Vascular Smooth Cell Migration Research Articles

WD Repeat Domain 1 Deficiency Inhibits Neointima Formation in Mice Carotid Artery by Modulation of Smooth Muscle Cell Migration and Proliferation.

The migration, dedifferentiation, and proliferation of vascular smooth muscle cells (VSMCs) are responsible for intimal hyperplasia, but the mechanism of this process has not been elucidated. WD repeat domain 1 (WDR1) promotes actin-depolymerizing factor (ADF)/cofilin-mediated depolymerization of actin filaments (F-actin). The role of WDR1 in neointima formation and progression is still unknown. A model of intimal thickening was constructed by ligating the left common carotid artery in Wdr1 deletion mice, and H&E staining showed that Wdr1 deficiency significantly inhibits neointima formation. We also report that STAT3 promotes the proliferation and migration of VSMCs by directly promoting WDR1 transcription. Mechanistically, we clarified that WDR1 promotes the proliferation and migration of VSMCs and neointima formation is regulated by the activation of the JAK2/STAT3/WDR1 axis.

GLP-1 attenuates Ang II-induced proliferation and migration in rat aorta smooth muscle cells via inhibition of the RhoA/ROCK2 signaling pathway.

Glucagon-like peptide 1 (GLP-1), a neuroendocrine hormone produced by the gastrointestinal tract, plays a significant role in blood glucose regulation; drugs derived from GLP-1 are currently used for the treatment of type 2 diabetes. In addition to regulating glucose homeostasis, the protective effects of GLP-1 on the cardiovascular system are also of interest. However, the vascular protective mechanisms of GLP-1 remain unclear. The present study was designed to evaluate the role of GLP-1 in the proliferation and migration of vascular smooth muscle cells, and the underlying mechanisms. In this study, proliferation, migration, cyclin D1 expression, and phosphorylation of MLC, as well as RhoA and Rho-associated coiled-coil forming protein kinase 2 (ROCK2) expression, were increased in rat aorta smooth muscle cells (RASMCs) following incubation with angiotensin II (Ang II). These effects were significantly attenuated by GLP-1, forskolin (a cAMP activator) and Y-27632 (a ROCK2 inhibitor). However, H89 (a PKA inhibitor) inhibited the action of GLP-1, both in terms of inhibition of RASMC proliferation and migration, and RHOA/ROCK2 expression. These results indicate that GLP-1 inhibits Ang II-induced RASMC proliferation and migration via the cAMP/PKA/RhoA/ROCK2 signaling pathway. Our data suggest that GLP-1 should be considered for use in the clinical treatment of cardiovascular diseases, in addition to its current use in the treatment of diabetes mellitus.

Effect of angiotensin1-7 on intimal hyperplasia in autogenous vein graft in rats

Objective Angiotensin1-7 (Ang1-7) and ITS inhibitor A779 were used to study the effect of Ang1-7 on intimal hyperplasia in autogenous vein graft. Methods The transplantation models of the external jugular vein to the abdominal aorta were established in 40 Specific Pathogen Free SD male rats. All models were randomly divided into control group, gel group, Ang1-7 group and A779 group (10 rats each). The adventitia of the transplanted veins was evenly sprayed with normal saline, Poloxamer 407, Poloxamer 407 mixed with Ang 1-7, Poloxamer 407 mixed with A779 respectively in each group forementioned. 5 rats form each group were sacrificed at 14th and 28th day respectively. The transplanted vein was intercepted and then stained with hematoxylin and eosin (HE). The thickness of the inner membrane (I) and medium membrane (M) of the vein grafts were measured by computer, and I/M ratio was calculated. The expression of proliferating cell nuclear antigen (PCNA), extracellular regulated protein kinase-1 (ERK-1) and angiotensin Ⅱ receptor 1 (AT1R) was determined by immunohistochemistry. Results (1)14th and 28th Day, the intimal hyperplasia of vein graft in Ang1-7 group [(59.26±7.51), (62.50±5.91) μm] was lighter than that in the other groups(P=0.001, respectively). (2)14th Day, the I/M ratio of vein graft in Ang1-7 group (0.67±0.05) was lower compare to that in other groups (P=0.001, 0.006, 0.001). 28th Day, the I/M ratio of vein graft in Ang1-7 group (0.71±0.14) was lower than that in the other groups(P=0.001, respectively). (3)There was statistically differences in the intimal thickness of vein graft between A779 group and control group at 14th, 28th day(P=0.007, 0.001). (4)14th Day, the expression of PCNA in the Ang1-7 group (21.80±7.05) was lower than that in other groups (P=0.011, 0.016, 0.001). 28th Day, the PCNA expression in the Ang1-7 group (12.80±5.31) was lower than that in the other groups (P=0.001, 0.002, 0.001). (5)14th Day, the express of ERK1 in vein graft Ang1-7 group (57.00±5.79) was lower than that in other groups (P=0.002, 0.003, 0.001). 28th Day, the express of ERK1 in vein graft Ang1-7 group(50.20±8.79) was lower than that in other groups (P=0.024, 0.017, 0.001). (6)14th Day, the express of AT1R in vein graft Ang1-7 group (27.20±6.87) was lower than that in other groups (P=0.001, 0.003, 0.001). 28th Day, the express of PCNA in vein graft Ang1-7 group(20.60±4.83) was lower than that in other groups (P=0.001, respectively). Conclusion The main cells resulting in intimal hyperplasia may be derived from the proliferation and migration of vascular smooth muscle cells in the medium membrane. Ang1-7 can significantly inhibit the expression of AT1R in the bridging vein, reduce its role in promoting endometrial proliferation, thereby reduce the proliferation and migration of VSMCs. Ang1-7 can also inhibit the expression of ERK in the bridging vein after vein transplantation; RAS and MAPK signal systems are involved in the intimal hyperplasia of vein graft. Key words: Angiotensin1-7; Vein graft; Intimal hyperplasia; Renin angiotensin system

Inhibition of intimal hyperplasia in murine aortic allografts by administration of a small-molecule TLR4 inhibitor TAK-242

Graft arteriosclerosis (GA) is the leading cause of late cardiac allograft dysfunction. The innate immune system plays a major role in GA, paprticularly Toll-like receptor 4 (TLR4) signaling. Here we characterized the role of TLR4 and its antagonist TAK-242 in a mouse model of GA. BALB/c (H-2d) donor aortas were transplanted into C57BL/6 (H-2b) recipients, and the mice received intraperitoneal injection of 3 or 10 mg/kg of TAK-242 or vehicle every other day for 1, 2, 4, 6, 8 and 12 weeks. With TAK-242 administration, intimal hyperplasia initially appeared at 2 weeks after transplantation, and TAK-242 postponed the progression of neointimal formation in allogeneic aortic grafts. TAK-242 treatment reduced CD68+ macrophage accumulation in the allografts, reduced the levels of ly-6Chi monocytes in peripheral blood, bone marrow and spleen, and downregulated proinflammatory cytokine and chemokine levels. Ex vivo we observed that TAK-242 could improve the graft microenvironment by interfering the Tck/Mφ IL12p70 and IFNγ axis, reducing CCL2-mediated migration of vascular smooth cells.

Effects of small interfering RNA silencing in epidermal growth factor receptor on proliferation and migration of vascular smooth muscle cells

Objective To investigate the effect of silencing epidermal growth factor receptor (EGFR) by small interfering RNA (siRNA) interference on the proliferation and migration of vascular smooth muscle cells (VSMCs), and the related mechanisms. Methods VSMCs of rats were isolated and cultured. VSMCs were infected with EGFR siRNA lentiviral vectors. The cells were divided into siRNA-EGFR group, siRNA-negative control group and blank control group. The cell proliferation abilities were detected by 5’-bromo-2’-deoxyuridine (5-BrdU) method. The cell migration abilities in different transfection groups were examined by Transwell chamber and scratch assays. The expression levels of EGFR, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 genes in different transfection groups were detected using real-time quantitative polymerase chain reaction (Real-time PCR) technology. The expression levels of EGFR, p-ERK, p-JNK and p-p38 proteins in different transfection groups were detected by Western blotting. Results The number of migration cells in the siRNA-EGFR group was 48.5±6.1, which were significantly lower than the siRNA-negative control group and blank control group, which were 86.2±7.2 and 88.3±7.6; the difference was statistically significant (F=37.261, P=0.000). Compared with 0 h, after 24 h, the migration rate of VSMC in the siRNA-EGFR group was (35.7±3.8)%, which was significantly lower than that of siRNA-EGFR group and blank group, which were (74.2±5.7)% and (75.5±6.1)%; the differences were statistically significant (F=40.315, P=0.000). The relative expression levels of ERK, JNK and p38 mRNA in the siRNA-EGFR group (0.53±0.09, 0.47±0.11, 0.41±0.07) were lower than the siRNA-negative control group (0.82±0.12, 0.64±0.10, 0.68±0.09) and blank control group (0.85±0.14, 0.66±0.13, 0.70±0.12), the differences were statistically significant (F=15.391, 12.817, 11.264, P=0.000, 0.000, 0.000). The relative expression levels of p-ERK, p-JNK and p-p38 proteins in the siRNA-EGFR group (0.46±0.09, 0.38±0.06, 0.33±0.06) were lower than the siRNA-negative control group (0.71±0.12, 0.57±0.08, 0.52±0.07) and blank control group (0.73±0.13, 0.56±0.07, 0.54±0.09), the differences were statistically significant (F=20.152, 14.375, 13.843, P=0.000, 0.000, 0.000). Conclusion Specific inhibition of EGFR could effectively reduce proliferation and migration capacities of VSMCs, which might be related to inhibition of mitogen-activated protein kinase signaling pathway. Key words: Vascular smooth muscle cells; Epidermal growth factor receptor; Small interference; Cell proliferation; Cell migration

Impact and related mechanism of exogenous receptor activity modifying protein 1 on calcitonin gene-related peptide modified bone marrow mesenchymal stem cells on the migration of vascular smooth muscle cells in vitro

To investigate the impact of calcitonin gene-related peptide (CGRP) modified bone marrow mesenchymal stem cell (MSC) on the migration of vascular smooth muscle cell (VSMC) and related mechanisms. The MSC and VSMC were isolated from rats and cultured, CGRP was transfected to MSC with the high expression lentivirus vector, VSMC was transfected with high expression lentivirus vector of receptor activity modifying protein 1 (RAMP1) and the silence expression lentivirus vector of RAMP1. Then MSC was co-cultured with VSMC. Experimental groups were as follows: (1) Ang II group (MSC + VSMC + Ang II); (2) MSC(CGRP+) group (MSC(CGRP+) + VSMC + Ang II); (3) MSC(CGRP+) RAMP1(-) group (MSC(CGRP+) + VSMC(RAMP1-) + Ang II); (4) MSC(CGRP+) RAMP1(+) group (MSC(CGRP+) + VSMC(RAMP1+) + Ang II); (5) RAMP1(+) group (MSC + VSMC(RAMP1+) + Ang II). Transwell assay was applied to detect the migration of smooth muscle cells, Western blot was applied to detect the protein expression of cells in various groups. VSMC migration number was significantly lower in MSC(CGRP+) group compared with Ang II group (50.8 ± 2.6 vs. 71.4 ± 2.3, P < 0.05), but higher than in MSC(CGRP+) RAMP1(+) group (50.8 ± 2.6 vs. 30.4 ± 3.0, P < 0.05). When RAMP1 expression reduced in VSMC, compared with MSC(CGRP+) RAMP1(+) group, VSMC migration increased in the MSC(CGRP+) RAMP1(-) group compared to MSC(CGRP+)RAMP1(+) (69.0 ± 5.6 vs. 30.4 ± 3.0, P < 0.05) and was similar to Ang II group (69.0 ± 5.6 vs. 71.4 ± 2.3, P > 0.05) and RAMP1(+) group (71.6 ± 3.4). According to the result of Western blot, P-P65 protein expression in MSC(CGRP+) group was lower than that in Ang II group (0.475 ± 0.022 vs.0.642 ± 0.035, P < 0.05). P-P65 protein expression in MSC(CGRP+)RAMP1(-) group was higher than that in MSC(CGRP+) RAMP1(+) group (0.670 ± 0.030 vs. 0.373 ± 0.041, P < 0.05), and there was no difference between MSC(CGRP+)RAMP1(-) group and Ang II group (P > 0.05). P-P65 protein expression was similar between RAMP1(+) group (0.643 ± 0.039) and Ang II group (P > 0.05). CGRP inhibits VSMC migration through RAMP1. NF-κB and RAMP1 play crucial role in the inhibiting effects of CGRP on VSMC migration. Thus, RAMP1-CGRP signaling inhibits VSMC migration through NF-κB signal pathways.

Effect of integrin signaling pathway on proliferation and migration of vascular smooth muscle cell

Many cardiovascular diseases, such as pulmonary hypertension, atherosclerotic plaque formation and restenosis after angioplasty, are charaterized by vascular remodeling resulting from migration and proliferation of vascular smooth muscle cells. Integin receptors can activate the expression of related genes through the corresponding signal pathway, and lead to vascular smooth muscle cell' s proliferation and migration and other biological behavioral change, which may be a new hotspot of pathogenesis and intervention of vascular remodeling. Key words: Integrin; Vascular smooth muscle cell; Proliferation; Migration

Letter to the editor: Is homocysteine the culprit molecule in vascular diseases or just a bystander?

to the editor: Jiang et al. ([3][1]) reported an important finding that vascular smooth cell migration was promoted by the resistin gene expression induced by ultrahigh concentrations of homocysteine (500 μM). The homocysteine concentrations used by Jiang et al. were markedly higher than the

The role of Syk in migration and proliferation of the vascular smooth muscle cells

Spleen tyrosine kinase(Syk) has been considered to be a signaling molecule of hematopoietic cells,which play an important role in lymphocyte maturation and activation of immune cells.Recent studies showed that Syk plays a crucial role in regulation of proliferation and migration in vascular smooth muscle cells.Prominent pathways include mitogen-activated protein kinases,phosphatidylinositol 3-kinases,Src and Rho-activated protein kinase.In this review,the role of Syk in those signal pathways is summarized. Key words: Spleen tyrosine kinase; Vascular smooth muscle cells; Signal transduction

The expression of resistin-like molecules-a in atherosclerotic plaque

Objective To explore the expression of resistin-like molecules-a (RELMa) in atherosclerotic plaque of ApoE-/- mouse, and to study the effects of RELMa on the proliferation and migration of vascular smooth musclee cells (VSMCs) . Method Nine ApoE-/- mice and nine C57BL/6J mice were fed with high fat diet. All mice were sacrificed 24 weeks after force feeding. Vessels were dissected from to abdominal aorta. Sections of aortic tissue were stained with HE dyeing and RELMa in aortic tissue was assayed by immunohistochemistry. The expression of RELMa mRNA in vessels was detected by RT-PCR. The effects of different concentration RELMa in different concentrations on the proliferation and migration of VSMCs were detected. Data was expressed as mean ± standard deviation. ANOVA were used for comparison in SPSS 11.0, and changes were considered as statistically significant if P value was less than 0.05. Results Atherosclerosis plaque formed in aortic root of ApoE-/- mice after they were fed with high fat diet for 24 weeks. RELMa protein and RELMa mRNA were found by immunohistochemistry and RT-PCR in atherosclerotic plaque of ApoE-/- mouse. RELMa protein didn't be found in vessels of control mouse. RELMa promoted the proliferation of VSMCs (RELMa groups: 2811. 21 ± 216. 89,4056. 87 ±220.65,5061.45 ± 335.86, vs. control 1609.58 ± 203.53, P < 0.01). RELMa promoted the migration of VSMCs (RELMa groups: 130.54±12.98,158.39±11.58,203.50± 17.37 vs. control:70.54± 11.92, P<0.01).Conclusions RELMa expresses in atherosclerotic plaque of ApoE-/- mouse. RELMa enhances the proliferation and migration of VSMCs of aorta. Key words: Artherosclerosis; RELMα; Vascular smooth muscles cells; Proliferation

Effects of the inhibitors of PI3K, P38 MAPK and ERK1/2 on the migration of EGF-induced vascular smooth muscle cells

Objective To study the effects of Wortmannin, inhibitor of PI3K and SB202190, inhibitor of P38 MAPK and PD98059, inhibitor of ERK1/2 on the migration of epidermal growth factor (EGF)-induced vascular smooth muscle cells (VSMCs). Methods There were fives groups in this experiment, including control group, EGF group, PD98059 (PD) group, SB202190(SB) group and Wortnannin (WT) group. The migration rate of the VSMCs was measured by wound healing assay. Results At the 24th hours after wounding, there was obvious migration in EGF group compared to control group. The migration of VSMCs was significantly inhibited in PD group, SB group and WT group compared to the EGF group, but there were no significant difference among three inhibitor groups. At the 30th hours after woun-ding, there was still obvious migration in EGF group compared to control group. The migration of VSMCs was significantly inhibited in the three iuhibitors group compared to the EGF group, and there were significant difference among three inhibitor groups. Furthermore, inhibiting effect on VSMCs in SB group was more obvious compared to PD group and WT group. Conclusion These results suggested that the migra-tion of EGF-induced VSMCs may play a role through PI3K, P38 MAPK and ERKI/2 signal pathways, and the effect of P38 MAPK signal pathway is very important. Key words: 1-phosphatidylinositel 3-kinase/AI ; p38 mitogen-activated protein kinases/M; Myocytes, smooth muscle/DE; Epider-mal growth factor

Experimental Studies and Numerical Analysis of the Inflation and Interaction of Vascular Balloon Catheter-Stent Systems

Balloon angioplasty with stenting is a well-established interventional procedure to treat stenotic arteries. Despite recent advances such as drug eluting stents, clinical studies suggest that stent design is linked to vascular injury. Additionally, dilation of the medical devices may trigger pathological responses such as growth and migration of vascular smooth cells, and may be a potent stimulus for neointimal hyperplasia. The purpose of this study is to experimentally investigate the mechanical characteristics of the transient expansion of six commercially available balloon-expandable stent systems, and to develop a robust finite element model based on the obtained experimental results. To reproduce the inflation of stent systems as in clinical practice, a pneumatic-hydraulic experimental setup is built, able to record loads and deformations. Characteristic pressure-diameter diagrams for the balloon-expandable stents and the detached balloons are experimentally obtained. Additionally, typical measures such as the burst opening pressure, the maximum dog-boning and foreshortening, and the elastic recoil are determined. The adopted constitutive models account for elastoplastic deformation of the stent, and for the nonlinear and anisotropic behavior of the balloon. The employed contact algorithm, based on a C(2)-continuous surface parametrization, efficiently simulates the interaction of the balloon and stent. The computational model is able to successfully capture the experimentally observed deformation mechanisms. Overall, the numerical results are in satisfactory agreement with experimental data.

Activation of IP and EP 3 receptors alters cAMP-dependent cell migration

Migration of vascular smooth cells from the media to the intima essentially contributes to neointima formation after percutaneous transluminal angioplasty and stent implantation. The stable prostacyclin mimetic iloprost has been shown to inhibit neointima formation in experimental restenosis, but it is currently unknown whether this may be caused by an antimigratory effect. Hence, the present study analyses (i) the influence of G s-coupled prostacyclin (IP) receptors on cell migration and (ii) verifies whether EP 3 receptors with opposite (i.e., G i) coupling may conversely stimulate cell migration. In a modified Boyden chamber model, it was shown that iloprost dose-dependently inhibits the migration of primary human arterial smooth muscle cells, which constitutively express the IP receptor. On the other hand, human arterial smooth muscle cell migration was stimulated by the EP 3 receptor agonist M&B 28.767. To independently study the effects of these receptors, IP or EP 3 receptors were stably overexpressed in chinese hamster ovary cells (CHO-IP and CHO-EP 3). Chemotaxis of CHO cells transfected with G s-coupled IP receptors was concentration-dependently inhibited by iloprost (2–100 nM), while there was no effect of iloprost on mock-transfected CHO. By contrast, CHO-cells that overexpressed EP 3 receptors showed a significant, concentration dependent (1–100 nM) increase of cell migration in presence of the selective EP 3 agonist M&B 28.767. It is concluded that the prostacyclin mimetic iloprost inhibits vascular cell migration, which probably depends on a G s-mediated increase of intracellular cAMP. EP 3 receptors conversely stimulate CHO migration.

Chronic Angiotensin II Augments Integrin-Dependent Cell Functions Via PKC and Map-Kinase Activation in Rat Vsmcs

P78 The activation of local and systemic renin-angiotensin-system is directly and indirectly involved in the mechanisms of vascular remodeling during chronic hypertension. The following study investigated the effect of chronic angiotensin II (AII) admininstration on cell- matrix interaction of VSMCs in vitro. Adhesion to vitronectin (VN) and collagen I (ColI) as well as migration of rat vascular smooth muscle cells (VSMCs) were significantly augmented by chronic treatment with AII for 48 hours and 72 hours. These changes were not due to an increase of the integrins on the cells surface known to be capable in binding vitronectin and collagen I: alpha1, alpha5, beta1 and beta5, as demonstrated by flow cytometry. Also the phosporylation of the focal adhesion kinase (FAK), known to play an important role in adhesion and spreading processes, was not affected after chronic stimulation with Ang II. Downregulation of PKC activity by phorbol-myristate acetate (PMA, 0.1μmol/L), or treatment with the PKC inhibitor calphostin C before treatment with AII significantly reduced the effect of AII on adhesion to VN and ColI (each p

Endothelial cells exposed to nicotine act as a chemoattractant for vascular smooth cell migration

Endothelial cells exposed to nicotine act as a chemoattractant for vascular smooth cell migration