Differential effects of NF-κB and p38 MAPK inhibitors and combinations thereof on TNF-α- and IL-1β-induced proinflammatory status of endothelial cells in vitro

Abstract

Endothelial cells actively participate in inflammatory events by regulating leukocyte recruitment via the expression of inflammatory genes such as E-selectin, VCAM-1, ICAM-1, IL-6, IL-8, and cyclooxygenase (COX)-2. In this study we showed by real-time RT-PCR that activation of human umbilical vein endothelial cells (HUVEC) by TNF-alpha and IL-1beta differentially affected the expression of these inflammatory genes. Combined treatment with TNF-alpha and IL-1beta resulted in nonadditive, additive, and even synergistic induction of expression of VCAM-1, IL-8, and IL-6, respectively. Overexpression of dominant-negative inhibitor kappaB protein blocking NF-kappaB signaling confirmed a major role of this pathway in controlling both TNF-alpha- and IL-1beta-induced expression of most of the genes studied. Although dexamethasone exerted limited effects at 1 muM, the thioredoxin inhibitor MOL-294, which regulates the redox state of NF-kappaB, mainly inhibited adhesion molecule expression. Its most pronounced effect was seen on VCAM-1 mRNA levels, especially in IL-1beta-activated endothelium. One micromolar RWJ-67657, an inhibitor of p38 MAPK activity, diminished TNF-alpha- and IL-1beta-induced expression of IL-6, IL-8, and E-selectin but had little effect on VCAM-1 and ICAM-1. Combined treatment of HUVEC with MOL-294 and RWJ-67657 resulted in significant blocking of the expression of E-selectin, IL-6, IL-8, and COX-2. The inhibitory effects were much stronger than those observed with single drug treatment. Application of combinations of drugs that affect multiple targets in activated endothelial cells may therefore be considered as a potential new therapeutic strategy to inhibit inflammatory disease activity.

S‐Allyl‐L‐Cysteine Sulfoxide Inhibits Tumor Necrosis Factor‐Alpha Induced Monocyte Adhesion and Intercellular Cell Adhesion Molecule‐1 Expression in Human Umbilical Vein Endothelial Cells

“…These findings are consistent with previous observation that the ERK1/2 and JNK, but not P38, is involved in the TNF-a-induced endothelial permeability (Kumar et al, 1998;Kuldo et al, 2005). The downregulation of ERK1/2 and JNK phosphorylation should negatively affect downstream JNK activation, which is a crucial regulator for cell activation.…”

“…TNF-a can induce the phosphorylation of IjB, through its receptor, and result in the activation and translocation of NF-jB to the nucleus, promoting the expression of downstream genes, such as ICAM-1. The activation of NF-jB is also regulated by JNK signaling through inducing b-TrCP and protecting NF-jB from degradation (Kuldo et al, 2005). Furthermore, activation of both NF-jB and JNK together induce a number of gene expressions (Min and Pober, 1997).…”

Delivery of the p38 MAPkinase Inhibitor SB202190 to Angiogenic Endothelial Cells: Development of Novel RGD-Equipped and PEGylated Drug−Albumin Conjugates Using Platinum(II)-Based Drug Linker Technology

“…TNFR induced a 20-fold upregulation in gene expression of hIL-8 and 200-fold upregulation of hEselectin (Figure 8). Free SB202190 inhibited this upregulation up to 50%, which is in agreement with previous studies (23). RGDPEG-SB-HSA was tested at 21 µg/mL and 70 µg/mL, which corresponds to a drug concentration of 3 µM and 10 µM, respectively.…”

Unique and redundant roles of α4 and β2 integrins in kinetics of recruitment of lymphoid vs myeloid cell subsets to the inflamed peritoneum revealed by studies of genetically deficient mice

“…6A). These data suggest the preferential usage of α4 integrins by lymphocytes for adhesion to bEND3 cells, since TNFα not only up-regulates VCAM-1 but also ICAM-1, a β2 integrin ligand on endothelial cells [27,28]. To determine whether ability to migrate per se is preserved in α4Δ/Δ lymphocytes, we performed migration through an uncoated Transwell insert (trans-well migration) not requiring engagement of α4 integrins.…”