Effect of TRIF on permeability and apoptosis in bovine microvascular endothelial cells exposed to lipopolysaccharide.
Bovine respiratory disease complex (BRDC) can be caused by several Gram negative bacteria. Lung endothelial cells may be damaged by the release of lipopolysaccharide (LPS) from these organisms. Toll-like receptor (TLR-4) signaling pathways include the myeloid differentiation primary response gene 88 (MyD88) and the Toll/interleukin (IL)-1 receptor (TIR) domain-containing adapter-inducing interferon-β (TRIF) pathways. The aim of this study was to determine which of these pathways is responsible for permeability changes, apoptosis and cytokine production in bovine lung microvascular cells exposed to LPS. Bovine lung endothelial cells were treated with a peptide to inhibit MyD88 signaling or small interfering RNA (siRNA) to inhibit TRIF signaling. Effects were measured using trans-well endothelial electrical resistance to determine cell monolayer permeability, annexin staining to estimate apoptosis and real-time PCR to measure levels of expression of IL-1β and tumor necrosis factor (TNF)-α mRNA. Inhibition of TRIF signaling reduced permeability changes and apoptosis in endothelial cells exposed to LPS. In contrast, MyD88 inhibition reduced expression of IL-1β and TNF-α mRNA in LPS treated cells, but had no effect on permeability. It was concluded that TRIF signaling in LPS-stimulated lung endothelial cells results in permeability changes and apoptosis.
Department of Biology, University of Northern Iowa, Cedar Falls, IA 50614, USA.
Jason
J
Dubbert
Amy
A
Bowers
Yutao
Y
Su
David
D
McClenahan
D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis; D002417:Cattle / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D008070:Lipopolysaccharides / Q000494:pharmacology; D010539:Permeability; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
D033942:Adaptor Proteins, Vesicular Transport; D016207:Cytokines; D008070:Lipopolysaccharides; D012333:RNA, Messenger
Bovine; Lipopolysaccharide; Microvascular endothelial cells; MyD88; TRIF; Toll-like receptor 4