1.Isolation and Preparation of Phloroglucinol Derivatives from Dryopteris Fragrans by Semi-preparation HPLC
Guoqiang SONG ; Guotong DENG ; Wenzhao DU ; Guanfeng DU ; Jiangchen CHEN ; Zhibin SHEN
China Pharmacist 2015;(4):545-547,548
Objective:To isolate and prepare the reference substances of phloroglucinol derivatives from Dryopteris fragrans by semi-preparation HPLC. Methods:After reflux extraction of Dryopteris fragrans with petroleum ether,the extracting solution was con-centrated and separated by silica gel column chromatography,and then isolated by semi-preparation HPLC. The isocratic elution was carried out using acetonitrile as the mobile phase at 3. 0 ml·min-1 and the injection volume was 0. 5 ml. Two phloroglucinol deriva-tives were isolated. Results:The chemical structure of the two phloroglucinol derivatives respectively was aspidin BB with the purity of 98. 81% and aspidin PB with the purity of 98. 57% by ultra high performance liquid chromatography. Conclusion:The isolation of as-pidin BB and aspidin PB by semi-preparation HPLC is simple and fast with the purity over 98%,which can be used to prepare the ref-erence substances.
2.In intro delivery of gene encoding neurotrophic (GDNF) into brain by gene transfer.
Acta Pharmaceutica Sinica 2005;40(1):87-90
AIMThe gene encoding neurotrophic factor was transfected into brain capillary endothelial cells with the aim of delivering the gene product extensively into the brain parenchyma by making use of the secretory function of BCECs.
METHODSPlasmid DNA encoding mouse glial cell-derived neurotrophic factor (mGDNF gene) was constructed and prepared. Then, mGDNF gene was transfected into cultured mouse brain capillary endothelial cells (BCECs) in vitro. The amount of mGDNF protein in the transfected cells and secreted from the transfected cells were determined by ELISA. The polarity of the secretion of mGDNF protein from BCECs was investigated in a bicameral culture system.
RESULTSThe mGDNF protein was detected out not only from the transfected cells but also the cultured media. And mGDNF protein was mainly found in the brain side of the culture compartment.
CONCLUSIONIt has been demonstrated that a secretory protein can be successfully delivered into brain parenchyma by utilizing the secretory pathway of BCECs.
Animals ; Blood-Brain Barrier ; Brain ; cytology ; metabolism ; Cells, Cultured ; Culture Media ; metabolism ; Endothelial Cells ; metabolism ; secretion ; Mice ; Plasmids ; Transfection
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