Role of p38 MAP kinase in endothelial cell alignment induced by fluid shear stress

2001 ◽  
Vol 280 (1) ◽  
pp. H189-H197 ◽  
Author(s):  
Nobuyoshi Azuma ◽  
Nobuyuki Akasaka ◽  
Hiroyuki Kito ◽  
Masataka Ikeda ◽  
Vivian Gahtan ◽  
...  
Keyword(s):  
Shear Stress ◽  
Map Kinase ◽  
Fluid Shear Stress ◽  
Morphological Changes ◽  
Critical Role ◽  
Specific Inhibitor ◽  
Orientation Angle ◽  
Fiber Formation ◽  
Dominant Negative Mutant ◽  
Sb 203580

The p38/mitogen-activated protein (MAP) kinase-activated protein kinase 2 (MAPKAP kinase 2)/heat shock protein (HSP)25/27 pathway is thought to play a critical role in actin dynamics. In the present study, we examined whether p38 was involved in the morphological changes seen in endothelial cells (EC) exposed to shear stress. Cultured bovine aortic EC were subjected to 14 dyn/cm2 laminar steady shear stress. Peak activation of p38, MAPKAP kinase 2, and HSP25 were sixfold at 5 min, sixfold at 5 min, and threefold at 30 min compared with static control, respectively. SB-203580 (1 μM), a specific inhibitor of p38, abolished the activation of MAPKAP kinase 2 and HSP25 as well as EC elongation and alignment in the direction of flow elicited by shear stress. The mean orientation angle of cells subjected to shear without SB-203580, with SB-203580, or static control were 17, 50, and 43°, respectively ( P < 0.05). EC transfected with the dominant negative mutant of p38-α aligned randomly with no stress fiber formation despite exposure to shear stress. These data suggests that the pathway of p38/MAPKAP kinase 2/HSP25/27 is activated in response to shear stress, and this pathway plays an important role in morphological changes induced by shear stress.

Fluid shear stress stimulates platelet-derived growth factor expression in endothelial cells

1993 ◽  
Vol 265 (1) ◽  
pp. H3-H8 ◽  
Author(s):  
M. Mitsumata ◽  
R. S. Fishel ◽  
R. M. Nerem ◽  
R. W. Alexander ◽  
B. C. Berk
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Growth Factor ◽  
Fluid Shear Stress ◽  
Critical Role ◽  
Fold Increase ◽  
Flow Chamber ◽  
Platelet Derived Growth Factor ◽  
Aortic Endothelial Cells ◽  
A Chain

Fluid flow and the associated shear stress play a critical role in vascular growth and remodeling. Recent data suggest that increased endothelial cell expression of platelet-derived growth factor (PDGF) A- and B-chain by flow may participate in these events. In the present study, we examined the mechanism for flow-induced PDGF expression, focusing on protein kinase C (PKC). Bovine aortic endothelial cells were exposed to flow (shear stress = 30 dyn/cm2) in a parallel-plate flow chamber. Increases in PDGF B-chain, but not PDGF A-chain, were observed within 3 h, maximal within 6 h (13-fold increase), and sustained for 24 h. PKC appeared to be involved because phorbol 12-myristate 13-acetate induced PDGF B-chain mRNA. Activation of PKC alone, however, was insufficient to induce PDGF mRNA because the selective PKC activator, 1-oleoyl-2-acetyl-sn-glycerol, did not induce PDGF expression. A PKC-independent pathway was suggested by the fact that inhibition of PKC (downregulation with phorbol 12,13-dibutyrate or exposure to staurosporine) failed to block PMA or flow-induced PDGF B-chain expression. These results demonstrate flow-induced PDGF B-chain expression in endothelial cells that appears to be mediated, in part, by a PKC-independent pathway.

Activation of p38 MAP Kinase and JNK But Not ERK Is Required for Erythropoietin-Induced Erythroid Differentiation

Blood ◽  
1998 ◽  
Vol 92 (6) ◽  
pp. 1859-1869 ◽  
Author(s):  
Yuka Nagata ◽  
Noriko Takahashi ◽  
Roger J. Davis ◽  
Kazuo Todokoro
Keyword(s):  
Cell Growth ◽  
Map Kinase ◽  
Antisense Oligonucleotides ◽  
Map Kinases ◽  
Physiological Function ◽  
Critical Role ◽  
Specific Inhibitor ◽  
Erythroid Differentiation ◽  
P38 Map Kinase ◽  
American Society

p38 MAP kinase (p38) and JNK have been described as playing a critical role in the response to a variety of environmental stresses and proinflammatory cytokines. It was recently reported that hematopoietic cytokines activate not only classical MAP kinases (ERK), but also p38 and JNK. However, the physiological function of these kinases in hematopoiesis remains obscure. We found that all MAP kinases examined, ERK1, ERK2, p38, JNK1, and JNK2, were rapidly and transiently activated by erythropoietin (Epo) stimulation in SKT6 cells, which can be induced to differentiate into hemoglobinized cells in response to Epo. Furthermore, p38-specific inhibitor SB203580 but not MEK-specific inhibitor PD98059 significantly suppressed Epo-induced differentiation and antisense oligonucleotides of p38, JNK1, and JNK2, but neither ERK1 nor ERK2 clearly inhibited Epo-induced hemoglobinization. However, in Epo-dependent FD-EPO cells, inhibition of either ERKs, p38, or JNKs suppressed cell growth. Furthermore, forced expression of a gain-of-function MKK6 mutant, which specifically activated p38, induced hemoglobinization of SKT6 cells without Epo. These results indicate that activation of p38 and JNKs but not of ERKs is required for Epo-induced erythroid differentiation of SKT6 cells, whereas all of these kinases are involved in Epo-induced mitogenesis of FD-EPO cells. © 1998 by The American Society of Hematology.

Polyamines regulate Rho-kinase and myosin phosphorylation during intestinal epithelial restitution

2003 ◽  
Vol 284 (4) ◽  
pp. C848-C859 ◽  
Author(s):  
Jaladanki N. Rao ◽  
Xin Guo ◽  
Lan Liu ◽  
Tongtong Zou ◽  
Karnam S. Murthy ◽  
...  
Keyword(s):  
Cell Migration ◽  
Kinase Activity ◽  
Rho Kinase ◽  
Specific Inhibitor ◽  
Fiber Formation ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Intestinal Epithelial ◽  
Epithelial Restitution ◽  
Mlc Phosphorylation

Polyamines are required for the early phase of mucosal restitution that occurs as a consequence of epithelial cell migration. Our previous studies have shown that polyamines increase RhoA activity by elevating cytosolic free Ca2+ concentration ([Ca2+]cyt) through controlling voltage-gated K+ channel expression and membrane potential ( E m) during intestinal epithelial restitution. The current study went further to determine whether increased RhoA following elevated [Ca2+]cyt activates Rho-kinase (ROK/ROCK) resulting in myosin light chain (MLC) phosphorylation. Studies were conducted in stable Cdx2-transfected intestinal epithelial cells (IEC-Cdx2L1), which were associated with a highly differentiated phenotype. Reduced [Ca2+]cyt, by either polyamine depletion or exposure to the Ca2+-free medium, decreased RhoA protein expression, which was paralleled by significant decreases in GTP-bound RhoA, ROCK-1, and ROKα proteins, Rho-kinase activity, and MLC phosphorylation. The reduction of [Ca2+]cyt also inhibited cell migration after wounding. Elevation of [Ca2+]cyt induced by the Ca2+ ionophore ionomycin increased GTP-bound RhoA, ROCK-1, and ROKα proteins, Rho-kinase activity, and MLC phosphorylation. Inhibition of RhoA function by a dominant negative mutant RhoA decreased the Rho-kinase activity and resulted in cytoskeletal reorganization. Inhibition of ROK/ROCK activity by the specific inhibitor Y-27632 not only decreased MLC phosphorylation but also suppressed cell migration. These results indicate that increase in GTP-bound RhoA by polyamines via [Ca2+]cytcan interact with and activate Rho-kinase during intestinal epithelial restitution. Activation of Rho-kinase results in increased MLC phosphorylation, leading to the stimulation of myosin stress fiber formation and cell migration.

Activation of p38 MAP Kinase and JNK But Not ERK Is Required for Erythropoietin-Induced Erythroid Differentiation

Blood ◽  
1998 ◽  
Vol 92 (6) ◽  
pp. 1859-1869 ◽  
Author(s):  
Yuka Nagata ◽  
Noriko Takahashi ◽  
Roger J. Davis ◽  
Kazuo Todokoro
Keyword(s):  
Cell Growth ◽  
Map Kinase ◽  
Antisense Oligonucleotides ◽  
Map Kinases ◽  
Physiological Function ◽  
Critical Role ◽  
Specific Inhibitor ◽  
Erythroid Differentiation ◽  
P38 Map Kinase ◽  
American Society

Abstract p38 MAP kinase (p38) and JNK have been described as playing a critical role in the response to a variety of environmental stresses and proinflammatory cytokines. It was recently reported that hematopoietic cytokines activate not only classical MAP kinases (ERK), but also p38 and JNK. However, the physiological function of these kinases in hematopoiesis remains obscure. We found that all MAP kinases examined, ERK1, ERK2, p38, JNK1, and JNK2, were rapidly and transiently activated by erythropoietin (Epo) stimulation in SKT6 cells, which can be induced to differentiate into hemoglobinized cells in response to Epo. Furthermore, p38-specific inhibitor SB203580 but not MEK-specific inhibitor PD98059 significantly suppressed Epo-induced differentiation and antisense oligonucleotides of p38, JNK1, and JNK2, but neither ERK1 nor ERK2 clearly inhibited Epo-induced hemoglobinization. However, in Epo-dependent FD-EPO cells, inhibition of either ERKs, p38, or JNKs suppressed cell growth. Furthermore, forced expression of a gain-of-function MKK6 mutant, which specifically activated p38, induced hemoglobinization of SKT6 cells without Epo. These results indicate that activation of p38 and JNKs but not of ERKs is required for Epo-induced erythroid differentiation of SKT6 cells, whereas all of these kinases are involved in Epo-induced mitogenesis of FD-EPO cells. © 1998 by The American Society of Hematology.

scholarly journals The activation of distinct mitogen-activated protein kinase cascades is required for the stimulation of 2-deoxyglucose uptake by interleukin-1 and insulin-like growth factor-1 in KB cells

1995 ◽  
Vol 311 (3) ◽  
pp. 735-738 ◽  
Author(s):  
G W Gould ◽  
A Cuenda ◽  
F J Thomson ◽  
P Cohen
Keyword(s):  
Growth Factor ◽  
Map Kinase ◽  
Interleukin 1 ◽  
Specific Inhibitor ◽  
Drug Binding ◽  
Insulin Like Growth Factor ◽  
Kb Cells ◽  
Mitogen Activated Protein ◽  
Sb 203580 ◽  
Stimulation Of

The uptake of 2-deoxyglucose into KB cells was stimulated about 2-fold by interleukin-1 (IL1), anisomycin or insulin-like growth factor-1 (IGF1). Stimulation by IL1 and anisomycin was prevented by SB 203580, a specific inhibitor of the mitogen-activated protein (MAP) kinase homologue termed ‘re-activating kinase’ [RK; also known as p38, p40 and CSBP (cytokine synthesis anti-inflammatory-drug-binding protein)], but was unaffected by PD 98059, a specific inhibitor of the activation of the classical MAP kinase pathway. In contrast, the stimulation of 2-deoxyglucose uptake by IGF1 was blocked by PD 98059 and unaffected by SB 203580. Consistent with these observations, IL1 and anisomycin were potent activators of MAP kinase-activated protein (MAPKAP) kinase-2, a physiological substrate of RK, whereas IGF1 was only a very weak activator of MAPKAP kinase-2. Conversely, IGF1 was a stronger activator of p42 MAP kinase than IL1 or anisomycin. These results imply that the activation of distinct MAP kinase pathways is required for the stimulation of glucose transport by IL1/anisomycin and IGF1 in KB cells, and suggest that the combined use of SB 203580 and PD 98059 is a powerful new approach to explore the roles of different MAP kinase cascades in cell regulation.

scholarly journals p38 MAP kinase-dependent regulation of endothelial cell permeability

2004 ◽  
Vol 287 (5) ◽  
pp. L911-L918 ◽  
Author(s):  
Talaibek Borbiev ◽  
Anna Birukova ◽  
Feng Liu ◽  
Saule Nurmukhambetova ◽  
William T. Gerthoffer ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Map Kinase ◽  
Light Chain ◽  
Myosin Light Chain ◽  
P38 Map Kinase ◽  
Fiber Formation ◽  
Dominant Negative ◽  
Cell Permeability ◽  
Endothelial Cell Permeability ◽  
Sb 203580

We have previously shown that thrombin induces endothelial cell barrier dysfunction via cytoskeleton activation and contraction and have determined the important role of endothelial cell myosin light chain kinase (MLCK) in this process. In the present study we explored p38 MAP kinase as a potentially important enzyme in thrombin-mediated endothelial cell contractile response and permeability. Thrombin induces significant p38 MAP kinase activation in a time-dependent manner with maximal effect at 30 min, which correlates with increased phosphorylation of actin- and myosin-binding protein, caldesmon. Both SB-203580 and dominant negative p38 adenoviral vector significantly attenuated thrombin-induced declines in transendothelial electrical resistance. Consistent with these data SB-203580 decreased actin stress fiber formation produced by thrombin in endothelium. In addition, dominant negative p38 had no effect on thrombin-induced myosin light chain diphosphorylation. Thrombin-induced total and site-specific caldesmon phosphorylation (Ser789) as well as dissociation of caldesmon-myosin complex were attenuated by SB-203580 pretreatment. These results suggest the involvement of p38 MAP kinase activities and caldesmon phosphorylation in the MLCK-independent regulation of thrombin-induced endothelial cell permeability.

Role of p120-catenin in the morphological changes of endothelial cells exposed to fluid shear stress

2010 ◽  
Vol 398 (3) ◽  
pp. 426-432 ◽  
Author(s):  
Naoya Sakamoto ◽  
Kei Segawa ◽  
Makoto Kanzaki ◽  
Toshiro Ohashi ◽  
Masaaki Sato
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Fluid Shear Stress ◽  
Morphological Changes ◽  
Fluid Shear ◽  
P120 Catenin

Endothelial Yin Yang 1 Phosphorylation at S118 Induces Atherosclerosis Under Flow

2021 ◽  
Author(s):  
Shu-Yi Wei ◽  
Yu-Tsung Shih ◽  
Hsin-Yi Wu ◽  
Wei-Li Wang ◽  
pei ling lee ◽  
...  
Keyword(s):  
Shear Stress ◽  
Expression Profiles ◽  
Critical Role ◽  
Vascular Endothelial Cell ◽  
Specific Inhibitor ◽  
Direct Interaction ◽  
Disturbed Flow ◽  
Yin Yang 1 ◽  
Yin Yang ◽  
Atherosclerosis Treatment

Rationale: Disturbed flow occurring in arterial branches and curvatures induces vascular endothelial cell (EC) dysfunction and atherosclerosis. We postulated that disturbed flow plays important roles in modulating phosphoprotein expression profiles to regulate endothelial functions and atherogenesis. Objective: The goal of this study is to discover novel site-specific phosphorylation alterations induced by disturbed flow in ECs to contribute to atherosclerosis. Methods and Results: Quantitative phosphoproteomics analysis of ECs exposed to disturbed flow with low and oscillatory shear stress (OS, 0.5plusminus4 dynes/cm 2 ) vs. pulsatile flow with high shear stress (PS, 124plusminus dynes/cm 2 ) revealed that OS induces serine (S)118 phosphorylation of Yin Yang 1 (phospho-YY1 S118 ) in ECs. Elevated phospho-YY1 S118 level in ECs was further confirmed to be present in the disturbed flow regions in experimental animals and human atherosclerotic arteries. This disturbed flow-induced EC phospho-YY1 S118 is mediated by casein kinase 2α (CK2α) through its direct interaction with YY1. Yeast two-hybrid library screening and in situ proximity ligation assays demonstrate that phospho-YY1 S118 directly binds zinc finger with KRAB and SCAN domains 4 (ZKSCAN4) to induce promoter activity and gene expression of human double minute 2 (HDM2), which consequently induces EC proliferation through down-regulations of p53 and p21 CIP1 . Administration of apolipoprotein E-deficient (ApoE -/- ) mice with CK2-specific inhibitor tetrabromocinnamic acid or atorvastatin inhibits atherosclerosis formation through down-regulations of EC phospho-YY1 S118 and HDM2. Generation of novel transgenic mice bearing EC-specific overexpression of S118-non-phosphorylatable mutant of YY1 in ApoE -/- mice confirms the critical role of phospho-YY1 S118 in promoting atherosclerosis through EC HDM2. Conclusions: Our findings provide new insights into the mechanisms by which disturbed flow induces endothelial phospho-YY1 S118 to promote atherosclerosis, thus indicating phospho-YY1 S118 as a potential molecular target for atherosclerosis treatment.

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