Abstract
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.
Similar content being viewed by others
References
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 275:37798–37806
Andres V (1998) Control of vascular smooth muscle cell growth and its implication in atherosclerosis and restenosis (review). Int J Mol Med 2:81–89
Chiu JJ, Chen LJ, Chen CN, Lee PL, Lee CI (2004) A model for studying the effect of shear stress on interactions between vascular endothelial cells and smooth muscle cells. J Biomech 37:531–539
Cucina A, Sterpetti AV, Borrelli V, Pagliei S, Cavallaro A, D’Angelo LS (1998) Shear stress induces transforming growth factor-beta 1 release by arterial endothelial cells. Surgery 123:212–217
Davenport C, Harper E, Forde H, Rochfort KD, Murphy RP, Smith D, Cummins PM (2016) RANKL promotes osteoblastic activity in vascular smooth muscle cells by upregulating endothelial BMP-2 release. Int J Biochem Cell Biol 77:171–180
Davies PF, Truskey GA, Warren HB, O’Connor SE, Eisenhaure BH (1985) Metabolic cooperation between vascular endothelial cells and smooth muscle cells in co-culture: changes in low density lipoprotein metabolism. J Cell Biol 101:871–879
Frösen J (2014) Smooth muscle cells and the formation, degeneration, and rupture of saccular intracranial aneurysm wall–a review of current pathophysiological knowledge. Transl Stroke Res 5:347–356
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)
Isakson BE, Duling BR (2005) Heterocellular contact at the myoendothelial junction influences gap junction organization. Circ Res 97:44–51
Nakazawa T, Yasuhara H, Shigematsu K, Shigematsu H (2000) Smooth muscle cell migration induced by shear-loaded platelets and endothelial cells. Enhanced platelet-derived growth factor production by shear-loaded platelets. Int Angiol 19:142–146
Penn DL, Witte SR, Komotar RJ, Sander Connolly E Jr (2014) The role of vascular remodeling and inflammation in the pathogenesis of intracranial aneurysms. J Clin Neurosci 21:28–32
Powell RJ, Bhargava J, Basson MD, Sumpio BE (1998) Coculture conditions alter endothelial modulation of TGF-beta 1 activation and smooth muscle growth morphology. Am J Physiol 274:H642–H649
Qi YX, Jiang J, Jiang XH, Wang XD, Ji SY, Han Y, Long DK, Shen BR, Yan ZQ, Chien S, Jiang ZL (2011) PDGF-BB and TGF-{beta}1 on cross-talk between endothelial and smooth muscle cells in vascular remodeling induced by low shear stress. Proc Natl Acad Sci USA 108:1908–1913
Sakamoto N, Kiuchi T, Sato M (2011) Development of an endothelial-smooth muscle cell coculture model using phenotype-controlled smooth muscle cells. Ann Biomed Eng 39:2750–2758
Starke RM, Chalouhi N, Ding D, Raper DM, Mckisic MS, Owens GK, Hasan DM, Medel R, Dumont AS (2014) Vascular smooth muscle cells in cerebral aneurysm pathogenesis. Transl Stroke Res 5:338–346
Tabas I (2017) 2016 Russell ross memorial lecture in vascular biology: molecular-cellular mechanisms in the progression of atherosclerosis. Arterioscler Thromb Vasc Biol 37:183–189
Uchida C, Haas TL (2013) Endothelial cell TIMP-1 is upregulated by shear stress via Sp-1 and the TGFβ1 signaling pathways. Biochem Cell Biol 92:77–83
Walshe TE, dela Paz NG, D’Amore PA (2013) The role of shear-induced transforming growth factor-β signaling in the endothelium. Arterioscler Thromb Vasc Biol 33:2608–2617
Wang HQ, Huang LX, Qu MJ, Yan ZQ, Liu B, Shen BR, Jiang ZL (2006a) Shear stress protects against endothelial regulation of vascular smooth muscle cell migration in a coculture system. Endothelium 13:171–180
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 26:1503–1509
Wang L, Han Y, Shen Y, Yan ZQ, Zhang P, Yao QP, Shen BR, Gao LZ, Qi YX, Jiang ZL (2014) Endothelial insulin-like growth factor-1 modulates proliferation and phenotype of smooth muscle cells induced by low shear stress. Ann Biomed Eng 42:776–786
Wu S, Wu X, Zhu W, Cai WJ, Schaper J, Schaper W (2010) Immunohistochemical study of the growth factors, aFGF, bFGF, PDGF-AB, VEGF-A and its receptor (Flk-1) during arteriogenesis. Mol Cell Biochem 343:223–229
Xu S, He Y, Vokurkova M, Touyz RM (2009) Endothelial cells negatively modulate reactive oxygen species generation in vascular smooth muscle cells: role of thioredoxin. Hypertension 54:427–433
Acknowledgements
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).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Han, X., Sakamoto, N., Tomita, N. et al. Influence of TGF-β1 expression in endothelial cells on smooth muscle cell phenotypes and MMP production under shear stress in a co-culture model. Cytotechnology 71, 489–496 (2019). https://doi.org/10.1007/s10616-018-0268-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10616-018-0268-7