Ductus arteriosus smooth muscle cell migration on collagen: dependence on laminin and its receptors

During permanent closure of the ductus arteriosus, smooth muscle cells migrate through the extracellular matrix (ECM) to form intimal mounds that occlude the vessel's lumen. Smooth muscle cells (SMC) migrate over surfaces coated with collagen in vitro. During the migration SMC also synthesize fibronectin (FN) and laminin (LN). Antibodies against FN and LN inhibit migration on collagen by 30% and 67%, respectively. Because of the apparent importance of LN in migration, we examined how SMC interact with LN and LN fragments (P1, E8, P1′, E1′, E3, E4, and G). Ductus SMC adhere to high concentrations of LN and two fragments of the molecule: P1 and E8. They use a unique set of integrin receptors to bind to LN (alpha 1 beta 1, alpha 6 beta 1 and alpha v beta 3), to P1 (alpha 1 beta 1, alpha v beta 3), and to E8 (alpha 6 beta 1, alpha v beta 3). The alpha v beta 3 integrin binds to the P1 fragment of LN in an RGD peptide-dependent manner, and to the E8 fragment in an RGD-independent manner; the RGD site on the P1 fragment probably is not available to the cell in intact LN. Antibodies against beta 1 integrins completely inhibit SMC adhesion to LN; antibodies against the alpha v beta 3 integrin do not block SMC adhesion to LN, but do prevent cell spreading. LN is also capable of interfering with SMC adhesion to other ECM components. The antiadhesive effect of LN is located in the E1′ domain. Both exogenous and endogenous LN increase SMC motility on collagen I. The locomotion-promoting activity of LN resides in the E1′ antiadhesive domain, and not in its adhesive (P1, E8) domains. LN causes a decrease in the number of focal contacts on collagen I. This might enable SMC to alter their mobility as they move through the extracellular matrix to occlude the ductus arteriosus lumen.

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Characterization and confocal imaging of calponin in gastrointestinal smooth muscle

AJP Cell Physiology ◽

10.1152/ajpcell.1993.265.5.c1371 ◽

1993 ◽

Vol 265 (5) ◽

pp. C1371-C1378 ◽

Cited By ~ 30

Author(s):

M. P. Walsh ◽

J. D. Carmichael ◽

G. J. Kargacin

Keyword(s):

Smooth Muscle ◽

Protein Kinase ◽

Smooth Muscle Cells ◽

Thin Filament ◽

Muscle Cells ◽

Biochemical Properties ◽

Confocal Imaging ◽

Isolated Cells ◽

Independent Manner

Calponin isolated from chicken gizzard smooth muscle binds in vitro to actin in a Ca(2+)-independent manner and thereby inhibits the actin-activated Mg(2+)-adenosinetriphosphatase of smooth muscle myosin. This inhibition is relieved when calponin is phosphorylated by protein kinase C or Ca2+/calmodulin-dependent protein kinase II, suggesting that calponin is involved in thin filament-associated regulation of smooth muscle contraction. To further examine this possibility, calponin was isolated from toad stomach smooth muscle, characterized biochemically, and localized in intact isolated cells. Toad stomach calponin had the same basic biochemical properties as calponin from other sources. Confocal immunofluorescence microscopy revealed that calponin in intact smooth muscle cells was localized to long filamentous structures that were colabeled by antibodies to actin or tropomyosin. Preservation of the basic biochemical properties of calponin from species to species suggests that these properties are relevant for its in vivo function. Its colocalization with actin and tropomyosin indicates that calponin is associated with the thin filament in intact smooth muscle cells.

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Effect of Paclitaxel+Hirudin on the TLR4-MyD88 Signaling Pathway During Inflammatory Activation of Human Coronary Artery Smooth Muscle Cells and Mechanistic Analysis

Cellular Physiology and Biochemistry ◽

10.1159/000494588 ◽

2018 ◽

Vol 50 (4) ◽

pp. 1301-1317 ◽

Cited By ~ 1

Author(s):

Hongmei Li ◽

Xian Wang ◽

Anlong Xu

Keyword(s):

Smooth Muscle ◽

Coronary Artery ◽

Smooth Muscle Cells ◽

Muscle Cells ◽

Dependent Manner ◽

Human Coronary Artery ◽

Post Intervention ◽

Inflammatory Activation ◽

Myd88 Pathway

Background/Aims: Approximately 10%-20% of patients with acute cardiovascular disease who have received coronary intervention suffer restenosis and high inflammation. The stent compound paclitaxel+hirudin was prepared for the treatment of post-intervention restenosis. This study aimed to explore the anti-inflammatory and anti-restenosis mechanisms of paclitaxel+hirudin with regard to the TLR4/MyD88/NF-κB pathway. Methods: Human coronary artery smooth muscle cells (HCASMCs) at 4-6 generations after in vitro culture were used as a model. Lipopolysaccharide (LPS) was used as an inducer to maximally activate the TLR4/MyD88/NF-κB inflammation pathway. After MyD88 knockdown and selective blocking of MyD88 degradation with epoxomicin, the effects of paclitaxel+hirudin stenting on key sites of the TLR4/MyD88/NF-κB pathway were detected using ELISA, Q-PCR, and western blot analysis. Results: LPS at 1 μg/mL for 48 h was the optimal modeling condition for inflammatory activation of HCASMCs. Paclitaxel+hirudin inhibited the levels of key proteins and the gene expression, except for that of the MyD88 gene, of the TLR4-MyD88 pathway. The trend of the effect of paclitaxel+hirudin on the pathway proteins was similar to that of MyD88 knockdown. After epoxomicin intervention, the inhibitory effects of paclitaxel+hirudin on the key genes and proteins of the TLR4-MyD88 pathway were significantly weakened, which even reached pre-intervention levels. Paclitaxel+hirudin affected the MyD88 protein in a dosage-dependent manner. Conclusion: The paclitaxel+hirudin compound promotes MyD88 degradation in the TLR4/MyD88/NF-κB pathway to reduce the activity of TLR4 and NF-κB p65 and to weaken the LPS-initiated inflammatory reactions of IL-1β, IL-6, and TNF-α.

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Protective Effects of Adiponectin against Cobalt Chloride-Induced Apoptosis of Smooth Muscle Cells via cAMP/PKA Pathway

BioMed Research International ◽

10.1155/2020/7169348 ◽

2020 ◽

Vol 2020 ◽

pp. 1-11

Author(s):

Jingjie Xiao ◽

Yingying Zhang ◽

Wei Zhang ◽

Liang Zhang ◽

Li Li ◽

...

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Cell Viability ◽

Cobalt Chloride ◽

Muscle Cells ◽

Camp Level ◽

Control Group ◽

Dependent Manner ◽

Sod Activity

Adiponectin (APN) is an adipokine secreted from adipose tissue and exhibits biological functions such as microcirculation-regulating, hearing-protective, and antiapoptotic. However, the effect of APN on the apoptosis of spiral arterial smooth muscle cells (SMCs) under hypoxic conditions in vitro is not clear. We used cobalt chloride (CoCl2) to simulate chemical hypoxia in vitro, and the SMCs were pretreated with APN and then stimulated with CoCl2. The viability of cells and apoptosis were assessed by CCK-8 and flow cytometry, respectively. Superoxide dismutase (SOD) activity, malondialdehyde (MDA) levels, cAMP level, and the activity of PKA were detected by ELISA. Protein expression and localization were studied by Western blot and immunofluorescence analysis. In the present study, we found that APN exhibits antiapoptosis effects. CoCl2 exhibited decreased cell viability, increased apoptosis and MDA levels, and decreased SOD activity in a concentration-dependent manner, compared with the control group. Moreover, CoCl2 upregulated the expression levels of Bax and cleaved caspase-3 and then downregulated Bcl-2 levels in a time-dependent manner. Compared with the CoCl2 group, the group pretreated with APN had increased cell viability, SOD activity, PKA activity, cAMP level, and PKA expression, but decreased MDA levels and apoptosis. Lastly, the protective effect of APN was blocked by cAMP inhibitor SQ22536 and PKA inhibitor H 89. These results showed that APN protected SMCs against CoCl2-induced hypoxic injury via the cAMP/PKA signaling pathway.

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Endothelium-independent, ouabain-sensitive relaxation of bovine coronary arteries by EETs

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.2001.280.3.h1113 ◽

2001 ◽

Vol 280 (3) ◽

pp. H1113-H1121 ◽

Cited By ~ 33

Author(s):

Phillip F. Pratt ◽

Pinlan Li ◽

Cecilia J. Hillard ◽

Jason Kurian ◽

William B. Campbell

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Potassium Chloride ◽

Coronary Arteries ◽

Muscle Tone ◽

Muscle Cells ◽

Resting Membrane Potential ◽

Dependent Manner ◽

Independent Manner ◽

Concentration Dependent Manner

Endothelium-derived hyperpolarizing factor (EDHF) is released in response to agonists such as ACh and bradykinin and regulates vascular smooth muscle tone. Several studies have indicated that ouabain blocks agonist-induced, endothelium-dependent hyperpolarization of smooth muscle. We have demonstrated that epoxyeicosatrienoic acids (EETs), cytochrome P-450 metabolites of arachidonic acid, function as EDHFs. To further test the hypothesis that EETs represent EDHFs, we have examined the effects of ouabain on the electrical and mechanical effects of 14,15- and 11,12-EET in bovine coronary arteries. These arteries are relaxed in a concentration-dependent manner to 14,15- and 11,12-EET (EC50 = 6 × 10−7 M), bradykinin (EC50 = 1 × 10−9 M), sodium nitroprusside (SNP; EC50 = 2 × 10−7 M), and bimakalim (BMK; EC50 = 1 × 10−7 M). 11,12-EET-induced relaxations were identical in vessels with and without an endothelium. Potassium chloride (1–15 × 10−3 M) inhibited [3H]ouabain binding to smooth muscle cells but failed to relax the arteries. Ouabain (10−5 to 10−4 M) increased basal tone and inhibited the relaxations to bradykinin, 11,12-EET, and 14,15-EET, but not to SNP or BMK. Barium (3 × 10−5 M) did not alter EET-induced relaxations and ouabain plus barium was similar to ouabain alone. Resting membrane potential ( E m) of isolated smooth muscle cells was −50.2 ± 0.5 mV. Ouabain (3 × 10−5 and 1 × 10−4 M) decreased E m(−48.4 ± 0.2 mV), whereas 11,12-EET (10−7 M) increased E m (−59.2 ± 2.2 mV). Ouabain inhibited the 11,12-EET-induced increase in E m. In cell-attached patch clamp studies, 11,12-EET significantly increased the open-state probability ( NP o) of a calcium-activated potassium channel compared with control cells (0.26 ± 0.06 vs. 0.02 ± 0.01). Ouabain did not change NP o but blocked the 14,15-EET-induced increase in NP o. These results indicate that: 1) EETs relax coronary arteries in an endothelium-independent manner, 2) unlike EETs, potassium chloride does not relax the coronary artery, and 3) ouabain inhibits bradykinin- and EET-induced relaxations as has been reported for EDHF. These findings provide further evidence that EETs are EDHFs.

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In vitro elastogenesis: instructing human vascular smooth muscle cells to generate an elastic fiber-containing extracellular matrix scaffold

Biomedical Materials ◽

10.1088/1748-6041/10/3/034102 ◽

2015 ◽

Vol 10 (3) ◽

pp. 034102 ◽

Cited By ~ 27

Author(s):

Svenja Hinderer ◽

Nian Shen ◽

Léa-Jeanne Ringuette ◽

Jan Hansmann ◽

Dieter P Reinhardt ◽

...

Keyword(s):

Extracellular Matrix ◽

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cells ◽

Vascular Smooth Muscle Cells ◽

Elastic Fiber ◽

Muscle Cells ◽

Extracellular Matrix Scaffold

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Cholesterol loading in vivo and in vitro alters extracellular matrix proteins production in smooth muscle cells

European Journal of Lipid Science and Technology ◽

10.1002/ejlt.201500287 ◽

2015 ◽

Vol 118 (9) ◽

pp. 1317-1325

Author(s):

Rogelio Palomino-Morales ◽

Sonia Perales ◽

Carolina Torres ◽

Ana Linares ◽

Maria Jose Alejandre

Keyword(s):

Extracellular Matrix ◽

Smooth Muscle ◽

Smooth Muscle Cells ◽

Extracellular Matrix Proteins ◽

Muscle Cells ◽

Matrix Proteins

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Extracellular matrix metalloproteinase inducer (EMMPRIN) is present in smooth muscle cells of human aneurysmal aorta and is induced by angiotensin II in vitro

Clinical Science ◽

10.1042/cs20080235 ◽

2009 ◽

Vol 116 (11) ◽

pp. 819-826 ◽

Cited By ~ 13

Author(s):

Xiao-feng Chen ◽

Jian-an Wang ◽

Jun Hou ◽

Chun Gui ◽

Li-jiang Tang ◽

...

Keyword(s):

Extracellular Matrix ◽

Smooth Muscle ◽

Angiotensin Ii ◽

Aortic Aneurysm ◽

Aortic Dissection ◽

Matrix Metalloproteinase ◽

Smooth Muscle Cells ◽

Muscle Cells ◽

Extracellular Matrix Metalloproteinase Inducer

The aim of the present study was to determine whether EMMPRIN (extracellular matrix metalloproteinase inducer) is present and is up-regulated in human aneurysmal aortas, and to assess a possible association with AngII (angiotensin II)-induced aneurysm formation. The presence of EMMPRIN was assessed in 41 surgical specimens from patients with a TAA (thoracic aortic aneurysm) (Type A aortic dissection, n=12; Type B aortic dissection, n=7; and TAA without dissection, n=7) or an AAA (abdominal aortic aneurysm, n=15) by immunohistochemistry. EMMPRIN expression in aortic aneurysm tissues was compared with 12 aortas obtained during autopsy (free of any vascular diseases), and scored for both staining intensity and the percentage of vascular cells stained. EMMPRIN protein levels in cultured human aortic SMCs (smooth muscle cells) following stimulation of AngII were analysed by Western blotting. Significant EMMPRIN immunoreactivity was detected in aortic aneurysm lesions from patients with TAAs and AAAs. In the aneurysmal wall, α-actin-positive SMCs were the main source of EMMPRIN. The frequency of EMMPRIN overexpression was significantly higher (P=0.026) in TAAs with dissection (68.4%) than TAAs without dissection (14.3%). AngII stimulation up-regulated the expression of EMMPIRN in cultured human aortic SMCs, which was suppressed by the addition of the AT1R (AngII type 1 receptor) antagonist losartan. In conclusion, the present study is the first to report the expression of EMMPRIN in aortic aneurysmal diseases, and we speculate that EMMPRIN may be important in the pathogenesis of these diseases. Whether these abnormalities are potential therapeutic targets deserve further investigation.

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