[Effect of propofol on the transcription activity of endothelial nitric oxide synthase gene promoter in human umbilic vein endothelial cells induced by lipopolysaccharide].
OBJECTIVE
To 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).


METHODS
heNOS 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.


RESULTS
Double 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.


CONCLUSIONS
Propofol 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.
Chen
Xu-gui
XG
Department of Anesthesiology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. xuguic2002@yahoo.com.cn
Gu
Miao-nin
MN
Wang
Zhuo-qiang
ZQ
D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015742:Propofol / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology
D008070:Lipopolysaccharides; D011993:Recombinant Fusion Proteins; D008156:Luciferases; D052250:Nitric Oxide Synthase Type III; D015742:Propofol