Abstract
Summary Background: The aim of this study was to characterize the effects of the coagulation proteinase thrombin on proteomic level in human hepatic stellate LX-2 cells. Methods: Proteomic analyses were performed using surface-enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI-TOF-MS). The protein profiles obtained from LX-2 cell lysates using strong anion exchanger Q10 ProteinChip arrays were statistically analyzed. Results: The peak intensities of 50 protein/peptide clusters were identified as being different between nonstimulated and LX-2 cells treated with thrombin for 6 h and 24 h, respectively. As the most significantly enhanced single signal in LX-2 cells stimulated with thrombin, a protein with a molecular mass of 13.560 kDa has been identified that corresponds exactly to calcium dependent phospholipase 2 (cPLA2). Thrombin-induced increase in the cPLA2 protein expression in LX-2 cells was confirmed by using the Western blotting technique. Conclusions: Together with the finding that thrombin induced phosphorylating activation of cPLA2 in LX-2 cells, our data point to an important function of the thrombin-mediated modulation of cytosolic phospholipase A2 in hepatic stellate cells.
Highlights
Beside its critical role in blood coagulation, the serine proteinase thrombin (EC 3.4.21.5) is known to evoke biological responses from a variety of cells, e.g. platelets, fibroblasts, vascular smooth muscle cells and monocytes [1,2,3], and may influence a number of cellular responses that play a role in subsequent proinflammatory and profibrotic processes in different organs including the liver [4].Recent data suggest a role for thrombin in the activation of hepatic stellate cells (HSCs) that has been recognized as a central event in the development of liver fibrosis and cirrhosis and malignancy
The aim of this study was to characterize the effects of the coagulation proteinase thrombin on proteomic level in human hepatic stellate LX-2 cells
As the most significantly enhanced single signal in LX-2 cells stimulated with thrombin, a protein with a molecular mass of 13.560 kDa has been identified that corresponds exactly to calcium dependent phospholipase 2
Summary
Recent data suggest a role for thrombin in the activation of hepatic stellate cells (HSCs) that has been recognized as a central event in the development of liver fibrosis and cirrhosis and malignancy. Thrombin has been shown to stimulate matrix synthesis and the regulation of MCP1-production of cultured hepatic stellate cells [6,7,8]. Hepatic stellate cells express the proteinase activated receptors 1 and 4 [8], members of a novel subfamily of G-protein coupled receptors mediating the cellular effects of thrombin in different cell types. We used SELDI-MS to evaluate the effect of thrombin on protein expression profiles in cells of the human stellate cell line LX-2 that has been characterized comprehensively as a suitable model for investigations on liver fibrosis [13]. Lysates from nonstimulated LX-2 cells and LX-2 cells stimulated with thrombin (1.0 NIH-U/mL) for 6 h and 24 h, respectively, were evaluated using strong anion exchanger Q10 ProteinChip arrays
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
