Chemical Constituents From Lycoris And Their Bioactivities Of Anti-Alzheimer's Disease

Traditional medicinal plants contribute a lot to the elimination of human diseases in the developed countries and developing countries.The plant resource of genus Lycoris(Amaryllidaceae)is very rich in China.There are more than 20 species in the world,in which more than about 15 species(12 endemic species)occur in China,relatively concentrated distribution in the south Yangtze River area of China.The Lycoris species are ornamental and used as folk medicine for a long history in China.There are abundant of wild Lycoris species,which are used for the treatment of rheumatoid arthritis,furuncle,edema and snake bites in the ethnic areas of China.This study systematically focuses on the ethnobotanical and ethnoecological investigations of Lycoris species,plant distribution,metabolomics,phytochemistry and their pharmacological activities of anti-Alzheimer's disease(anti-AD),for the aims to 1)discover the natural and potential medicinal plants for treating AD,2)contribute to the scientific evaluation of the traditional medicinal knowledge of Lycoris species,and 3)provide the basis for further research and development of these plant resources.1.The ethnobotanical and ethnoecological investigations and studies of Lycoris species in the local peopled society were carried out in Zhejiang,Jiangsu,Anhui,Guangxi and Guizhou provinces.In total 54 voucher specimens,and 60 samples of 12 Lycoris species were collected and obtained the traditional utilization of their horticultural and medicinal values were obtained in the different ethnic areas.The local people had the unique cognition of the classification and application of Lycoris species,and we recorded the biological characteristics,distributions and habitats of the wild and cultivated Lycoris species.2.The chemical constituents of L.hayvardii bulbs had been investigated.Serial of chromatography methods by the guidance of UPLC-QTOF-MS were used to isolate,purify and identify their chemical constituents.Their chemical structures were elucidated by MS and IH,13C NMR.20 alkaloid compounds were speculated by UPLC-QTOF-MS,and 15 pure compounds were isolated,and 8 compounds were firstly isolated and identified from L.haywardii,respectively.It provided basis for furtherly studying the isolation and identification of the chemical constitutes from L.haywardii,in order to discover new compounds and their pharmacological activity.3.(l)Twelve species including L.aurea,L.caldwellii,L.chinensis,L.incarnata,L.longituba,L.radiata,L.sprengeri,L.squamigera,L.straminea,L.x houdyshelii,L.haywardii,L.hybrid were collected and identified from wild.The crude extracts of these 12 species were analyzed by UPLC-QTOF-MS and run in the appropriate method of UPLC.(2)It is the first time to use the metabonomics method to analyze the chemical constituents and their differences among Lycoris species by UPLC-ESI-MS.The ion fraction were identified by the co-injection of known standards or using accurate ion fragments by QTOF-ESI-MS and searching SciFinder and related literatures.Twelve compounds were identified(galanthamine,lycoramine,lycorannine N-oxide,lycorine,dihydrolycorine,hippeastrine,oduline,haemanthamine?crinamidine,vittatine,narciclasine and dihydro-latifaliumin C).(3)Metabolic profiles of Lycoris species using UPLC-QTOF-MS were used to investigate the chemical metabolics and the chemical fingerprints of different species.PC A,PLS and OPLS-DA models were performed to analyze the total of 12 species and resulted in two clusters,which showed the similar trend to the classical taxonomy.The same cluster showed the similar metabolic profiles among these species.OPLS-DA analysis was used to find 20 markers,in which 16 markers were confirmed to explore the interspecific differences among these species.(4)The contents of lycorine,lycoramine and galanthamine were analyzed among 12 species for showing the compound differences.This study provided the scientific data basis of different germplasm breeding and drug discovery.4.Twelve crude extracts of Lycoris species bulbs were tested for anti-AD in C.elegans CL2355.5-HT sensitivity analysis and acetyl cholinesterase ace-2 tested by qRT-PCR technology were used in this study.The results showed that all of these crude extracts delayed the paralysis of CL2355 under the exogenous 5-HT,and these crude extracts effecting to reduce the ace-2 gene expression had some connections with acetylcholinesterase inhibitors(ace-2).Therefore,we can make sense clearly the mechanism of Amyloid-? expression.All of the crude extracts were effective,especially L.incarnata,L.chinensis,L.straminea,L.x houdyshelii and L.radiata.So the screened alkaloids have potential value or can provide some scientific reference to new drug discovery for anti-AD.This project was built an interdisciplinary technological platform of integrating ethnobotany,ethnoecology,metabolomics,pharmacology,and molecular biology,in the order to explore a new research mode of the moden ethnobotany.Twelve Lycoris species were collected as a research object for l)analyzing their chemical constitutes and differences of Amaryllis alkaloids,2)exploring the existence and content of the effective ingredients of anti-AD,and 3)studying on intra-and interspecific interactions among Lycoris species.It was an effective strategy to discover the bioactive compounds and evaluate the traditional knowledge of local people.Our work is meaningful in exploring the ethno-oriented drug sereening,and providing a scientifie basis for sustainable development and utilization of genus Lycoris.