Fleabane Brain Targeted Nerve Protection Pharmacodynamic Material Basis For Research

Cerebral ischemia,known as the "stroke" in Chinese medicine,is a complex mechanism of cascade of pathological,and its core problem is nerval dysfunction caused by neural damage.Therefore,the neuroprotection is of great necessity to the treatment.Because of the overall concept,the complexity,and the diversity of the mechanism,Chinese medicine has great advantages in the therapy of cerebral ischemic on the neuroprotection.But the active components from Chinese medicine and natural medicine have low bioavailability and are difficult through the blood-brain barrier,so they are difficult to clinical application.In this paper,regarding exist preparation as the breakthrough point,the research idea is "traditional Chinese medicine→the blood-brain barrier+neuroprotection→brain-targeting effective components of neuroprotective"Erigeron breviscapus(Vant.) Hand.-Mazz.was firstly recorded in Yunnan Materia Medica.Erigeron breviscapus and Erigeron injection are contained in "People's Republic of China Pharmacopoeia"(2005).Erigeron injection,erigeron injection,tablets,et al.have been widely used in clinical.At present,the improved blood rheology,ischemia-reperfusion injury protection and anti-cerebral thrombosis are mainly studied on Erigeron breviscapus,however,the effective components of cerebral neuroprotective and the blood-brain barrier permeability are not yet clear.Objective:To study on the clinic efficacy of preparations,based on the blood-brain barrier permeability and nerve protection mechanism,the effective substance of the treatment of cerebral ischemia,of E.breviscapus has been studied.Methods:1.To establish HPLC fingerprints of the different preparations of erigeron was analyzed the effective components of antioxidant activity of erigeron injection by CE-CL.2.To extract and isolate the compounds of E.breviscapus by silica gel,ODS, polyamide column chromatography and HPLC,their structures were characterized by ¹H-NMR,¹³C-NMR and MS.With the method of esterifying and acetyling, derivatives were synthesized.3.To cultivate rat cerebral conical neurons cell in vitro and determine the survival rate by MTT,the neuroprotection activity of erigeron preparations and E. breviscapus was evaluated,with neuroprotection active ingredients screened.4.To co-culture BMVEC and AC in vitro,blood-brain barrier model(BBBM) was established with the active components analysized by HPLC.Result:1.The main chemical group of Erigeron preparation are contain with scutellarin, caffeic acid,double caffeoyl quinic acid such as flavonoids and caffeoyl-constituents. And erigeron injections had four effective ingredients.2.12 compounds were isolated,with 9 identified,as follows:scutellarin, scutellarein,apigenin,apigenin-7-O-glucuronic acid,1,5-di-O-caffeoyl quinic acid, 3,4-di-O-caffeoyl quinic acid,friedelin,stigmasterol,fourteen acids.At the same time,after structurally modified,synthesis caffeoyl methyl ester,caffeoyl ethyl ester, 3,4-diacetyl-caffeic,scutellarin and glycolamide were gained.3.The preparations,such as erigeron injection,scutellarin injection,and so on, could promote the survival of nerve cell in vitro,so did the flavonoids of scutellarin, apigenin-7-O-glucuronic acid,the caffeoyl constituents of caffeic acid,double caffeoyl quinic acid and the derivatives of caffeic acid and scutellarin.4.The blood-brain barrier model in vitro was successful.1,5-DCQ,3,5-DCQ, 3,4-DCQ,apigenin-7-O-glucuronic,could pass through BBBM,so did coffee methyl ester,3,4-diacetyl-caffeic acid,glycolamide ester of scutellarin.Conclusion:1.Erigeron preparation and E.breviscapus mainly contain flavonoids and double caffeoyl quinic acids.The neuroprotective compounds are scutellarin,caffeic acid, 1,5-DCQ,3,5-DCQ,3,4-DCQ and so on.The neuroprotection of scutellarin are related on the antioxidant activity.2.The brain-targeting effective components of neuroprotective from Erigeron are 1,5-DCQ,3,5-DCQ,3,4-DCQ,apigenin-7-O-glucuronic.3.The products of the structural modification of caffeic acid and scutellarin have changed from hydrophilicity to lipophilicify,which the oil-water partition coefficient increases about 10³ï½ž10⁵.Coffee methyl ester,3,4-diacetyl-caffeic acid,glycolamide ester of scutellarin are also the brain-targeting effective components of neuroprotective.Innovation:1.The research idea is "Traditional Chinese medicine→through the blood-brain barrier+neuroprotection→brain-targeting effective components of neuroprotective"2.The effective components of neuroprotective of E.breviscapus has essentially evaluated by co-cultured in vitro rat brain microvascular endothelial cells and brain microvascular endothelial cells.