Regulation of inducible nitricoxide synthase gene transcription by p38 mitogen-activated protein kinase in human embryonic kidney 293 cells.
OBJECTIVE
To study the transcriptional regulation of inducible nitric oxide synthase (iNOS) gene by p38 mitogen-activated protein kinase (MAPK).


METHODS
With human embryonic kidney (HEK) 293 cells as the target and the assistance of lipofectamine-mediated co-transfection techniques and luciferase reporter gene systems, FLAG-tagged p38 isoforms (namely FLAG-p38 alpha, FLAG-p38 beta;, FLAG-p38 gamma and FLAG-p38 phi;) in pcDNA3, pcDNA3, piNOS-Luc and pCMV-beta; were transfected into HEK 293 cells, and the relative activity of luciferase was subsequently tested.


RESULTS
Highest luciferase activity occurred only in p38 alpha group compared with the other three isoform groups under no stimulation. Under the stimulation by lipopolysaccharide (LPS), the luciferase activity of each group was obviously increased and the highest activity occurred in p38 beta group.


CONCLUSIONS
LPS can induce transcription and activation of iNOS gene, and p38 MAPK is involved in the transcription regulation of iNOS gene in HEK 293 cells.
Guo
Ai-hua
AH
Key Laboratory for Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China.
Gong
Xiao-wei
XW
Kan
Wen-hong
WH
Qin
Qing-he
QH
Deng
Peng
P
Wang
Jing-zhen
JZ
Jiang
Yong
Y
D002478:Cells, Cultured; D004622:Embryo, Mammalian; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007668:Kidney / Q000166:cytology; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D014158:Transcription, Genetic; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases
C496319:NOS2 protein, human; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases