RESUMEN
Abstract The renin-angiotensin-aldosterone system (RAAS) plays a key role in diabetic nephropathy (DN). Angiotensin-II secreted during the RAAS pathway increases nephropathy. It stimulates oxidative stress which can quench nitric oxide. Reduced nitric oxide level aggravates Ang-II-induced vasoconstriction. Ang-II has also emerged as a central mediator of the glomerular hemodynamic changes that are associated with renal injury. Deletion of ACE2 is also noted due to increased Ang-II level which leads to the development of DN. We hypothesize that nephropathy caused by Ang-II in the periphery may be controlled by brain RAAS. ACE inhibitors and ARBs may show the renoprotective effect when administered through ICV without crossing the blood-brain barrier. DN was observed after 8 weeks of diabetes induction through alloxan. Administration of captopril and valsartan once and in combined therapy for 2 weeks, significantly reduced urine output, blood urea nitrogen, total protein in the urine, serum cholesterol, serum creatinine, serum triglycerides, and kidney/body weight ratio as compared to diabetic control rats. Further, combination therapy significantly increased the body weight and serum nitrate level as compared to diabetic control animals. However, increased ACE2 levels in the brain may reduce the sympathetic outflow and might have decreased the peripheral activity of Ang-II which shows beneficial effects in DN.
Asunto(s)
Animales , Masculino , Femenino , Ratas , Sistema Renina-Angiotensina/inmunología , Angiotensina II/análisis , Nefropatías Diabéticas/patología , Heridas y Lesiones/clasificación , Inhibidores de la Enzima Convertidora de Angiotensina/administración & dosificación , Peptidil-Dipeptidasa A/administración & dosificaciónAsunto(s)
Betacoronavirus/fisiología , Infecciones por Coronavirus/fisiopatología , Modelos Animales de Enfermedad , Virus de la Encefalitis Equina Venezolana/genética , Peptidil-Dipeptidasa A/administración & dosificación , Neumonía Viral/fisiopatología , Replicón , Enzima Convertidora de Angiotensina 2 , Animales , COVID-19 , Línea Celular Transformada , Infecciones por Coronavirus/virología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Pandemias , Peptidil-Dipeptidasa A/genética , Neumonía Viral/virología , SARS-CoV-2 , Transducción Genética , Replicación ViralRESUMEN
COVID-19 is a new zoonotic disease caused by the SARS-CoV-2 virus. Since its emergence in Wuhan City, China, the virus has rapidly spread across the globe causing calamitous health, economic and societal consequences. It causes disproportionately severe disease in the elderly and those with co-morbidities, such as hypertension and diabetes. There is currently no proven treatment for COVID-19 and a safe and effective vaccine is at least a year away. The virus gains access to the respiratory epithelium through cell surface angiotensin converting enzyme 2 (ACE2). The receptor binding domain (RBD) of the virus is unlikely to mutate without loss of pathogenicity and thus represents an attractive target for antiviral treatment. Inhaled modified recombinant human ACE2, may bind SARS-CoV-2 and mitigate lung damage. This decoy strategy is unlikely to provoke an adverse immune response and may reduce morbidity and mortality in high-risk groups.
Asunto(s)
Administración por Inhalación , Infecciones por Coronavirus/tratamiento farmacológico , Peptidil-Dipeptidasa A/uso terapéutico , Neumonía Viral/tratamiento farmacológico , Enzima Convertidora de Angiotensina 2 , Betacoronavirus , COVID-19 , Humanos , Pulmón/virología , Pandemias , Peptidil-Dipeptidasa A/administración & dosificación , Unión Proteica , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/uso terapéutico , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus , Tratamiento Farmacológico de COVID-19RESUMEN
The COVID-19 pandemic is associated with neurological symptoms and complications including stroke. There is hypercoagulability associated with COVID-19 that is likely a "sepsis-induced coagulopathy" and may predispose to stroke. The SARS-CoV-2 virus binds to angiotensin-converting enzyme 2 (ACE2) present on brain endothelial and smooth muscle cells. ACE2 is a key part of the renin angiotensin system (RAS) and a counterbalance to angiotensin-converting enzyme 1 (ACE1) and angiotensin II. Angiotensin II is proinflammatory, is vasoconstrictive, and promotes organ damage. Depletion of ACE2 by SARS-CoV-2 may tip the balance in favor of the "harmful" ACE1/angiotensin II axis and promote tissue injury including stroke. There is a rationale to continue to treat with tissue plasminogen activator for COVID-19-related stroke and low molecular weight heparinoids may reduce thrombosis and mortality in sepsis-induced coagulopathy.
Asunto(s)
Infecciones por Coronavirus/complicaciones , Neumonía Viral/complicaciones , Accidente Cerebrovascular/virología , Enzima Convertidora de Angiotensina 2 , Betacoronavirus/metabolismo , COVID-19 , Infecciones por Coronavirus/sangre , Infecciones por Coronavirus/tratamiento farmacológico , Infecciones por Coronavirus/epidemiología , Coagulación Intravascular Diseminada/tratamiento farmacológico , Coagulación Intravascular Diseminada/virología , Femenino , Humanos , Masculino , Pandemias , Peptidil-Dipeptidasa A/administración & dosificación , Peptidil-Dipeptidasa A/metabolismo , Neumonía Viral/sangre , Neumonía Viral/tratamiento farmacológico , Neumonía Viral/epidemiología , Proteínas Recombinantes/administración & dosificación , SARS-CoV-2 , Accidente Cerebrovascular/tratamiento farmacológico , Activador de Tejido Plasminógeno/sangre , Activador de Tejido Plasminógeno/uso terapéuticoRESUMEN
Angiotensin-converting enzyme 2 (ACE2) is a monocarboxypeptidase in the renin-angiotensin system that catalyzes the breakdown of angiotensin II to angiotensin 1-7. We have reported that ACE2 expression in the kidney is reduced in experimental Alport syndrome but the impact of this finding on disease progression has not been studied. Accordingly, we evaluated effects of murine recombinant ACE2 treatment in Col4a3 knockout mice, a model of Alport syndrome characterized by proteinuria and progressive renal injury. Murine recombinant ACE2 (0.5 mg/kg/day) was administered from four to seven weeks of age via osmotic mini-pump. Pathological changes were attenuated by murine recombinant ACE2 treatment which ameliorated kidney fibrosis as shown by decreased expression of COL1α1 mRNA, less accumulation of extracellular matrix proteins, and inhibition of transforming growth factor-ß signaling. Further, increases in proinflammatory cytokine expression, macrophage infiltration, inflammatory signaling pathway activation, and heme oxygenase-1 levels in Col4a3 knockout mice were also reduced by murine recombinant ACE2 treatment. Lastly, murine recombinant ACE2 influenced the turnover of renal ACE2, as it suppressed the expression of tumor necrosis factor-α converting enzyme, a negative regulator of ACE2. Thus, treatment with exogenous ACE2 alters angiotensin peptide metabolism in the kidneys of Col4a3 knockout mice and attenuates the progression of Alport syndrome nephropathy.
Asunto(s)
Riñón/efectos de los fármacos , Nefritis Hereditaria/tratamiento farmacológico , Peptidil-Dipeptidasa A/administración & dosificación , Albuminuria/tratamiento farmacológico , Albuminuria/etiología , Albuminuria/metabolismo , Enzima Convertidora de Angiotensina 2 , Angiotensinas/metabolismo , Animales , Autoantígenos/genética , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Cadena alfa 1 del Colágeno Tipo I , Colágeno Tipo IV/deficiencia , Colágeno Tipo IV/genética , Citocinas/metabolismo , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Proteínas de la Matriz Extracelular/genética , Proteínas de la Matriz Extracelular/metabolismo , Fibrosis , Predisposición Genética a la Enfermedad , Mediadores de Inflamación/metabolismo , Riñón/metabolismo , Riñón/patología , Masculino , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Ratones Noqueados , Nefritis Hereditaria/complicaciones , Nefritis Hereditaria/genética , Nefritis Hereditaria/metabolismo , Estrés Oxidativo/efectos de los fármacos , Fenotipo , Proteínas Recombinantes/administración & dosificación , Transducción de Señal/efectos de los fármacos , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
Blockers of the renin-angiotensin system are effective in the treatment of experimental and clinical diabetic nephropathy. An approach different from blocking the formation or action of angiotensin II (1-8) that could also be effective involves fostering its degradation. Angiotensin-converting enzyme 2 (ACE2) is a monocarboxypeptidase that cleaves angiotensin II (1-8) to form angiotensin (1-7). Therefore, we examined the renal effects of murine recombinant ACE2 in mice with streptozotocin-induced diabetic nephropathy as well as that of amplification of circulating ACE2 using minicircle DNA delivery prior to induction of experimental diabetes. This delivery resulted in a long-term sustained and profound increase in serum ACE2 activity and enhanced ability to metabolize an acute angiotensin II (1-8) load. In mice with streptozotocin-induced diabetes pretreated with minicircle ACE2, ACE2 protein in plasma increased markedly and this was associated with a more than 100-fold increase in serum ACE2 activity. However, minicircle ACE2 did not result in changes in urinary ACE2 activity as compared to untreated diabetic mice. In both diabetic groups, glomerular filtration rate increased significantly and to the same extent as compared to non-diabetic controls. Albuminuria, glomerular mesangial expansion, glomerular cellularity, and glomerular size were all increased to a similar extent in minicircle ACE2-treated and untreated diabetic mice, as compared to non-diabetic controls. Recombinant mouse ACE2 given for 4 weeks by intraperitoneal daily injections in mice with streptozotocin-induced diabetic nephropathy also failed to improve albuminuria or kidney pathology. Thus, a profound augmentation of ACE2 confined to the circulation failed to ameliorate the glomerular lesions and hyperfiltration characteristic of early diabetic nephropathy. These findings emphasize the importance of targeting the kidney rather than the circulatory renin angiotensin system to combat diabetic nephropathy.
Asunto(s)
Diabetes Mellitus Experimental/complicaciones , Nefropatías Diabéticas/etiología , Riñón/enzimología , Peptidil-Dipeptidasa A/sangre , Albuminuria/enzimología , Albuminuria/etiología , Albuminuria/genética , Angiotensina I/metabolismo , Angiotensina II/metabolismo , Enzima Convertidora de Angiotensina 2 , Animales , Autoantígenos/genética , Colágeno Tipo IV/deficiencia , Colágeno Tipo IV/genética , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Experimental/enzimología , Nefropatías Diabéticas/sangre , Nefropatías Diabéticas/enzimología , Nefropatías Diabéticas/fisiopatología , Femenino , Tasa de Filtración Glomerular , Riñón/patología , Riñón/fisiopatología , Ratones Endogámicos C57BL , Ratones Noqueados , Fragmentos de Péptidos/metabolismo , Peptidil-Dipeptidasa A/administración & dosificación , Peptidil-Dipeptidasa A/genética , Proteínas Recombinantes/administración & dosificación , Factores de TiempoRESUMEN
BACKGROUND/AIMS: Angiotensin converting enzyme 2 (ACE2) treatment suppresses the severity of acute lung injury (ALI), through antagonizing hydrolyzing angiotensin II (AngII) and the ALI-induced apoptosis of pulmonary endothelial cells. Nevertheless, the effects of ACE2 on vessel permeability and its relationship with vascular endothelial growth factor a (VEGFa) remain ill-defined. In the current study, we examined the relationship between ACE2 and VEGFa in ALI model in mice. METHODS: Here, we used a previously published bleomycin method to induce ALI in mice, and treated the mice with ACE2. We analyzed the levels of VEGFa in these mice. The mouse lung vessel permeability was determined by a fluorescence pharmacokinetic assay following i.v. injection of 62.5µg/kg Visudyne. VEGFa pump or SU5416 pump was given to increase or decrease VEGFa effects, respectively. The long-term effects on lung function were determined by measurement of lung resistance using methacholine. RESULTS: ACE2 treatment did not alter VEGFa levels in lung, but antagonized the effects of VEGFa on increases of lung vessel permeability. Ectogenic VEGFa abolished the antagonizing effects of ACE2 on the vessel permeability against VEGFa. On the other hand, suppression of VEGF signaling mimicked the effects of ACE2 on the vessel permeability against VEGFa. The suppression of vessel permeability resulted in improvement of lung function after ALI. CONCLUSION: ACE2 may antagonize the VEGFa-mediated increases in lung vessel permeability during ALI, resulting in improvement of lung function after ALI.
Asunto(s)
Lesión Pulmonar Aguda/dietoterapia , Permeabilidad Capilar/efectos de los fármacos , Peptidil-Dipeptidasa A/administración & dosificación , Factor A de Crecimiento Endotelial Vascular/metabolismo , Lesión Pulmonar Aguda/inducido químicamente , Lesión Pulmonar Aguda/metabolismo , Enzima Convertidora de Angiotensina 2 , Animales , Bleomicina/efectos adversos , Células Endoteliales , Regulación de la Expresión Génica , Indoles/administración & dosificación , Indoles/farmacología , Ratones , Peptidil-Dipeptidasa A/farmacología , Porfirinas/administración & dosificación , Porfirinas/farmacocinética , Pirroles/administración & dosificación , Pirroles/farmacología , VerteporfinaRESUMEN
BACKGROUND: Hypertension is considered the most serious risk factor for cardiovascular disease. Angiotensin-converting enzyme inhibitory peptides (ACEIPs), which are made from tuna frame protein (TFP) and yellow fin sole frame protein (YFP), have been used previously to treat hypertension. However, the production of these short peptides is usually dependent on enzymatic hydrolysis, resulting in a digested mixture that makes it difficult to purify the ACEIPs. Although it has been reported that ACEIPs could be produced in recombinant Escherichia coli strains, the use of lactic acid bacteria in the production of ACEIPs has not been demonstrated. RESULTS: In this study, the ACEIP coding sequences from TFP and YFP were joined through an arginine linker and expressed in the Lactobacillus plantarum (Lb. plantarum) NC8 strain by an inducible vector pSIP-409. Then, the antihypertensive effects were determined in the model of spontaneously hypertensive rats (SHRs) by measuring the blood pressure, hematology, blood biochemistry and nitric oxide (NO), endothelin (ET) and angiotensin II (Ang II) levels. The results showed that oral administration of recombinant Lb. plantarum NC8 (RLP) significantly decreased systolic blood pressure (P < 0.01) during treatment, which lasted for at least 10 days after the last dose. Furthermore, the presence of RLP resulted in an increased level of NO, as well as decreased levels of ET and Ang II in plasma, heart, and kidney. In addition, a dramatically decreased triglyceride level was also observed even though there was no significant change in hematology or blood biochemistry. Although some drawbacks were still observed, such as the presence of an antibiotic selection marker, no obvious side effects or bacterial translocation were observed in vivo, indicating the potential application of RLP in the treatment of hypertension. CONCLUSION: These results demonstrated the effectiveness and safety of RLP on the treatment of hypertension.
Asunto(s)
Antihipertensivos/uso terapéutico , Hipertensión/tratamiento farmacológico , Lactobacillus plantarum/química , Peptidil-Dipeptidasa A/uso terapéutico , Animales , Antihipertensivos/administración & dosificación , Humanos , Masculino , Ratones , Peptidil-Dipeptidasa A/administración & dosificación , RatasRESUMEN
BACKGROUND/AIMS: Angiotensin converting enzyme 2 (ACE2) treatment suppresses the severity of acute lung injury (ALI). The effects of ACE2 in ALI have been shown to not only result from its antagonizing hydrolyzing angiotensin II (AngII), which is responsible for reduction in the vascular tension and pulmonary accumulation of inflammatory cells, but also result from a role of ACE2 in suppressing the ALI-induced apoptosis of pulmonary endothelial cells (PECs). Nevertheless, the underlying mechanisms of the role of ACE2 on PEC apoptosis are not completely understood. METHODS: Here, we used a bleomycin-induced mouse model for ALI that has been published in our previous studies. We analyzed the mRNA and protein levels of an anti-apoptotic protein Bcl-2 in the ALI-mice that have been treated w/o ACE2. We analyzed miR-4262 levels in the mouse lung in these mice. Bcl-2-targeting miRNAs were predicted using bioinformatics algorithms and a luciferase reporter assay was applied to examine the effects of miR-4262 on the Bcl-2 protein translation upon their binding to 3'-UTR of Bcl-2 mRNA. Adeno-associated viruses carrying either miR-4262 mimics or antisense were injected into ALI-mice without ACE2, and their effects on the apoptosis in mouse lung cells were analyzed by Western blot. RESULTS: ACE2 inhibited the ALI-induced apoptosis of pulmonary cells in vivo partially through upregulation of Bcl-2 protein, but not Bcl-2 mRNA. ACE2 appeared to significantly suppress the upregulation of miR-4262 in mouse lung after ALI. MiR-4262 was found to target 3'-UTR of Bcl-2 mRNA to inhibit its protein translation in PECs. In vivo administration of antisense of miR-4262 decreased apoptosis of pulmonary cells and severity of the ALI in mice. CONCLUSION: ACE2-induced suppression of miR-4262 partially contribute to the inhibition of the PEC apoptosis after ALI through Bcl-2. MiR-4262 may be a novel promising treatment target for ALI and ARDS.
Asunto(s)
Lesión Pulmonar Aguda/tratamiento farmacológico , Bleomicina/efectos adversos , Células Endoteliales/efectos de los fármacos , MicroARNs/genética , Peptidil-Dipeptidasa A/administración & dosificación , Lesión Pulmonar Aguda/inducido químicamente , Enzima Convertidora de Angiotensina 2 , Animales , Apoptosis/efectos de los fármacos , Línea Celular , Modelos Animales de Enfermedad , Células Endoteliales/citología , Células HEK293 , Humanos , Pulmón/citología , Pulmón/patología , Ratones , MicroARNs/metabolismo , Peptidil-Dipeptidasa A/farmacología , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismoAsunto(s)
Angiotensina I/administración & dosificación , Presión Sanguínea/efectos de los fármacos , Hipertensión Pulmonar/tratamiento farmacológico , Fragmentos de Péptidos/administración & dosificación , Peptidil-Dipeptidasa A/administración & dosificación , Sistema Renina-Angiotensina/fisiología , Enzima Convertidora de Angiotensina 2 , Animales , MasculinoRESUMEN
Emerging evidences indicate that diminished activity of the vasoprotective axis of the renin-angiotensin system, constituting angiotensin-converting enzyme 2 (ACE2) and its enzymatic product, angiotensin-(1-7) [Ang-(1-7)] contribute to the pathogenesis of pulmonary hypertension (PH). However, long-term repetitive delivery of ACE2 or Ang-(1-7) would require enhanced protein stability and ease of administration to improve patient compliance. Chloroplast expression of therapeutic proteins enables their bioencapsulation within plant cells to protect against gastric enzymatic degradation and facilitates long-term storage at room temperature. Besides, fusion to a transmucosal carrier helps effective systemic absorption from the intestine on oral delivery. We hypothesized that bioencapsulating ACE2 or Ang-(1-7) fused to the cholera nontoxin B subunit would enable development of an oral delivery system that is effective in treating PH. PH was induced in male Sprague Dawley rats by monocrotaline administration. Subset of animals was simultaneously treated with bioencapsulaed ACE2 or Ang-(1-7) (prevention protocol). In a separate set of experiments, drug treatment was initiated after 2 weeks of PH induction (reversal protocol). Oral feeding of rats with bioencapsulated ACE2 or Ang-(1-7) prevented the development of monocrotaline-induced PH and improved associated cardiopulmonary pathophysiology. Furthermore, in the reversal protocol, oral ACE2 or Ang-(1-7) treatment significantly arrested disease progression, along with improvement in right heart function, and decrease in pulmonary vessel wall thickness. In addition, a combination therapy with ACE2 and Ang-(1-7) augmented the beneficial effects against monocrotaline-induced lung injury. Our study provides proof-of-concept for a novel low-cost oral ACE2 or Ang-(1-7) delivery system using transplastomic technology for pulmonary disease therapeutics.
Asunto(s)
Angiotensina I/administración & dosificación , Presión Sanguínea/efectos de los fármacos , Hipertensión Pulmonar/tratamiento farmacológico , Fragmentos de Péptidos/administración & dosificación , Peptidil-Dipeptidasa A/administración & dosificación , Sistema Renina-Angiotensina/fisiología , Administración Oral , Enzima Convertidora de Angiotensina 2 , Animales , Antihipertensivos/administración & dosificación , Cloroplastos , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Portadores de Fármacos , Quimioterapia Combinada , Hipertensión Pulmonar/metabolismo , Hipertensión Pulmonar/fisiopatología , Masculino , Ratas , Ratas Sprague-Dawley , Sistema Renina-Angiotensina/efectos de los fármacosRESUMEN
Hyperactivity of the renin-angiotensin system (RAS) resulting in elevated Angiotensin II (Ang II) contributes to all stages of inflammatory responses including ocular inflammation. The discovery of angiotensin-converting enzyme 2 (ACE2) has established a protective axis of RAS involving ACE2/Ang-(1-7)/Mas that counteracts the proinflammatory and hypertrophic effects of the deleterious ACE/AngII/AT1R axis. Here we investigated the hypothesis that enhancing the systemic and local activity of the protective axis of the RAS by oral delivery of ACE2 and Ang-(1-7) bioencapsulated in plant cells would confer protection against ocular inflammation. Both ACE2 and Ang-(1-7), fused with the non-toxic cholera toxin subunit B (CTB) were expressed in plant chloroplasts. Increased levels of ACE2 and Ang-(1-7) were observed in circulation and retina after oral administration of CTB-ACE2 and Ang-(1-7) expressing plant cells. Oral feeding of mice with bioencapsulated ACE2/Ang-(1-7) significantly reduced endotoxin-induced uveitis (EIU) in mice. Treatment with bioencapsulated ACE2/Ang-(1-7) also dramatically decreased cellular infiltration, retinal vasculitis, damage and folding in experimental autoimmune uveoretinitis (EAU). Thus, enhancing the protective axis of RAS by oral delivery of ACE2/Ang-(1-7) bioencapsulated in plant cells provide an innovative, highly efficient and cost-effective therapeutic strategy for ocular inflammatory diseases.
Asunto(s)
Angiotensina I/administración & dosificación , Cloroplastos/genética , Modelos Animales de Enfermedad , Fragmentos de Péptidos/administración & dosificación , Peptidil-Dipeptidasa A/administración & dosificación , Plantas Modificadas Genéticamente/metabolismo , Retinitis/terapia , Uveítis/terapia , Administración Oral , Enzima Convertidora de Angiotensina 2 , Animales , Cloroplastos/metabolismo , Humanos , Ratones , Ratones Endogámicos C57BL , Plantas Modificadas Genéticamente/genética , Vasculitis Retiniana , Retinitis/inducido químicamente , Retinitis/inmunología , Uveítis/inducido químicamente , Uveítis/inmunologíaRESUMEN
BACKGROUND AND OBJECTIVES: Angiotensin-converting enzyme 2 (ACE2) converts angiotensin II (Ang1-8) to angiotensin 1-7 (Ang1-7), a functional antagonist of Ang1-8, with vasodilatory, antiproliferative, antiangiogenic, and anti-inflammatory properties. In conditions with an unbalanced renin-angiotensin-aldosterone system with elevated Ang1-8, administration of ACE2 has shown promising effects in a variety of animal models. Enhancing ACE2 activity by exogenous administration of ACE2 might also be beneficial in human diseases with pathologically elevated Ang1-8. As a first step we performed a first-in-man study to determine pharmacokinetics, pharmacodynamics, safety, and tolerability of recombinant ACE2 in healthy volunteers. METHODS: Recombinant human ACE2 (rhACE2) was administered intravenously to healthy human subjects in a randomized, double-blind, placebo-controlled, single-dose, dose-escalation study followed by an open-label multiple-dose study. ACE2 concentrations were determined by quantifying ACE2 activity and ACE2 content in plasma samples. Concentrations of the angiotensin system effector peptides Ang1-8, Ang1-7, and Ang1-5 were determined using a liquid chromatography-tandem mass spectrometry method. RESULTS: Single rhACE2 doses of 100-1,200 µg/kg caused a dose-dependent increase of systemic exposure with biphasic elimination and a dose-independent terminal half-life of 10 h. In all single-dose cohorts, Ang1-8 decreased within 30 min postinfusion, angiotensin 1-7 (Ang1-7) either increased (100 and 200 µg/kg doses), decreased, or remained unchanged (400-1,200 µg/kg doses), whereas angiotensin 1-5 (Ang1-5) transiently increased for all doses investigated. With the exception of the lowest rhACE2 dose, the decrease in Ang1-8 levels lasted for at least 24 h. Repeated dosing (400 µg/kg for 3 or 6 days) caused only minimal accumulation of ACE2, and Ang1-8 levels were suppressed over the whole application period. CONCLUSIONS: Administration of rhACE2 was well tolerated by healthy human subjects. Exposure was dose dependent with a dose-independent terminal elimination half-life in the range of 10 h. Despite marked changes in angiotensin system peptide concentrations, cardiovascular effects were absent, suggesting the presence of effective compensatory mechanisms in healthy volunteers.
Asunto(s)
Peptidil-Dipeptidasa A/farmacocinética , Proteínas Recombinantes/farmacocinética , Angiotensina I/sangre , Angiotensina II/sangre , Enzima Convertidora de Angiotensina 2 , Método Doble Ciego , Femenino , Humanos , Masculino , Fragmentos de Péptidos/sangre , Peptidil-Dipeptidasa A/administración & dosificación , Proteínas Recombinantes/administración & dosificaciónRESUMEN
Resumen: La enzima convertidora de angiotensina I (ECA2) a través de Angiotensina (Ang)-(1-9) más que Ang-(1-7) contrarresta los efectos deletéreos de ECA y Ang II. Se desconoce si Ang-(1-9) es efectiva en el tratamiento del remodelamiento cardiovascular (RMCV) hipertensivo, en ratas con polimorfismo del gen de la ECA. Objetivo: Determinar el efecto de Ang-(1-9) en el tratamiento del RMCV hipertensivo en ratas con niveles genéticamente determinados de ECA y Ang II. Métodos: Ratas normotensas homocigotas, Lewis (LL) y Brown Norway (BN), se les indujo HTA a través del modelo Goldblatt (GB, 2 riñones-1 pinzado). Después de 4 semanas, las ratas hipertensas se rando-mizaron para recibir Ang-(1-9) (602 ng/Kg min) o una coadministración de Ang-(1-9)+A779 (100 ng/Kg min, antagonista del receptor MAS de Ang-(1-7)) durante 14 días mediante una minibomba. Como controles se usaron ratas sometidas a operación ficticia (Sham). Se determinó masa corporal (MC), presión arterial sistólica (PAS), masa ventricular (MV), área de cardiomiocitos (AC), área y grosor de la túnica media (ATM, GTM), fracción volumétrica de colágeno total (FVCT) en el ventrículo izquierdo (VI), niveles proteicos de colágeno tipo I (Col I) en la aorta (Ao) y la infiltración de macrófagos en Ao y VI, por medio de su molécula especifica ED1 (ED1-Ao, ED1-VI). Resultados: La administración de Ang-(1-9) disminuyó significativamente PAS, MV, AC, FVCT, Col I, ATM, GTM, ED1-Ao (-) y ED1-VI, en las ratas hipertensas LL y BN respecto a las ratas GB sin tratamiento, respectivamente. Este efecto no fue inhibido por el antagonista A779. El polimorfismo de la ECA no modificó la respuesta al tratamiento. Conclusión: Ang-(1-9) redujo eficazmente la HTA y el RMCV secundario, independiente al polimorfismo en el gen de la ECA. Este efecto posiblemente es directo ya que no fue mediado por Ang-(1-7). Fondecyt 1100874.
Background: The angiotensin I converting enzyme 2 (ACE2) counteracts the deleterious effects of ACE and Ang II through angiotensin (Ang) -(1-9) rather than Ang-(1-7). In addition, it is not clear whether Ang-(1-9) is effective in the reversal of hypertensive cardiovascular remodeling (CVRM) in rats with ACE gene polymorphism. Objective: To determine the effect of Ang-(1-9) in the prevention of hypertensive CVRM in rats with genetically determined levels of ACE and Ang II. Methods: In normotensive homozygous Lewis (LL) and Brown Norway (BN) rats hypertension was induced by the Goldblatt 2 kidney-1 pinch model. After 4 weeks, rats were randomized to receive Ang- (1-9) (602 ng / Kg min) or the co administration of Ang- (19) + A779 (100 ng / kg min, a MAS receptor antagonist of Ang- (1-7)) for 14 days. Sham operated rats were used as controls. We determined body mass (BM), systolic blood pressure (SBP), ventricular mass (VM), cardiomyocyte area (CA), area and thickness of the aortic media (ATM, TTM), LV total collagen volume fraction (FVCT), type I collagen protein levels (Col I) in the aorta (Ao) and macrophage infiltration in LV and Ao, through its specific molecule ED1 (ED1-Ao, ED1-VI). Results: Continuous administration of Ang- (1-9) significantly decreased SBP, VM, CA, TCVF, Col I, TTM, and ED1 in the aorta and left ventricle of hypertensive rats. This effect was not inhibited by the antagonist A779. ACE polymorphism did not modify the response to treatment. Conclusion: Ang- (1-9) effectively reduced hypertension induced CVRM independent of ACE gene polymorphism. This effect was not mediated by Ang- (1-7).
Asunto(s)
Animales , Ratas , Hipertensión/inducido químicamente , Peptidil-Dipeptidasa A/administración & dosificación , Peptidil-Dipeptidasa A/genética , Sistema Cardiovascular/patología , Polimorfismo Genético , Sistema Cardiovascular/enzimologíaRESUMEN
BACKGROUND: Right ventricular (RV) dysfunction is a complication of pulmonary hypertension and portends a poor prognosis. Pharmacological therapies targeting RV function in pulmonary hypertension may reduce symptoms, improve hemodynamics, and potentially increase survival. We hypothesize that recombinant human angiotensin-converting enzyme 2 (rhACE2) will improve RV function in a pressure overload model. RESULTS: rhACE2 administered at 1.8 mg/kg/day improved RV systolic and diastolic function in pulmonary artery banded mice as measured by in vivo hemodynamics. Specifically, rhACE2 increased RV ejection fraction and decreased RV end diastolic pressure and diastolic time constant (p<0.05). In addition, rhACE2 decreased RV hypertrophy as measured by RV/LV+S ratio (p<0.05). There were no significant negative effects of rhACE2 administration on LV function. rhACE2 had no significant effect on fibrosis as measured by trichrome staining and collagen1α1 expression. In pulmonary artery banded mice, rhACE2 increased Mas receptor expression and normalized connexin 37 expression. CONCLUSION: In a mouse RV load-stress model of early heart failure, rhACE2 diminished RV hypertrophy and improved RV systolic and diastolic function in association with a marker of intercellular communication. rhACE2 may be a novel treatment for RV failure.
Asunto(s)
Peptidil-Dipeptidasa A/metabolismo , Presión , Función Ventricular Derecha/fisiología , Enzima Convertidora de Angiotensina 2 , Animales , Diástole/efectos de los fármacos , Modelos Animales de Enfermedad , Fibrosis , Hemodinámica/efectos de los fármacos , Humanos , Hipertrofia Ventricular Derecha/patología , Hipertrofia Ventricular Derecha/fisiopatología , Inmunohistoquímica , Masculino , Ratones , Ratones Endogámicos C57BL , Microvasos/efectos de los fármacos , Microvasos/patología , Peptidil-Dipeptidasa A/administración & dosificación , Peptidil-Dipeptidasa A/farmacología , Proteínas Recombinantes/farmacología , Transducción de Señal/efectos de los fármacos , Sístole/efectos de los fármacos , Función Ventricular Derecha/efectos de los fármacosRESUMEN
BACKGROUND: Angiotensin-converting enzyme 2 (ACE2) is a pleiotropic monocarboxypeptidase capable of metabolizing several peptide substrates. We hypothesized that ACE2 is a negative regulator of angiotensin II (Ang II)-mediated signaling and its adverse effects on the cardiovascular system. METHODS AND RESULTS: Ang II infusion (1.5 mg x kg(-1) x d(-1)) for 14 days resulted in worsening cardiac fibrosis and pathological hypertrophy in ACE2 knockout (Ace2(-/y)) mice compared with wild-type (WT) mice. Daily treatment of Ang II-infused wild-type mice with recombinant human ACE2 (rhACE2; 2 mg x kg(-1) x d(-1) IP) blunted the hypertrophic response and expression of hypertrophy markers and reduced Ang II-induced superoxide production. Ang II-mediated myocardial fibrosis and expression of procollagen type I alpha 1, procollagen type III alpha 1, transforming growth factor-beta1, and fibronectin were also suppressed by rhACE2. Ang II-induced diastolic dysfunction was inhibited by rhACE2 in association with reduced plasma and myocardial Ang II and increased plasma Ang 1-7 levels. rhACE2 treatment inhibited Ang II-mediated activation of protein kinase C-alpha and protein kinase C-beta1 protein levels and phosphorylation of the extracellular signal-regulated 1/2, Janus kinase 2, and signal transducer and activator of transcription 3 signaling pathways in wild-type mice. A subpressor dose of Ang II (0.15 mg . kg(-1) . d(-1)) resulted in a milder phenotype that was strikingly attenuated by rhACE2 (2 mg x kg(-1) x d(-1) IP). In adult ventricular cardiomyocytes and cardiofibroblasts, Ang II-mediated superoxide generation, collagen production, and extracellular signal-regulated 1/2 signaling were inhibited by rhACE2 in an Ang 1-7-dependent manner. Importantly, rhACE2 partially prevented the development of dilated cardiomyopathy in pressure-overloaded wild-type mice. CONCLUSIONS: Elevated Ang II induced hypertension, myocardial hypertrophy, fibrosis, and diastolic dysfunction, which were exacerbated by ACE2 deficiency, whereas rhACE2 attenuated Ang II- and pressure-overload-induced adverse myocardial remodeling. Hence, ACE2 is an important negative regulator of Ang II-induced heart disease and suppresses adverse myocardial remodeling.
Asunto(s)
Cardiomegalia/enzimología , Cardiomegalia/prevención & control , Regulación hacia Abajo/fisiología , Hipertrofia Ventricular Izquierda/enzimología , Hipertrofia Ventricular Izquierda/prevención & control , Miocardio/enzimología , Miocardio/patología , Peptidil-Dipeptidasa A/deficiencia , Angiotensina II/administración & dosificación , Angiotensina II/antagonistas & inhibidores , Angiotensina II/biosíntesis , Enzima Convertidora de Angiotensina 2 , Animales , Células CHO , Cardiomegalia/patología , Cardiomegalia/fisiopatología , Células Cultivadas , Cadena alfa 1 del Colágeno Tipo I , Cricetinae , Cricetulus , Fibrosis , Humanos , Hipertrofia Ventricular Izquierda/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Peptidil-Dipeptidasa A/administración & dosificación , Peptidil-Dipeptidasa A/fisiología , Proteínas Recombinantes/administración & dosificaciónRESUMEN
Angiotensin (Ang)-converting enzyme 2 (ACE2) cleaves Ang II to form Ang-(1-7). Here we examined whether soluble human recombinant ACE2 (rACE2) can efficiently lower Ang II and increase Ang-(1-7) and whether rACE2 can prevent hypertension caused by Ang II infusion as a result of systemic versus local mechanisms of ACE2 activity amplification. rACE2 was infused via osmotic minipumps for 3 days in conscious mice or acutely in anesthetized mice. rACE2 caused a dose-dependent increase in serum ACE2 activity but had no effect on kidney or cardiac ACE2 activity. After Ang II infusion (40 pmol/min), rACE2 (1 mg/kg per day) resulted in normalization of systolic blood pressure and plasma Ang II. In acute studies, rACE2 (1 mg/kg) prevented the rapid hypertensive effect of Ang II (0.2 mg/kg), and this was associated with both a decrease in Ang II and an increase in Ang-(1-7) in plasma. Moreover, during infusion of Ang II, the effect of rACE2 on blood pressure was unaffected by a specific Ang-(1-7) receptor blocker, A779 (0.2 mg/kg), and infusing supraphysiologic levels of Ang-(1-7) (0.2 mg/kg) had no effect on blood pressure. We conclude that, during Ang II infusion, rACE2 effectively degrades Ang II and, in the process, normalizes blood pressure. The mechanism of rACE2 action results from an increase in systemic, not tissue, ACE2 activity and the lowering of plasma Ang II rather than the attendant increase in Ang-(1-7). Increasing ACE2 activity may provide a new therapeutic target in states of Ang II overactivity by enhancing its degradation, an approach that differs from the current focus on blocking Ang II formation and action.
Asunto(s)
Angiotensina II/metabolismo , Hipertensión/metabolismo , Peptidil-Dipeptidasa A/metabolismo , Proteínas Recombinantes/metabolismo , Anestesia , Angiotensina I/sangre , Angiotensina I/metabolismo , Angiotensina II/administración & dosificación , Angiotensina II/análogos & derivados , Angiotensina II/sangre , Bloqueadores del Receptor Tipo 1 de Angiotensina II/antagonistas & inhibidores , Enzima Convertidora de Angiotensina 2 , Animales , Presión Sanguínea/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Hipertensión/inducido químicamente , Hipertensión/prevención & control , Bombas de Infusión , Riñón/efectos de los fármacos , Riñón/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fragmentos de Péptidos/administración & dosificación , Fragmentos de Péptidos/sangre , Fragmentos de Péptidos/metabolismo , Peptidil-Dipeptidasa A/administración & dosificación , Peptidil-Dipeptidasa A/genética , Proteínas Recombinantes/administración & dosificaciónRESUMEN
Antecedentes: La sobreexpresion génica de la enzima convertidora de angiotensina I homologa (ECA2) se asocia con prevención de la hipertrofia y fibrosis cardiaca dependiente de angiotensina (Ang) II. Sin embargo se desconoce si su sobreexpresion reduce la hipertensión arterial (HTA) y revierte el consecuente remodelado miocárdico (RM) dependiente de Ang II. Objetivo: Determinar si la sobreexpresion adenoviral (Ad) del gen de la ECA2 en el miocardio disminuye la HTA y RM experimental en ratas con niveles genéticamente determinados de ECA y Ang II. Métodos: Ratas homocigotas normotensas Lewis (LL) y Brown Borway (BN), con menores y mayores niveles circulantes de ECA y Ang II, respectivamente, se hicieron hipertensas por el procedimiento Goldblatt (GB). Como controles se usaron ratas seudo-operadas (sham). A la semana 5 post cirugía y con HTA establecida > 140 mmHg, las ratas se randomizaron a inyección intramiocárdica con un AdECA2 o Ad proteína fluorescente verde (GFP) como controles de infección. A la semana post infección adenoviral, los ratas se sacrificaron y se determinaron peso corporal (PC, g), masa cardiaca (MC, mg), presión arterial sistólica (RAS, mmHg), área (AC, um2) y perímetro (PERC, um) de cardiomiocitos y contenido de colágeno ( por ciento) miocárdico (CM), sub-endocárdico (CS)ytotal(CT). Resultados: La HTA aumentó significativamente la MC, MCR, AC, PERC como también el CM, CS y CT en las ratas GB vs Sham, sin diferencias en el PC ni por efecto del polimorfismo de la ECA. La sobreexpresion de ECA2 disminuyó significativamente la RAS (15 por ciento y 27 por ciento), AC (25 por ciento y 25 por ciento ) y PERC (17 por ciento y 18 por ciento) en las ratas LL y BN vs ratas hipertensas, respectivamente. Estos resultados se asociaron a una disminución significativa del CS (LL = 37 por ciento, BN = 39 por ciento), CM (LL = 54 por ciento) y CT (LL = 42 por ciento, BN = 22 por ciento) respecto a las ratas GB. Conclusión: En ratas con HTA...
Objective Background. Over expression of the gene for homologous angiotensin I converting enzyme (ACE) is associated with prevention of angiotensin II dependent cardiac hypertrophy and fibrosis. Whether this over expression is able to reduce hypertension and revert cardiac remodeling is unknown. Aim: To determine whether adenoviral ACE2 gene over expression in the myocardium decreases experimental hypertension and cardiac remodeling in rats with genetically determined levels of ACE and ANGII. Methods: Homozygous normotensive Lewis (LL) and Brown Norway (BN) rats with decreased or increased levéis of ACE and Ang II, respectively, were made hy-pertensive using the Goldblatt procedure. Sham opera-ted rats were used as control s. 5 weeks after surgery, when systolic blood pressure reached > 140 mmHg, rats were randomized to receive intramyocardial injec-tion of either AdACE2 or green fluorescent Ad protein (GFP) as infection controlling agents. One week after adenoviral infection, rats were sacrificed. Body weight (BW, Gr), relative cardiac mass (RCM, mG), systolic blood pressure (SBP, mmHg), cardiomyocite área (MA um2) and perimeter (MPER, uM), and myocardial co-llagen content, both subendocardial (SC, percent) and total (TC percent) were measured. Results: Hypertension was associated to significant in-creases in total and RCM, MA, MPER as well as SC and TC in Goldblatt vs normal rats. This was independent of BW and not affected by ACE polymorphism. ACE II over expression significantly decreased SBP (27 vs 15 percent), MA (25 vs 25 percent) and MPER (18 vs 17 percent) hy-pertensive vs normal rats, respectively. Conclusion: Over expression of ACE2 decreased hypertension, hypertrophy and fibrosis in rats with experimental hypertension and different levels of ACE and Ang II. (Fondecyt 1070662).
Asunto(s)
Masculino , Animales , Ratas , Corazón , Hipertensión/tratamiento farmacológico , Peptidil-Dipeptidasa A/administración & dosificación , Remodelación Ventricular , Peso Corporal , Cardiomegalia/prevención & control , Fibrosis , Fibrosis Endomiocárdica/prevención & control , Expresión Génica , Terapia Genética , Miocitos Cardíacos , Miocardio/patología , Presión Arterial , Ratas Endogámicas , Tamaño de los ÓrganosRESUMEN
Angiotensin converting enzyme inhibitors (ACEis) are widely used anti-hypertensive agents that are also reported to have positive effects on mood and cognition. The present study examined the influence of the ACEi, perindopril, on cognitive performance and anxiety measures in rats. Two groups of rats were treated orally for one week with the ACEi, perindopril, at doses of 0.1 and 1.0mg/kg/day. Learning was assessed by the reference memory task in the water maze, comparing treated to control rats. Over five training days both perindopril-treated groups learnt the location of the submerged platform in the water maze task significantly faster than control rats. A 60s probe trial on day 6 showed that the 1.0mg/kg/day group spent significantly longer time in the training quadrant than control rats. This improved performance in the swim maze task was not due to the effect of perindopril on motor activity or the anxiety levels of the rats as perindopril-treated and control animals behaved similarly in activity boxes and on the elevated+maze. These results confirm the anecdotal human studies that ACEis have a positive influence on cognition and provide possibilities for ACEis to be developed into therapies for memory loss.
Asunto(s)
Memoria/efectos de los fármacos , Peptidil-Dipeptidasa A/administración & dosificación , Perindopril/administración & dosificación , Conducta Espacial/efectos de los fármacos , Administración Oral , Análisis de Varianza , Animales , Conducta Animal/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Actividad Motora/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Tiempo de Reacción/efectos de los fármacosRESUMEN
El efecto favorable de la administración de los inhibidores de la enzima de conversión de la angiotensina (IECA) a largo plazo en pacientes con insuficiencia cardiaca crónica y fracción de eyección deprimida está bien establecido, pero no hay tanta evidencia en pacientes con disfunción sistólica secundaria a miocarditis aguda que normalizan la contracción ventricular. Hemos seguido a 35 pacientes con miocarditis aguda y fracción de eyección 50% en los 35 casos. De esos 35, en 15 se suspendieron los IECA, mientras que 20 continuaron tomándolos. A los 3 años de seguimiento no hubo ninguna muerte, pero los pacientes en los que se suspendieron los IECA tuvieron una mayor incidencia de nuevos episodios de insuficiencia cardiaca con fracción de eyección < 45% (el 33 frente al 5%; p = 0,064) y menor fracción de eyección (47 ± 12 frente a 57 ± 11%; p = 0,002), lo que indica que estos fármacos se deben mantener a largo plazo también en estos pacientes
It is well established that long-term administration of angiotensin-converting enzyme (ACE) inhibitors has a favorable effect in patients with chronic heart failure and dilated cardiomyopathy. However, less information is available on patients whose left ventricular ejection fraction normalizes after an episode of systolic dysfunction secondary to acute myocarditis. We followed 35 patients who were diagnosed at our center between 1987 and 1995 with acute myocarditis and an ejection fraction 50% in all 35 patients. Treatment with ACE inhibitors was discontinued in 15 of the 35 patients, while the other 20 continued ACE inhibitor therapy. After 3 years of follow-up, no death had occurred, but the incidence of new episodes of heart failure with a left ventricular ejection fraction <45% was higher in patients who stopped taking ACE inhibitors (33% vs 5%, P=.064), and their ejection fraction was lower (47 [12%] vs 57 [11%], P=.002). These results suggest that ACE inhibitors should be continued over the long term in these patients