[Activation of NF-kappa B and its effect on induction of platelet-derived growth factor-B transcription by angiotensin II in ECV304 cell line].
OBJECTIVE
The renin-angiotensin system may contribute to the pathogenesis of atherosclerosis. The transcription factor nuclear factor-kappa B (NF-kappa B) participates in most signal pathways involved in the inflammatory process. In this project the effect of angiotensin II (Ang II) on NF-kappa B activation and the promotion of PDGF-B mRNA expression in human endothelial cell line ECV304 was studied.


METHODS
Electrophoretic mobility shift assay, immunofluorescence and immunoelectronic microscope techniques, including confocal microscopy and gold particle labelled electronic microscopy were applied to investigate the mechanism by which Ang II activates NF-kappa B, ECV304 cells were transiently transfected with an NF-kappa B/luciferase reporter gene and catalytically inactive NIK, IKK alpha, IKK beta mutants respectively. Northern blot was carried out to detect PDGF-B mRNA.


RESULTS
By the findings of immunofluorescence confocal microscopy, immunoelectronic microscopy and Northern blot, Ang II was effective in stimulating NF-kappa B activation and there was definited cytoplasmic-to-nuclear translocation of NF-kappa B subunits p50 and p65 and overexpression of PDGF-B mRNA expression. Over-expression of the transiently transfected IKK alpha-KM, IKK beta-KM and NIK-KM mutant genes enabled to block the reporter gene activation induced by ang II.


CONCLUSIONS
Ang II is effective to activate NF-kappa B through a pathway dependent on NIK, IKK alpha and IKK beta, and induces PDGF-B transcription in the endothelial cell line ECV304.
Li
H
H
Department of Pathology, Institute of Basic Medical Sciences, Chinese-Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China.
Yin
H
H
Zhang
H
H
Wang
Z
Z
She
M
M
D000804:Angiotensin II / Q000494:pharmacology*; D015152:Blotting, Northern; D002460:Cell Line; D024202:Electrophoretic Mobility Shift Assay; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D016328:NF-kappa B / Q000378:metabolism*; D020574:Proto-Oncogene Proteins c-sis / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects*
D016328:NF-kappa B; D020574:Proto-Oncogene Proteins c-sis; D000804:Angiotensin II