OBJECTIVETo observe the effect of propofol on the transcription activity of endothelial nitric oxide synthase (heNOS) gene promoter in human umbilic vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS).

METHODSheNOS gene promoter sequence (from-1 to -1600 bp) was subcloned into the Bgl II/Hind III sites of the firefly luciferase reporter gene vector, pGL2-Basic, to obtain the recombinant plasmid peNOS-Luc. peNOS-Luc, pGL2-Basic and pCMV-beta were cotransfected into HUVECs, which were treated subsequently with LPS, LPS+propofol and LPS+transforming growth factor beta1 (TGF beta 1), respectively. The relative activities (Luc/beta-gal) were determined in the cell lysates to evaluate the activity of heNOS gene promoter.

RESULTSDouble restriction enzyme digestion and sequencing both confirmed successful construction of the recombinant plasmid peNOS-Luc, which could be effectively expressed in HUVECs. Upon LPS stimulation, the luciferase activity was obviously decreased, contrary to the effects of propofol and TGFb1 treatment, and between the latter two agents, TGF beta 1 produced higher transcription activity.

CONCLUSIONPropofol can up-regulate the activity of heNOS gene promoter in HUVECs and affect the nitrogen monoxide production and release at the transcriptional level, which is probably one of mechanisms for propofol to ameliorate LPS-induced inflammatory reaction.

Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; pharmacology ; Luciferases ; genetics ; metabolism ; Nitric Oxide Synthase Type III ; genetics ; Promoter Regions, Genetic ; genetics ; Propofol ; pharmacology ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transcription, Genetic ; drug effects ; Transfection ; Umbilical Veins ; cytology