RESUMO
BACKGROUND: To characterize how prior incisional glaucoma surgery affects the intraocular pressure (IOP) elevation immediately following intravitreal anti-VEGF injections (IVI). METHODS: Single institution, experimental study. Patients with a history of incisional glaucoma surgery who were receiving anti-VEGF injections were recruited as well as control eyes. Pre- and post-injection IOP measurements were compared as well as time to recovery to within 5 and 10 mmHg of baseline IOP. RESULTS: Ten eyes with a history of glaucoma surgery and 29 control eyes receiving anti-VEGF injections were included. The most common indication for intravitreal anti-VEGF injection was proliferative diabetic retinopathy in both surgical and control eyes (50% vs 45%, p = 1.00). Post-injection IOP was significantly decreased compared to baseline IOP after anti-VEGF injection in surgical versus control eyes (26.5 ± 8.9 mmHg vs 44.2 ± 8.5 mmHg, respectively, p < 0.001). The mean change in IOP following intravitreal anti-VEGF injection was lower in surgical eyes (10.7 ± 6.6 mmHg vs 28.6 ± 8.3 mmHg, p < 0.001). The mean time for the IOP to return to within 10 mmHg of pre-injection IOP was less in surgical eyes (5.2 ± 4.1 min vs 13.3 ± 7.6 min, p = 0.002). CONCLUSIONS: Eyes with prior incisional glaucoma surgery demonstrated a significantly lower post-injection IOP elevation and a faster recovery to within 10 mmHg of their pre-injection IOP. Incisional glaucoma surgery may be considered for patients where the attenuation of post-injection IOP elevation is needed and other less invasive measures have failed.
Assuntos
Bevacizumab/administração & dosagem , Cirurgia Filtrante/métodos , Glaucoma/terapia , Pressão Intraocular/fisiologia , Ranibizumab/administração & dosagem , Idoso , Inibidores da Angiogênese/administração & dosagem , Feminino , Seguimentos , Glaucoma/fisiopatologia , Humanos , Injeções Intravítreas , Masculino , Pessoa de Meia-Idade , Período Pré-Operatório , Estudos Prospectivos , Tonometria Ocular , Resultado do Tratamento , Fator A de Crescimento do Endotélio VascularRESUMO
AIMS: We investigated the mechanisms of diabetic bladder dysfunction (BD) through analysis of the roles of L- and T-type voltage-gated calcium channels (VGCCs), with the ultimate goal of identifying potential drug targets for diabetic BD. METHODS: Bladder function of db/db (type 2 diabetes) and wild type (Wt) mice was evaluated by behavioral tests and in vivo cystometry. Contractile responses of bladder strips to carbachol were measured with or without pre-treatment with nifedipine (a L-type VGCC blocker) or mibefradil (a T-type VGCC blocker). Furthermore, the effects of mibefradil and nifedipine on the proliferation of human bladder smooth muscle cells (BSMCs) were studied. RESULTS: db/db mice had significantly increased voiding frequency, bladder weight, bladder compliance and capacity, and heightened contractile response to carbachol, compared to Wt mice. Nifedipine, but not mibefradil, dramatically suppressed bladder tissue contraction in Wt mice. Whereas nifedipine nearly completely inhibited bladder contraction in db/db mice, mibefradil "normalized" the heightened bladder contractility of db/db mice to the level of Wt mice. In culture, mibefradil, but not nifedipine, inhibited the proliferation of human BSMCs. CONCLUSION: Our results indicate that while L-type VGCCs play a major role in the contraction of both diabetic and non-diabetic bladders, T-type VGCCs are involved in the contraction of diabetic bladders and mediate BSMC proliferation. This study provides support for further investigations on the effect of blockade of T-type VGCC or combined blockade of both types of VGCCs in the treatment of diabetic BD.
Assuntos
Canais de Cálcio Tipo L/metabolismo , Canais de Cálcio Tipo T/metabolismo , Diabetes Mellitus Tipo 2/complicações , Contração Muscular , Doenças da Bexiga Urinária/etiologia , Bexiga Urinária/metabolismo , Acetilcolina/farmacologia , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Canais de Cálcio Tipo L/efeitos dos fármacos , Canais de Cálcio Tipo L/genética , Canais de Cálcio Tipo T/efeitos dos fármacos , Canais de Cálcio Tipo T/genética , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Humanos , Masculino , Mibefradil/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Contração Muscular/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , Nifedipino/farmacologia , RNA Mensageiro/metabolismo , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/patologia , Bexiga Urinária/fisiopatologia , Doenças da Bexiga Urinária/metabolismo , Doenças da Bexiga Urinária/patologia , Doenças da Bexiga Urinária/fisiopatologiaRESUMO
CONTEXT: Inhalation of fine particulate matter (PM2.5) is associated with acute pulmonary inflammation and impairments in cardiovascular function. In many regions, PM2.5 is largely derived from diesel exhaust (DE), and these pathophysiological effects may be due in part to oxidative stress resulting from DE inhalation. The antioxidant glutathione (GSH) is important in limiting oxidative stress-induced vascular dysfunction. The rate-limiting enzyme in GSH synthesis is glutamate cysteine ligase and polymorphisms in its catalytic and modifier subunits (GCLC and GCLM) have been shown to influence vascular function and risk of myocardial infarction in humans. OBJECTIVE: We hypothesized that compromised de novo synthesis of GSH in Gclmâ»/⺠mice would result in increased sensitivity to DE-induced lung inflammation and vascular effects. MATERIALS AND METHODS: WT and Gclmâ»/⺠mice were exposed to DE via inhalation (300 µg/m³) for 6 h. Neutrophil influx into the lungs, plasma GSH redox potential, vascular reactivity of aortic rings and aortic nitric oxide (NOâ¢) were measured. RESULTS: DE inhalation resulted in mild bronchoalveolar neutrophil influx in both genotypes. DE-induced effects on plasma GSH oxidation and acetylcholine (ACh)-relaxation of aortic rings were only observed in Gclmâ»/⺠mice. Contrary to our hypothesis, DE exposure enhanced ACh-induced relaxation of aortic rings in Gclmâ»/⺠mice. DISCUSSION AND CONCLUSION: THESE data support the hypothesis that genetic determinants of antioxidant capacity influence the biological effects of acute inhalation of DE. However, the acute effects of DE on the vasculature may be dependent on the location and types of vessels involved. Polymorphisms in GSH synthesis genes are common in humans and further investigations into these potential gene-environment interactions are warranted.
Assuntos
Poluentes Atmosféricos/toxicidade , Glutamato-Cisteína Ligase/genética , Emissões de Veículos/toxicidade , Administração por Inalação , Animais , Aorta/fisiologia , Líquido da Lavagem Broncoalveolar/imunologia , Feminino , Glutationa/sangue , Dissulfeto de Glutationa/sangue , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neutrófilos/imunologia , Óxido Nítrico/metabolismo , Oxirredução , Pneumonia/induzido quimicamente , Pneumonia/imunologia , Pneumonia/metabolismo , Pneumonia/fisiopatologia , VasodilataçãoRESUMO
OBJECTIVE: ⢠To examine functional and molecular changes of the bladders from elastin-haploinsufficient mice (Eln(+/-) ) at baseline as well as in response to partial bladder outlet obstruction (pBOO). MATERIALS AND METHODS: ⢠Female Eln(+/-) and wild type (Wt) mice (3-4 months old) were studied. ⢠The bladder elastin content was quantified by measuring desmosine. ⢠Mice were divided into two groups to undergo surgery to create pBOO or to undergo sham surgery. Three days after surgery, bladder function was evaluated by in vivo cystometry, and the contractile response of bladder strips exposed to electrical field stimulation (EFS) and carbachol was examined by ex vivo myography. RESULTS: ⢠The Eln(+/-) -sham mice had a 33.6% decrease in bladder elastin compared with Wt-sham mice. ⢠Cystometry showed significantly decreased bladder compliance and capacity in Eln(+/-) -sham vs Wt-sham mice; pBOO increased bladder compliance and capacity to a greater extent in Eln(+/-) mice compared with Wt mice. ⢠Bladder strips from Eln(+/-) -sham mice showed a significantly heightened contractile response to both EFS and carbachol compared with Wt-sham mice. ⢠A significantly increased contractile response to carbachol was detected in Wt-pBOO vs Wt-sham but not between Eln(+/-) -pBOO and Eln(+/-) -sham mice. CONCLUSION: ⢠The results that elastin-deficient mice had decreased bladder compliance and capacity and increased bladder contractility; and that Wt-pBOO mice showed an enhanced contractile response to carbachol, but Eln(+/-) -pBOO mice did not, suggest that elastin is critical for normal bladder function and is involved in bladder response to pBOO.
Assuntos
Elastina/deficiência , Obstrução do Colo da Bexiga Urinária/fisiopatologia , Bexiga Urinária/fisiopatologia , Animais , Colágeno Tipo III/metabolismo , Desmosina/metabolismo , Estimulação Elétrica , Feminino , Hidroxiprolina/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Contração Muscular/fisiologia , Tamanho do Órgão , RNA Mensageiro/metabolismo , Tropoelastina/metabolismoRESUMO
OBJECTIVE: Vascular gene transfer is a powerful tool for investigating and treating vascular diseases; however, its utility is limited by brevity of transgene expression and vector-associated inflammation. Helper-dependent adenovirus (HDAd), an advanced-generation adenovirus that lacks all viral genes, is superior to first-generation adenovirus (FGAd) in normal rabbit arteries. We compared HDAd to FGAd in arteries of cholesterol-fed rabbits, a model of early atherogenesis in which transgene expression might be decreased, and inflammation increased. METHODS AND RESULTS: Carotid arteries of chow- and cholesterol-fed rabbits were infused with FGAd, HDAd, or medium. HDAd expressed a transgene at least as well in arteries of cholesterol-fed rabbits as in arteries of chow-fed rabbits and expressed more durably than FGAd. In arteries of cholesterol-fed rabbits, HDAd stimulated less intimal growth, lipid deposition, and inflammation than FGAd. Neither vector affected phenylephrine-induced contraction or nitroprusside-mediated relaxation; however, both vectors decreased maximal acetylcholine-stimulated vasorelaxation. The relative absence of intimal growth in HDAd arteries could interfere with the utility of this model for testing atheroprotective genes; however, both coinfusion of FGAd and extension of cholesterol feeding yielded larger intimal lesions, on which atheroprotective genes could be tested. CONCLUSION: HDAd is superior to FGAd for expression of transgenes in atherosclerosis-prone arteries.
Assuntos
Adenoviridae/genética , Artérias/metabolismo , Aterosclerose/etiologia , Vírus Auxiliares/fisiologia , Transgenes , Animais , Molécula 1 de Adesão Intercelular/análise , Metabolismo dos Lipídeos , Macrófagos/fisiologia , Masculino , Coelhos , Túnica Íntima/patologia , Molécula 1 de Adesão de Célula Vascular/análiseRESUMO
INTRODUCTION: Elastin fibers confer passive recoil to many tissues including the lung, skin, and arteries. In the penis, elastin is present in sinusoids, arterioles, and in the tunica albuginea. Although decreased penile elastin has been reported in men with erectile dysfunction, the exact role of elastin in physiologic processes integral to erection remains speculative. AIM: The aim of this study was to characterize erectile function in elastin-deficient mice. METHODS: Elastin haploinsufficient mice (Eln(+/-) ) and aged match Eln(+/+) (Wt) mice were used. Cavernosum was removed from some mice for quantification of elastin, collagen, and smooth muscle actin. Ex vivo assessment of contractile force generation was performed by myography. In vivo assessment of intracorporal pressure normalized to mean arterial pressure in response to electrical stimulation of the cavernosal nerve was measured. Veno-occlusive function was determined by cavernosography. MAIN OUTCOME MEASURES: The main outcome measures of this study were the in vitro and in vivo assessment of cavernosal vasoreactivity, veno-occlusive function and erection in mice deficient in elastin. RESULTS: Eln (+/-) mice exhibited â¼33% less penile elastin than Wt mice, with no change in collagen. Cavernosal tissue from Eln(+/-) mice has a significantly heightened contractile response, explained in part by increased smooth muscle cell content. Veno-occlusive function was significantly altered in Eln(+/-) mice. Interestingly, erectile function was impaired only at submaximal voltage (1 V) stimulation (there was no impairment during the higher 2-V stimulus). CONCLUSIONS: Eln (+/-) mice display a cavernosal phenotype consistent with developmental changes attributable to the loss of elastin. These alterations confer a degree of altered erectile function that is able to be overridden by maximal stimulatory input. Altogether, these data suggest that elastin is important for erectile function.
Assuntos
Elastina/deficiência , Haploinsuficiência/fisiologia , Ereção Peniana/fisiologia , Actinas/análise , Actinas/fisiologia , Animais , Western Blotting , Colágeno/análise , Colágeno/fisiologia , Elastina/análise , Elastina/genética , Haploinsuficiência/genética , Masculino , Camundongos , Músculo Liso Vascular/química , Músculo Liso Vascular/fisiologia , Ereção Peniana/genética , Pênis/anatomia & histologia , Pênis/química , Pênis/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Vasodilatação/fisiologiaRESUMO
OBJECTIVE: Diet-induced obesity (DIO) in mice causes vascular inflammation and insulin resistance that are accompanied by decreased endothelial-derived NO production. We sought to determine whether reduced NO-cGMP signaling contributes to the deleterious effects of DIO on the vasculature and, if so, whether these effects can be blocked by increased vascular NO-cGMP signaling. METHODS AND RESULTS: By using an established endothelial cell culture model of insulin resistance, exposure to palmitate, 100 micromol/L, for 3 hours induced both cellular inflammation (activation of IKK beta-nuclear factor-kappaB) and impaired insulin signaling via the insulin receptor substrate-phosphatidylinositol 3-kinase pathway. Sensitivity to palmitate-induced endothelial inflammation and insulin resistance was increased when NO signaling was reduced using an endothelial NO synthase inhibitor, whereas endothelial responses to palmitate were blocked by pretreatment with either an NO donor or a cGMP analogue. To investigate whether endogenous NO-cGMP signaling protects against vascular responses to nutrient excess in vivo, adult male mice lacking endothelial NO synthase were studied. As predicted, both vascular inflammation (phosphorylated I kappaB alpha and intercellular adhesion molecule levels) and insulin resistance (phosphorylated Akt [pAkt] and phosphorylated eNOS [peNOS] levels) were increased in endothelial NO synthase(-/-) (eNOS(-/-)) mice, reminiscent of the effect of DIO in wild-type controls. Next, we asked whether the vascular response to DIO in wild-type mice can be reversed by a pharmacological increase of cGMP signaling. C57BL6 mice were either fed a high-fat diet or remained on a low-fat diet for 8 weeks. During the final 2 weeks of the study, mice on each diet received either placebo or the phosphodiesterase-5 inhibitor sildenafil, 10 mg/kg per day orally. In high-fat diet-fed mice, vascular inflammation and insulin resistance were completely prevented by sildenafil administration at a dose that had no effect in mice fed the low-fat diet. CONCLUSIONS: Reduced signaling via the NO-cGMP pathway is a mediator of vascular inflammation and insulin resistance during overnutrition induced by high-fat feeding. Therefore, phosphodiesterase-5, soluble guanylyl cyclase, and other molecules in the NO-cGMP pathway (eg, protein kinase G) constitute potential targets for the treatment of vascular dysfunction in the setting of obesity.
Assuntos
Aorta Torácica/metabolismo , Doenças da Aorta/metabolismo , GMP Cíclico/metabolismo , Células Endoteliais/metabolismo , Inflamação/metabolismo , Resistência à Insulina , Óxido Nítrico/metabolismo , Transdução de Sinais , Animais , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/fisiopatologia , Doenças da Aorta/etiologia , Doenças da Aorta/fisiopatologia , Moléculas de Adesão Celular/metabolismo , Células Cultivadas , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Gorduras na Dieta , Modelos Animais de Doenças , Regulação para Baixo , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/enzimologia , Inibidores Enzimáticos/farmacologia , Humanos , Quinase I-kappa B/metabolismo , Inflamação/etiologia , Inflamação/fisiopatologia , Inflamação/prevenção & controle , Mediadores da Inflamação/metabolismo , Insulina/metabolismo , Proteínas Substratos do Receptor de Insulina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico Sintase Tipo III/antagonistas & inibidores , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo III/metabolismo , Ácido Palmítico/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores da Fosfodiesterase 5 , Inibidores de Fosfodiesterase/farmacologia , Fosforilação , Piperazinas/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Purinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Citrato de Sildenafila , Sulfonas/farmacologiaRESUMO
BACKGROUND/AIMS: To compare intraocular pressure (IOP) measurements using a prototype smartphone tonometer with other tonometers used in clinical practice. METHODS: Patients from an academic glaucoma practice were recruited. The smartphone tonometer uses fixed force applanation and in conjunction with a machine-learning computer algorithm is able to calculate the IOP. IOP was also measured using Goldmann applanation tonometry (GAT) in all subjects. A subset of patients were also measured using ICare, pneumotonometry (upright and supine positions) and Tono-Pen (upright and supine positions) and the results were compared. RESULTS: 92 eyes of 81 subjects were successfully measured. The mean difference (in mm Hg) for IOP measurements of the smartphone tonometer versus other devices was +0.24 mm Hg for GAT, -1.39 mm Hg for ICare, -3.71 mm Hg for pneumotonometry and -1.30 mm Hg for Tono-Pen. The 95% limits of agreement for the smartphone tonometer versus other devices was -4.35 to 4.83 mm Hg for GAT, -6.48 to 3.70 mm Hg for ICare, -7.66 to -0.15 mm Hg for pneumotonometry and -5.72 to 3.12 mm Hg for Tono-Pen. Overall, the smartphone tonometer results correlated best with GAT (R2=0.67, p<0.001). Of the 92 videos, 90 (97.8%) were within ±5 mm Hg of GAT and 58 (63.0%) were within ±2 mm Hg of GAT. CONCLUSIONS: Preliminary IOP measurements using a prototype smartphone-based tonometer was grossly equivalent to the reference standard.
Assuntos
Glaucoma de Ângulo Fechado/diagnóstico , Glaucoma de Ângulo Aberto/diagnóstico , Pressão Intraocular/fisiologia , Glaucoma de Baixa Tensão/diagnóstico , Aprendizado de Máquina , Smartphone/instrumentação , Tonometria Ocular/instrumentação , Adulto , Idoso , Feminino , Glaucoma de Ângulo Fechado/fisiopatologia , Glaucoma de Ângulo Aberto/fisiopatologia , Humanos , Glaucoma de Baixa Tensão/fisiopatologia , Masculino , Pessoa de Meia-Idade , Hipertensão Ocular/diagnóstico , Projetos Piloto , Reprodutibilidade dos TestesRESUMO
Vascular dysfunction is a major complication of metabolic disorders such as diabetes and obesity. The current studies were undertaken to determine whether inflammatory responses are activated in the vasculature of mice with diet-induced obesity, and if so, whether Toll-Like Receptor-4 (TLR4), a key mediator of innate immunity, contributes to these responses. Mice lacking TLR4 (TLR4(-/-)) and wild-type (WT) controls were fed either a low fat (LF) control diet or a diet high in saturated fat (HF) for 8 weeks. In response to HF feeding, both genotypes displayed similar increases of body weight, body fat content, and serum insulin and free fatty acid (FFA) levels compared with mice on a LF diet. In lysates of thoracic aorta from WT mice maintained on a HF diet, markers of vascular inflammation both upstream (IKKbeta activity) and downstream of the transcriptional regulator, NF-kappaB (ICAM protein and IL-6 mRNA expression), were increased and this effect was associated with cellular insulin resistance and impaired insulin stimulation of eNOS. In contrast, vascular inflammation and impaired insulin responsiveness were not evident in aortic samples taken from TLR4(-/-) mice fed the same HF diet, despite comparable increases of body fat mass. Incubation of either aortic explants from WT mice or cultured human microvascular endothelial cells with the saturated FFA, palmitate (100 micromol/L), similarly activated IKKbeta, inhibited insulin signal transduction and blocked insulin-stimulated NO production. Each of these effects was subsequently shown to be dependent on both TLR4 and NF-kappaB activation. These findings identify the TLR4 signaling pathway as a key mediator of the deleterious effects of palmitate on endothelial NO signaling, and are the first to document a key role for TLR4 in the mechanism whereby diet-induced obesity induces vascular inflammation and insulin resistance.
Assuntos
Mediadores da Inflamação/imunologia , Resistência à Insulina/imunologia , Obesidade/imunologia , Receptor 4 Toll-Like/imunologia , Animais , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/metabolismo , Aorta Torácica/patologia , Composição Corporal/genética , Peso Corporal/genética , Células Cultivadas , Gorduras na Dieta , Modelos Animais de Doenças , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/imunologia , Ácidos Graxos não Esterificados/sangue , Ácidos Graxos não Esterificados/farmacologia , Humanos , Proteínas I-kappa B/metabolismo , Mediadores da Inflamação/metabolismo , Insulina/sangue , Insulina/farmacologia , Resistência à Insulina/genética , Quinases Associadas a Receptores de Interleucina-1/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Inibidor de NF-kappaB alfa , Obesidade/genética , Obesidade/patologia , Técnicas de Cultura de Órgãos , Ácido Palmítico/farmacologia , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Receptor 4 Toll-Like/genéticaRESUMO
PURPOSE: To use optical coherence tomography angiography (OCTA) to evaluate the changes in optic nerve head perfusion following intravitreal antivascular endothelial growth factor injections. METHODS: Preinjection and postinjection intraocular pressure (IOP) and OCTA images were taken of both the injected and uninjected fellow eyes. RESULTS: Mean preinjection IOP was 16.6±4.7 mm Hg, which increased to a mean of 40.3±13.0 mm Hg (P<0.0001) during the first postinjection image and remained elevated at 36.1±11.5 mm Hg (P<0.0001) during the second postinjection image. Although no significant change was observed in flux, vessel area density, or normalized flux when comparing the OCTA preinjection and first postinjection images, a significant decrease at the second postinjection image was observed (P=0.03, 0.02, and 0.03, respectively). No significant change was observed in the uninjected fellow eye during the same time period (P=0.47, 0.37, and 0.38, respectively). CONCLUSIONS: Following an antivascular endothelial growth factor injection, mean IOP increased significantly and OCTA imaging of the optic nerve demonstrated a mild but significant decrease in optic nerve head perfusion parameters. Clinicians performing these injections should be aware of these findings and monitor the status of the optic nerve in patients undergoing injections.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Disco Óptico/irrigação sanguínea , Fluxo Sanguíneo Regional/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Angiofluoresceinografia , Humanos , Pressão Intraocular/fisiologia , Injeções Intravítreas , Masculino , Pessoa de Meia-Idade , Disco Óptico/diagnóstico por imagem , Índice de Perfusão , Doenças Retinianas/tratamento farmacológico , Tomografia de Coerência Óptica , Tonometria Ocular , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidoresRESUMO
INTRODUCTION: With the large-scale availability of transgenic and knockout mouse models, the use of mice may greatly facilitate the examination of the mechanisms underlying diabetic erectile dysfunction (ED). Although in vitro studies of the mouse cavernosum show impairment of vasoreactivity, to date, no studies have demonstrated the in vivo impairment of erectile function in diabetic mice. AIM: To establish whether mouse models of type I diabetes exhibit in vivo ED. METHODS: Hyperglycemia was induced by injection with streptozotocin (STZ, 125 mg/kg x 2 days) in two mouse strains, C57BLKS (BKS) and BALB/c. Six weeks after injection, the cavernosum was removed from some mice for the in vitro assessment of the endothelium and nerve-mediated dilatory responses of the cavernosal strips. The in vivo assessment of intracorporal pressure normalized to mean arterial pressure, in response to the electrical stimulation of the cavernosal nerve, was performed in the remaining mice. MAIN OUTCOME MEASURES: The main outcome measure of this study was the in vivo assessment of erectile function following diabetic induction in mice. RESULTS: Despite similar levels of sustained hyperglycemia following STZ injection, the phenotype of diabetic ED was observed only in BKS and not BALB/c mice. The cavernosum from diabetic BKS mice showed decreased endothelium-dependent dilation in response to acetylcholine (ACh), as well as impaired parasympathetic nerve-mediated relaxation. There was no change in ACh or nerve-mediated relaxation in the cavernousum from diabetic vs. control BALB/c mice. Further, in vivo physiologic assessment of erectile activity revealed a significant decrease in erectile function in diabetic BKS but not in BALB/c mice. CONCLUSION: Together these data first established in vivo ED in a mouse model of type I diabetes (BKS mouse) and importantly demonstrated that certain inbred strains may be protected from hyperglycemia-induced erectile impairment. Further study of the strain-dependent effects may offer important clues into the mechanisms of ED as it relates to type I diabetes.
Assuntos
Diabetes Mellitus Experimental/fisiopatologia , Disfunção Erétil/fisiopatologia , Camundongos Endogâmicos , Animais , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ereção Peniana/fisiologiaRESUMO
To investigate the full range of molecular changes associated with erectile dysfunction (ED) in Type 1 diabetes, we examined alterations in penile gene expression in streptozotocin-induced diabetic rats and littermate controls. With the use of Affymetrix GeneChip arrays and statistical filtering, 529 genes/transcripts were considered to be differentially expressed in the diabetic rat cavernosum compared with control. Gene Ontology (GO) classification indicated that there was a decrease in numerous extracellular matrix genes (e.g., collagen and elastin related) and an increase in oxidative stress-associated genes in the diabetic rat cavernosum. In addition, PubMatrix literature mining identified differentially expressed genes previously shown to mediate vascular dysfunction [e.g., ceruloplasmin (Cp), lipoprotein lipase, and Cd36] as well as genes involved in the modulation of the smooth muscle phenotype (e.g., Kruppel-like factor 5 and chemokine C-X3-C motif ligand 1). Real-time PCR was used to confirm changes in expression for 23 relevant genes. Further validation of Cp expression in the diabetic rat cavernosum demonstrated increased mRNA levels of the secreted and anchored splice variants of Cp. CP protein levels showed a 1.9-fold increase in tissues from diabetic rats versus controls. Immunohistochemistry demonstrated localization of CP protein in cavernosal sinusoids of control and diabetic animals, including endothelial and smooth muscle layers. Overall, this study broadens the scope of candidate genes and pathways that may be relevant to the pathophysiology of diabetes-induced ED as well as highlights the potential complexity of this disorder.
Assuntos
Diabetes Mellitus Experimental/complicações , Disfunção Erétil/complicações , Disfunção Erétil/genética , Regulação da Expressão Gênica , Análise de Sequência com Séries de Oligonucleotídeos , Animais , Glicemia/metabolismo , Pressão Sanguínea , Ceruloplasmina/genética , Ceruloplasmina/metabolismo , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/genética , Estimulação Elétrica , Masculino , Tecido Nervoso/metabolismo , Transporte Proteico , Ratos , Ratos Endogâmicos F344 , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima/genéticaRESUMO
Oxidative stress has been implicated in the development of vascular disease and in the promotion of endothelial dysfunction via the reduction in bioavailable nitric oxide (NO()). Glutathione (GSH) is a tripeptide thiol antioxidant that is utilized by glutathione peroxidase (GPx) to scavenge reactive oxygen species such as hydrogen peroxide and phospholipid hydroperoxides. Relatively frequent single-nucleotide polymorphisms (SNPs) within the 5' promoters of the GSH synthesis genes GCLC and GCLM are associated with impaired vasomotor function, as measured by decreased acetylcholine-stimulated coronary artery dilation, and with increased risk of myocardial infarction. Although the influence of genetic knockdown of GPx on vascular function has been investigated in mice, no work to date has been published on the role of genetic knockdown of GSH synthesis genes on vascular reactivity. We therefore investigated the effects of targeted disruption of Gclm in mice and the subsequent depletion of GSH on vascular reactivity, NO() production, aortic nitrotyrosine protein modification, and whole-genome transcriptional responses as measured by DNA microarray. Gclm(-/+) and Gclm(-/-) mice had 72 and 12%, respectively, of wild-type (WT) aortic GSH content. Gclm(-/+) mice had a significant impairment in acetylcholine (ACh)-induced relaxation in aortic rings as well as increased aortic nitrotyrosine protein modification. Surprisingly, Gclm(-/-) aortas showed enhanced relaxation compared to Gclm(-/+) aortas, as well as increased NO() production. Although aortic rings from Gclm(-/-) mice had enhanced ACh relaxation, they had a significantly increased sensitivity to phenylephrine (PE)-induced contraction. Alternatively, the PE response of Gclm(-/+) aortas was nearly identical to that of their WT littermates. To examine the role of NO() or other potential endothelium-derived factors in differentially regulating vasomotor activity, we incubated aortic rings with the NO() synthase inhibitor L-NAME or physically removed the endothelium before PE treatment. L-NAME treatment and endothelium removal enhanced PE-induced contraction in WT and Gclm(-/+) mice, but this effect was severely diminished in Gclm(-/-) mice, indicating a potentially unique role for GSH in mediating vessel contraction. Whole-genome assessment of aortic mRNA in Gclm(-/-) and WT mice revealed altered expression of genes within the canonical Ca(2+) signaling pathway, which may have a role in mediating these observed functional effects. These findings provide additional evidence that the de novo synthesis of GSH can influence vascular reactivity and provide insights regarding possible mechanisms by which SNPs within GCLM and GCLC influence the risk of developing vascular diseases in humans.
Assuntos
Aorta/fisiologia , Glutamato-Cisteína Ligase/genética , Glutationa/metabolismo , Acetilcolina/farmacologia , Agonistas de Receptores Adrenérgicos alfa 1/farmacologia , Animais , Aorta/enzimologia , Aorta/metabolismo , Agonistas Colinérgicos/farmacologia , GMP Cíclico/metabolismo , Glutamato-Cisteína Ligase/metabolismo , Técnicas In Vitro , Masculino , Redes e Vias Metabólicas/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Fenilefrina/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Transcriptoma , Tirosina/análogos & derivados , Tirosina/metabolismo , Vasoconstrição/efeitos dos fármacos , Vasodilatação/efeitos dos fármacosRESUMO
Urokinase-type plasminogen activator (uPA) is expressed at increased levels in stenotic, atherosclerotic human arteries. However, the biological roles of uPA in the artery wall are poorly understood. Previous studies associate uPA with both acute vasoconstriction and chronic vascular remodeling and attribute uPA-mediated vasoconstriction to the kringle - not the catalytic - domain of uPA. We used an in-vivo uPA overexpression model to test the hypothesis that uPA-induced vasoconstriction is a reversible vasomotor process that can be prevented - and uPA fibrinolytic activity preserved - by: 1) removing the growth factor and kringle domains; or 2) anchoring uPA to the endothelial surface. To test this hypothesis we constructed adenoviral vectors that express: wild-type rabbit uPA (AduPA); a uPA mutant lacking the NH(2)-terminal growth-factor and kringle domains (AduPAdel); a mutant lacking catalytic activity (AduPAS-->A), and a cell-surface anchored mutant (AdTMuPA). uPA mutants were expressed and characterised in vitro and in carotid arteries in vivo. uPAS-->A had no plasminogen activator activity. Activity was similar for uPA and uPAdel, whereas AdTMuPA had only cell-associated activity. AduPAS-->A arteries were not constricted. AduPA, AduPAdel, and AdTM-uPA arteries were constricted (approximately 30% smaller lumens; p< or =0.008 vs. AdNull arteries). Papaverine reversed constriction of AduPA arteries. uPA-mediated arterial constriction is a vasomotor process that is mediated by uPA catalytic activity, not by the NH(2)-terminal domains. Anchoring uPA to the endothelial surface does not prevent vasoconstriction. uPA catalytic activity, generated by artery wall cells, may contribute to lumen loss in human arteries. Elimination of uPA vasoconstrictor activity requires concomitant loss of fibrinolytic activity.
Assuntos
Artéria Carótida Primitiva/efeitos dos fármacos , Ativador de Plasminogênio Tipo Uroquinase/farmacologia , Vasoconstrição/efeitos dos fármacos , Adenoviridae/genética , Animais , Catálise , Domínio Catalítico , Células Endoteliais/química , Fibrinólise/efeitos dos fármacos , Vetores Genéticos/farmacologia , Humanos , Técnicas In Vitro , Masculino , Músculo Liso Vascular/citologia , Mutagênese Sítio-Dirigida , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/efeitos dos fármacos , Estrutura Terciária de Proteína , Coelhos , Proteínas Recombinantes de Fusão/farmacologia , Deleção de Sequência , Organismos Livres de Patógenos Específicos , Ativador de Plasminogênio Tipo Uroquinase/química , Ativador de Plasminogênio Tipo Uroquinase/genéticaRESUMO
The number of men with type II diabetes-associated erectile dysfunction (ED) continues to grow rapidly; however, the majority of basic science studies has examined mechanisms of ED in animal models of type I diabetes. In this study, we first establish an in vivo mouse model of type II diabetic ED using the leptin receptor mutated db/db and wild-type control BKS mouse. Furthermore, we hypothesized that dual mechanistic impairments contribute to the impaired erectile function in the type II diabetic mouse, altered vasoreactivity, and venoocclusive disorder. In vivo erectile function was measured as intracavernosal pressure (ICP) normalized to mean arterial pressure (MAP) following electrical stimulation of the cavernosal nerve. Venoocclusion was assessed by the maintenance of elevated in vivo ICP following intracorporal saline infusion. Vasoreactivity of isolated cavernosum in response to contractile and dilatory stimulation was examined in vitro by myography. Collagen and elastin content were evaluated by quantification of hydroxyproline and desmosine, respectively, as well as by quantitative PCR and histological analysis of isolated cavernosum. Erectile function was significantly decreased in db/db vs. BKS mice in a manner consistent with impairments in venoocclusive ability and decreased inflow. Heightened vasoconstriction and attenuated dilation in cavernosum of db/db vs. BKS mice suggest an overall lowered relaxation ability and thus impaired filling of the cavernosal spaces. A decrease in desmosine and hydroxyproline as well as lowered mRNA levels for tropoelastin, fibrillin-1, and alpha1(I) collagen were detected. These vasoreactive and sinusoidal matrix alterations may alter tissue compliance dispensability, preventing the normal expansion necessary for erection.