Date on Master's Thesis/Doctoral Dissertation
Senior Honors Thesis
Department of Pharmacology and Toxicology
Sinusoidal endothelial cells; Kupffer cells; Cell culture; Extracellular matrix; Inflammatory liver injury
Fatty liver disease, be it from alcohol (ALD), obesity (NAFLD) or other sources (e.g. viral infection), share similar mechanisms of disease progression. One potential mechanism is that altered sinusoidal endothelial cell (SEC) extracellular matrix (ECM) favors a more inflammatory phenotype in resident macrophages (i.e., Kupffer cells). Here, the hypothesis was tested that SEC-derived matrices directly affect the inflammatory response of macrophages. Methods. Transformed sinusoidal endothelial cells (TSECs) were cultured for 24 or 72 hours. Culture plates were then washed with a solution that selectively removed the cells, but preserved the ECM. Cultured macrophages (RAW 264.7 cells) were then seeded on the matrix and cultured for 24 hours; the cells were then stimulated with LPS for 0, 3, 6, 12, or 24 hours (100 ng/mL) ± CycloRGDfV. Real time RT-PCR was used to measure mRNA expression of proinflammatory mediators (IL-6, IL-1β, TNF-α, and INOS) and anti-inflammatory mediators (IL-10 and TGF-β). Results. LPS stimulated the production of all mediators by macrophages; when plated on ECM from TSECS, this response was attenuated for IL-6 and IL-1β. The presence of TSEC-derived ECM increased expression of TGF-β. The addition of the small peptide antagonist CycloRGDfV produced varying effects on the expression of inflammatory mediators in the presence or absence of TSEC-derived ECM. Conclusions. These data serve as first proof-of-concept that macrophage 4 activation can be modulated by ECM produced by TSECs and identifies a new interaction between these cells that may contribute to inflammatory liver disease.
Poole, Lauren G., "Sinusoidal endothelial cell-derived extracellular matrix regulates basal and stimulated macrophage activation." (2013). College of Arts & Sciences Senior Honors Theses. Paper 21.
Retrieved from http://ir.library.louisville.edu/honors/21