| Blood-brain barrier (BBB) is a dynamic adjustive interface which exists between capillary vessel on brain and spinal cord and nerve tissue, prevents most exogenous material from permeating into BBB, pumps the noxious or excess substance out of brain selectively in order to keep the microenvironment stable. However, the permeability of many central nervous system medicine is low, impacts the therapeutic effect of medicine seriously. Therefore, more and more attention is paid to the research of BBB.The action of aromatic Chinese traditional medicine is to restore consciousness, Acorus tatarinowii Schott is one of traditional "kaiqiao" drugs, used in folk alone or compatibility with other drugs, the effect is indeed, which cues us that acorus tatarinowii schott could increase permeability of BBB, so we research it to solve neck-problem in central nervous system drugs development, and set up new rational and experimental evidence for traditional application of aromatice Chinese medicine.We extracted volatile oil according to Chinese pharmacopeia (2005 edition), extraction ratio was 1.58%, decoction was condensed to 1g/ml.Volatile oil of Acorus tatarinowii Schott samples were analyzed by analytical technique. α-Asarone, β-asarone, guaiene, acorenone were detected in volatile oil with GC/MS and α-asarone, β-asarone was detected in rat cerebrospinal fluid barrier with GC/MS under the same condition after gastric infusion of volatile oil.Decoction of acorus tatarinowii schott samples were analyzed with HPLC/MS, there're three compounds was detected.The ultramicrostructure of BBB in rat cerebral cortex was observed under electron microscope, and acorus tatarinowii schott could relax the tight junction(TJ), but BBB was still integrity.Primarily study the promotion of evan's blue (EB) penetration into BBB administrated acorus tatarinowii schott and borneol which was control drug, namely the content of EB in brain reflected the permeability of BBB. Statistical analysis showed after administrating borneol 2.5h the content of EB in brain was high compared with blank group significantly(P<0.01), that is borneol could enhance permeability administrated 2.5h later, but acorus tatarinowii schott couldn't.Prolong action time of acorus tatarinowii schott, observe effect to BBB with the high dose (23.4g/kg), middle dose (11.7g/kg), low dose (1.17g/kg) volatile oil. Statistical analysis indicated middle and high dose could promote the permeability of BBB compared with blank group significantly (P<0.05), and showed dose-dependent; at middle dose volatile oil and decoction could promote the permeability of BBB (P<0.05), the whole drag could promote the permeability of BBB compared with blank group significantly (P<0.01).Administrated acorus tatarinowii schott a week, intraperitoneal injected phenytoin sodium, and the concentration of phenytoin sodium in brain was detected used by HPLC. At 4h after injected, the concentration of phenytoin sodium was increased significantly (P<0.01). In word, acorus tatarinowii schott appeared to be a prospecting assisting agent with auxo-action on the penetration of other substances across BBB.Concentration of 5-HT was detected used by electrochemical detector, compared with blank group, the content of 5-HT in volatile oil group increased significantly (P<0.05), as well as decoction and whole drag (P<0.01), so we presumed one of molecular pathways which acorus tatarinowii schott influenced permeability of BBB was to regulate content of 5-HT in brain.Cultivated Hela cells as career to evaluate the activity of P-glycoprotein, the result was that all parts of acorus tatarinowii could reinforce the cytotoxicity caused by VCR significantly(P<0.01) at 0.5h; after 1h, only volatile oil couldreinforce the cytotoxicity significantly (P<0.01) ; after 24h, neither control drug nor all parts of acorus tatarinowii schott could affect the cytotoxicity caused by VCR(P>0.05).Cerebral microvessel fragments were obtained by two steps of enzyme digestions and gradient centrifugation with BSA and Percoll, and cultivated primary rat cerebral microvascular endothelial cells, we established a BBB model in vitro initially.In short, the resuscitative effect of acorus tatarinowii schott is resulted from the comprehensive action of multiple components; It is of great importance to improve the blood concentration in brain and bioavailability of central nervous system drugs; Acorus tatarinowii Schott could open BBB by increasing the level of 5-HT in brain, or P-glycoprotein inhibitor can get drugs across the blood-brain barrier; relatively pure primary culture of RCMEC was successfully established using this method and the model system could be used to develop an in vitro model of the rat blood-brain barrier.
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