Original articleInvolvement of connexin 43 in angiotensin II-induced migration and proliferation of saphenous vein smooth muscle cells via the MAPK-AP-1 signaling pathway
Introduction
Intimal hyperplasia is a proliferative vasculopathy that causes stenosis in the venous bypass graft, a pathophysiologic feature leading to occlusion of the vessel lumen [1], resulting in failure rates of 20% and 50% at 5years and 10years, respectively [2]. Proliferation and migration of vascular smooth muscle cells (VSMCs) into the sub-intimal space play an important role in intimal thickening in atherosclerosis and restenosis and influences the long-term patency of the venous graft [3]. Initiation and progression of the atherosclerotic plaque and restenotic vessel involve complex patterns of interaction between the cells of the arterial wall, in which cytokines, chemokines, and growth factors are known to play a critical role [4], [5]. For example, angiotensin II (Ang II), a potent growth factor, stimulates proliferation and migration of VSMCs, and induces the accumulation and deposition of collagen through Ang II type 1 (AT1) receptor; all factors contributing to the pathogenesis of atherosclerosis [6].
Another form of cell-to-cell interaction involves direct exchange of small cytosolic components via gap junctions, which are composed of connexon hemichannels, each of which are made of 6 connexin (Cx) molecules [7]. Three subtypes of Cx, Cx37, Cx40, and Cx43, are found in the cells of the vasculature, with Cx43 being the predominant one [7], [8]. Changes in connexin expression and function have been found in association with phenotypic changes in VSMCs during the progression of atherosclerotic lesions and following injury in large arteries [9]. Also, Ang II can activate the expression of Cx43 in the myocardial cells of rats in vitro[10]. However, little is known about the relationship between Ang II and Cx43 expression in VSMCs of the human saphenous vein (hSV). In this study, we examined the Cx43 expression and its role in the proliferation and migration of VSMCs. Further, we delineated the signaling pathways involved in the proliferation of smooth muscle cells in hSV bypass conduits following stimulation with Ang II.
Section snippets
Specimen collection, processing and culture of smooth muscle cell
The experimental protocol for this study was approved by the Institutional Review Board of Creighton University. The hSV specimens (mean patient age 71.5 ± 3.7years) left over from the bypass surgical procedure were used for the study. Specimens were collected fresh in ice-cold University of Wisconsin solution, which is used to maintain cellular and tissue integrity in the organs for transplantation [11]. The hSV conduits were immediately brought to the laboratory where all of the procedures
Ang II-induced Cx43 expression through AT1 receptors in the SMCs of human venous bypass vessels
Cx43 protein expression in hSV SMCs was evaluated in a time- and dose-dependent manner. After treatment with Ang II (10− 7mol/L), the Cx43 expression was increased significantly at 0.5h, peaked at 1 h and remained at an elevated level even after 3h in the SV samples (Fig. 1A). Also, Ang II (10− 7and 10− 6mol/L) induced a significant maximal response in SV at 1 h in a dose-dependent manner (Fig. 1B).
To determine which AT receptor subtype is involved in Ang II-induced Cx43 protein expression in hSV
Discussion
The hSV is the most commonly employed CABG conduit. Unfortunately, intimal hyperplasia leading to stenosis and occlusion of hSV commonly limit long-term results following coronary artery bypass graft (CABG) surgery [15]. The mechanisms involved in the intimal hyperplasia are proliferation and migration of medial smooth muscle cells towards the sub-intimal space [16]. There is increasing evidence that connexins are involved in the development of intimal hyperplasia and restenosis involving mouse
Acknowledgments
This study was supported by National Institutes of Health grants R01HL070885 (D.K.A.) and R01HL073349 (D.K.A.), and the State of Nebraska Tobacco Settlement Funds (D.K.A.). We thank Nebraska Heart Institute for providing us the venous bypass specimens.
References (43)
- et al.
Increased connexin43 expression in human saphenous veins in culture is associated with intimal hyperplasia
J Vasc Surg
(2005) - et al.
Angiotensin II and IGF-1 regulate connexin43 expression via ERK and p38 signaling pathways in vascular smooth muscle cells of coronary artery bypass conduits
J Surg Res
(2007) - et al.
Gap junctional communication in tissue inflammation and repair
Biochim Biophys Acta
(2005) - et al.
Upregulation of connexin43 gap junctions between neointimal smooth muscle cells
Eur J Cell Biol
(2004) - et al.
Effects of angiotensin II on expression of the gap junction channel protein connexin43 in neonatal rat ventricular myocytes
J Am Coll Cardiol
(1998) - et al.
Endothelin-1 decreases gap junctional intercellular communication by inducing phosphorylation of connexin 43 in human ovarian carcinoma cells
J Biol Chem
(2003) - et al.
Tyrosine kinase inhibitors suppress alpha1-adrenoceptor mediated contraction in human radial, internal mammary arteries and saphenous vein
Neurosci Lett
(2002) - et al.
Modulation of intercellular communication between smooth muscle cells by growth factors and cytokines
Eur J Pharmacol
(1996) - et al.
Statins reduce connexin40 and connexin43 expression in atherosclerotic aorta of rabbits
Int J Cardiol
(2005) - et al.
Regulation of epidermal growth factor-induced connexin43 gap junction communication by big mitogen-activated protein kinase1/ERK5 but not ERK1/2kinase activation
J Biol Chem
(2003)
Differential effects of insulin-like growth factor-1 and atheroma-associated cytokines on cell proliferation and apoptosis in plaque smooth muscle cells of symptomatic and asymptomatic patients with carotid stenosis
Immunol Cell Biol
Autophagy of vascular smooth muscle cells in atherosclerotic lesions
Autophagy
Altered pattern of vascular connexin expression in atherosclerotic plaques
Arterioscler Thromb Vasc Biol
Angiotensin II induces smooth muscle cell proliferation in the normal and injured rat arterial wall
Circ Res
Altered pattern of vascular connexin expression in atherosclerotic plaques
Arterioscler Thromb Vasc Biol
Functional and morphological assessment of rat aorta stored in University of Wisconsin and Eurocollins solutions
Transplantation
Up-regulation of gap junctional intercellular communication and connexin43 expression by retinoic acid in human endometrial stromal cells
J Clin Endocrinol Metab
Matrix metalloproteinase 2 and tissue inhibitors of metalloproteinases regulate human aortic smooth muscle cell migration during in vitro aging
FASEB J
Contrasting structure of the saphenous vein and internal mammary artery used as coronary bypass vessels
Cardiovasc Res
Dual benefit of reduced Cx43 on atherosclerosis in LDL receptor-deficient mice
Cell Commun Adhes
Involvement of myoendothelial gap junctions in the actions of endothelium-derived hyperpolarizing factor
Circ Res
Cited by (57)
The promoting role of Cx43 on the proliferation and migration of arterial smooth muscle cells for angiotensin II-dependent hypertension
2021, Pulmonary Pharmacology and TherapeuticsRutaecarpine prevented ox-LDL-induced VSMCs dysfunction through inhibiting overexpression of connexin 43
2019, European Journal of PharmacologyCitation Excerpt :It was also reported that overexpression of Cx43 by transfection markedly promoted the proliferation of saphenous VSMCs (Rama et al., 2006). Conversely, silencing of Cx43 expression by siRNA prevented angiotensin II-induced proliferation of VSMCs (Jia et al., 2008). Multiple risk factors of atherosclerosis, such as hyperlipidemia and angiotensin II (Ang II) (Alonso et al., 2010; Jia et al., 2008), were reported to upregulate the vascular expression of Cx43, which may contribute to the development of atherosclerosis.
The pivotal role of extracellular signal-regulated kinase in gap junction-mediated regulation of TXNIP
2017, Cellular SignallingCitation Excerpt :The molecular mechanisms involved in the regulation of cellular redox status by GJs are still incompletely understood. It is thought to be either related to channel-dependent transmission and propagation of intercellular signals, or communication-independent regulation of several key signaling molecules, such as AMPK, Src, and NF-κb, etc. [9,12–14]. TXNIP stands for thioredoxin-interacting protein.
MicroRNA-221 sponge therapy attenuates neointimal hyperplasia and improves blood flows in vein grafts
2016, International Journal of CardiologyCitation Excerpt :Vein grafts is ideally suited for gene therapy due to the bypass grafts can be direct local delivery of recombinant viral vectors ex vivo or in situ without the need for delivery devices and minimizes risk of systemic exposure there during a short clinical window (approximately 40–60 min) prior to grafting. Previous studies have demonstrated that genetic interventions designed to inhibit VSMC proliferation and migration at the genetic level has significant effect on prevention of vein graft failure [27–29]. Multiple lines of experimental in vitro and vivo studies have showed that miRNAs play key roles in the control of VSMC function and plasticity response to both direct and indirect stimuli through targeting transcriptional factors or key signal molecules in VSMC proliferation, migration and apoptosis [26].