In situ Evidence of Collagen V and Interleukin-6/Interleukin-17 Activation in Vascular Remodeling of Experimental Pulmonary Hypertension

Several studies have reported the pathophysiologic and molecular mechanisms responsible for pulmonary arterial hypertension (PAH). However, the in situ evidence of collagen V (Col V) and interleukin-17 (IL-17)/interleukin-6 (IL-6) activation in PAH has not been fully elucidated. We analyzed the effects of collagen I (Col I), Col V, IL-6, and IL-17 on vascular remodeling and hemodynamics and its possible mechanisms of action in monocrotaline (MCT)-induced PAH. Twenty male Wistar rats were randomly divided into two groups. In the PAH group, animals received MCT 60 mg/kg intraperitoneally, whereas the control group (CTRL) received saline. On day 21, the pulmonary blood pressure (PAP) and right ventricular systolic pressure (RVSP) were determined. Lung histology (smooth muscle cell proliferation [α-smooth muscle actin; α-SMA] and periadventitial fibrosis), immunofluorescence (Col I, Col V, and α-SMA), immunohistochemistry (IL-6, IL-17, and transforming growth factor-beta [TGF-β]), and transmission electron microscopy to detect fibronexus were evaluated. The RVSP (40 ± 2 vs. 24 ± 1 mm Hg, respectively; <i>p</i> < 0.0001), right ventricle hypertrophy index (65 ± 9 and 25 ± 5%, respectively; <i>p</i> < 0.0001), vascular periadventitial Col I and Col V, smooth muscle cell α-SMA+, fibronexus, IL-6, IL-17, and TGF-β were higher in the MCT group than in the CTRL group. In conclusion, our findings indicate in situ evidence of Col V and IL-6/IL-17 activation in vascular remodeling and suggest that increase of Col V may yield potential therapeutic targets for treating patients with PAH.

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The Role of Ubiquitin E3 Ligase in Atherosclerosis

Current Medicinal Chemistry ◽

10.2174/0929867327666200306124418 ◽

2020 ◽

Vol 28 (1) ◽

pp. 152-168

Author(s):

Zhi-Xiang Zhou ◽

Zhong Ren ◽

Bin-Jie Yan ◽

Shun-Lin Qu ◽

Zhi-Han Tang ◽

...

Keyword(s):

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Smooth Muscle Cell ◽

Protein Modification ◽

Molecular Mechanisms ◽

Cholesterol Metabolism ◽

Atherosclerotic Cardiovascular Disease ◽

E3 Ligases

: Atherosclerosis is a chronic inflammatory vascular disease. Atherosclerotic cardiovascular disease is the main cause of death in both developed and developing countries. Many pathophysiological factors, including abnormal cholesterol metabolism, vascular inflammatory response, endothelial dysfunction and vascular smooth muscle cell proliferation and apoptosis, contribute to the development of atherosclerosis and the molecular mechanisms underlying the development of atherosclerosis are not fully understood. Ubiquitination is a multistep post-translational protein modification that participates in many important cellular processes. Emerging evidence suggests that ubiquitination plays important roles in the pathogenesis of atherosclerosis in many ways, including regulation of vascular inflammation, endothelial cell and vascular smooth muscle cell function, lipid metabolism and atherosclerotic plaque stability. This review summarizes important contributions of various E3 ligases to the development of atherosclerosis. Targeting ubiquitin E3 ligases may provide a novel strategy for the prevention of the progression of atherosclerosis.

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Contribution of transcription factor EB to adipoRon-induced inhibition of arterial smooth muscle cell proliferation and migration

AJP Cell Physiology ◽

10.1152/ajpcell.00294.2019 ◽

2019 ◽

Vol 317 (5) ◽

pp. C1034-C1047 ◽

Cited By ~ 1

Author(s):

Yun-Ting Wang ◽

Jiajie Chen ◽

Xiang Li ◽

Michihisa Umetani ◽

Yang Chen ◽

...

Keyword(s):

Cell Proliferation ◽

Transcription Factor ◽

Smooth Muscle ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Remodeling ◽

Therapeutic Effects ◽

Transcription Factor Eb ◽

And Migration ◽

Proliferation And Migration

Abnormal vascular smooth muscle cell (SMC) dedifferentiation with increased proliferation and migration during pathological vascular remodeling is associated with vascular disorders, such as atherosclerosis and in-stent restenosis. AdipoRon, a selective agonist of adiponectin receptor, has been shown to protect against vascular remodeling by preventing SMC dedifferentiation. However, the molecular mechanisms that mediate adipoRon-induced SMC differentiation are not well understood. The present study aimed to elucidate the role of transcription factor EB (TFEB), a master regulator of autophagy, in mediating adipoRon’s effect on SMCs. In cultured arterial SMCs, adipoRon dose-dependently increased TFEB activation, which is accompanied by upregulated transcription of genes involved in autophagy pathway and enhanced autophagic flux. In parallel, adipoRon suppressed serum-induced cell proliferation and caused cell cycle arrest. Moreover, adipoRon inhibited SMC migration as characterized by wound-healing retardation, F-actin reorganization, and matrix metalloproteinase-9 downregulation. These inhibitory effects of adipoRon on proliferation and migration were attenuated by TFEB gene silencing. Mechanistically, activation of TFEB by adipoRon is dependent on intracellular calcium, but it is not associated with changes in AMPK, ERK1/2, Akt, or molecular target of rapamycin complex 1 activation. Using ex vivo aortic explants, we demonstrated that adipoRon inhibited sprouts that had outgrown from aortic rings, whereas lentiviral TFEB shRNA transduction significantly reversed this effect of adipoRon on aortic rings. Taken together, our results indicate that adipoRon activates TFEB signaling that helps maintain the quiescent and differentiated status of arterial SMCs, preventing abnormal SMC dedifferentiation. This study provides novel mechanistic insights into understanding the therapeutic effects of adipoRon on TFEB signaling and pathological vascular remodeling.

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Matrix Metalloproteinase 2 as a Potential Mediator of Vascular Smooth Muscle Cell Migration and Chronic Vascular Remodeling in Hypertension

Journal of Vascular Research ◽

10.1159/000441621 ◽

2015 ◽

Vol 52 (4) ◽

pp. 221-231 ◽

Cited By ~ 59

Author(s):

V.A. Belo ◽

Danielle A. Guimarães ◽

Michele Mazzaron Castro

Keyword(s):

Smooth Muscle ◽

Cell Migration ◽

Matrix Metalloproteinase ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Remodeling ◽

Matrix Metalloproteinase 2 ◽

Potential Mediator ◽

Smooth Muscle Cell Migration

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Exploring the molecular mechanisms of OSU-03012 on vascular smooth muscle cell proliferation

Molecular and Cellular Biochemistry ◽

10.1007/s11010-010-0531-5 ◽

2010 ◽

Vol 344 (1-2) ◽

pp. 81-89 ◽

Cited By ~ 1

Author(s):

Wei-Wen Kuo ◽

Jing-Ru Weng ◽

Chih-Yang Huang ◽

Chang-Hai Tsai ◽

Wei-Hung Liu ◽

...

Keyword(s):

Cell Proliferation ◽

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Smooth Muscle Cell ◽

Molecular Mechanisms ◽

Smooth Muscle Cell Proliferation

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Abstract 126: Differential Effects of Respectively Blocking Two BET Bromodomains on Vascular Smooth Muscle Cell Phenotype Transformation

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.36.suppl_1.126 ◽

2016 ◽

Vol 36 (suppl_1) ◽

Author(s):

Mengxue Zhang ◽

Bowen Wang ◽

Craig Kent ◽

Lian-Wang Guo

Keyword(s):

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Smooth Muscle Cell ◽

Molecular Mechanisms ◽

Cell Phenotype ◽

Dependent Manner ◽

Stat3 Phosphorylation ◽

Phenotype Transformation

Introduction: Intimal hyperplasia (IH) occurs primarily due to vascular smooth muscle cell (SMC) transformation from quiescent to pathogenic phenotypes (e.g. proliferation and inflammation). Identification and effective targeting of key epigenetic factors governing SMC pathogenic transformation may lead to novel therapeutic methods for prevention of IH. We previously found that globally blocking the bromo- and extra-terminal (BET) epigenetic “reader” family abrogated SMC phenotype transformation and IH. We further investigated the functions of the two BET bromodomains (Bromo1 and Bromo2). Hypothesis: Bromo1 and Bromo2 play different roles in SMC pathogenic transformation. Methods and Results: We pre-treated rat primary aortic SMCs (for 2h) with Olinone or RVX208, inhibitors specific for Bromo1 and Bromo2 respectively, and then stimulated SMC phenotype transformation. Whereas RVX208 abrogated PDGF-BB-stimulated SMC proliferation (BrdU assay) in a dose dependent manner, Olinone enhanced SMC proliferation at high concentrations (>20 μM). RVX208 at 50 μM reduced TNFα-induced SMC inflammation (MCP-1 ELISA) by 80%,but Olinone at the same concentration slightly increased MCP-1. Furthermore, whereas RVX208 abolished PDGF-BB or TNFα-induced STAT3 phosphorylation (Western blotting), Olinone slightly increased phospho-STAT3. Conclusions: Our results reveal that blocking two BET bromodomains respectively produces distinct effects on SMC phenotype transformation, suggesting their differential epigenetic functions. Further elucidation of the underlying molecular mechanisms should contribute to precise targeting of the BET family for optimal mitigation of IH.

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Long Non-Coding RNA MEG3 Downregulation Triggers Human Pulmonary Artery Smooth Muscle Cell Proliferation and Migration via the p53 Signaling Pathway

Cellular Physiology and Biochemistry ◽

10.1159/000480218 ◽

2017 ◽

Vol 42 (6) ◽

pp. 2569-2581 ◽

Cited By ~ 47

Author(s):

Zengxian Sun ◽

Xiaowei Nie ◽

Shuyang Sun ◽

Shumin Dong ◽

Chunluan Yuan ◽

...

Keyword(s):

Cell Proliferation ◽

Smooth Muscle ◽

Pulmonary Artery ◽

Smooth Muscle Cells ◽

Smooth Muscle Cell ◽

Muscle Cell ◽

Vascular Remodeling ◽

Pulmonary Artery Smooth Muscle ◽

And Migration ◽

Proliferation And Migration

Background/Aims: Increasing evidence has demonstrated a significant role of long non-coding RNAs (lncRNAs) in diverse biological processes, and many of which are likely to have functional roles in vascular remodeling. However, their functions in pulmonary arterial hypertension (PAH) remain largely unknown. Pulmonary vascular remodeling is an important pathological feature of PAH, leading to increased vascular resistance and reduced compliance. Pulmonary artery smooth muscle cells (PASMCs) dysfunction is involved in vascular remodeling. Long noncoding RNAs are potential regulators of PASMCs function. Herein, we determined whether long noncoding RNA–maternally expressed gene 3 (MEG3) was involved in PAH-related vascular remodeling. Methods: The arterial wall thickness was examined by hematoxylin and eosin (H&E) staining in distal pulmonary arteries (PAs) isolated from lungs of healthy volunteers and PAH patients. The expression level of MEG3 was analyzed by qPCR. The effects of MEG3 on human PASMCs were assessed by cell counting Kit-8 assay, BrdU incorporation assay, flow cytometry, scratch-wound assay, immunofluorescence, and western blotting in human PASMCs. Results: We revealed that the expression of MEG3 was significantly downregulated in lung and PAs of patients with PAH. MEG3 knockdown affected PASMCs proliferation and migration in vitro. Moreover, inhibition of MEG3 regulated the cell cycle progression and made more smooth muscle cells from the G0/G1 phase to the G2/M+S phase and the process could stimulate the expression of PCNA, Cyclin A and Cyclin E. In addition, we found that the p53 pathway was involved in MEG3–induced smooth muscle cell proliferation. Conclusions: This study identified MEG3 as a critical regulator in PAH and demonstrated the potential of gene therapy and drug development for treating PAH.

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Abstract 315: Krüppel-like Factor 5 Promotes Vascular Remodeling in a Biphasic Manner by Inhibiting Vascular Smooth Muscle Cell Apoptosis and Stimulating Cell Growth

Circulation ◽

10.1161/circ.116.suppl_16.ii_44-d ◽

2007 ◽

Vol 116 (suppl_16) ◽

Author(s):

Daigo Sawaki ◽

Toru Suzuki ◽

Kenichi Aizawa ◽

Takayoshi Matsumura ◽

Nanae Kada ◽

...

Keyword(s):

Cell Proliferation ◽

Smooth Muscle ◽

Cell Growth ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cell ◽

Cyclin D1 ◽

Muscle Cell ◽

Vascular Smooth Muscle Cell ◽

Vascular Remodeling ◽

Neointimal Formation

Introduction: Vascular remodeling is characterized by cell proliferation and/or apoptosis with further phenotypic change of vascular cells. Vascular smooth muscle cell (VSMC)s, in particular, play a major role in the proliferative process such as neointimal formation and restenosis after angioplasty. In deciphering the transcriptional regulatory mechanisms in cardiovascular remodeling, Krüppel-like factor 5 (KLF5) was originally isolated as a regulatory factor of phenotypically modulated VSMCs. Past studies collectively have shown that KLF5 can induce cell growth pathologically in non-cardiovascular cells. However, how KLF5 contributes to vascular remodeling, notably its effects on apoptosis in the vascular lesion, had yet to be addressed. In the present study, we have aimed to address the effects of KLF5 not only on VSMC growth but also on apoptosis in vascular remodeling. Methods&Results: We performed adenoviral overexpression of KLF5 and other related factors after rat carotid balloon injury. In the early phase (48 hours after injury), KLF5 administered animals showed significantly decreased TUNEL positive cells in the medial layer. In the chronic phase (14 days after injury), apoptotic cells were recognized neither in the KLF5 animals nor in the others. While, neointimal formation and PCNA labeling index significantly increased in the KLF5 animals. Rat VSMCs transfected with KLF5 showed marked increase in cell proliferation and BrdU uptake. Additionally, cleavage of caspase-3 recognized in the quiescent VSMCs was attenuated after transfection of KLF5. Even under apoptotic stimulation using anisomysin, KLF5 overexpression resulted in significant inhibition of apoptosis induction. Further, KLF5 up-regulated gene expression of cell cycle factors such as cyclin D1, and conversely, knockdown of KLF5 by RNA interference showed down-regulation of cyclin D1 and impairment of VSMC proliferation. Conclusion: These findings taken together suggest that KLF5 plays a central role in VSMC proliferative pathologies such as vascular remodeling through biphasic contribution; inhibition of apoptosis and growth stimulation. Therapeutic intervention targeted against KLF5 may be potentially exploitable for VSMC proliferative pathology.

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