Effect of PP60c-Src on angiotensin II-induced signal transduction in rat vascular smooth muscle cells.
- Author:
Mei-xiang XIANG
1
;
Dong LIU
;
Peng CHEN
;
Jun-Sheng LU
Author Information
- Publication Type:Journal Article
- MeSH: Angiotensin II; pharmacology; Animals; Male; Mitogen-Activated Protein Kinases; metabolism; Muscle, Smooth, Vascular; cytology; drug effects; metabolism; Oligonucleotides, Antisense; pharmacology; Proto-Oncogene Proteins pp60(c-src); analysis; physiology; Rats; Rats, Sprague-Dawley; Signal Transduction; drug effects
- From: Journal of Zhejiang University. Medical sciences 2003;32(2):126-130
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo further clarify the mechanism of Ang II-induced intracellular signal transduction in vascular smooth muscle cells(VSMCs) proliferation by observing the effect of c-Src on Ang II-mediated MAPK activation and c-fos protein expressions in rat VSMCs.
METHODSAortic VSMCs from SD rats were cultured primarily and subcultured, which were transfected with anti-sense c-Src oligodeoxynucleotides(ODNs) wrapped with lipofectin to inhibit c-Src activity and protein production. Untransfected VSMCs were used as control, to observe the role of Ang II stimulation in MAPK activation and c-fos protein expression in VSMC. Protein immunoprecipitation and kinase phosphorylation were employed to measure c-Src kinase activity; MAPK kinase activity was assessed by the phosphorylation rate of the substrate MBP(Myelin Basic Protein); Western blot was used to assess the protein expression of c-Src and c-fos.
RESULTSc-Src protein expressions in VSMC, which were transfected with different concentrations of anti-sense c-Src ODNs, were significantly decreased in a negative dose-effect manner (0.2 microm, 0.5 microm, 1.0 microm and 2.0 microm were 68.2%, 34.7%, 30.3% and 15.8% respectively compared with control). c-Src kinase activity was also obviously inhibited. Following stimulation of Ang II on VSMC transfected with anti-sense c-Src ODNs, the increase of c-Src activity was only 8.7% of control,the activity of MAPK only 1.6% compared with control, and the increase in c-fos protein expression 30.3% as control.
CONCLUSIONAng II can induce c-Src activation and intracellular signal transduction in VSMC which depend on c-Src activation, indicating that c-Src is a pivotal signal factor in VSMC proliferation.