Marinobufagenin regulates gene expression in human brain microvascular endothelial cells in vitro.

To understand the mechanism by which MBG increases permeability of human brain microvascular endothelial cell (HBMEC) monolayers, the primary cultures were treated with 10nM MBG for 12 h. RNA was isolated and analysed for gene expression on Agilent one color microarrays. Triplicate wells of HBMEC were treated with 10 nM MBG in DMSO or with just the drug vehicle for 12 h. RNA was isolated from each well and used to reverse transcribe and generate Cy3-labelled cRNA. Microarrays were hybridized, washed and scanned. Data were normalized and assessed for differential gene expression between MBG treated and control HBMEC.