Influence of TGF-β1 expression in endothelial cells on smooth muscle cell phenotypes and MMP production under shear stress in a co-culture model
- 16 Downloads
Recently, our group has contrasted an endothelial cell-smooth muscle cell (EC-SMC) co-culture model with 3D-cultured SMCs and found that SMCs could respond to high shear stress (SS), which has not been explored before. SMCs were not directly exposed to the flow but were under an EC monolayer; therefore, it is necessary to explore the influence of EC on SMC behaviors under high SS for understanding the mechanism of SMC response to various magnitudes of SS. In the present study, TGF-β1 expression in ECs in an EC-SMC co-culture model was suppressed by an siRNA transfection method. Next, phenotypic changes were observed and MMP-2 and -9 productions were measured in SMCs in the co-culture model after 72-h flow exposure to different SS levels. We confirmed that TGF-β1 expression in ECs could influence SMC phenotypic change under SS conditions and that TGF-β1 expression in ECs could also change MMP-2 production but not MMP-9 production in SMCs under SS conditions in the co-culture model. These results could be useful for understanding the mechanisms of SMC response to SS, particularly for understanding signal transduction emanating from ECs.
KeywordsCo-culture model Smooth muscle cell Endothelial cell Shear stress TGF-β1 siRNA transfection
The authors would like to thank Dr. Jennifer M. Dolan for technical advices on cell culturing and flow-exposure experiment. We also thank Prof. Takehiko Sato, Prof. Makoto Kanzaki and Dr. Daisuke Yoshino for support during the experiments. This work was partly supported by grants from the JSPS Core-to-Core Program, A. Advanced Research Networks, ‘International research core on smart layered materials and structures for energy saving’ and the Ministry of Education, Science, Sports and Culture, Grant-in-Aid for Scientific Research (B), 2013-2015 (Grant No. 25282140 awarded to Makoto Ohta).
- Adam PJ, Regan CP, Hautmann MB, Owens GK (2000) Positive- and negative-acting Kruppel-like transcription factors bind a transforming growth factor beta control element required for expression of the smooth muscle cell differentiation marker SM22alpha in vivo. J Biol Chem 275:37798–37806CrossRefGoogle Scholar
- Han X, Sakamoto N, Tomita N, Meng H, Sato M, Ohta M (2017). Influence of shear stress on phenotype and MMP production of smooth muscle cells in a co-culture model. J Biorheol (accepted)Google Scholar
- Wang M, Zhao D, Spinetti G, Zhang J, Jiang LQ, Pintus G, Monticone R, Lakatta EG (2006b) Matrix metalloproteinase 2 activation of transforming growth factor-beta1 (TGF-beta1) and TGF-beta1-type II receptor signaling within the aged arterial wall. Arterioscler Thromb Vasc Biol 26:1503–1509CrossRefGoogle Scholar