Advertisement

Human Cell

, Volume 31, Issue 3, pp 242–250 | Cite as

Acetylshikonin attenuates angiotensin II-induced proliferation and motility of human brain smooth muscle cells by inhibiting Wnt/β-catenin signaling

  • Zequn Li
  • Zhiyuan Yan
  • Chunbo Xu
  • Yiqun Dong
  • Ye Xiong
  • Yongyue Dai
Research Article
  • 31 Downloads

Abstract

Cerebrovascular smooth muscle cell proliferation and migration contribute to hyperplasia in case of cerebrovascular remodeling and stroke. In the present study, we investigated the effects of acetylshikonin, the main ingredient of a Chinese traditional medicine Zicao, on human brain vascular smooth muscle cell (HBVSMCs) proliferation and migration induced by angiotensin II (AngII), and the underlying mechanisms. We found that acetylshikonin treatment significantly inhibited AngII-induced HBVSMCs proliferation and cell cycle transition from G1 to S phase. Wound-healing assay and Transwell assay showed that AngII-induced cell migration and invasion were markedly attenuated by acetylshikonin. In addition, AngII challenge significantly induced Wnt/β-catenin signaling activation, as evidenced by increased β-catenin phosphorylation and nuclear translocation and GSK-3β phosphorylation. However, acetylshikonin treatment inhibited the activation of Wnt/β-catenin signaling. Consequently, western blotting analysis revealed that acetylshikonin effectively reduced the expression of downstream target genes in AngII-treated cells, including c-myc, survivin and cyclin D1, which contributed to the inhibitory effect of acetylshikonin on HBVSMCs proliferation. Further, stimulation with recombinant Wnt3a dramatically reversed acetylshikonin-mediated inhibition of proliferation and cell cycle transition in HBVSMCs. Our study demonstrates that acetylshikonin prevents AngII-induced cerebrovascular smooth muscle cells proliferation and migration through inhibition of Wnt/β-catenin pathway, indicating that acetylshikonin may present a potential option for the treatment of cerebrovascular remodeling.

Keywords

Cerebrovascular smooth muscle cells Proliferation Angiotensin II Acetylshikonin Wnt/β-catenin pathway 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Lemarie CA, Tharaux PL, Lehoux S. Extracellular matrix alterations in hypertensive vascular remodeling. J Mol Cell Cardiol. 2010;48(3):433–9.  https://doi.org/10.1016/j.yjmcc.2009.09.018.CrossRefPubMedGoogle Scholar
  2. 2.
    Yu ZL, Wang JN, Wu XH, Xie HJ, Han Y, Guan YT, Qin Y, Jiang JM. Tanshinone IIA prevents rat basilar artery smooth muscle cells proliferation by inactivation of PDK1 during the development of hypertension. J Cardiovasc Pharmacol Ther. 2015;20(6):563–71.  https://doi.org/10.1177/1074248415574743.CrossRefPubMedGoogle Scholar
  3. 3.
    Mancia G, Messerli F, Bakris G, Zhou Q, Champion A, Pepine CJ. Blood pressure control and improved cardiovascular outcomes in the International Verapamil SR-Trandolapril Study. Hypertension. 2007;50(2):299–305.  https://doi.org/10.1161/HYPERTENSIONAHA.107.090290.CrossRefPubMedGoogle Scholar
  4. 4.
    Bihl JC, Zhang C, Zhao Y, Xiao X, Ma X, Chen Y, Chen S, Zhao B, Chen Y. Angiotensin-(1-7) counteracts the effects of Ang II on vascular smooth muscle cells, vascular remodeling and hemorrhagic stroke: role of the NFsmall ka, CyrillicB inflammatory pathway. Vascul Pharmacol. 2015;73:115–23.  https://doi.org/10.1016/j.vph.2015.08.007.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Chan SH, Chan JY. Angiotensin-generated reactive oxygen species in brain and pathogenesis of cardiovascular diseases. Antioxid Redox Signal. 2013;19(10):1074–84.  https://doi.org/10.1089/ars.2012.4585.CrossRefPubMedGoogle Scholar
  6. 6.
    Wang XM, Xiao H, Liu LL, Cheng D, Li XJ, Si LY. FGF21 represses cerebrovascular aging via improving mitochondrial biogenesis and inhibiting p53 signaling pathway in an AMPK-dependent manner. Exp Cell Res. 2016;346(2):147–56.  https://doi.org/10.1016/j.yexcr.2016.06.020.CrossRefPubMedGoogle Scholar
  7. 7.
    Wang Y, Ma TT, Gao NN, Zhou XL, Jiang H, Guo R, Jia LN, Chang H, Gao Y, Gao ZM, Pan L. Effect of Tongxinluo on pulmonary hypertension and pulmonary vascular remodeling in rats exposed to a low pressure hypoxic environment. J Ethnopharmacol. 2016;194:668–73.  https://doi.org/10.1016/j.jep.2016.10.004.CrossRefPubMedGoogle Scholar
  8. 8.
    Tsai FJ, Ho TJ, Cheng CF, Liu X, Tsang H, Lin TH, Liao CC, Huang SM, Li JP, Lin CW, Lin JG, Lin JC, Lin CC, Liang WM, Lin YJ. Effect of Chinese herbal medicine on stroke patients with type 2 diabetes. J Ethnopharmacol. 2017;200:31–44.  https://doi.org/10.1016/j.jep.2017.02.024.CrossRefPubMedGoogle Scholar
  9. 9.
    Chen X, Yang L, Oppenheim JJ, Howard MZ. Cellular pharmacology studies of shikonin derivatives. PTR. 2002;16(3):199–209.  https://doi.org/10.1002/ptr.1100.PubMedGoogle Scholar
  10. 10.
    Li Q, Zeng J, Su M, He Y, Zhu B. Acetylshikonin from Zicao attenuates cognitive impairment and hippocampus senescence in d-galactose-induced aging mouse model via upregulating the expression of SIRT1. Brain Res Bull. 2018;137:311–8.  https://doi.org/10.1016/j.brainresbull.2018.01.007.CrossRefPubMedGoogle Scholar
  11. 11.
    Su ML, He Y, Li QS, Zhu BH. Efficacy of acetylshikonin in preventing obesity and hepatic steatosis in db/db mice. Molecules. 2016.  https://doi.org/10.3390/molecules21080976.Google Scholar
  12. 12.
    Su M, Huang W, Zhu B. Acetylshikonin from zicao prevents obesity in rats on a high-fat diet by inhibiting lipid accumulation and inducing lipolysis. PLoS One. 2016;11(1):e0146884.  https://doi.org/10.1371/journal.pone.0146884.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Zhang X, Hu W, Wu F, Yuan X, Hu J. Shikonin inhibits TNF-alpha-induced growth and invasion of rat aortic vascular smooth muscle cells. Can J Physiol Pharmacol. 2015;93(8):615–24.CrossRefPubMedGoogle Scholar
  14. 14.
    Zhang ZQ, Cao XC, Zhang L, Zhu WL. Effect of shikonin, a phytocompound from Lithospermum erythrorhizon, on rat vascular smooth muscle cells proliferation and apoptosis in vitro. Zhonghua Yi Xue Za Zhi. 2005;85(21):1484–8.PubMedGoogle Scholar
  15. 15.
    Rao TP, Kuhl M. An updated overview on Wnt signaling pathways: a prelude for more. Circ Res. 2010;106(12):1798–806.  https://doi.org/10.1161/CIRCRESAHA.110.219840.CrossRefPubMedGoogle Scholar
  16. 16.
    Tsaousi A, Mill C, George SJ. The Wnt pathways in vascular disease: lessons from vascular development. Curr Opin Lipidol. 2011;22(5):350–7.  https://doi.org/10.1097/MOL.0b013e32834aa701.CrossRefPubMedGoogle Scholar
  17. 17.
    Clevers H, Nusse R. Wnt/beta-catenin signaling and disease. Cell. 2012;149(6):1192–205.  https://doi.org/10.1016/j.cell.2012.05.012.CrossRefPubMedGoogle Scholar
  18. 18.
    Handeli S, Simon JA. A small-molecule inhibitor of Tcf/beta-catenin signaling down-regulates PPARgamma and PPARdelta activities. Mol Cancer Ther. 2008;7(3):521–9.  https://doi.org/10.1158/1535-7163.MCT-07-2063.CrossRefPubMedGoogle Scholar
  19. 19.
    Bao XL, Song H, Chen Z, Tang X. Wnt3a promotes epithelial-mesenchymal transition, migration, and proliferation of lens epithelial cells. Mol Vis. 2012;18:1983–90.PubMedPubMedCentralGoogle Scholar
  20. 20.
    Marchand A, Atassi F, Gaaya A, Leprince P, Le Feuvre C, Soubrier F, Lompre AM, Nadaud S. The Wnt/beta-catenin pathway is activated during advanced arterial aging in humans. Aging Cell. 2011;10(2):220–32.  https://doi.org/10.1111/j.1474-9726.2010.00661.x.CrossRefPubMedGoogle Scholar
  21. 21.
    Wu X, Liu W, Jiang H, Chen J, Wang J, Zhu R, Li B. Kindlin-2 siRNA inhibits vascular smooth muscle cell proliferation, migration and intimal hyperplasia via Wnt signaling. Int J Mol Med. 2016;37(2):436–44.  https://doi.org/10.3892/ijmm.2015.2429.CrossRefPubMedGoogle Scholar
  22. 22.
    Cui M, Cai Z, Chu S, Sun Z, Wang X, Hu L, Yi J, Shen L, He B. Orphan nuclear receptor Nur77 inhibits angiotensin II-induced vascular remodeling via downregulation of beta-catenin. Hypertension. 2016;67(1):153–62.  https://doi.org/10.1161/HYPERTENSIONAHA.115.06114.CrossRefPubMedGoogle Scholar
  23. 23.
    Hua JY, He YZ, Xu Y, Jiang XH, Ye W, Pan ZM. Emodin prevents intima thickness via Wnt4/Dvl-1/beta-catenin signaling pathway mediated by miR-126 in balloon-injured carotid artery rats. Exp Mol Med. 2015;47:e170.  https://doi.org/10.1038/emm.2015.36.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Tsaousi A, Williams H, Lyon CA, Taylor V, Swain A, Johnson JL, George SJ. Wnt4/beta-catenin signaling induces VSMC proliferation and is associated with intimal thickening. Circ Res. 2011;108(4):427–36.  https://doi.org/10.1161/CIRCRESAHA.110.233999.CrossRefPubMedGoogle Scholar
  25. 25.
    Novellasdemunt L, Antas P, Li VS. Targeting Wnt signaling in colorectal cancer. A review in the theme: cell signaling: proteins, pathways and mechanisms. Am J Physiol Cell Physiol. 2015;309(8):C511–21.  https://doi.org/10.1152/ajpcell.00117.2015.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    He TC, Sparks AB, Rago C, Hermeking H, Zawel L, da Costa LT, Morin PJ, Vogelstein B, Kinzler KW. Identification of c-MYC as a target of the APC pathway. Science. 1998;281(5382):1509–12.CrossRefPubMedGoogle Scholar
  27. 27.
    Altieri DC. Survivin, cancer networks and pathway-directed drug discovery. Nat Rev Cancer. 2008;8(1):61–70.  https://doi.org/10.1038/nrc2293.CrossRefPubMedGoogle Scholar
  28. 28.
    Chiron D, Martin P, Di Liberto M, Huang X, Ely S, Lannutti BJ, Leonard JP, Mason CE, Chen-Kiang S. Induction of prolonged early G1 arrest by CDK4/CDK6 inhibition reprograms lymphoma cells for durable PI3Kdelta inhibition through PIK3IP1. Cell Cycle. 2013;12(12):1892–900.  https://doi.org/10.4161/cc.24928.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Wang X, Xiao Y, Mou Y, Zhao Y, Blankesteijn WM, Hall JL. A role for the beta-catenin/T-cell factor signaling cascade in vascular remodeling. Circ Res. 2002;90(3):340–7.CrossRefPubMedGoogle Scholar

Copyright information

© Japan Human Cell Society and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  • Zequn Li
    • 1
  • Zhiyuan Yan
    • 1
  • Chunbo Xu
    • 1
  • Yiqun Dong
    • 1
  • Ye Xiong
    • 1
  • Yongyue Dai
    • 2
  1. 1.Department of Neurosurgery, First Affiliated Hospital of Wenzhou Medical UniversityWenzhou Medical UniversityWenzhouChina
  2. 2.Department of PathophysiologyWenzhou Medical UniversityWenzhouChina

Personalised recommendations