Liuwei Dihuang, a traditional Chinese medicinal formula, inhibits proliferation and migration of vascular smooth muscle cells via modulation of estrogen receptors

Zhang,Yayun; Qian,Xing; Sun,Xin; Lin,Chao; Jing,Yi; Yao,Yuan; Ma,Zhi; Kuai,Meiyu; Lu,Ying; Kong,Xueyun; Chen,Qi; Wu,Xiang; Zhao,Xuan; Li,Yu; Bian,Huimin

doi:10.3892/ijmm.2018.3622

July-2018 Volume 42 Issue 1

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July-2018 Volume 42 Issue 1

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Liuwei Dihuang, a traditional Chinese medicinal formula, inhibits proliferation and migration of vascular smooth muscle cells via modulation of estrogen receptors

Authors:
- Yayun Zhang
- Xing Qian
- Xin Sun
- Chao Lin
- Yi Jing
- Yuan Yao
- Zhi Ma
- Meiyu Kuai
- Ying Lu
- Xueyun Kong
- Qi Chen
- Xiang Wu
- Xuan Zhao
- Yu Li
- Huimin Bian
View Affiliations / Copyright

Affiliations: School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, P.R. China
Pages: 31-40
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Published online on: April 16, 2018

https://doi.org/10.3892/ijmm.2018.3622
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Abstract

The phenotypic modulation of vascular smooth muscle cells (VSMCs) serves an important role in atherosclerosis‑induced vascular alterations, including vascular remodeling. However, the precise mechanisms underlying VSMC phenotypic modulation remain to be elucidated. Our previous study demonstrated that Liuwei Dihuang formula (LWDHF) could improve menopausal atherosclerosis by upregulating the expression of estrogen receptors (ERs). The present study examined the role of ERs in the effects of LWDHF on VSMC phenotypic modulation. VSMC proliferation and cell cycle progression were examined by MTT assay and flow cytometry, respectively. The expression levels of α‑smooth muscle actin, osteopontin and ERs were determined using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis. Cell ultrastructure was observed under an electron microscope. F‑actin polymerization was detected by fluorescein isothiocyanate‑phalloidin staining using fluorescence microscopy. A modified Boyden chamber assay was employed to assess VSMCs migration. Small interfering (si)RNA technology was used to examine the role of ERα in the effects of LWDHF on phenotypic modulation. The results indicated that LWDHF (3‑12 µg/ml) inhibited proliferation and induced a cell cycle arrest in VSMCs treated with angiotensin II (Ang II; 100 nM) in a concentration‑dependent manner. In addition, Ang II‑stimulated migration of VSMCs and reorganization of actin were markedly inhibited by treatment with 12 µg/ml LWDHF. Results of RT‑qPCR and western blotting demonstrated that LWDHF markedly stimulated transcription and expression of ERα and ERβ, and inhibited VSMC synthetic phenotype. Furthermore, LWDHF‑induced inhibition of phenotypic switching was partially suppressed by tamoxifen, and transfection with ERα siRNA markedly abolished the effects of LWDHF on VSMC phenotypic switching. In conclusion, these results revealed that ERα served an important role in LWDHF‑induced regulation of the VSMC phenotype, including proliferation and migration.

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1	Rivard A and Andrés V: Vascular smooth muscle cell proliferation in the pathogenesis of atherosclerotic cardiovascular diseases. Histol Histopathol. 15:557–571. 2000.PubMed/NCBI
2	Lacolley P, Regnault V, Nicoletti A, Li Z and Michel JB: The vascular smooth muscle cell in arterial pathology: a cell that can take on multiple roles. Cardiovasc Res. 95:194–204. 2012. View Article : Google Scholar : PubMed/NCBI
3	Deatrick KB, Eliason JL, Lynch EM, Moore AJ, Dewyer NA, Varma MR, Pearce CG, Upchurch GR Jr, Wakefield TW and Henke PK: Vein wall remodeling after deep vein thrombosis involves matrix metalloproteinases and late fibrosis in a mouse model. J Vasc Surg. 42:140–148. 2005. View Article : Google Scholar : PubMed/NCBI
4	Owens GK, Kumar MS and Wamhoff BR: Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev. 84:767–801. 2004. View Article : Google Scholar : PubMed/NCBI
5	Rensen SS, Doevendans PA and van Eys GJ: Regulation and characteristics of vascular smooth muscle cell phenotypic diversity. Neth Heart J. 15:100–108. 2007. View Article : Google Scholar : PubMed/NCBI
6	Schwartz SM: Perspectives series: cell adhesion in vascular biology. Smooth muscle migration in atherosclerosis and restenosis. J Clin Invest. 99:2814–2816. 1997. View Article : Google Scholar : PubMed/NCBI
7	Doran AC, Meller N and McNamara CA: Role of smooth muscle cells in the initiation and early progression of atherosclerosis. Arterioscler Thromb Vasc Biol. 28:812–819. 2008. View Article : Google Scholar : PubMed/NCBI
8	Hao H, Gabbiani G and Bochaton-Piallat ML: Arterial smooth muscle cell heterogeneity: implications for atherosclerosis and restenosis development. Arterioscler Thromb Vasc Biol. 23:1510–1520. 2003. View Article : Google Scholar : PubMed/NCBI
9	Babapulle MN and Eisenberg MJ: Coated stents for the prevention of restenosis: Part I. Circulation. 106:2734–2740. 2002. View Article : Google Scholar : PubMed/NCBI
10	Zhao Y, Liu YX, Xie SL, Deng BQ, Wang JF and Nie RQ: Increased expression of granulocyte colony-stimulating factor mediates mesenchymal stem cells recruitment after vascular injury. Chin Med J (Engl). 124:4286–4292. 2011.
11	Watanabe T, Pakala R, Katagiri T and Benedict CR: Synergistic effect of urotensin II with mildly oxidized LDL on DNA synthesis in vascular smooth muscle cells. Circulation. 104:16–18. 2001. View Article : Google Scholar : PubMed/NCBI
12	Intengan HD and Schiffrin EL: Vascular remodeling in hypertension: roles of apoptosis, inflammation, and fibrosis. Hypertension. 38:581–587. 2001. View Article : Google Scholar : PubMed/NCBI
13	Liu JP, Feng L, Zhang MH, Ma DY, Wang SY, Gu J, Fu Q, Qu R and Ma SP: Neuroprotective effect of Liuwei Dihuang decoction on cognition deficits of diabetic encephalopathy in streptozotocin-induced diabetic rat. J Ethnopharmacol. 150:371–381. 2013. View Article : Google Scholar : PubMed/NCBI
14	Kalu DN: The ovariectomized rat model of postmenopausal bone loss. Bone Miner. 15:175–191. 1991. View Article : Google Scholar : PubMed/NCBI
15	Wu Y, L Y and Zhang QC: The effect of LWDHF on menopausal atherosclerosis model in ovariectomized rats. Chinese Traditional Patent Medicine. 34:553–556. 2012.In Chinese.
16	Yin QY, Guo J and Meng QH: Effects of Liuweidihuang formula mediated serum on H₂O₂ -injured human umbilical vascular endothelial cells. Chin Pharmacol Bull. 29:1753–1757. 2013.In Chinese.
17	Mendelsohn ME and Karas RH: Molecular and cellular basis of cardiovascular gender differences. Science. 308:1583–1587. 2005. View Article : Google Scholar : PubMed/NCBI
18	Farhat MY, Lavigne MC and Ramwell PW: The vascular protective effects of estrogen. FASEB J. 10:615–624. 1996. View Article : Google Scholar : PubMed/NCBI
19	Kawagoe J, Ohmichi M, Tsutsumi S, Ohta T, Takahashi K and Kurachi H: Mechanism of the divergent effects of estrogen on the cell proliferation of human umbilical endothelial versus aortic smooth muscle cells. Endocrinology. 148:6092–6099. 2007. View Article : Google Scholar : PubMed/NCBI
20	Sivritas D, Becher MU, Ebrahimian T, Arfa O, Rapp S, Bohner A, Mueller CF, Umemura T, Wassmann S, Nickenig G, et al: Antiproliferative effect of estrogen in vascular smooth muscle cells is mediated by Kruppel-like factor-4 and manganese superoxide dismutase. Basic Res Cardiol. 106:563–575. 2011. View Article : Google Scholar : PubMed/NCBI
21	Liu HM, Zhao XF, Guo LN, Tan Z and Wang TH: Effects of caveolin-1 on the 17β-estradiol-mediated inhibition of VSMC proliferation induced by vascular injury. Life Sci. 80:800–812. 2007. View Article : Google Scholar
22	Yang S, Zhou W, Zhang Y, Yan C and Zhao Y: Effects of Liuwei Dihuang decoction on ion channels and synaptic transmission in cultured hippocampal neuron of rat. J Ethnopharmacol. 106:166–172. 2006. View Article : Google Scholar : PubMed/NCBI
23	Rodríguez A, Fortuño A, Gómez-Ambrosi J, Zalba G, Díez J and Frühbeck G: The inhibitory effect of leptin on angiotensin II-induced vasoconstriction in vascular smooth muscle cells is mediated via a nitric oxide-dependent mechanism. Endocrinology. 148:324–331. 2007. View Article : Google Scholar
24	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Method Methods. 25:402–408. 2001. View Article : Google Scholar
25	Hedin U and Thyberg J: Plasma fibronectin promotes modulation of arterial smooth-muscle cells from contractile to synthetic phenotype. Differentiation. 33:239–246. 1987. View Article : Google Scholar : PubMed/NCBI
26	Rousseau S, Houle F, Landry J and Huot J: p38 MAP kinase activation by vascular endothelial growth factor mediates actin reorganization and cell migration in human endothelial cells. Oncogene. 15:2169–2177. 1997. View Article : Google Scholar : PubMed/NCBI
27	Gunst SJ and Zhang W: Actin cytoskeletal dynamics in smooth muscle: a new paradigm for the regulation of smooth muscle contraction. Am J Physiol Cell Physiol. 295:C576–C587. 2008. View Article : Google Scholar : PubMed/NCBI
28	Shanahan CM, Weissberg PL and Metcalfe JC: Isolation of gene markers of differentiated and proliferating vascular smooth muscle cells. Circ Res. 73:193–204. 1993. View Article : Google Scholar : PubMed/NCBI
29	Hultgårdh-Nilsson A, Lövdahl C, Blomgren K, Kallin B and Thyberg J: Expression of phenotype-and proliferation-related genes in rat aortic smooth muscle cells in primary culture. Cardiovascular research. 34:418–430. 1997. View Article : Google Scholar

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Liuwei Dihuang, a traditional Chinese medicinal formula, inhibits proliferation and migration of vascular smooth muscle cells via modulation of estrogen receptors

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