TY - JOUR
T1 - Metal-catalyzed oxidation of extracellular matrix components perturbs hepatocyte survival with activation of intracellular signaling pathways
AU - Giri, Ranjit K.
AU - Malhi, Harmeet
AU - Joseph, Brigid
AU - Kandimalla, Jithender
AU - Gupta, Sanjeev
N1 - Funding Information:
The work was supported in part by NIH Grants R01 DK46952, P33 DK41296, and P30 CA13330. We thank Dr. J. Mattana for helpful advice.
PY - 2003/12/1
Y1 - 2003/12/1
N2 - To investigate whether oxidative manipulation of extracellular matrix components could affect cell survival, we studied primary rat hepatocytes cultured on dishes coated with collagen type 1, which was oxidized with a metal-based system. Culture of hepatocytes on oxidized collagen led to decreased cellular catalase activity along with impaired cell survival. The fraction of polyploid hepatocytes decreased early followed by greater reaccumulation of polyploid cells. Cells cultured on oxidized collagen showed greater susceptibility to additional oxidant stress induced by tert.-butyl-hydroperoxide. The capacity of hepatocytes for growth factor-induced DNA synthesis was unaffected by culture on oxidized collagen. In response to culture on oxidized matrix, AP-1, Egr-1, CREB, and NF-κB transcription factor activity was rapidly increased. This change in transcription factor activity was ameliorated by treatment of collagen with a free radical spin trap, N-tert.-butyl-α-phenylnitrone, prior to oxidation. Moreover, culture of hepatocytes with aminoguanidine, an antioxidant drug, decreased cell injury. These findings established that exposure of primary hepatocytes to oxidized extracellular matrix components rapidly activates cell signaling events with loss of hepatocyte subpopulations. Such cell-extracellular matrix interactions may play roles in organ homeostasis and oncogenetic progression.
AB - To investigate whether oxidative manipulation of extracellular matrix components could affect cell survival, we studied primary rat hepatocytes cultured on dishes coated with collagen type 1, which was oxidized with a metal-based system. Culture of hepatocytes on oxidized collagen led to decreased cellular catalase activity along with impaired cell survival. The fraction of polyploid hepatocytes decreased early followed by greater reaccumulation of polyploid cells. Cells cultured on oxidized collagen showed greater susceptibility to additional oxidant stress induced by tert.-butyl-hydroperoxide. The capacity of hepatocytes for growth factor-induced DNA synthesis was unaffected by culture on oxidized collagen. In response to culture on oxidized matrix, AP-1, Egr-1, CREB, and NF-κB transcription factor activity was rapidly increased. This change in transcription factor activity was ameliorated by treatment of collagen with a free radical spin trap, N-tert.-butyl-α-phenylnitrone, prior to oxidation. Moreover, culture of hepatocytes with aminoguanidine, an antioxidant drug, decreased cell injury. These findings established that exposure of primary hepatocytes to oxidized extracellular matrix components rapidly activates cell signaling events with loss of hepatocyte subpopulations. Such cell-extracellular matrix interactions may play roles in organ homeostasis and oncogenetic progression.
KW - Culture
KW - Extracellular matrix
KW - Hepatocyte
KW - Injury
KW - Oxidation
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U2 - 10.1016/S0014-4827(03)00405-1
DO - 10.1016/S0014-4827(03)00405-1
M3 - Article
C2 - 14644166
AN - SCOPUS:0344873274
SN - 0014-4827
VL - 291
SP - 451
EP - 462
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 2
ER -