Mechanisms of induction of endothelial cell E-selectin expression by smooth muscle cells and its inhibition by shear stress.
E-selectin is a major adhesion molecule expressed by endothelial cells (ECs), which are exposed to shear stress and neighboring smooth muscle cells (SMCs). We investigated the mechanisms underlying the modulation of EC E-selectin expression by SMCs and shear stress. SMC coculture induced rapid and sustained increases in expression of E-selectin and phosphorylation of interleukin-1 (IL-1) receptor-associated kinase glycoprotein-130, as well as the downstream mitogen-activated protein kinases (MAPKs) and Akt. By using specific inhibitors, dominant-negative mutants, and small interfering RNA, we demonstrated that activations of c-Jun-NH(2)-terminal kinase (JNK) and p38 of the MAPK pathways are critical for the coculture-induced E-selectin expression. Gel shifting and chromatin immunoprecipitation assays showed that SMC coculture increased the nuclear factor-kappaB (NF-kappaB)-promoter binding activity in ECs; inhibition of NF-kappaB activation by p65-antisense, lactacystin, and N-acetyl-cysteine blocked the coculture-induced E-selectin promoter activity. Protein arrays and blocking assays using neutralizing antibodies demonstrated that IL-1beta and IL-6 produced by EC/SMC cocultures are major contributors to the coculture induction of EC signaling and E-selectin expression. Preshearing of ECs at 12 dynes/cm(2) inhibited the coculture-induced EC signaling and E-selectin expression. Our findings have elucidated the molecular mechanisms underlying the SMC induction of EC E-selectin expression and the shear stress protection against this SMC induction.
Chiu
Jeng-Jiann
JJ
Division of Medical Engineering Research, National Health Research Institutes, Miaoli, Taiwan, Republic of China. jjchiu@nhri.org.tw
Chen
Li-Jing
LJ
Lee
Chih-I
CI
Lee
Pei-Ling
PL
Lee
Ding-Yu
DY
Tsai
Min-Chien
MC
Lin
Chia-Wen
CW
Usami
Shunichi
S
Chien
Shu
S
D000818:Animals; D018920:Coculture Techniques; D019040:E-Selectin / Q000235:genetics*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D005786:Gene Expression Regulation; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology*; D015398:Signal Transduction; D013314:Stress, Mechanical; D014471:Umbilical Veins
D019040:E-Selectin; D007375:Interleukin-1; D020928:Mitogen-Activated Protein Kinases