PMID- 16972260
OWN - NLM
STAT- MEDLINE
DCOM- 20070207
LR  - 20131121
IS  - 0021-9541 (Print)
IS  - 0021-9541 (Linking)
VI  - 209
IP  - 3
DP  - 2006 Dec
TI  - Homocysteine-induced myofibroblast differentiation in mouse aortic endothelial 
      cells.
PG  - 767-74
AB  - Differentiation of myofibroblast, as evidenced by alpha-smooth muscle actin 
      (alpha-SMA) expression, is largely mediated by transforming growth factor-beta1 
      (TGF-beta1). This mechanism often follows inflammatory events such as endothelial 
      damage due to oxidative stress, which can further leads to vascular thickening, 
      stiffness, and fibrosis. We hypothesized that hyperhomocysteinemia (HHcy)-induced 
      oxidative stress lead to vascular stiffness, in part due to 
      endothelial-myofibroblast differentiation and alteration of collagen homeostasis 
      in the extracellular matrix (ECM). We tested our hypothesis in vitro using mouse 
      aortic endothelial cells (MAEC). Our result shows that Hcy induces alpha-SMA and 
      collagen type-1 expression in MAEC as evidenced by immunoblot and confocal 
      imaging. RT-PCR shows robust increase of alpha-SMA and collagen type-1 mRNA level 
      in Hcy-induced condition. We demonstrated that Hcy induces autophosphorylation of 
      focal adhesion kinase (FAK) (a member of the protein tyrosine kinase (PTK) 
      family) at Tyr-397. PP2 (general PTK inhibitor) as well as FAK siRNA abrogates 
      Hcy-mediated alpha-SMA formation. In addition to that, Hcy-mediated TGF-beta1 
      induction was inhibited by TGF-beta R1 kinase inhibitor II (ALK5 inhibitor II) 
      and attenuated FAK phosphorylation and alpha-SMA expression. Furthermore, we 
      showed that Hcy activates ERK-44/42 (extracellular signal-regulated kinase) 
      pathway and augments collagen type-1 deposition. Studies with pharmacological ERK 
      blocker, PD98059 and ERK siRNA attenuated ERK-44/42 phosphorylation and collagen 
      type-1 synthesis. Taken together our results demonstrate that Hcy-mediated 
      TGF-beta1 upregulation triggers endothelial-myofibroblast differentiation 
      secondary to FAK phosphorylation and that Hcy-induced ERK activation is involved 
      in ECM remodeling by altering collagen type-1 homeostasis.
CI  - (c) 2006 Wiley-Liss, Inc.
FAU - Sen, Utpal
AU  - Sen U
AD  - Department of Physiology & Biophysics, HSC, University of Louisville School of 
      Medicine, 500 South Preston St., Louisville, KY 40202, USA.
FAU - Moshal, Karni S
AU  - Moshal KS
FAU - Tyagi, Neetu
AU  - Tyagi N
FAU - Kartha, Ganesh K
AU  - Kartha GK
FAU - Tyagi, Suresh C
AU  - Tyagi SC
LA  - eng
GR  - HL 71010/HL/NHLBI NIH HHS/United States
GR  - HL 74185/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PL  - United States
TA  - J Cell Physiol
JT  - Journal of cellular physiology
JID - 0050222
RN  - 0 (Actins)
RN  - 0 (Collagen Type I)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Transforming Growth Factor beta1)
RN  - 0LVT1QZ0BA (Homocysteine)
RN  - EC 2.7.10.2 (Focal Adhesion Kinase 1)
RN  - EC 2.7.10.2 (Ptk2 protein, mouse)
RN  - EC 2.7.11.24 (Extracellular Signal-Regulated MAP Kinases)
SB  - IM
MH  - Actins/genetics/metabolism
MH  - Animals
MH  - Aorta/*cytology
MH  - Cell Differentiation/*drug effects
MH  - Cells, Cultured
MH  - Collagen Type I/genetics/metabolism
MH  - Endothelial Cells/cytology/drug effects/*physiology
MH  - Enzyme Induction
MH  - Enzyme Inhibitors/metabolism
MH  - Extracellular Signal-Regulated MAP Kinases/metabolism
MH  - Fibroblasts/cytology/drug effects/*physiology
MH  - Focal Adhesion Kinase 1/metabolism
MH  - *Homocysteine/metabolism/pharmacology
MH  - Mice
MH  - RNA, Small Interfering/genetics/metabolism
MH  - Transforming Growth Factor beta1/metabolism
EDAT- 2006/09/15 09:00
MHDA- 2007/02/08 09:00
CRDT- 2006/09/15 09:00
PHST- 2006/09/15 09:00 [pubmed]
PHST- 2007/02/08 09:00 [medline]
PHST- 2006/09/15 09:00 [entrez]
AID - 10.1002/jcp.20752 [doi]
PST - ppublish
SO  - J Cell Physiol. 2006 Dec;209(3):767-74. doi: 10.1002/jcp.20752.