RESUMO
Vascular remodeling and restenosis are common complications after percutaneous coronary intervention. Excessive proliferation and migration of vascular smooth muscle cells (VSMCs) play important roles in intimal hyperplasia-induced vascular restenosis. NK2 Homeobox 3 (Nkx2-3), a critical member of Nkx family, is involved in tissue differentiation and organ development. However, the role of Nkx2-3 in VSMCs proliferation and migration remains unknown. In this study, we used carotid balloon injury model and platelet-derived growth factor-BB (PDGF)-treated VSMCs as in vivo and in vitro experimental models. EdU assay and CCK-8 assay were used to detect cell proliferation. Migration was measured by scratch test. Hematoxylin and eosin staining and immunohistochemistry staining were used to evaluate the intimal hyperplasia. The autophagy level was detected by fluorescent mRFP-GFP-LC3 in vitro and by transmission electron microscopy in vivo. It was shown that Nkx2-3 was upregulated both in balloon injured carotid arteries and PDGF-stimulated VSMCs. Adenovirus-mediated Nkx2-3 overexpression inhibited intimal hyperplasia after balloon injury, and suppressed VSMCs proliferation and migration induced by PDGF. Conversely, silencing of Nkx2-3 by small interfering RNA exaggerated proliferation and migration of VSMCs. Furthermore, we found that Nkx2-3 enhanced autophagy level, while the autophagy inhibitor 3-MA eliminated the inhibitory effect of Nkx2-3 on VSMCs proliferation and migration both in vivo and in vitro. Moreover, Nkx2-3 promoted autophagy in VSMCs by activating the AMP-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) signaling pathway. These results demonstrated for the first time that Nkx2-3 inhibited VSMCs proliferation and migration through AMPK/mTOR-mediated autophagy.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Autofagia , Lesões das Artérias Carótidas/enzimologia , Movimento Celular , Proliferação de Células , Proteínas de Homeodomínio/fisiologia , Músculo Liso Vascular/enzimologia , Miócitos de Músculo Liso/enzimologia , Serina-Treonina Quinases TOR/metabolismo , Fatores de Transcrição/fisiologia , Animais , Autofagia/efeitos dos fármacos , Becaplermina/farmacologia , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/patologia , Lesões das Artérias Carótidas/prevenção & controle , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Modelos Animais de Doenças , Proteínas de Homeodomínio/genética , Masculino , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/ultraestrutura , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/ultraestrutura , Neointima , Ratos Sprague-Dawley , Transdução de Sinais , Fatores de Transcrição/genética , Remodelação VascularRESUMO
Dysfunction of late endothelial progenitor cells (EPCs) has been suggested to be associated with hypertension. ß2-Adrenergic receptor (ß2AR) is a novel and key target for EPC homing. Here, we proposed that attenuated ß2AR signaling contributes to EPCs dysfunction, whereas enhanced ß2AR signaling restores EPCs' functions in hypertension. EPCs derived from hypertensive patients exhibited reduced cell number, impaired in vitro migratory and adhesion abilities, and impaired re-endothelialization after transplantation in nude mice with carotid artery injury. ß2AR expression of EPCs from hypertensive patients was markedly downregulated, whereas the phosphorylation of the p38 mitogen-activated protein kinase (p38-MAPK) was elevated. The cleaved caspase-3 levels were elevated in EPCs. The overexpression of ß2AR in EPCs from hypertensive patients inhibited p38-MAPK signaling, whereas it enhanced in vitro EPC proliferation, migration, and adhesion and in vivo re-endothelialization. The ß2AR-mediated effects were attenuated by treating the EPCs with a neutralizing monoclonal antibody against ß2AR, which could be partially antagonized by the p38-MAPK inhibitor SB203580. Moreover, shear stress stimulation, a classic nonpharmacological intervention, increased the phosphorylation levels of ß2AR and enhanced the in vitro and in vivo functions of EPCs from hypertensive patients. Collectively, the current investigation demonstrated that impaired ß2AR/p38-MAPK/caspase-3 signaling at least partially reduced the re-endothelialization capacity of EPCs from hypertensive patients. Restoration of ß2AR expression and shear stress treatment could improve their endothelial repair capacity by regulating the p38-MAPK/caspase-3 signaling pathway. The clinical significance of ß2AR in endothelium repair still requires further investigation.NEW & NOTEWORTHY Impaired ß2-adrenergic receptor (ß2AR) expression with an elevation of p38-MAPK/caspase-3 signaling at least partially contributes to the decline of re-endothelialization capacity of late endothelial progenitor cells (EPCs) from hypertensive patients. ß2AR gene transfer and shear stress treatment improve the late EPC-mediated enhancement of the re-endothelialization capacity in hypertensive patients through activating ß2AR/p38-MAPK/caspase-3 signaling. The present study is the first to reveal the potential molecular mechanism of the impaired endothelium-reparative capacity of late EPCs in hypertension after vascular injury and strongly suggests that ß2AR is a novel and crucial therapeutic target for increasing EPC-mediated re-endothelialization capacity in hypertension.
Assuntos
Lesões das Artérias Carótidas/prevenção & controle , Proliferação de Células , Células Progenitoras Endoteliais/metabolismo , Hipertensão/metabolismo , Reepitelização , Receptores Adrenérgicos beta 2/metabolismo , Animais , Apoptose , Lesões das Artérias Carótidas/metabolismo , Lesões das Artérias Carótidas/patologia , Estudos de Casos e Controles , Caspase 3/metabolismo , Adesão Celular , Movimento Celular , Células Cultivadas , Técnicas de Cocultura , Modelos Animais de Doenças , Células Progenitoras Endoteliais/patologia , Células Progenitoras Endoteliais/transplante , Feminino , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Hipertensão/patologia , Masculino , Camundongos Nus , Pessoa de Meia-Idade , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Knowledge of variations of the internal carotid artery is significant to surgeons and radiologists. The internal carotid artery normally runs a straight course in the neck. Its anomalies can lead to its iatrogenic injuries. We report a case of a large loop of the internal carotid artery in a male cadaver aged about 75 years. The common carotid artery terminated by dividing it into the external carotid artery and internal carotid arteries at the level of the upper border of the thyroid cartilage. From the level of origin, the internal carotid artery coursed upwards, backwards and laterally, and formed a large loop behind the internal jugular vein. The variation was found on the left side of the neck and was unilateral. The uncommon looping of the internal carotid artery might result in altered blood flow to the brain and may lead to misperceptions in surgical, imaging, and invasive procedures.
Assuntos
Variação Anatômica , Artéria Carótida Interna/anormalidades , Idoso , Angiografia/métodos , Região Branquial/embriologia , Cadáver , Lesões das Artérias Carótidas/etiologia , Lesões das Artérias Carótidas/prevenção & controle , Artéria Carótida Interna/embriologia , Desenvolvimento Embrionário , Humanos , Veias Jugulares/anatomia & histologia , Masculino , Esvaziamento Cervical/efeitos adversos , Esvaziamento Cervical/métodosRESUMO
The Cordyceps extract exhibits antiproliferative potential in vascular smooth muscle cells (SMCs) through the mitogen-activated protein kinase signaling pathway. In this study, we aimed to identify the active compounds in the Cordyceps extract and analyze their role in remodeling the arterial wall. On investigation, we discovered the following active compound: 4-methoxyphenyl (E)-3-(furan-3-yl) acrylate and synthesized it. We performed antiproliferation and antimigration assays in addition to an in vivo vessel wall remodeling experiment. Investigation of the mechanism adopted by the active compound to remodel the vessel was performed. The newly synthesized compound inhibited the proliferation and migration of SMCs. Treatment with the synthesized compound reduced neointima formation in the balloon-injured Sprague-Dawley rat model. In addition, this compound inhibited the activation of matrix metalloproteinase-2 and matrix metalloproteinase-9 in type I collagen-activated SMCs. Moreover, this compound suppressed the expression of cycloxygenase-2 (COX-2) in SMCs. Therefore, this compound can exert potential antiarteriosclerotic effects by modulating vessel wall remodeling. In conclusion, the newly synthesized 4-methoxyphenyl (E)-3-(furan-3-yl) acrylate might be an alternative therapeutic intervention for the treatment of atherosclerosis.
Assuntos
Lesões das Artérias Carótidas/prevenção & controle , Proliferação de Células/efeitos dos fármacos , Cordyceps , Furanos/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , Neointima , Remodelação Vascular/efeitos dos fármacos , Animais , Lesões das Artérias Carótidas/metabolismo , Lesões das Artérias Carótidas/patologia , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Cordyceps/química , Ciclo-Oxigenase 2/metabolismo , Modelos Animais de Doenças , Furanos/síntese química , Furanos/isolamento & purificação , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Ratos Sprague-DawleyRESUMO
PURPOSE: Periosteal releasing incision (PRI) techniques are often used with guided bone regeneration procedures. As complications such as intra- and postoperative bleeding have been noticed, we aimed to study and clarify these as related to the PRI, especially on the mandibular buccal periosteum. METHODS: Fourteen sides from seven fresh-frozen Caucasian cadaveric heads were used in this study. The seven cadavers were derived from two females and five males. The mean age at the time of death was 75.9 ± 10.8 years. The PRI was made using a no. 15c blade using a surgical microscope. Subsequently, the fat tissue lateral to the periosteum was slightly dissected. The diameter of the facial artery (or its branch) and closest relationship between the tooth and position of the artery was recorded. Finally, the artery was traced back proximally to clarify its origin. RESULTS: On all sides, the inferior labial artery (ILA) was identified in the fat tissue lateral to and close to the periosteum. The ILA was closest to the periosteum at the midpoint of the PRI (approximately between the first and second molar teeth area or 10 mm mesial to the apex of the retromolar pad). The mean diameter of the ILA was 2.72 ± 0.26 mm. CONCLUSION: This anatomical finding should encourage dentists to make the PRI incision without invading the tissue underneath the periosteum.
Assuntos
Lesões das Artérias Carótidas/etiologia , Artéria Carótida Externa/anatomia & histologia , Regeneração Tecidual Guiada Periodontal/efeitos adversos , Dente Molar/anatomia & histologia , Periósteo/anatomia & histologia , Periósteo/cirurgia , Idoso , Idoso de 80 Anos ou mais , Cadáver , Lesões das Artérias Carótidas/prevenção & controle , Feminino , Regeneração Tecidual Guiada Periodontal/métodos , Humanos , Masculino , Mandíbula/anatomia & histologia , Dente Molar/cirurgia , Retalhos CirúrgicosRESUMO
OBJECTIVE: Recently, we have demonstrated that acute glucosamine-induced augmentation of protein O-linked ß-N-acetylglucosamine (O-GlcNAc) levels inhibits inflammation in isolated vascular smooth muscle cells and neointimal formation in a rat model of carotid injury by interfering with NF-κB (nuclear factor-κB) signaling. However, the specific molecular target for O-GlcNAcylation that is responsible for glucosamine-induced vascular protection remains unclear. In this study, we test the hypothesis that increased A20 (also known as TNFAIP3 [tumor necrosis factor α-induced protein 3]) O-GlcNAcylation is required for glucosamine-mediated inhibition of inflammation and vascular protection. APPROACH AND RESULTS: In cultured rat vascular smooth muscle cells, both glucosamine and the selective O-linked N-acetylglucosaminidase inhibitor thiamet G significantly increased A20 O-GlcNAcylation. Thiamet G treatment did not increase A20 protein expression but did significantly enhance binding to TAX1BP1 (Tax1-binding protein 1), a key regulatory protein for A20 activity. Adenovirus-mediated A20 overexpression further enhanced the effects of thiamet G on prevention of TNF-α (tumor necrosis factor-α)-induced IκB (inhibitor of κB) degradation, p65 phosphorylation, and increases in DNA-binding activity. A20 overexpression enhanced the inhibitory effects of thiamet G on TNF-α-induced proinflammatory cytokine expression and vascular smooth muscle cell migration and proliferation, whereas silencing endogenous A20 by transfection of specific A20 shRNA significantly attenuated these inhibitory effects. In balloon-injured rat carotid arteries, glucosamine treatment markedly inhibited neointimal formation and p65 activation compared with vehicle treatment. Adenoviral delivery of A20 shRNA to the injured arteries dramatically reduced balloon injury-induced A20 expression and inflammatory response compared with scramble shRNA and completely abolished the vascular protection of glucosamine. CONCLUSIONS: These results suggest that O-GlcNAcylation of A20 plays a key role in the negative regulation of NF-κB signaling cascades in TNF-α-treated vascular smooth muscle cells in culture and in acutely injured arteries, thus protecting against inflammation-induced vascular injury.
Assuntos
Anti-Inflamatórios/farmacologia , Lesões das Artérias Carótidas/prevenção & controle , Proteínas de Ligação a DNA/metabolismo , Glucosamina/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , NF-kappa B/metabolismo , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/metabolismo , Acetilglucosamina/metabolismo , Animais , Anti-Inflamatórios/metabolismo , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/metabolismo , Lesões das Artérias Carótidas/patologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Proteínas de Ligação a DNA/genética , Modelos Animais de Doenças , Glucosamina/metabolismo , Glicosilação , Masculino , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Neointima , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/genéticaRESUMO
OBJECTIVE: Phenotypic modulation of vascular smooth muscle cells represents a hallmark event in vascular injury. The underlying mechanism is not completely sorted out. We investigated the involvement of angiogenic factor with G patch and FHA domains 1 (Aggf1) in vascular injury focusing on the transcriptional regulation of vascular smooth muscle cell signature genes. APPROACH AND RESULTS: We report here that Aggf1 expression was downregulated in several different cell models of phenotypic modulation in vitro and in the vessels after carotid artery ligation in mice. Adenovirus-mediated Aggf1 overexpression dampened vascular injury and normalized vascular smooth muscle cell signature gene expression. Mechanistically, Aggf1 interacted with myocardin and was imperative for the formation of a serum response factor-myocardin complex on gene promoters. In response to injurious stimuli, kruppel-like factor 4 was recruited to the Aggf1 promoter and enlisted histone deacetylase 11 to repress Aggf1 transcription. In accordance, depletion of kruppel-like factor 4 or histone deacetylase 11 restored Aggf1 expression and abrogated vascular smooth muscle cell phenotypic modulation. Finally, treatment of a histone deacetylase 11 inhibitor attenuated vascular injury in mice. CONCLUSIONS: Therefore, we have unveiled a previously unrecognized role for Aggf1 in regulating vascular injury.
Assuntos
Proteínas Angiogênicas/metabolismo , Lesões das Artérias Carótidas/metabolismo , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Proteínas Angiogênicas/genética , Animais , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/patologia , Lesões das Artérias Carótidas/prevenção & controle , Linhagem Celular , Modelos Animais de Doenças , Inibidores de Histona Desacetilases/farmacologia , Histona Desacetilases/genética , Histona Desacetilases/metabolismo , Humanos , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Masculino , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/patologia , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fenótipo , Interferência de RNA , Ratos Sprague-Dawley , Fator de Resposta Sérica/genética , Fator de Resposta Sérica/metabolismo , Transdução de Sinais , Transativadores/genética , Transativadores/metabolismo , TransfecçãoRESUMO
The aim of this study was to measure the relative location between foramen ovale and internal carotid artery. The results can guide surgeons to avoid damaging internal carotid artery in cavernous sinus biopsy. One hundred twenty people (73 men and 47 women) were involved in this study anonymously. Five parameters of both sides were measured on 2 planes. This study provides data for safer cavernous sinus biopsy.
Assuntos
Biópsia/métodos , Lesões das Artérias Carótidas/prevenção & controle , Artéria Carótida Interna , Seio Cavernoso/diagnóstico por imagem , Seio Cavernoso/patologia , Cirurgia Assistida por Computador/métodos , Tomografia Computadorizada por Raios X/métodos , Adulto , Idoso , Lesões das Artérias Carótidas/diagnóstico por imagem , Artéria Carótida Interna/diagnóstico por imagem , Artéria Carótida Interna/patologia , Feminino , Forame Oval/diagnóstico por imagem , Forame Oval/patologia , Humanos , Doença Iatrogênica/prevenção & controle , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
OBJECTIVE: Intimal hyperplasia (IH) contributes to the failure of vascular interventions. While many investigational therapies inhibit the development of IH in animal models, few of these potential therapies can reverse established lesions. Inhaled carbon monoxide (CO) dramatically inhibits IH in both rats and pigs when given perioperatively. It also prevented the development of pulmonary arterial hypertension in rodents. Interestingly, CO could reverse pulmonary artery structural changes and right heart hemodynamic changes when administered after the establishment of pulmonary hypertension. Thus, we hypothesize that inhaled CO may mediate the regression of established neointimal lesions. METHODS: Rats underwent carotid artery balloon angioplasty injury. Carotid arteries were collected at 2 and 4 weeks after injury for morphometric analysis of the neointima. Another group was treated with inhaled CO (250 parts per million) for 1 hour daily from week 2 until week 4. Additional rats were sacrificed 3 days after initiating CO treatment, and the carotid arteries were examined for apoptosis by terminal deoxynucleotidyl transferase dUTP nick end-labeling, proliferation by Ki67 staining, and autophagy by microtubule-associated protein light chain 3 I/II staining. RESULTS: At 2 weeks following injury, sizable neointimal lesions had developed (intimal/media = 0.92 ± 0.22). By 4 weeks, lesion size remained stable (0.80 ± 0.09). Delayed inhaled CO treatment greatly reduced neointimal lesion size vs the 2- and 4-week control mice (0.38 ± 0.05; P < .05). Arteries from the CO-treated rats exhibited significantly reduced apoptosis compared with control vessels (3.18% ± 1.94% vs 16.26% ± 5.91%; P = .036). Proliferation was also dramatically reduced in the CO-treated animals (2.98 ± 1.55 vs 10.37 ± 2.80; P = .036). No difference in autophagy between control and CO-treated rats was detected. CONCLUSIONS: Delayed administration of inhaled CO reduced established neointimal lesion size. This effect was mediated by the antiproliferative effect of CO on medial and intimal smooth muscle cells without increases in arterial wall apoptosis or autophagy. Future studies will examine additional time points to determine if there is temporal variation in the rates of apoptosis and autophagy.
Assuntos
Monóxido de Carbono/administração & dosagem , Lesões das Artérias Carótidas/prevenção & controle , Proliferação de Células/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Neointima , Administração por Inalação , Angioplastia com Balão , Animais , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Lesões das Artérias Carótidas/etiologia , Lesões das Artérias Carótidas/patologia , Artéria Carótida Externa/efeitos dos fármacos , Artéria Carótida Externa/patologia , Modelos Animais de Doenças , Esquema de Medicação , Hiperplasia , Masculino , Músculo Liso Vascular/patologia , Ratos Sprague-Dawley , Fatores de TempoRESUMO
BACKGROUND: Overexpression of matrix metalloproteinase (MMP) has been implicated in the incidence of restenosis after vascular angioplasty. Reversion-inducing cysteine-rich protein with kazal motifs (RECK) is a membrane-anchored glycoprotein that negatively regulates the activity of MMPs, such as MMP-9 and MMP-2, which play a key role in the angiogenesis during tumor growth. This study was designed to investigate the potential association between RECK and restenosis after vascular angioplasty. METHODS: Balloon-injured rabbit carotid arterial models were established. Arterial morphology was assessed by hematoxylin-eosin staining. The area of intimal hyperplasia was measured using image microscopy and image analyzer. The messenger RNA (mRNA) expression levels of RECK, MMP-9, and MMP-2 were detected using reverse transcription-polymerase chain reaction (RT-PCR) at 7, 14, and 21 days. Vascular smooth muscle cells (VSMCs) were transfected with RECK small interfering RNA (siRNA). VSMC proliferation rate was detected by MTT assay at 24, 48 and 72 hr. The protein expression of RECK, MMP-9, and MMP-2 was determined by Western blot. RESULTS: MMP-2 and MMP-9 in carotid artery of rats were significantly overexpressed in the injured-artery group, compared with unmanipulated control and contralateral uninjured groups (P < 0.05). With the time of the injury extended, MMP-2 and MMP-9 mRNA levels gradually increased. RECK showed a marked peak of mRNA level at 7 days after injury, compared with unmanipulated control and contralateral uninjured groups (P < 0.001). However, the increasing trend gradually decreased at 14 days after the balloon surgery. RECK mRNA was still detectable at 21 days postoperatively, but the expression level of RECK mRNA in injured and contralateral uninjured groups was significantly lower than that in unmanipulated control group (P < 0.001). The expression level of RECK protein in VSMCs in transfected group was significantly lower compared with that in untransfected group, whereas the expression of MMP-2 and MMP-9 proteins in transfected group was significantly higher compared with that in untransfected group. Over the extension of transfection time, the proliferation of VSMCs in transfected group was increased gradually, compared with negative and blank plasmid controls (P < 0.05). CONCLUSIONS: RECK, as siRNA-mediated RECK silencing regulation of MMP-9 and MMP-2, plays an important role in intimal hyperplasia, which provides a new target for prevention and treatment of restenosis after vascular angioplasty.
Assuntos
Angioplastia com Balão/efeitos adversos , Lesões das Artérias Carótidas/metabolismo , Proteínas Ligadas por GPI/metabolismo , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Neointima , Animais , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/patologia , Lesões das Artérias Carótidas/prevenção & controle , Artéria Carótida Primitiva/metabolismo , Artéria Carótida Primitiva/patologia , Linhagem Celular , Modelos Animais de Doenças , Proteínas Ligadas por GPI/genética , Humanos , Hiperplasia , Masculino , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/patologia , Interferência de RNA , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Fatores de Tempo , TransfecçãoRESUMO
RATIONALE: Innate and adaptive immune responses alter numerous homeostatic processes that are controlled by nuclear hormone receptors. NR4A1 is a nuclear receptor that is induced in vascular pathologies, where it mediates protection. OBJECTIVE: The underlying mechanisms that regulate the activity of NR4A1 during vascular injury are not clear. We therefore searched for modulators of NR4A1 function that are present during vascular inflammation. METHODS AND RESULTS: We report that the protein encoded by interferon stimulated gene 12 (ISG12), is a novel interaction partner of NR4A1 that inhibits the transcriptional activities of NR4A1 by mediating its Crm1-dependent nuclear export. Using 2 models of vascular injury, we show that ISG12-deficient mice are protected from neointima formation. This effect is dependent on the presence of NR4A1, as mice deficient for both ISG12 and NR4A1 exhibit neointima formation similar to wild-type mice. CONCLUSIONS: These findings identify a previously unrecognized feedback loop activated by interferons that inhibits the vasculoprotective functions of NR4A nuclear receptors, providing a potential new therapeutic target for interferon-driven pathologies.
Assuntos
Lesões das Artérias Carótidas/prevenção & controle , Artéria Femoral/metabolismo , Inflamação/prevenção & controle , Proteínas de Membrana/metabolismo , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Proteínas/metabolismo , Lesões do Sistema Vascular/prevenção & controle , Transporte Ativo do Núcleo Celular , Animais , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/imunologia , Lesões das Artérias Carótidas/metabolismo , Lesões das Artérias Carótidas/patologia , Células Cultivadas , Modelos Animais de Doenças , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Retroalimentação Fisiológica , Artéria Femoral/lesões , Artéria Femoral/patologia , Regulação da Expressão Gênica , Inflamação/genética , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/patologia , Interferons/metabolismo , Carioferinas/metabolismo , Proteínas de Membrana/genética , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Músculo Liso Vascular/lesões , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Domínios e Motivos de Interação entre Proteínas , Proteínas/genética , Interferência de RNA , Receptores Citoplasmáticos e Nucleares/metabolismo , Fatores de Tempo , Transcrição Gênica , Transfecção , Lesões do Sistema Vascular/genética , Lesões do Sistema Vascular/imunologia , Lesões do Sistema Vascular/metabolismo , Lesões do Sistema Vascular/patologia , Proteína Exportina 1RESUMO
OBJECTIVE: The lipid phosphate phosphatase 3 (LPP3) degrades bioactive lysophospholipids, including lysophosphatidic acid and sphingosine-1-phosphate, and thereby terminates their signaling effects. Although emerging evidence links lysophosphatidic acid to atherosclerosis and vascular injury responses, little is known about the role of vascular LPP3. The goal of this study was to determine the role of LPP3 in the development of vascular neointima formation and smooth muscle cells (SMC) responses. METHODS AND RESULTS: We report that LPP3 is expressed in vascular SMC after experimental arterial injury. Using gain- and loss-of-function approaches, we establish that a major function of LPP3 in isolated SMC cells is to attenuate proliferation (extracellular signal-regulated kinases) activity, Rho activation, and migration in response to serum and lysophosphatidic acid. These effects are at least partially a consequence of LPP3-catalyzed lysophosphatidic acid hydrolysis. Mice with selective inactivation of LPP3 in SMC display an exaggerated neointimal response to injury. CONCLUSIONS: Our observations suggest that LPP3 serves as an intrinsic negative regulator of SMC phenotypic modulation and inflammation after vascular injury, in part, by regulating lysophospholipid signaling. These findings may provide a mechanistic link to explain the association between a PPAP2B polymorphism and coronary artery disease risk.
Assuntos
Lesões das Artérias Carótidas/prevenção & controle , Proliferação de Células , Músculo Liso Vascular/enzimologia , Miócitos de Músculo Liso/enzimologia , Fosfatidato Fosfatase/metabolismo , Animais , Lesões das Artérias Carótidas/enzimologia , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/patologia , Artéria Carótida Primitiva/enzimologia , Artéria Carótida Primitiva/patologia , Movimento Celular , Modelos Animais de Doenças , Ativação Enzimática , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Regulação da Expressão Gênica , Genótipo , Células HEK293 , Humanos , Hidrólise , Hiperplasia , Lisofosfolipídeos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/patologia , Neointima , Fenótipo , Fosfatidato Fosfatase/deficiência , Fosfatidato Fosfatase/genética , Transdução de Sinais , Fatores de Tempo , Transfecção , Quinases Associadas a rho/metabolismoRESUMO
OBJECTIVE: Myocardin, a potent transcriptional coactivator of serum response factor, is involved in vascular development and promotes a contractile smooth muscle phenotype. Myocardin levels are reduced during vascular injury, in association with phenotypic switching of smooth muscle cells (SMCs). However, the direct role of myocardin in vascular disease is unclear. APPROACH AND RESULTS: We show that re-expression of myocardin prevents the vascular injury response in murine carotid arteries, with reduced neointima formation due to decreased SMC migration and proliferation. Myocardin reduced SMC migration by downregulating platelet-derived growth factor receptor-ß (PDGFRB) expression. Pdgfrb was regulated by myocardin-induced miR-24 and miR-29a expression, and antagonizing these microRNAs restored SMC migration. Furthermore, using miR-24 and miR-29a mimics, we demonstrated that miR-29a directly regulates Pdgfrb expression at the 3' untranslated region while miR-24 has an indirect effect on Pdgfrb levels. Myocardin heterozygous-null mice showed an augmented neointima formation with increased SMC migration and proliferation, demonstrating that endogenous levels of myocardin are a critical regulator of vessel injury responses. CONCLUSIONS: Our results extend the function of myocardin from a developmental role to a pivotal regulator of SMC phenotype in response to injury, and this transcriptional coactivator may be an attractive target for novel therapeutic strategies in vascular disease.
Assuntos
Lesões das Artérias Carótidas/metabolismo , Estenose das Carótidas/metabolismo , MicroRNAs/metabolismo , Músculo Liso Vascular/metabolismo , Proteínas Nucleares/metabolismo , Receptor beta de Fator de Crescimento Derivado de Plaquetas/metabolismo , Transativadores/metabolismo , Regiões 3' não Traduzidas , Animais , Apolipoproteínas E/deficiência , Apolipoproteínas E/genética , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/patologia , Lesões das Artérias Carótidas/prevenção & controle , Artéria Carótida Primitiva/metabolismo , Artéria Carótida Primitiva/patologia , Estenose das Carótidas/genética , Estenose das Carótidas/patologia , Estenose das Carótidas/prevenção & controle , Movimento Celular , Proliferação de Células , Células Cultivadas , Modelos Animais de Doenças , Regulação da Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Músculo Liso Vascular/lesões , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Neointima , Proteínas Nucleares/deficiência , Proteínas Nucleares/genética , Ratos , Ratos Wistar , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Transdução de Sinais , Fatores de Tempo , Transativadores/deficiência , Transativadores/genética , TransfecçãoRESUMO
OBJECTIVE: Vascular remodeling as a result of smooth muscle cell (SMC) proliferation and neointima formation is a major medical challenge in cardiovascular intervention. However, antineointima drugs often indistinguishably block re-endothelialization, an essential step toward successful vascular repair, because of their nonspecific effect on endothelial cells (ECs). The objective of this study is to identify a therapeutic target that differentially regulates SMC and EC proliferation. APPROACH AND RESULTS: Using both rat balloon injury and mouse wire injury models, we identified CTP synthase 1 (CTPS1) as one of the potential targets that may be used for developing therapeutics for treating neointima-related disorders. CTPS1 was induced in proliferative SMCs in vitro and neointima SMCs in vivo. Blockade of CTPS1 expression by small hairpin RNA or activity by cyclopentenyl cytosine suppressed SMC proliferation and neointima formation. Surprisingly, cyclopentenyl cytosine had much less effect on EC proliferation. Of importance, blockade of CTPS1 in vivo sustained the re-endothelialization as a result of induction of CTP synthesis salvage pathway enzymes nucleoside-diphosphate kinase A and B in ECs. Diphosphate kinase B seemed to preserve EC proliferation via use of extracellular cytidine to synthesize CTP. Indeed, blockade of both CTPS1 and diphosphate kinase B suppressed EC proliferation in vitro and the re-endothelialization in vivo. CONCLUSIONS: Our study uncovered a fundamental difference in CTP biosynthesis between SMCs and ECs during vascular remodeling, which provided a novel strategy by using cyclopentenyl cytosine or other CTPS1 inhibitors to selectively block SMC proliferation without disturbing or even promoting re-endothelialization for effective vascular repair after injury.
Assuntos
Carbono-Nitrogênio Ligases/antagonistas & inibidores , Lesões das Artérias Carótidas/prevenção & controle , Citidina/análogos & derivados , Inibidores Enzimáticos/farmacologia , Terapia Genética , Terapia de Alvo Molecular , Músculo Liso Vascular/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , Lesões do Sistema Vascular/prevenção & controle , Angioplastia com Balão/efeitos adversos , Animais , Carbono-Nitrogênio Ligases/genética , Carbono-Nitrogênio Ligases/metabolismo , Lesões das Artérias Carótidas/enzimologia , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/patologia , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citidina/farmacologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/enzimologia , Células Endoteliais/patologia , Terapia Genética/métodos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Liso Vascular/enzimologia , Músculo Liso Vascular/lesões , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/enzimologia , Miócitos de Músculo Liso/patologia , Nucleosídeo NM23 Difosfato Quinases/metabolismo , Neointima , Interferência de RNA , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Lesões do Sistema Vascular/enzimologia , Lesões do Sistema Vascular/genética , Lesões do Sistema Vascular/patologiaRESUMO
PURPOSE: In high-risk head and neck cases treated with tumor resection and associated radical neck dissection, orocutaneous fistulas and wound breakdowns in the neck are relatively frequent and can have serious consequences, such as carotid blowout syndrome (CBS), the need for salvage reoperations, and prolonged recovery time. The authors present the application of a prophylactic chimeric anterolateral thigh (ALT) and vastus lateralis (VL) flap to prevent complications. MATERIALS AND METHODS: A retrospective review was performed of a historical group (96 patients) of patients with head and neck cancer treated with tumor resection, radical neck dissection, and microsurgical reconstruction of the tumor site only and a prospective cohort (21 patients) in which a chimeric ALT-VL flap was used to simultaneously reconstruct the tumor site and sternocleidomastoid muscle to fill dead space and protect the carotid artery. RESULTS: The rate of complications was higher in the historical group: CBS occurred in 4.1% and orocutaneous fistulas in 11.5% of patients; 5.2% of patients required major salvage surgery for a wound complication. In the cohort group, no CBS or orocutaneous fistula occurred and no major salvage surgical procedure was needed. CONCLUSIONS: Prophylactic ALT-VL flaps in high-risk head and neck cancers provide adequate and long-lasting soft tissue coverage for the carotid artery, with minimal additional morbidity, and could be beneficial in preventing serious and life-threatening wound complications and the need for reoperation.
Assuntos
Neoplasias de Cabeça e Pescoço/cirurgia , Músculo Esquelético/transplante , Procedimentos de Cirurgia Plástica/métodos , Complicações Pós-Operatórias/prevenção & controle , Retalhos Cirúrgicos/transplante , Adulto , Idoso , Perda Sanguínea Cirúrgica/prevenção & controle , Lesões das Artérias Carótidas/prevenção & controle , Quimiorradioterapia Adjuvante , Estudos de Coortes , Fístula Cutânea/etiologia , Feminino , Seguimentos , Humanos , Complicações Intraoperatórias/prevenção & controle , Masculino , Microcirurgia/métodos , Pessoa de Meia-Idade , Esvaziamento Cervical/reabilitação , Músculos do Pescoço/cirurgia , Terapia Neoadjuvante , Fístula Bucal/etiologia , Estudos Prospectivos , Reoperação , Estudos Retrospectivos , Terapia de Salvação , Deiscência da Ferida Operatória/etiologiaRESUMO
The present review encompasses the publications concerning peculiarities of blood supply of palatal tonsils, the frequency and variants of the anomalous structure of internal carotid artery. Analysis of the literature data has shown that the anomalous vascular structure is one of the important risk factors of the development of intra- and postoperative hemorrhage, besides redundant blood supply of the oropharynx, inflammatory changes in the tissue of tonsils and peritonsillar region. Special attention is given to the peculiarities of diagnostics of such conditions. The emphasis is laid on the importance of timely diagnostics of the anomalous structural changes in the main vessels of the neck prior to the surgical intervention on the pharynx. The analysis of the anomalous structure of internal carotid artery is presented.
Assuntos
Lesões das Artérias Carótidas/complicações , Artéria Carótida Interna/anormalidades , Tonsila Palatina/irrigação sanguínea , Hemorragia Pós-Operatória/etiologia , Tonsilectomia/métodos , Tonsilite/cirurgia , Lesões das Artérias Carótidas/diagnóstico , Lesões das Artérias Carótidas/prevenção & controle , Humanos , Masculino , Pessoa de Meia-Idade , Tonsila Palatina/cirurgia , Hemorragia Pós-Operatória/diagnóstico , Hemorragia Pós-Operatória/prevenção & controle , Tonsilectomia/efeitos adversos , Tonsilite/patologiaRESUMO
BACKGROUND: Interleukin-8 (IL8) receptors IL8RA and IL8RB on neutrophil membranes bind to IL8 and direct neutrophil recruitment to sites of inflammation, including acutely injured arteries. This study tested whether administration of IL8RA- and/or IL8RB-transduced rat aortic endothelial cells (ECs) accelerates adhesion of ECs to the injured surface, thus suppressing inflammation and neointima formation in balloon-injured rat carotid arteries. We tested the hypothesis that targeted delivery of ECs by overexpressing IL8RA and IL8RB receptors prevents inflammatory responses and promotes structural recovery of arteries after endoluminal injury. METHODS AND RESULTS: Young adult male rats received balloon injury of the right carotid artery and were transfused intravenously with ECs (total, 1.5×10(6) cells at 1, 3, and 5 hours after injury) transduced with adenoviral vectors carrying IL8RA, IL8RB, and IL8RA/RB (dual transduction) genes, AdNull (empty vector), or vehicle (no EC transfusion). ECs overexpressing IL8Rs inhibited proinflammatory mediators expression significantly (by 60% to 85%) and reduced infiltration of neutrophils and monocytes/macrophages into injured arteries at 1 day after injury, as well as stimulating a 2-fold increase in reendothelialization at 14 days after injury. IL8RA-EC, IL8RB-EC, and IL8RA/RB-EC treatment reduced neointima formation dramatically (by 80%, 74%, and 95%) at 28 days after injury. CONCLUSIONS: ECs with overexpression of IL8RA and/or IL8RB mimic the behavior of neutrophils that target and adhere to injured tissues, preventing inflammation and neointima formation. Targeted delivery of ECs to arteries with endoluminal injury provides a novel strategy for the prevention and treatment of cardiovascular disease.
Assuntos
Lesões das Artérias Carótidas/genética , Células Endoteliais/metabolismo , Regulação da Expressão Gênica , Neointima/metabolismo , Receptores de Interleucina-8A/biossíntese , Receptores de Interleucina-8B/biossíntese , Animais , Lesões das Artérias Carótidas/patologia , Lesões das Artérias Carótidas/prevenção & controle , Células Endoteliais/patologia , Células Endoteliais/transplante , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Inflamação/metabolismo , Inflamação/patologia , Masculino , Neointima/patologia , Ratos , Ratos Sprague-Dawley , Receptores de Interleucina-8A/genética , Receptores de Interleucina-8A/fisiologia , Receptores de Interleucina-8B/genética , Receptores de Interleucina-8B/fisiologiaRESUMO
OBJECTIVE: Abnormal proliferation and migration of vascular smooth muscle cells (SMCs) are the key events in the progression of neointima formation in response to vascular injury. The goal of this study is to investigate the functional role of a potent oncogene yes-associated protein (YAP) in SM phenotypic modulation in vitro and in vivo. METHODS AND RESULTS: In vitro cell culture and in vivo in both mouse and rat arterial injury models YAP expression is significantly induced and correlated with the vascular SMC synthetic phenotype. Overexpression of YAP promotes SMC migration and proliferation while attenuating SM contractile gene expression. Conversely, knocking down endogenous YAP in SMCs upregulates SM gene expression but attenuates SMC proliferation and migration. Consistent with this, knocking down YAP expression in a rat carotid balloon injury model and genetic deletion of YAP, specifically, in vascular SMCs in mouse after carotid artery ligation injury attenuates injury-induced SM phenotypic switch and neointima formation. CONCLUSIONS: YAP plays a novel integrative role in SM phenotypic modulation by inhibiting SM-specific gene expression while promoting SM proliferation and migration in vitro and in vivo. Blocking the induction of YAP would be a potential therapeutic approach for ameliorating vascular occlusive diseases.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas Reguladoras de Apoptose/metabolismo , Lesões das Artérias Carótidas/enzimologia , Músculo Liso Vascular/enzimologia , Miócitos de Músculo Liso/enzimologia , Fosfoproteínas/metabolismo , Lesões do Sistema Vascular/enzimologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Aorta/enzimologia , Aorta/patologia , Proteínas Reguladoras de Apoptose/genética , Artérias Carótidas/enzimologia , Artérias Carótidas/patologia , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/patologia , Lesões das Artérias Carótidas/prevenção & controle , Proteínas de Ciclo Celular , Movimento Celular , Proliferação de Células , Células Cultivadas , Modelos Animais de Doenças , Regulação da Expressão Gênica , Técnicas de Silenciamento de Genes , Genótipo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/patologia , Neointima , Fenótipo , Fosfoproteínas/genética , Interferência de RNA , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Transfecção , Lesões do Sistema Vascular/genética , Lesões do Sistema Vascular/patologia , Lesões do Sistema Vascular/prevenção & controle , Proteínas de Sinalização YAPRESUMO
AIM: To determine military-specific cervical neurovascular and external anthropometric data to scale future numerical injury models of the neck and improve body armour design with a view to prevention or mitigation of combat neck injury. MATERIALS AND METHODS: Contrast-enhanced computed tomography (CT) angiograms of 50 UK servicemen were analysed. Mean diameters and distances from the skin surface were determined for the carotid artery (CA), internal jugular vein (IJV), vertebral artery (VA) and spinal cord (SC) at the three surgical neck zones. Horizontal neck circumference at C6 and three potential vertical cervical anthropometric measurements were analysed to determine which had the least variability between subjects. RESULTS: The diameters of cervical vascular structures are greater and the vessels more superficial as the anatomical plane moves caudally. The SC and VA are better protected than the IJV and CA due to their greater depth and bony coverage, except for the VA in zone 1. CONCLUSION: Future cervical anthropometric assessments should use the vertical angle of mandible to mid-claviclular distance in combination with the horizontal neck circumference as these demonstrated the least variability. Cervical neurovascular structures are least vulnerable posterosuperiorly and therefore extending the posterior aspect of a ballistic helmet inferiorly or adding a nape protector would appear to be less justified. Cervical vessels are most vulnerable in zone 1 and a circumferential collar of ballistic material at least 75 mm high would cover this area in 95% of this population.
Assuntos
Militares , Lesões do Pescoço/prevenção & controle , Traumatismos do Sistema Nervoso/prevenção & controle , Lesões do Sistema Vascular/prevenção & controle , Ferimentos Penetrantes/prevenção & controle , Adulto , Antropometria , Artérias Carótidas/diagnóstico por imagem , Lesões das Artérias Carótidas/diagnóstico por imagem , Lesões das Artérias Carótidas/prevenção & controle , Meios de Contraste , Desenho de Equipamento , Humanos , Veias Jugulares/diagnóstico por imagem , Lesões do Pescoço/diagnóstico por imagem , Roupa de Proteção , Medula Espinal/diagnóstico por imagem , Traumatismos da Medula Espinal/diagnóstico por imagem , Traumatismos da Medula Espinal/prevenção & controle , Tomografia Computadorizada por Raios X/métodos , Traumatismos do Sistema Nervoso/diagnóstico por imagem , Lesões do Sistema Vascular/diagnóstico por imagem , Artéria Vertebral/diagnóstico por imagem , Ferimentos Penetrantes/diagnóstico por imagemRESUMO
Common carotid artery (CCA) injury is a serious complication of internal jugular vein (IJV) cannulation. To minimize unintentional CCA puncture, the anatomic relationship between the IJV and the CCA and the size of IJV were compared under different head positions. Ultrasound analyses of the IJV and the CCA were performed in 103 consecutive patients. Overlapping angle (OA), real puncture angle (RPA) and diameter of IJV (D IJV) were evaluated with 30° and 60° left rotation and with 30° left flexion. When the head position was changed from 30° left rotation to 60° left rotation, OA increased significantly from 6.5° ± 7.7° to 14.5° ± 7.4° at the cricoid cartilage level (Cricoid-level) and from 14.4° ± 8.4° to 20.6° ± 6.9° at the middle triangle level (Triangle-level), whereas RPA decreased significantly at these levels (from 49.7° ± 11.9° to 43.5° ± 13.1° and from 51.1° ± 14.4° to 44.3° ± 13.9°, respectively; P < 0.01 for both). When the head position was changed from 30° left rotation to 30° left flexion, neither OA nor RPA significantly changed (OA: 6.3° ± 6.1° and 15.0° ± 7.2°, RPA: 48.5° ± 12.4° and 51.8° ± 13.6°, P not significant vs 30° left rotation). There was no difference in D IJV when comparing 30° left rotation and 30° left flexion, although D IJV was largest at 60° left rotation. RPA positively correlated with age, and D IJV positively correlated with body mass index. In conclusion, excessive left rotation should be avoided to minimize the probability of unintentional CCA puncture during IJV cannulation. When 30° left rotation is not feasible, the head-flexion position should be utilized.