A model for studying the effect of shear stress on interactions between vascular endothelial cells and smooth muscle cells.
Vascular endothelial cells (ECs) are constantly subjected to blood flow-induced shear stress and the influences of neighboring smooth muscle cells (SMCs). In the present study, a coculture flow system was developed to study the effect of shear stress on EC-SMC interactions. ECs and SMCs were separated by a porous membrane with only the EC side subjected to the flow condition. When ECs were exposed to a shear stress of 12 dynes/cm2 for 24 h, the cocultured SMCs tended to orient perpendicularly to the flow direction. This perpendicular orientation of the cocultured SMCs to flow direction was not observed when ECs were exposed to a shear stress of 2 dynes/cm2. Under the static condition, long and parallel actin bundles were observed in the central regions of the cocultured SMCs, whereas the actin filaments localized mainly at the periphery of the cocultured ECs. After 24 h of flow application, the cocultured ECs displayed very long, well-organized, parallel actin stress fibers aligned with the flow direction in the central regions of the cells. Immunostaining of platelet endothelial cell adhesion molecule-1 confirmed the elongation and alignment of the cocultured ECs with the flow direction. Coculture with SMCs under static condition induced EC gene expressions of growth-related oncogene-alpha and monocyte chemotactic protein-1, and shear stress was found to abolish these SMC-induced gene expressions. Our results suggest that shear stress may serve as a down-regulator for the pathophysiologically relevant gene expression in ECs cocultured with SMCs.
Division of Medical Engineering Research, National Health Research Institutes, Rm 8324, 8F, No. 161, Sec. 6, Min-Chuan East Road, 114 Taipei, Taiwan, ROC. jjchiu@nhri.org.tw
Jeng-Jiann
JJ
Chiu
Li-Jing
LJ
Chen
Cheng-Nan
CN
Chen
Pei-Ling
PL
Lee
Chih-I
CI
Lee
D008841:Actin Cytoskeleton / Q000502:physiology; D018932:Chemokine CCL2 / Q000037:antagonists & inhibitors / Q000235:genetics; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D007158:Immunologic Techniques; D009131:Muscle, Smooth, Vascular / Q000378:metabolism / Q000502:physiology*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000502:physiology*; D009857:Oncogenes / Q000235:genetics; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013194:Staining and Labeling; D013314:Stress, Mechanical; D013997:Time Factors
D018932:Chemokine CCL2; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D012333:RNA, Messenger