Mechanistic study of angiogenesis induced by shh-transfected BMMSC transplantation after myocardial infarction


Objective: To investigate the changes and mechanism of angiogenesis in myocardium induced by transplantation of the sonic hedgehog (shh) gene transfected in bone marrow mesenchymal stem cells (BMMSC) after myocardial infarction. Methods: A rat model of acute myocardial infarction was made by coronary artery ligation. The rats were randomly divided into five groups of 40 rats each. These were further subdivided into groups of 10 rats. The peripheral regions of the infarcts were injected with either BMMSCSHH (transfection group), equivalent BMMSC (cell only group), BMMSC and pcDNA3.1-Shh DNA mixture (mixture group), pcDNA3.1-shh DNA alone (gene only group), or equal volumes of low-sugar DMEM medium (control group). One, two, four, and eight weeks after transplantation, specimens were harvested from the transplantation site to determine the expression of SHH signaling pathway downstream genes Ptc1, Gli-2, COUP-TF II, angiogenesis promoting factor VEGF, and Ang-1 using RT-PCR. Results: Seven days after transplantation, the expression of SHH signaling pathway downstream genes, Ptc1, Gli-2, and COUP-TF II was significantly more pronounced in the transfection group than in the control group, cell only group, gene only group, or mixture group (Ptc1: P < 0.01, P < 0.01, P < 0.05, and P < 0.05, respectively; COUP-TF II: P < 0.01, P < 0.01, P < 0.05, and P < 0.05, respectively; Gli-2: P < 0.01, P < 0.01, P < 0.05, and P < 0.05, respectively). The expression of angiogenesis-promoting genes Vegf and Ang-1 was significantly more pronounced than in the control group, cell only group, or gene only group (Vegf: P < 0.01, P < 0.05, P < 0.05 respectively; Ang-1: P < 0.01, P < 0.05, and P < 0.05, respectively). Conclusion: Transplantation of the shh-gene-transfected BMMSCs to the peripheral regions of myocardial infarcts promoted angiogenesis by upregulating downstream gene expression.

Share and Cite:

Song, F. , Wu, M. , Tang, T. , Wu, X. , Zhou, W. , Shi, J. , Zhang, W. , Tan, Z. and Yang, J. (2013) Mechanistic study of angiogenesis induced by shh-transfected BMMSC transplantation after myocardial infarction. Stem Cell Discovery, 3, 83-89. doi: 10.4236/scd.2013.32013.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Siminiak, T. and Kurpisz, M. (2003) Myocardial replacement therapy. Circulation, 108, 1167-1171. doi:10.1161/01.CIR.0000086628.42652.8D
[2] Tomita, S., Li, R.K., Weisel, R.D., Mickle, D.A., Kim, E.J., Sakai, T., et al. (1999) Autologous transplantation of bone marrow cells improves damaged heart function. Circulation, 100, II-247-II-256.
[3] Wang, J.S., Slum-Tim, D., Galipeau, J., Chedrawy, E., Eliopoulos, N. and Chiu, R.C. (2000) Marrow stromal cells for cellular cardiomyoplasty: Feasibility and potential clinical advantages. The Journal of Thoracic and Cardiovascular Surgery, 120, 999-1005. doi:10.1067/mtc.2000.110250
[4] Pola, R., Ling, L.E., Aprahamian, T.R., Barban, E., Bosch-Marce, M., Curry, C., et al. (2003) Postnatal recapitulation of embryonic hedgehog pathway in response to skeletal muscle ischemia. Circulation, 108, 479-485. doi:10.1161/01.CIR.0000080338.60981.FA
[5] Pola, R., Ling, L.E., Silver, M., Corbley, M.J., Kearney, M., Blake, P.R., et al. (2001) The morphogen sonic hedgehog is an indirect angiogenic agent upent upregulating two families of angiogenic growth factors. Nature Medicine, 7, 706-711. doi:10.1038/89083
[6] Kusano, K.F., Pola, R., Murayama, T., Curry, C., Kawamoto, A., Iwakura, A., et al. (2005) Sonic hedgehog myocardial gene therapy: Tissue repair through transient reconstitution of embryonic signaling. Nature Medicine, 11, 1197-1204. doi:10.1038/nm1313
[7] Kusano, K.F., Allendoerfer, K.L., Munger, W., Pola, R., Bosch-Marce, M., Kirchmair, R., et al. (2004) Sonic hedgehog induces arteriogenesis in diabetic vasa nervorum and restores function in diabetic neuropathy. Arteriosclerosis, Thrombosis, and Vascular Biology, 24, 1-7. doi:10.1161/01.ATV.0000144813.44650.75
[8] Cohen Jr., M. M. (2003) The hedgehog signaling network. American Journal of Medical Genetics, 123, 5-28. doi:10.1002/ajmg.a.20495
[9] Krishnan, V., Pereira, F.A., Qiu, Y., Chen, C.H., Beachy, P.A., Tsai, S.Y., et al. (1997) Mediation of sonic hedgehog-induced expression of COUP-TF II by a protein phosphatase. Science, 278, 1947-1950.
[10] Sisson, J.C., Ho, K.S., Suyama, K. and Scott, M.P. (1997) Costal2, a novel kinesin-related protein in the hedgehog signaling pathway. Cell, 90, 235-245. doi:10.1016/S0092-8674(00)80332-3
[11] Robbins, D.J., Nybakken, K.E., Kobayashi, R., Sisson, J.C., Bishop, J.M. and Thérond, P.P. (1997) Hedgehog elicits signal transduction by means of a large complex containing the kinesis-related protein costal2. Cell, 90, 225-234. doi:10.1016/S0092-8674(00)80331-1
[12] Marigo, V., Johnson, R.L., Vortkamp, A. and Tabin, C.J. (1996) Sonic hedgehog differentially regulates expression of GLI and GLI3 during limb development. Developmental Biology; 180, 273-283. doi:10.1006/dbio.1996.0300
[13] Marigo, V. and Tabin, C. (1996) Regulation of patched by sonic hedgehog in the developing neural tube. Proceedings of the National Academy of Sciences of the United States of America, 93, 9346-9351. doi:10.1073/pnas.93.18.9346
[14] Kinnaird, T., Stabile, E., Burnett, M.S., Lee, C.W., Barr, S., Fuchs, S., et al. (2004) Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms. Circulation Research, 94, 678-685. doi:10.1161/01.RES.0000118601.37875.AC
[15] Ziegelhoeffer, T., Fernandez, B., Kostin, S., Heil, M., Voswinckel, R., Helisch, A., et al. (2004) Bone marrow-derived cells do not incorporate into adult growing vasculature. Circulation Research, 94, 230-238. doi:10.1161/01.RES.0000110419.50982.1C

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.