Simvastatin induces the apoptosis of normal vascular smooth muscle through the disruption of actin integrity via the impairment of RhoA/ Rac-1 activity

Seojin Kang; Keunyoung Kim; Ji-Yoon Noh; Yeryeon Jung; Ok-Nam Bae; Kyung-Min Lim; Jin-Ho Chung

doi:10.1160/TH15-11-0858

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2016; 116(03): 496-505
DOI: 10.1160/TH15-11-0858

Cellular Haemostasis and Platelets

Schattauer GmbH

Simvastatin induces the apoptosis of normal vascular smooth muscle through the disruption of actin integrity via the impairment of RhoA/ Rac-1 activity

Authors

Seojin Kang

¹College of Pharmacy, Seoul National University, Seoul, Korea
Keunyoung Kim

¹College of Pharmacy, Seoul National University, Seoul, Korea
Ji-Yoon Noh

¹College of Pharmacy, Seoul National University, Seoul, Korea
Yeryeon Jung

¹College of Pharmacy, Seoul National University, Seoul, Korea
Ok-Nam Bae

²College of Pharmacy, Hanyang University, Ansan, Korea
Kyung-Min Lim

³College of Pharmacy, Ewha Womans University, Seoul, Korea
Jin-Ho Chung

¹College of Pharmacy, Seoul National University, Seoul, Korea

Abstract

Summary

Statins, lipid-lowering agents for the prevention of atherosclerosis and fatal coronary heart diseases, have pleiotropic modalities on the function and physiology of vascular smooth muscle that include anti-contractile and pro-apoptotic effects. These effects were suggested to stem from the inhibition of small GTPase Rho A, but they are largely regarded as distinct and unrelated. Recently, we discovered that simvastatin causes both contractile dysfunction and apoptosis of vascular smooth muscle cells (VSMCs), reflecting that they may be closely related, yet their connecting link remains unexplained. Here, we elaborated the mechanism underlying simvastatin-induced apoptosis of normal VSMCs in connection with contractile dysfunction. Repeated oral administration of simvastatin to rats in vivo resulted in contractile dysfunction and apoptosis of vascular smooth muscle, of which pattern was well reproduced in rat VSMCs in vitro. Of note, contractile dysfunction and apoptosis occurred in concerted manners both in vivo and in vitro in the aspects of time course and dose of exposure. In rat VSMCs, simvastatin impaired the activation of small GTPases, RhoA along with Rac-1, which resulted in the disruption of actin integrity, a pivotal factor both for the generation of contractile force and survival of VSMCs. In line with the disruption of actin integrity, Bmf, a pro-apoptotic factor bound to intact actin, dissociated and translocated into mitochondria, which corresponded well with the dissipation of mitochondrial membrane potential, caspase-3 activation and ultimately apoptosis. These events were all rescued by an actin stabilisation agent, jasplakinolide as well as geranylgeraniol, indicating that damages of the actin integrity from disrupted activation of RhoA/ Rac-1 lies at the center of simvastatin-induced contractile dysfunction and apoptosis in vascular smooth muscle.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Statin - smooth muscle cells - VSMC - apoptosis

Full Text

References

References
1 Nissen SE, Tuzcu EM, Schoenhagen P. et al. Statin therapy, LDL cholesterol, C-reactive protein, and coronary artery disease. N Engl J Med 2005; 352: 29-38.
2 Brahm AJ, Hegele RA.. Incident Diabetes With Statins: Biology, Artifact, or Both?. Can J Cardiol 2015; 31: 963-965.
3 Joy TR, Hegele RA.. Narrative review: statin-related myopathy. Ann Intern Med 2009; 150: 858-868.
4 Dirks AJ, Jones KM.. Statin-induced apoptosis and skeletal myopathy. Am J Physiol Cell Physiol 2006; 291: C1208-1212.
5 Sauzeau V, Le Jeune H, Cario-Toumaniantz C. et al. Cyclic GMP-dependent protein kinase signaling pathway inhibits RhoA-induced Ca2+ sensitisation of contraction in vascular smooth muscle. J Biol Chem 2000; 275: 21722-21729.
6 Seko T, Ito M, Kureishi Y. et al. Activation of RhoA and inhibition of myosin phosphatase as important components in hypertension in vascular smooth muscle. Circ Res 2003; 92: 411-418.
7 Sauzeau V, Le Mellionnec E, Bertoglio J. et al. Human urotensin II-induced contraction and arterial smooth muscle cell proliferation are mediated by RhoA and Rho-kinase. Circ Res 2001; 88: 1102-1104.
8 Mack CP, Somlyo AV, Hautmann M. et al. Smooth muscle differentiation marker gene expression is regulated by RhoA-mediated actin polymerisation. J Biol Chem 2001; 276: 341-347.
9 Wirth A.. Rho kinase and hypertension. Biochim Biophys Acta 2010; 1802: 1276-1284.
10 Guijarro C, Blanco-Colio LM, Ortego M. et al. 3-Hydroxy-3-methylglutaryl coenzyme a reductase and isoprenylation inhibitors induce apoptosis of vascular smooth muscle cells in culture. Circ Res 1998; 83: 490-500.
11 de Godoy MA, Rattan S. Role of rho kinase in the functional and dysfunctional tonic smooth muscles. Trends Pharmacol Sci 2011; 32: 384-393.
12 De Pinieux G, Chariot P, Ammi-Said M. et al. Lipid-lowering drugs and mitochondrial function: effects of HMG-CoA reductase inhibitors on serum ubiquinone and blood lactate/pyruvate ratio. Br J Clin Pharmacol 1996; 42: 333-337.
13 Kang S, Woo HH, Kim K. et al. Dysfunction of vascular smooth muscle and vascular remodelling by simvastatin. Toxicol Sci 2014; 138: 446-556.
14 Etienne-Manneville S, Hall A. Rho GTPases in cell biology. Nature 2002; 420: 629-635.
15 Gourlay CW, Ayscough KR.. The actin cytoskeleton: a key regulator of apoptosis and ageing?. Nat Rev Mol Cell Biol 2005; 6: 583-589.
16 Reho JJ, Zheng X, Fisher SA.. Smooth muscle contractile diversity in the control of regional circulations. Am J Physiol Heart Circ Physiol 2014; 306: H163-172.
17 Lomonosova E, Chinnadurai G.. BH3-only proteins in apoptosis and beyond: an overview. Oncogene 2008; 27 (Suppl. 01) S2-19.
18 Pinon JD, Labi V, Egle A. et al. Bim and Bmf in tissue homeostasis and malignant disease. Oncogene 2008; 27 (Suppl. 01) S41-52.
19 Wei MC, Zong WX, Cheng EH. et al. Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 2001; 292: 727-730.
20 Takai Y, Sasaki T, Matozaki T.. Small GTP-binding proteins. Physiol Rev 2001; 81: 153-208.
21 Puthalakath H, Villunger A, O’Reilly LA. et al. Bmf: a proapoptotic BH3-only protein regulated by interaction with the myosin V actin motor complex, activated by anoikis. Science 2001; 293: 1829-1832.
22 Aoki T, Nishimura H, Nakagawa S. et al. Pharmacological profile of a novel synthetic inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A reductase. Arzneimittelforschung 1997; 47: 904-909.
23 Xu QY, Liu YH, Zhang Q. et al. Metabolomic analysis of simvastatin and fenofibrate intervention in high-lipid diet-induced hyperlipidemia rats. Acta Pharmacol Sin 2014; 35: 1265-1273.
24 Meador BM, Huey KA.. Statin-associated myopathy and its exacerbation with exercise. Muscle Nerve 2010; 42: 469-479.
25 Varkuti BH, Yang Z, Kintses B. et al. A novel actin binding site of myosin required for effective muscle contraction. Nat Struct Mol Biol 2012; 19: 299-306.
26 Posey SC, Bierer BE.. Actin stabilisation by jasplakinolide enhances apoptosis induced by cytokine deprivation. J Biol Chem 1999; 274: 4259-4265.
27 Rubtsova SN, Kondratov RV, Kopnin PB. et al. Disruption of actin microfilaments by cytochalasin D leads to activation of p53. FEBS Lett 1998; 430: 353-357.
28 Konishi H, Kikuchi S, Ochiai T. et al. Latrunculin a has a strong anticancer effect in a peritoneal dissemination model of human gastric cancer in mice. Anticancer Res 2009; 29: 2091-2097.
29 Anderl J, Echner H, Faulstich H.. Chemical modification allows phallotoxins and amatoxins to be used as tools in cell biology. Beilstein J Org Chem 2012; 8: 2072-2084.
30 Leeper NJ, Raiesdana A, Kojima Y. et al. Loss of CDKN2B promotes p53-dependent smooth muscle cell apoptosis and aneurysm formation. Arterioscler Thromb Vasc Biol 2013; 33: e1-e10.
31 Guijarro C, Blanco-Colio LM, Ortego M. et al. 3-Hydroxy-3-methylglutaryl coenzyme a reductase and isoprenylation inhibitors induce apoptosis of vascular smooth muscle cells in culture. Circ Res 1998; 83: 490-500.
32 Min J, Reznichenko M, Poythress RH. et al. Src modulates contractile vascular smooth muscle function via regulation of focal adhesions. J Cell Physiol 2012; 227: 3585-3592.
33 Wall BT, Dirks ML, van Loon LJ.. Skeletal muscle atrophy during short-term disuse: implications for age-related sarcopenia. Age Res Rev 2013; 12: 898-906.
34 Powers SK, Wiggs MP, Duarte JA. et al. Mitochondrial signaling contributes to disuse muscle atrophy. Am J Physiol Endocrinol Metab 2012; 303: E31-39.
35 Jackman RW, Kandarian SC.. The molecular basis of skeletal muscle atrophy. Am J Physiol Cell Physiol 2004; 287: C834-843.
36 Zhu S, Nagashima M, Khan MA. et al. Lack of caspase-3 attenuates immobilisation-induced muscle atrophy and loss of tension generation along with mitigation of apoptosis and inflammation. Muscle Nerve 2013; 47: 711-721.
37 Fanin M, Nascimbeni AC, Angelini C.. Muscle atrophy, ubiquitin-proteasome, and autophagic pathways in dysferlinopathy. Muscle Nerve 2014; 50: 340-347.
38 VanBrocklin MW, Verhaegen M, Soengas MS. et al. Mitogen-activated protein kinase inhibition induces translocation of Bmf to promote apoptosis in melanoma. Cancer Res 2009; 69: 1985-1994.
39 Kutuk O, Letai A.. Displacement of Bim by Bmf and Puma rather than increase in Bim level mediates paclitaxel-induced apoptosis in breast cancer cells. Cell Death Differ 2010; 17: 1624-1635.
40 Kang Y, Nian H, Rajendran P. et al. HDAC8 and STAT3 repress BMF gene activity in colon cancer cells. Cell Death Dis 2014; 5: e1476.
41 Geneste O, Copeland JW, Treisman R.. LIM kinase and Diaphanous cooperate to regulate serum response factor and actin dynamics. J Cell Biol 2002; 157: 831-838.
42 Sit ST, Manser E.. Rho GTPases and their role in organizing the actin cytoskeleton. J Cell Sci 2011; 124: 679-683.
43 Korobova F, Svitkina T.. Arp2/3 complex is important for filopodia formation, growth cone motility, and neuritogenesis in neuronal cells. Mol Biol Cell 2008; 19: 1561-1574.

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