraction process of essential oil from aerial parts of Salvia miltiorrhiza Bunge.with steam distillation method.The optimum extraction conditions were as follows:immersing for 6h,distillation for 8h,the ratio of material to liquid was 1:18g·ml^-1,and the extraction rate was about 0.0935%.The chemical compositions of the essential oil were analyzed through GC/GC-MS.The area normalized method was used to calculate the total terpenoids which accounted for about 53.88% of the total volatile of essential oil.The terpene components with higher contentinclude ?-caryophyllene,?-caryophylleneoxide,dihydro-neoproene,germacrene-D and ?-caryophyllene.UV spectrophotometry was applied to measure total terpenoids content which was up to 72.38%of essential oil from aerial parts of Salvia miltiorrhiza Bunge.,and met the National 5th new drug standards.The results in vitro antioxidant test show that compared with a considerable concentration of VC,essential oil from aerial parts of Salvia miltiorrhiza Bunge.has a stronger antioxidant capacity.The anti-AD effect of essential oil from aerial parts of Salvia miltiorrhiza Bunge.were tested based on individual behavioral level,molecular biology level,protein level and gene level.The results show that the essential could significantly improve the learning ability and memory ability of mice,significantly inhibit the expression of A?,BACE-1 and GFAP which associated with AD in hippocampus and cortex regions of mice,and promote NEP expression in hippocampus and cortex.The content of Ach,GSH and the activity of antioxidant enzymes SOD,CAT were obviously increased,and the gene expression levels of inflammatory cytokines that promote the brain inflammation,oxidative damage and A?-related enzyme in mice brain have also been reduced.Conclusion The pharmacological experiments in vitro and in vivo prove that essential oil from aerial parts of Salvia miltiorrhiza Bunge.has strong anti-inflammatory and anti-oxidation ability and could alleviate the symptoms of AD mouse model which induced by D-gal and AlCl₃.This study provides the experiment basis,ideas and methods for searching the natural anti-AD drugs,and has great significance for comprehensive development and utilization of Salvia miltiorrhiza Bunge.resources.