Anti-Inflammatory And Antioxidant Activities Of The Nonlipid (Aqueous) Components Of Sesame Oil: Potential Use In Atherosclerosis

Dietary intervention to prevent inflammation and atherosclerosis has been a major focus in recent years. We previously reported that sesame oil (SO) was effective in inhibiting atherosclerosis in low-density lipoprotein-receptor negative mice. We also noted that the levels of many proinflammatory markers were lower in the SO-treated animals. In this study we tested whether the non-lipid, aqueous components associated with SO would have anti-inflammatory and antioxidant effects. Polymerase chain reaction array data indicated that sesame oil aqueous extract (SOAE) was effective in reducing lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. Expression of inflammatory cytokines such as interleukin (IL)-1α, IL-6, and tumor necrosis factor α (TNF-α) was also analyzed independently in cells pretreated with SOAE followed by inflammatory assault. Effect of SOAE on TNF-α-induced MCP-1 and VCAM1 expression was also tested in human umbilical vein endothelial cells. We observed that SOAE significantly reduced inflammatory markers in both macrophages and endothelial cells in a concentration-dependent manner. SOAE was also effective in inhibiting LPS-induced TNF-α and IL-6 levels in vivo at different concentrations. We also noted that in the presence of SOAE, transcription and translocation of NF-kappaB was suppressed. SOAE was also effective in inhibiting oxidation of lipoproteins in vitro. These results suggest the presence of potent anti-inflammatory and antioxidant compounds in SOAE. Furthermore, SOAE differentially regulated expression of scavenger receptors and increased ATP-binding cassette A1 (ABCA1) mRNA expression by activating liver X receptors (LXRs), suggesting additional effects on lipid metabolism. Thus, SOAE appears multipotent and may serve as a valuable nonpharmacological agent in atherosclerosis and other inflammatory diseases

Proinflammatory Responses Induced by CD40 in Retinal Endothelial and Müller Cells are Inhibited by Blocking CD40-Traf2,3 or CD40-Traf6 Signaling

Citation: Portillo J-AC, Schwartz I, Zarini S, et al. Proinflammatory responses induced by CD40 in retinal endothelial and Müller cells are inhibited by blocking CD40-TRAF2,3 or CD40-TRAF6 signaling. Invest Ophthalmol Vis Sci. 2014;55:8590-8597. DOI:10.1167/iovs.14-15340 PURPOSE. The cell surface receptor CD40 is required for the development of retinopathies induced by diabetes and ischemia/reperfusion. The purpose of this study was to identify signaling pathways by which CD40 triggers proinflammatory responses in retinal cells, since this may lead to pharmacologic targeting of these pathways as novel therapy against retinopathies. METHODS. Retinal endothelial and Müller cells were transduced with vectors that encode wildtype CD40 or CD40 with mutations in sites that recruit TNF receptor associated factors (TRAF): TRAF2,3 (DT2,3), TRAF6 (DT6), or TRAF2,3 plus TRAF6 (DT2,3,6). Cells also were incubated with CD40-TRAF2,3 or CD40-TRAF6 blocking peptides. We assessed intercellular adhesion molecule-1 (ICAM-1), CD40, monocyte chemoattractant protein-1 (MCP-1), VEGF, and prostaglandin E 2 (PGE 2 ) by fluorescence-activated cell sorting (FACS), ELISA, or mass spectrometry. Mice (B6 and CD40 À/À ) were made diabetic using streptozotocin. The MCP-1 mRNA was assessed by real-time PCR. RESULTS. The CD40-mediated ICAM-1 upregulation in endothelial and Müller cells was markedly inhibited by expression of CD40 DT2,3 or CD40 DT6. The CD40 was required for MCP-1 mRNA upregulation in the retina of diabetic mice. The CD40 stimulation of endothelial and Müller cells enhanced MCP-1 production that was markedly diminished by CD40 DT2,3 or CD40 DT6. Similar results were obtained in cells incubated with CD40-TRAF2,3 or CD40-TRAF6 blocking peptides. The CD40 ligation upregulated PGE 2 and VEGF production by Müller cells, that was inhibited by CD40 DT2,3 or CD40 DT6. All cellular responses tested were obliterated by expression of CD40 DT2,3,6. CONCLUSIONS. Blockade of a single CD40-TRAF pathway was sufficient to impair ICAM-1, MCP-1, PGE 2 , and VEGF upregulation in retinal endothelial and/or Müller cells. Blockade of CD40-TRAF signaling may control retinopathies