Modulating alignment and contractile protein expression in vascular smooth muscle cell sheets: Using microcontact printing and mechanical conditioning

Vascular smooth muscle cell (SMC) de-differentiation with subsequent migration and proliferation into the subendothelial space is central to the progression of cardiovascular diseases. The Nox4 NADPH oxidase (Nox4) is implicated in maintaining the differentiated phenotype of SMC in part through myocardin, a master regulator of SMC gene expression. However, this process is poorly understood. We hypothesized that microRNAs (miR)-mediate changes in Nox4 expression and regulate SMC differentiation. Treatment of human SMCs with a miR-9 or miR-25 mimic silenced Nox4 mRNA through binding to the Nox4 3’UTR. However, only miR-25 was sufficient to downregulate Nox4 protein levels. We found that miR-25 induced the expression of miR-9 through a novel mechanism involving demethylation of the miR-9 promoter by Tet methylcytosine dioxygenase 2 (TET2). Inhibition of miR-9 induction by miR-25 with a miR-9 inhibitor restored Nox4 protein expression to basal levels. Furthermore, the miR-25-mediated decrease in Nox4 protein was ameliorated by inhibiting the proteasome with MG132. These data suggest a novel mechanism wherein miR-9 and miR-25 regulate Nox4 through both translational suppression and proteosomal degradation. Overexpression of miR-9 or miR-25 in human SMCs (1) suppressed myocardin mRNA and protein expression; (2) decreased expression of multiple SMC differentiation genes; and (3) was sufficient to induce cell migration. Thrombin and tumor necrosis factor increased the expression of miR-9 and miR-25 in human SMCs and inhibition of miR-9 prevented thrombin-mediated decrease in myocardin and SMC migration. Mir-9 and miR-25 levels were increased in SMCs derived from balloon injured rat aorta as compared to medial SMCs and in murine carotid artery ten days post carotid injury. A miR-9 inhibitor decreased neointimal formation by more than 50% in following partial carotid ligation in mice. These findings identify miR-9/Nox4 as a novel regulatory pathway of SMC differentiation and a potential therapeutic target in vascular disease.

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The cooperative effects of micro-grooved topography and TGF-β1 on the vascular smooth muscle cell contractile protein expression of the mesenchymal stem cells

Differentiation ◽

10.1016/j.diff.2020.06.003 ◽

2020 ◽

Vol 115 ◽

pp. 22-29

Author(s):

Sakhavat Abolhasani ◽

Masoumeh Rajabibazl ◽

Mohammad-Mehdi Khani ◽

Azim Parandakh ◽

Reyhaneh Hoseinpoor

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Protein Expression ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Smooth Muscle Cell ◽

Cooperative Effects ◽

Tgf Β1

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Indoxyl sulfate promotes vascular smooth muscle cell senescence with upregulation of p53, p21, and prelamin A through oxidative stress

AJP Cell Physiology ◽

10.1152/ajpcell.00329.2011 ◽

2012 ◽

Vol 303 (2) ◽

pp. C126-C134 ◽

Cited By ~ 62

Author(s):

Gulinuer Muteliefu ◽

Hidehisa Shimizu ◽

Atsushi Enomoto ◽

Fuyuhiko Nishijima ◽

Masahide Takahashi ◽

...

Keyword(s):

Oxidative Stress ◽

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Protein Expression ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Smooth Muscle Cell ◽

Cell Senescence ◽

Hypertensive Rats ◽

Prelamin A

We previously demonstrated that indoxyl sulfate (IS), a uremic toxin, induces aortic calcification in hypertensive rats and induces oxidative stress and the expression of osteoblast-specific proteins in vascular smooth muscle cells. This study aimed to clarify whether IS stimulates senescence of cultured human aortic smooth muscle cells (HASMCs) and aorta in Dahl salt-sensitive hypertensive rats and whether AST-120, an oral sorbent, prevents senescence of aorta in subtotally nephrectomized uremic rats. IS increased the mRNA expression of p53 and p21 in HASMCs, whereas it did not change that of p16 and retinoblastoma protein (pRb). The IS-induced expression of p53 and p21 was suppressed by N-acetylcysteine, an antioxidant. IS promoted protein expression of p53, p21, and senescence-associated β-galactosidase (SA-β-gal) activity in HASMCs, and N-acetylcysteine and pifithrin-α, p-nitro, a p53 inhibitor, blocked these effects. IS upregulated prelamin A, a hallmark of vascular smooth muscle cell senescence, and downregulated FACE1/Zempste24 protein expression in HASMCs, and N-acetylcysteine suppressed these effects. Administration of IS to hypertensive rats increased expression of SA-β-gal, p53, p21, prelamin A, and oxidative stress markers such as 8-hydroxyl-2′-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) in the cells embedded in the calcification area of arcuate aorta. Further, the uremic rat model showed positive staining for SA-β-gal, p53, p21, prelamin A, 8-OHdG, and MDA in the cells embedded in the calcification area of arcuate aorta, whereas AST-120 reduced the expression of these biomarkers. Taken together, IS accelerates vascular smooth muscle cell senescence with upregulation of p53, p21, and prelamin A and downregulation of FACE1 through oxidative stress.

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W01.11 Non-lipid lowering effect of statins: Inhibition of human aortic vascular smooth muscle cell (VSMC) migration

Atherosclerosis ◽

10.1016/s0021-9150(04)90011-1 ◽

2004 ◽

Vol 5 (1) ◽

pp. 3

Author(s):

M CAMPBELL

Keyword(s):

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Smooth Muscle Cell ◽

Lipid Lowering ◽

Lowering Effect ◽

Lipid Lowering Effect

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Involvement of Cyclooxygenase- and Lipoxygenase-Mediated Conversion of Arachidonic Acid in Controlling Human Vascular Smooth Muscle Cell Proliferation

Thrombosis and Haemostasis ◽

10.1055/s-0038-1645212 ◽

1990 ◽

Vol 63 (02) ◽

pp. 291-297 ◽

Cited By ~ 28

Author(s):

Herm-Jan M Brinkman ◽

Marijke F van Buul-Worteiboer ◽

Jan A van Mourik

Keyword(s):

Cell Proliferation ◽

Arachidonic Acid ◽

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cells ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Smooth Muscle Cell ◽

Muscle Cells ◽

Smooth Muscle Cell Proliferation

SummaryWe observed that the growth of human umbilical arterysmooth muscle cells was inhibited by the phospholipase A2 inhibitors p-bromophenacylbromide and mepacrine. Thesefindings suggest that fatty acid metabolism might be integrated in the control mechanism of vascular smooth muscle cell proliferation. To identify eicosanoids possibly involved in this process, we studied both the metabolism of arachidonic acid of these cells in more detail and the effect of certain arachidonic acid metabolites on smooth muscle cells growth. We found no evidence for the conversion of arachidonic acid via the lipoxygenase pathway. In contrast, arachidonic acid was rapidly converted via the cyclooxy-genase pathway. The following metabolites were identified: prostaglandin E2 (PGE2), 6-keto-prostaglandin F1α (6-k-PGF1α), prostaglandin F2α (PGF2α), 12-hydroxyheptadecatrienoic acid (12-HHT) and 11-hydroxyeicosatetetraenoic acid (11-HETE). PGE2 was the major metabolite detected. Arachidonic acid metabolites were only found in the culture medium, not in the cell. After synthesis, 11-HETE was cleared from the culture medium. We have previously reported that PGE2 inhibits the serum-induced [3H]-thymidine incorporation of growth-arrested human umbilical artery smooth muscle cells. Here we show that also 11-HETEexerts this inhibitory property. Thus, our data suggeststhat human umbilical artery smooth muscle cells convert arachidonic acid only via the cyclooxygenase pathway. Certain metabolites produced by this pathway, including PGE2 and 11-HETE, may inhibit vascular smooth muscle cell proliferation.

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