Low Intensity Shear Stress Increases Endothelial ELR+ CXC Chemokine Production via a Focal Adhesion Kinase-p38β MAPK-NF-κB Pathway

Abstract

CXC chemokines with a glutamate-leucine-arginine (ELR) tripeptide motif (ELR؉ CXC chemokines) play an important role in leukocyte trafficking into the tissues. For reasons that are not well elucidated, circulating leukocytes are recruited into the tissues mainly in small vessels such as capillaries and venules. Because ELR ؉ CXC chemokines are important mediators of endothelial-leukocyte interaction, we compared chemokine expression by microvascular and aortic endothelium to investigate whether differences in chemokine expression by various endothelial types could, at least partially, explain the microvascular localization of endothelial-leukocyte interaction. Both in vitro and in vivo models indicate that ELR ؉ CXC chemokine expression is higher in microvascular endothelium than in aortic endothelial cells. These differences can be explained on the basis of the preferential activation of endothelial chemokine production by low intensity shear stress. Low shear activated endothelial ELR ؉ CXC chemokine production via cell surface heparan sulfates, ␤ 3 -integrins, focal adhesion kinase, the mitogen-activated protein kinase p38␤, mitogen-and stress-associated protein kinase-1, and the transcription factor.

Smad7 inhibits autocrine expression of TGF-β2in intestinal epithelial cells in baboon necrotizing enterocolitis

“…TGF-␤ 2, Smad7, and SnoN/SKIL in intestinal tissues and IEC6 cells, and histone H3 dimethyl lysine 9 (H3K9me2), H3K27me, and H4K20me (Cell Signaling Technology, Boston, MA) in IEC6 cells were measured as described previously (41). We used ␤-actin as the housekeeping control because, consistent with existing information (18), it provided better interassay consistency at the protein level in our hands than GAPDH.…”

“…Primers were designed using the Beacon Design software (Bio-Rad, Hercules, CA) and are shown in Table 1. We used our previously described SYBR green protocol to measure mRNA expression; data were normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and group-wise comparisons were made by the 2 Ϫ⌬⌬CT method (3,41). GAPDH was used as housekeeping gene because it shows minimal regional variation at the mRNA level in the gastrointestinal tract, does not change significantly with gender, and provides excellent consistency between donor replicates (1).…”

Amniotic fluid-borne hepatocyte growth factor protects rat pups against experimental necrotizing enterocolitis

“…IEC-6 cells were immunostained by using our previously described protocol (33). Briefly, IEC6 cells were grown on glass coverslips and fixed with ice-cold 4% paraformaldehyde ϫ 5 min.…”

“…We used our previously described reverse transcriptase-quantitative PCR (RT-qPCR) protocol (2,33) to measure cytokine expression in intestinal tissue. Primers were designed by use of the Beacon Design software (Bio-Rad, Hercules, CA) and are shown in Table 1.…”

Kidney Injury Accelerates Cystogenesis via Pathways Modulated by Heme Oxygenase and Complement

“…Efficient tamoxifeninduced Kif3a deletion was confirmed by immunoblotting and by loss of primary cilia by immunofluorescence ( Figure 4, A and B). Postconfluent cultures of Kif3a-deficient (Kif3a2) and control Kif3a sufficient cells (Kif3a+) were exposed to fluid flow for 12 hours in 10-cm dishes using a cone-plate viscometer, 66,67 an approach that allowed generation of adequate amount of cells for immunoblotting studies (although only at moderate-high forces). Such treatment did not significantly alter cell viability (by trypan blue exclusion).…”