RESUMO
BACKGROUND: Increased reactive oxygen species (ROS) generation in diabetes mellitus (DM) is an important mechanism leading to diabetic cardiomyopathy. Apocynin, a drug isolated from the herb Picrorhiza kurroa, is considered an antioxidant agent by inhibiting NADPH oxidase activity and improving ROS scavenging. This study analyzed the influence of apocynin on cardiac remodeling in diabetic rats. METHODS: Six-month-old male Wistar rats were assigned into 4 groups: control (CTL, n = 15), control + apocynin (CTL + APO, n = 20), diabetes (DM, n = 20), and diabetes + apocynin (DM + APO, n = 20). DM was induced by streptozotocin. Seven days later, apocynin (16 mg/kg/day) or vehicle was initiated and maintained for 8 weeks. Left ventricular (LV) histological sections were used to analyze interstitial collagen fraction. NADPH oxidase activity was evaluated in LV samples. Comparisons between groups were performed by ANOVA for a 2 × 2 factorial design followed by the Bonferroni post hoc test. RESULTS: Body weight (BW) was lower and glycemia higher in diabetic animals. Echocardiogram showed increased left atrial diameter, LV diastolic diameter, and LV mass indexed by BW in both diabetic groups; apocynin did not affect these indices. LV systolic function was impaired in DM groups and unchanged by apocynin. Isovolumic relaxation time was increased in DM groups; transmitral E/A ratio was higher in DM + APO compared to DM. Myocardial functional evaluation through papillary muscle preparations showed impaired contractile and relaxation function in both DM groups at baseline conditions. After positive inotropic stimulation, developed tension (DT) was lower in DM than CTL. In DM + APO, DT had values between those in DM and CTL + APO and did not significantly differ from either group. Myocardial interstitial collagen fraction was higher in DM than CTL and did not differ between DM + APO and CTL + APO. Serum activity of antioxidant enzymes glutathione peroxidase, superoxide dismutase (SOD), and catalase was lower in DM than CTL; apocynin restored catalase and SOD levels in DM + APO. Myocardial NADPH oxidase activity did not differ between groups. CONCLUSION: Apocynin restores serum antioxidant enzyme activity despite unchanged myocardial NADPH oxidase activity in diabetic rats.
Assuntos
Acetofenonas/farmacologia , Diabetes Mellitus Experimental/tratamento farmacológico , Cardiomiopatias Diabéticas/prevenção & controle , Sequestradores de Radicais Livres/farmacologia , Ventrículos do Coração/efeitos dos fármacos , Estreptozocina , Função Ventricular Esquerda/efeitos dos fármacos , Remodelação Ventricular/efeitos dos fármacos , Animais , Catalase/sangue , Colágeno , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/complicações , Cardiomiopatias Diabéticas/sangue , Cardiomiopatias Diabéticas/etiologia , Cardiomiopatias Diabéticas/fisiopatologia , Glutationa Peroxidase/sangue , Ventrículos do Coração/metabolismo , Ventrículos do Coração/fisiopatologia , Masculino , NADPH Oxidases/metabolismo , Ratos Wistar , Superóxido Dismutase/sangueRESUMO
PURPOSE: Although increased oxidative stress is a major component of diabetic hypertensive cardiomyopathy, research into the effects of antioxidants on cardiac remodeling remains scarce. The actions of antioxidant apocynin include inhibiting reactive oxygen species (ROS) generation by nicotinamide adenine dinucleotide phosphate (NADPH) oxidases and ROS scavenging. We evaluated the effects of apocynin on cardiac remodeling in spontaneously hypertensive rats (SHR) with diabetes mellitus (DM). METHODS: Male SHR were divided into four groups: control (SHR, n = 16); SHR treated with apocynin (SHR-APO; 16 mg/kg/day, added to drinking water; n = 16); diabetic SHR (SHR-DM, n = 13); and SHR-DM treated with apocynin (SHR-DM-APO, n = 14), for eight weeks. DM was induced by streptozotocin (40 mg/kg, single dose). Statistical analyzes: ANOVA and Tukey or Mann-Whitney. RESULTS: Echocardiogram in diabetic groups showed higher left ventricular and left atrium diameters indexed for body weight, and higher isovolumetric relaxation time than normoglycemic rats; systolic function did not differ between groups. Isolated papillary muscle showed impaired contractile and relaxation function in diabetic groups. Developed tension was lower in SHR-APO than SHR. Myocardial hydroxyproline concentration was higher in SHR-DM than SHR, interstitial collagen fraction was higher in SHR-DM-APO than SHR-APO, and type III collagen protein expression was lower in SHR-DM and SHR-DM-APO than their controls. Type I collagen and lysyl oxidase expression did not differ between groups. Apocynin did not change collagen tissue. Myocardial lipid hydroperoxide concentration was higher in SHR-DM than SHR and SHR-DM-APO. Glutathione peroxidase activity was lower and catalase higher in SHR-DM than SHR. Apocynin attenuated antioxidant enzyme activity changes in SHR-DM-APO. Advanced glycation end-products and NADPH oxidase activity did not differ between groups. CONCLUSION: Apocynin reduces oxidative stress independently of NADPH oxidase activity and does not change ventricular or myocardial function in spontaneously hypertensive rats with diabetes mellitus. The apocynin-induced myocardial functional impairment in SHR shows that apocynin actions need to be clarified during sustained chronic pressure overload.
Assuntos
Acetofenonas/farmacologia , Antioxidantes/farmacologia , Diabetes Mellitus Experimental/tratamento farmacológico , Cardiomiopatias Diabéticas/prevenção & controle , Hipertensão/tratamento farmacológico , Hipertrofia Ventricular Esquerda/prevenção & controle , Miocárdio/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Remodelação Ventricular/efeitos dos fármacos , Animais , Catalase/metabolismo , Colágeno Tipo III/metabolismo , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/fisiopatologia , Cardiomiopatias Diabéticas/etiologia , Cardiomiopatias Diabéticas/metabolismo , Cardiomiopatias Diabéticas/fisiopatologia , Glutationa Peroxidase/metabolismo , Hipertensão/complicações , Hipertensão/metabolismo , Hipertensão/fisiopatologia , Hipertrofia Ventricular Esquerda/etiologia , Hipertrofia Ventricular Esquerda/metabolismo , Hipertrofia Ventricular Esquerda/fisiopatologia , Peroxidação de Lipídeos/efeitos dos fármacos , Peróxidos Lipídicos/metabolismo , Masculino , Contração Miocárdica/efeitos dos fármacos , Miocárdio/patologia , Ratos Endogâmicos SHR , Estreptozocina , Função Ventricular Esquerda/efeitos dos fármacosRESUMO
Systemic lupus erythematosus (SLE) is a heterogeneous disease involving several immune cell types and pro-inflammatory signals, including the one triggered by binding of CD40L to the receptor CD40. Peroxisome-proliferator activated receptor gamma (PPARγ) is a transcription factor with anti-inflammatory properties. Here we investigated whether CD40 and PPARγ could exert opposite effects in the immune response and the possible implications for SLE. Increased PPARγ mRNA levels were detected by real-time PCR in patients with active SLE, compared to patients with inactive SLE PPARγ/GAPDH mRNA = 2.21 ± 0.49 vs. 0.57 ± 0.14, respectively (p < 0.05) or patients with infectious diseases and healthy subjects (p < 0.05). This finding was independent of the corticosteroid therapy. We further explored these observations in human THP1 and in SLE patient-derived macrophages, where activation of CD40 by CD40L promoted augmented PPARγ gene transcription compared to non-stimulated cells (PPARγ/GAPDH mRNA = 1.14 ± 0.38 vs. 0.14 ± 0.01, respectively; p < 0.05). This phenomenon occurred specifically upon CD40 activation, since lipopolysaccharide treatment did not induce a similar response. In addition, increased activity of PPARγ was also detected after CD40 activation, since higher PPARγ-dependent transcription of CD36 transcription was observed. Furthermore, CD40L-stimulated transcription of CD80 gene was elevated in cells treated with PPARγ-specific small interfering RNA (small interfering RNA, siRNA) compared to cells treated with CD40L alone (CD80/GAPDH mRNA = 0.11 ± 0.04 vs. 0.05 ± 0.02, respectively; p < 0.05), suggesting a regulatory role for PPARγ on the CD40/CD40L pathway. Altogether, our findings outline a novel mechanism through which PPARγ regulates the inflammatory signal initiated by activation of CD40, with important implications for the understanding of immunological mechanisms underlying SLE and the development of new treatment strategies.
Assuntos
Antígenos CD40/metabolismo , Ligante de CD40/metabolismo , Lúpus Eritematoso Sistêmico/imunologia , PPAR gama/genética , Adulto , Estudos de Casos e Controles , Linhagem Celular Tumoral , Humanos , Lúpus Eritematoso Sistêmico/genética , Macrófagos/metabolismo , Pessoa de Meia-Idade , Monócitos/metabolismo , PPAR gama/metabolismo , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/administração & dosagem , Transdução de Sinais , Transcrição Gênica , Adulto JovemRESUMO
Impaired relaxation induced by the new nitric oxide (NO) donor [Ru(NH.NHq)(terpy)NO(+)](3+) (TERPY) has been observed in the aortic rings from renal hypertensive rats (2K-1C). An increased production of reactive oxygen species (ROS) in the aortas from 2K-1C rats are capable of reducing NO bioavailability. Therefore, this study aimed at investigating the effects of an antioxidant (vitamin C) on the relaxant effect of NO released from TERPY on the 2K-1C rat aorta. As for vascular reactivity, the potency of TERPY is greater in the control rats (2K) than in 2K-1C whereas the maximum relaxation (ME) is not significantly different between the 2K and 2K-1C rat aortas. The relaxation of TERPY is potentiated only in the 2K-1C aortic ring treated with vitamin C. TERPY has a lower effect in decreasing cytosolic Ca(2+) concentration ([Ca(2+)]c) in vascular smooth muscle cells (VSMCs) from 2K-1C rats. This effect is also potentiated in 2K-1C aortic cells treated with vitamin C, but it is not altered in 2K cells. The basal cytosolic NO concentration ([NO]c) is lower in 2K-1C than in 2K cells, and the bioavailability of the NO released from TERPY is larger in 2K than in 2K-1C VSMCs. The superoxide radical concentration ([O(2)(*-)]) is higher in the 2K-1C aorta, and vitamin C reduces the [O(2)(*-)] in the 2K-1C aorta. Taken together, these results show that in the aortas of renal hypertensive 2K-1C rats, released NO from the new NO donor is not available to produce a similar effect in 2K aorta due to increased [O(2)(*-)].
Assuntos
Aorta/efeitos dos fármacos , Ácido Ascórbico/farmacologia , Hipertensão Renal/fisiopatologia , Músculo Liso Vascular/efeitos dos fármacos , Doadores de Óxido Nítrico/farmacologia , Compostos Organometálicos/farmacologia , Animais , Aorta/patologia , Cálcio/análise , Cálcio/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Rim/irrigação sanguínea , Masculino , Músculo Liso Vascular/patologia , Músculo Liso Vascular/fisiopatologia , Óxido Nítrico/análise , Óxido Nítrico/metabolismo , Técnicas de Cultura de Órgãos , Fenilefrina/farmacologia , Ratos , Ratos Wistar , Rutênio/química , Superóxidos/análise , Superóxidos/metabolismo , Fatores de Tempo , Vasodilatação/efeitos dos fármacosRESUMO
Coronary artery calcification (CAC) is associated with atherosclerotic complications. However, elevated CAC may not always imply a worse prognosis. Herein, we report the clinical evolution of long-term red wine (RW) drinkers in relation to CAC. We followed 200 healthy male habitual RW drinkers and compared them to 154 abstainers for a period of 5.5 years. The initial evaluation included coronary computed tomography angiography (CTA), clinical, demographics, and laboratory data. CAC was quantified by the Agatston score. The follow-up process was conducted by telephone calls and/or hospital record review. The composite end-point of total death, acute myocardial infarction (AMI), or coronary revascularization (or major adverse cardiac event - MACE) was assessed. The RW drinkers ingested 28.9±15 g of alcohol/day for 23.4±12.3 years. They had higher high-density lipoprotein and low-density lipoprotein, but lower C-reactive protein than abstainers. Age, total cholesterol, triglycerides, glucose, and liver enzymes were similar. History of diabetes was lower among drinkers, but other risk factors were similar. However, drinkers had higher CAC than abstainers; the mean value was 131.5±362 in drinkers vs 40.5±320 in abstainers (P<0.001). The median and interquartile range were 15 (0.0-131.5) in RW drinkers and 1 (0.0-40.5) in abstainers (P=0.003). During the follow-up, MACE was significantly lower in drinkers than in abstainers, despite their higher CAC. The difference was driven mainly by AMI (0 vs 6; P<0.03). Greater CAC values in this setting did not predict worse prognosis. A possible underlying mechanism is lesion calcification, which leads to plaque stabilization and less clinical events.
Assuntos
Consumo de Bebidas Alcoólicas , Doença da Artéria Coronariana/prevenção & controle , Calcificação Vascular/prevenção & controle , Vinho , Idoso , Angiografia por Tomografia Computadorizada , Doença da Artéria Coronariana/diagnóstico por imagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Calcificação Vascular/diagnóstico por imagemRESUMO
Essentials Inhibitors of protein disulfide isomerase (PDI) have been considered a new antithrombotic class. CxxC is a PDI-targeted peptide that has been previously shown to inhibit its reductase activity. CxxC binds to surface PDI and inhibits ADP- and thrombin-evoked platelet activation and aggregation. CxxC binds to Cys400 on CGHC redox motif of PDI a' domain, a site for PDI prothrombotic activity. SUMMARY: Background Protein disulfide isomerase (PDI) plays a major role in platelet aggregation, and its inhibitors have emerged as novel antithrombotic drugs. In previous work, we designed a peptide based on a PDI redox motif (CGHC) that inhibited both PDI reductase activity and PDI-modulated superoxide generation by neutrophil Nox2. Thus, we hypothesized that this peptide would also inhibit platelet aggregation by association with surface PDI. Methods Three peptides were used: CxxC, containing the PDI redox motif; Scr, presenting a scrambled sequence of the same residues and AxxA, with cysteines replaced by alanine. These peptides were tested under platelet aggregation and flow cytometry protocols to identify their possible antiplatelet activity. We labeled membrane free thiol and electrospray ionization liquid chromatography tandem mass spectrometry to test for an interaction. Results CxxC decreased platelet aggregation in a dose-dependent manner, being more potent at lower agonist concentrations, whereas neither AxxA nor Scr peptides exerted any effect. CxxC decreased aIIbb3 activation, but had no effect on the other markers. CxxC also decreased cell surface PDI pulldown without interfering with the total thiol protein content. Finally, we detected the addition of one CxxC molecule to reduced PDI through binding to Cys400 through mass spectrometry. Interestingly, CxxC did not react with oxidized PDI. Discussion CxxC has consistently shown its antiplatelet effects, both in PRP and washed platelets, corroborated by decreased aIIbb3 activation. The probable mechanism of action is through a mixed dissulphide bond with Cys400 of PDI, which has been shown to be essential for PDI's actions. Conclusion In summary, our data support antiplatelet activity for CxxC through binding to Cys400 in the PDI a0 domain, which can be further exploited as a model for sitedriven antithrombotic agent development.
Assuntos
Inibidores da Agregação Plaquetária/química , Pró-Colágeno-Prolina Dioxigenase/química , Isomerases de Dissulfetos de Proteínas/química , Alanina/química , Motivos de Aminoácidos , Plaquetas/metabolismo , Domínio Catalítico , Cisteína/química , Dissulfetos , Humanos , Oxirredução , Peptídeos/química , Ativação Plaquetária , Agregação Plaquetária , Ligação Proteica , Domínios Proteicos , Dobramento de ProteínaRESUMO
The complete spectrum of estrogen vascular effects remains unclear. In particular, estrogen effects in the vascular response to profound injury in males have not been explored in detail. Therefore, we submitted 44 male New Zealand rabbits weighing 3.4 +/- 0.6 kg to overdistention balloon injury of the right iliac artery. Rabbits were given 17beta-estradiol (5.45 micromol/day, sc) or vehicle for 7 days before and 14 days after injury, when the arteries were examined by post-mortem histomorphometry. Arteriographic caliber was assessed in vivo at baseline and before sacrifice. On day 14 after injury, in vivo arteriographic caliber (baseline = 2.44 +/- 0.43 mm) was decreased by 23.1 +/- 0.1% in controls and by 44.5 +/- 0.1% in estrogen-treated rabbits (P < 0.001). Neither the neointimal area nor the neointima/media area ratio changed after estrogen treatment. Collagen fraction was increased in the media and neointima of estrogen-treated rabbits vs control (1.38 +/- 1.30 vs 0.35 +/- 0.67, respectively, P = 0.01). Taken together, these findings suggest that estrogen increased negative vascular remodeling. Transcription of endothelial and inducible nitric oxide synthases (eNOS and iNOS) was analyzed by RT-PCR. eNOS mRNA expression was marginally increased after estrogen (P = 0.07) and injury. iNOS mRNA was increased 2- to 3-fold on day 14 after injury. With estrogen treatment, iNOS mRNA increased in uninjured arteries and exhibited a further 5.5-fold increase after injury. We concluded that estrogen increased lumen loss after balloon injury in male rabbits, likely by increased negative remodeling, which may be related to increased iNOS transcriptional rates.
Assuntos
Estradiol/farmacologia , Artéria Ilíaca/lesões , Óxido Nítrico Sintase Tipo III/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Túnica Íntima/efeitos dos fármacos , Angiografia , Angioplastia com Balão , Animais , Colágeno/efeitos dos fármacos , Artéria Ilíaca/efeitos dos fármacos , Artéria Ilíaca/enzimologia , Masculino , RNA Mensageiro/análise , Coelhos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Túnica Íntima/enzimologiaRESUMO
OBJECTIVE: Our aim was to assess whether exposure to oxidized thiols--a known usual consequence of oxidant stress--has the potential to affect the vascular repair response to angioplasty-induced injury. In addition, we also assessed the role of redox active metals in disulfide effects. METHODS: In 82 rabbits submitted to overdistention of iliac arteries, the following variables were analyzed: neointimal thickening, immunoreactivity to Proliferating Cell Nuclear Antigen, and cellular and collagen densities. RESULTS: A single intraarterial challenge of oxidized glutathione (GSSG, 6.5 mumol/kg) during and immediately after injury triggered a marked increase of the vascular repair reaction, as follows: (A) at day 7 after injury, there was a 2.7-fold increase in proliferation (p < 0.001 vs. control); (B) at day 14, there was increase of intimal/medial area ratio to 1.35 +/- 0.14, vs. 0.56 +/- 0.08 in controls. Proliferating cells increased to 5.5 +/- 0.8 cells/mm2, vs. 2.2 +/- 0.5 in controls (p < 0.002 for both variables). Overall cellularity was enhanced 2.2-fold; (C) at day 28, there was ongoing vessel wall proliferation, contrarily to controls. All GSSG effects were completely prevented by co-infusion of reduced glutathione (GSH) and were mimicked by cystine (6.5 mumol/kg). The uninjured artery showed no response to disulfides. To assess the role of redox active metals in GSSG action, the effects of 1,10-phenanthroline or N-CBZ-Pro-Leu-Gly hydroxamic acid (HXA), metal chelators with metalloproteinase inhibitor properties, were evaluated. Both compounds totally blocked the GSSG-induced amplification of vascular responses. In rabbits not exposed to GSSG, HXA decreased neointimal thickening by 50% (p < 0.05). CONCLUSIONS: Exposure to excess disulfide levels early after vascular balloon injury markedly amplified the late cellular response through interaction with redox active metals. These pathways can potentially mediate noxious effects of oxidative stress in vessels.
Assuntos
Dissulfeto de Glutationa/farmacologia , Glutationa/farmacologia , Artéria Ilíaca/lesões , Compostos de Sulfidrila/farmacologia , Animais , Cateterismo , Divisão Celular/efeitos dos fármacos , Quelantes/farmacologia , Colágeno/metabolismo , Inibidores Enzimáticos/farmacologia , Glutationa/sangue , Ácidos Hidroxâmicos/farmacologia , Artéria Ilíaca/efeitos dos fármacos , Artéria Ilíaca/metabolismo , Artéria Ilíaca/patologia , Imuno-Histoquímica , Masculino , Metaloendopeptidases/antagonistas & inibidores , Oxirredução , Fenantrolinas/farmacologia , Antígeno Nuclear de Célula em Proliferação/análise , Coelhos , Fatores de Tempo , Túnica Íntima/efeitos dos fármacos , Túnica Íntima/patologiaRESUMO
The prominent role of redox processes in tissue injury and in vascular cell signaling suggest their involvement in the repair reaction to vessel injury, which is a key determinant of restenosis post-angioplasty. Experimental studies showed a protective effect of superoxide dismutase or antioxidants on vasospasm, neointimal thickening or remodeling after balloon injury. It was also shown that oxidized thiols induce chelatable metal-dependent amplification of the vascular repair reaction. Ongoing or completed clinical trials show a promising effect of the antioxidant probucol against restenosis. However, few studies addressed the molecular physiological mechanisms underlying the redox hypothesis of restenosis. We recently showed evidence for marked oxidative stress early after balloon injury, with superoxide production mediated primarily by non-endothelial NAD(P)H oxidase-type flavoenzyme(s). This effect was closely related to the degree of injury. There is evidence supporting a role for such early redox processes in apoptotic cell loss and NF-kappa B activation. We present new data on the time course of oxidative stress after balloon injury of intact rabbit iliac arteries. Our data show that despite substantial neointimal growth and lumen narrowing, superoxide production and glutathione levels are unaltered at day 14 and 28 after balloon injury. At day 7 after injury, the peak neointimal proliferation in this model, there was significant decrease of vascular superoxide dismutase activity, without clear evidence of spontaneous superoxide production. Thus, oxidative stress after injury is likely to be an early transient event, which parallels the inflammatory and proliferative phases of the vascular response. We propose that such early redox processes act as dose-dependent signal transducers of gene programs that affect the final repair.
Assuntos
Doença das Coronárias/metabolismo , Endotélio Vascular/metabolismo , Estresse Oxidativo , Transdução de Sinais , Angioplastia Coronária com Balão/efeitos adversos , Animais , Antioxidantes/uso terapêutico , Divisão Celular , Doença das Coronárias/patologia , Doença das Coronárias/terapia , Endotélio Vascular/lesões , Endotélio Vascular/patologia , Humanos , Masculino , Oxirredução , Coelhos , Ensaios Clínicos Controlados Aleatórios como Assunto , Recidiva , Túnica Íntima/lesões , Túnica Íntima/metabolismo , Túnica Íntima/patologiaRESUMO
Vascular NAD(P)H oxidase activity contributes to oxidative stress. Thiol oxidants inhibit leukocyte NADPH oxidase. To assess the role of reactive thiols on vascular oxidase, rabbit iliac/carotid artery homogenates were incubated with distinct thiol reagents. NAD(P)H-driven enzyme activity, assessed by lucigenin (5 or 250 microM) luminescence, was nearly completely (> 97%) inhibited by the oxidant diamide (1mM) or the alkylator p-chloromercuryphenylsulfonate (pCMPS, 0.5mM). Analogous inhibition was also shown with EPR spectroscopy using DMPO as a spin trap. The oxidant dithionitrobenzoic acid (0.5mM) inhibited NADPH-driven signals by 92% but had no effect on NADH-driven signals. In contrast, the vicinal dithiol ligand phenylarsine oxide (PAO, 1 microM) induced minor nonsignificant inhibition of NADPH-driven activity, but significant stimulation of NADH-triggered signals. The alkylator N-ethyl maleimide (NEM, 0.5mM) or glutathione disulfide (GSSG, 3mM) had no effect with each substrate. Coincubation of N-acetylcysteine (NAC, 3mM) with diamide or pCMPS reversed their inhibitory effects by 30-60%, whereas NAC alone inhibited the oxidase by 52%. Incubation of intact arterial rings with the above reagents disclosed similar results, except that PAO became inhibitor and NAC stimulator of NADH-driven signals. Notably, the cell-impermeant reagent pCMPS was also inhibitory in whole rings, suggesting that reactive thiol(s) affecting oxidase activity are highly accessible. Since lack of oxidase inhibition by NEM or GSSG occurred despite significant cellular glutathione depletion, change in intracellular redox status is not sufficient to account for oxidase inhibition. Moreover, the observed differences between NADPH and NADH-driven oxidase activity point to complex or multiple enzyme forms.
Assuntos
Vasos Sanguíneos/efeitos dos fármacos , Vasos Sanguíneos/metabolismo , Glutationa/metabolismo , NADH NADPH Oxirredutases/antagonistas & inibidores , Reagentes de Sulfidrila/farmacologia , Acridinas , Animais , Vasos Sanguíneos/enzimologia , Artérias Carótidas/efeitos dos fármacos , Artérias Carótidas/metabolismo , Espectroscopia de Ressonância de Spin Eletrônica , Artéria Ilíaca/efeitos dos fármacos , Artéria Ilíaca/metabolismo , Técnicas In Vitro , Oxirredução , Estresse Oxidativo , CoelhosRESUMO
Available evidence for oxidative stress after angioplasty is indirect or ambiguous. We sought to characterize the pattern, time course, and possible sources of free radical generation early after arterial balloon injury. Ex vivo injury performed in arterial rings in buffer with lucigenin yielded a massive oxygen-dependent peak of luminescence that decayed exponentially and was proportional to the degree of injury. Signals for injured vs. control arteries were 207. 1 +/- 17.9 (n = 13) vs 4.1 +/- 0.7 (n = 22) cpm x 10(3)/mg/min (p <. 001). Data obtained with 0.25 mmol/l lucigenin were validated with 0. 005-0.05 mmol/l lucigenin or the novel superoxide-sensitive probe coelenterazine (5 micromol/l). Gentle removal of endothelium prior to injury scarcely affected the amount of luminescence. Lucigenin signals were amplified 5- to 20-fold by exogenous NAD(P)H, and were >85% inhibited by diphenyliodonium (DPI, a flavoenzyme inhibitor). Antagonists of several other potential free radical sources, including xanthine oxidase, nitric oxide synthase, and mitochondrial electron transport, were without effect. Overdistension of intact rabbit iliac arteries in vivo (n = 7) induced 72% fall in intracellular reduced glutathione and 68% increase in oxidized glutathione, so that GSH/GSSG ratio changed from 7.93 +/- 2.14 to 0. 81 +/- 0.16 (p <.005). There was also 28.7% loss of the glutathione pool. Further studies were performed with electron paramagnetic resonance spectroscopy. Rabbit aortas submitted to ex vivo overdistension in the presence of the spin trap DEPMPO (5-diethoxy-phosphoryl-5-methyl-1-pyrroline-N-oxide, 100 mmol/l, n = 5) showed formation of radical adduct spectra, abolished by DPI or superoxide dismutase. Computer simulation indicated a mixture of hydroxyl and carbon-centered radical adducts, likely due to decay of superoxide adduct. Electrical mobility shift assays for NF-kappaB activation were performed in nuclear protein extracts from intact or previously injured rabbit aortas. Balloon injury induced early NF-kappaB activation, which was decreased by DPI. In conclusion, our data show unambiguously that arterial injury induces an immediate profound vascular oxidative stress. Such redox imbalance is likely accounted for by activation of vessel wall NAD(P)H oxidoreductase(s), generating radical species potentially involved in tissue repair.
Assuntos
Cateterismo/efeitos adversos , Endotélio Vascular/lesões , Imidazóis , NADH NADPH Oxirredutases/metabolismo , NADPH Oxidases , Espécies Reativas de Oxigênio , Acridinas/metabolismo , Animais , Compostos de Bifenilo/farmacologia , Óxidos N-Cíclicos , Inibidores de Ciclo-Oxigenase/farmacologia , Espectroscopia de Ressonância de Spin Eletrônica , Transporte de Elétrons , Endotélio Vascular/enzimologia , Radicais Livres , Glutationa/metabolismo , Inibidores de Lipoxigenase/farmacologia , Medições Luminescentes , Masculino , Metaloporfirinas/farmacologia , NAD/metabolismo , NADP/metabolismo , NF-kappa B/metabolismo , Óxido Nítrico Sintase/farmacologia , Oniocompostos/farmacologia , Oxirredução , Estresse Oxidativo , Consumo de Oxigênio , Pirazinas/farmacologia , Coelhos , Proteínas Recombinantes de Fusão/metabolismo , Marcadores de Spin , Superóxido Dismutase/farmacologia , Superóxidos/metabolismo , Transcrição Gênica , Cicatrização , Xantina Oxidase/farmacologiaRESUMO
The incidence of fatal acute myocardial infarction (AMI) after valve replacement has decreased with use of cold potassium-induced cardioplegia. Despite this method of myocardial preservation, 12 of 662 consecutive patients submitted to valve replacement had this complication. This study retrospectively analyzes, in those 12 patients, the etiologic profile of fatal perioperative AMI, together with its morphologic aspects. The clinical picture in 11 patients was a refractory low cardiac output state. In only 3 cases was AMI diagnosis confirmed during life. Six patients either had a technical complication or a coronary embolus; in these patients AMI was localized in the vascular bed of a single occluded coronary artery, and its morphologic picture resembled that of usual AMI. The 6 other patients did not have a defined cause for AMI and coronary occlusion was not present. In 4 such patients, there was massive circumferential necrosis, mainly in the subendocardium; comparatively, there was a greater prevalence of hemorrhage, contraction bands and necrosis of the layer of subendocardial cells adjacent to the left ventricular cavity. The findings for this group suggest myocardial necrosis due to cell damage during cardiopulmonary bypass; no predisposing factor for perioperative AMI was identified.
Assuntos
Próteses Valvulares Cardíacas , Infarto do Miocárdio/etiologia , Adolescente , Adulto , Idoso , Feminino , Humanos , Complicações Intraoperatórias , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/patologiaRESUMO
1. The effects of balloon injury on the reactivity of ipsilateral and contralateral carotid arteries were compared to those observed in arteries from intact animals (control arteries). 2. Carotid arteries were obtained from Wistar rats 2, 4, 7, 15, 30 or 45 days after injury and mounted in an isolated organ bath. Reactivity to angiotensin II (Ang II), phenylephrine (Phe) and bradykinin (BK) was studied. Curves were constructed in the absence or presence of endothelium or after incubation with 10 microm indomethacin, 500 microm valeryl salicylate or 0.1 microm celecoxib. 3. Phe, Ang II and BK maximum effects (Emax) were decreased in ipsilateral arteries when compared to control arteries. No differences were observed among pD2 or Hill coefficient. 4. Emax to Phe (4 and 7 days) and to Ang II (15 and 30 days) increased in the contralateral artery. In addition, Phe or Ang II reactivity was not significantly different in aorta rings from control or carotid-injured animals. 5. The increased responsiveness of contralateral artery was not due to changes in carotid blood flow or resting membrane potential. The endothelium-dependent inhibitory component is not present in the contraction of contralateral arteries and it is not related to superoxide anion production. 6. Indomethacin decreased contralateral artery responsiveness to Phe and Ang II. Valeryl salicylate reduced the Ang II response in contralateral and control arteries. Celecoxib decreased the Phe Emax of contralateral artery. 7. In conclusion, decreased endothelium-derived factors and increased prostanoids appear to be responsible for the increased reactivity of contralateral arteries after injury.
Assuntos
Angiotensina II/farmacologia , Lesões das Artérias Carótidas/fisiopatologia , Artéria Carótida Primitiva/efeitos dos fármacos , Cateterismo/efeitos adversos , Fenilefrina/farmacologia , Animais , Artéria Carótida Primitiva/fisiologia , Relação Dose-Resposta a Droga , Técnicas In Vitro , Masculino , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Ratos , Ratos WistarRESUMO
Although intracoronary thrombosis often occurs after angioplasty and may affect its outcome, the accuracy of arteriography for identification of mural thrombi is unclear. This study analyzed the relationship between arteriographic abnormalities immediately before death and the histologic extent of thrombosis in 77 dogs submitted to balloon injury of intact left anterior descending coronary arteries. Survival time after angioplasty was 120 min. The incidence of mural thrombosis, defined on serial histologic sections, was 65.0 percent. A positive diagnosis of intracoronary thrombus at arteriography (AT+) was based on the presence of any of the following signs: filling defects, retention of contrast material, and slowed or interrupted flow. Seventeen dogs were AT+, and 60 were AT-. The overall sensitivity of arteriography was 34 percent, and the specificity was 100 percent. Even considering as significant only thrombi greater than 25.0 percent of the arterial lumen area, 11 of 27 dogs were AT- despite thrombus sizes between 27 percent and 75 percent of lumen area (sensitivity, 59 percent); arteriography consistently missed smaller thrombi (22 of 23 dogs were AT-). Arterial diameters and balloon-induced injury were similar between AT- and AT+ dogs. Scanning electron microscopy depicted a fibrin-poor thrombus in 14 of 19 AT+ dogs and a fibrin-rich thrombus in five, whereas all seven AT+ dogs had fibrin-rich thrombi. Logistic regression analysis showed a correlation between thrombus size and arteriographic positivity, whereas the presence of fibrin and slowed flow of contrast material did not independently predict positive arteriographic results. Thus, arteriography is inaccurate for identification of mural thrombosis after angioplasty, mostly because of its poor sensitivity.
Assuntos
Angioplastia Coronária com Balão/efeitos adversos , Angiografia Coronária , Trombose Coronária/diagnóstico por imagem , Animais , Plaquetas/patologia , Cinerradiografia , Trombose Coronária/patologia , Vasoespasmo Coronário/patologia , Vasos Coronários/patologia , Cães , Eritrócitos/patologia , Feminino , Fibrina , Fluoroscopia , Masculino , Microscopia Eletrônica de Varredura , Fluxo Sanguíneo Regional , Sensibilidade e EspecificidadeRESUMO
Platelet-activating factor (PAF) is a phospholipid mediator that induces cardiovascular collapse and release of the secondary mediator thromboxane A2 (TxA2). To clarify mechanisms involved in this collapse and, specifically, the relative contribution of left ventricular and right ventricular dysfunction, we studied 12 open-chest pigs. PAF infusion (0.04-0.28 nmol.kg-1.min-1) induced a 5- to 120-fold increase in pulmonary vascular resistance, a 75-98% fall in cardiac output, and systemic arterial hypotension. Right ventricular failure was indicated by chamber enlargement, decreased shortening, and increased right atrial pressures. In contrast, left ventricular dysfunction was accompanied by decreases in chamber dimensions and filling pressures that were unresponsive to volume expansion. U 46619 (a stable TxA2 analogue) and mechanical pulmonary artery constriction induced changes similar to PAF. In 11 additional closed-chest pigs, TxA2 blockade with indomethacin attenuated the PAF-induced rise in pulmonary vascular resistance, right ventricular dysfunction, and systemic hypotension. A specific TxA2 synthase inhibitor, OKY-046, also diminished hemodynamic effects of PAF in six other pigs. Tachyphylaxis was not observed in five pigs repeatedly given PAF. We conclude that acute right ventricular failure as the result of severe increase in pulmonary vascular resistance is the primary mechanism early in the course of PAF-induced shock in the pig. PAF-induced release of TxA2 may contribute significantly to these events.
Assuntos
Hipotensão/induzido quimicamente , Fator de Ativação de Plaquetas/farmacologia , Tromboxano A2/farmacologia , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico , Animais , Feminino , Hipotensão/fisiopatologia , Indometacina/farmacologia , Masculino , Metacrilatos/farmacologia , Endoperóxidos Sintéticos de Prostaglandinas/farmacologia , Suínos , Tromboxano-A Sintase/antagonistas & inibidoresRESUMO
Calcitonin gene-related peptide (CGRP) has been identified in nerve fibers innervating cardiovascular elements and particularly in coronary arteries (CA). To investigate its potential role in modulating coronary blood flow, we injected rat-CGRP into the CA of pentobarbital anesthetized, open chest pigs. Significant dose-related increments in coronary flow were observed. The rise in coronary flow was characterized by unusually, slow onset, late peak and prolonged duration. Arterial pressure, heart rate (HR) and myocardial contractility were unchanged, except at the highest dose (3.0 nmol), which produced mild systemic hypotension and sinus tachycardia. Coronary levels of catecholamines and 6-keto-PGF1 alpha were unchanged by CGRP. The direct, sustained, and potent dilatory activity of CGRP, in coronary arteries of the pig together with anatomical CGRP localization in this site suggest a role for this neuropeptide in hemodynamic regulation.
Assuntos
Circulação Coronária/efeitos dos fármacos , Neuropeptídeos/farmacologia , Vasodilatadores/farmacologia , 6-Cetoprostaglandina F1 alfa/sangue , Animais , Vasos Sanguíneos/efeitos dos fármacos , Peptídeo Relacionado com Gene de Calcitonina , Catecolaminas/sangue , Feminino , Hemodinâmica/efeitos dos fármacos , Masculino , Contração Miocárdica/efeitos dos fármacos , Substância P/farmacologia , SuínosRESUMO
In pentobarbital anesthetized, open chest pigs, coronary blood flow (CBF), myocardial contractility and systemic hemodynamic variables were monitored during intracoronary injections of substance P (SP), neurokinin A (NA) or neurokinin B (NB). SP was most potent in increasing CBF although NA was also active in high doses while NB had absolutely no effect. SP was also more potent than NA in producing systemic hypotension. The data suggests that SP and its receptors might be potentially important modulators of CBF.
Assuntos
Circulação Coronária/efeitos dos fármacos , Proteínas do Tecido Nervoso/farmacologia , Animais , Feminino , Hemodinâmica/efeitos dos fármacos , Masculino , Contração Miocárdica/efeitos dos fármacos , Neurocinina A , Neurocinina B , Oligopeptídeos/farmacologia , Substância P/farmacologia , Suínos , TaquicininasRESUMO
Sudden release of platelet-activating factor (PAF) into the circulation can cause hypotension, tachycardia, and circulatory collapse. To further examine this response, we performed detailed studies of cardiovascular function after PAF administration to young domestic pigs and newborn piglets. Our results indicate that circulatory dysfunction after PAF reflects severe constriction of pulmonary resistance vessels and consequent acute right ventricular failure. Although PAF-induced coronary artery constriction and contractile depression may be complicating problems, left ventricular underperfusion and dysfunction after PAF are mainly the result of systemic arterial hypotension and diminished left ventricular filling. The adverse hemodynamic effects of PAF are accompanied by substantial release of thromboxane A2 (TxA2). These effects are mimicked by the TxA2 agonist U-46619 and partially blocked by specific and nonspecific inhibitors of TxA2 synthesis (OKY-046 and indomethacin). Even more potent blockade of PAF action is exerted by the TxA2 receptor blocker, SQ 29,548. Taken together, these findings indicate that severe pulmonary vascular constriction and hemodynamic collapse soon after intravenous PAF are at least partially mediated by PAF-induced TxA2 release. Tachyphylaxis to PAF influence has been observed in studies of leukocyte and platelet function. We hypothesized that tachyphylaxis to PAF might also occur in our studies of constrictor responses in pulmonary vessels. Recently, we have examined the capacity of PAF to produce sustained pulmonary vasoconstriction in open-chested, anesthetized newborn piglets. Infusions sufficient to produce 100% increase in mean pulmonary artery pressure after 3 min showed no loss of efficacy when sustained for 30 min.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Sistema Cardiovascular/efeitos dos fármacos , Hemodinâmica/efeitos dos fármacos , Fator de Ativação de Plaquetas/farmacologia , Animais , Fenômenos Fisiológicos Cardiovasculares , Fator de Ativação de Plaquetas/toxicidade , Suínos , Tromboxano A2/antagonistas & inibidoresRESUMO
To investigate the cellular reactions to arterial injuries and the influence of a cholesterol-rich diet, 18 rabbits underwent endothelial denudation of the abdominal aorta with a balloon catheter. Fourteen animals were fed a 2% cholesterol diet and 4 were fed normal rabbit chow for 8 weeks. In the cholesterol-fed group, 6 animals had only the expected intimal lesions; however, 8 animals exhibited different degrees of balloon-induced medial layer injury, with fibrous healing. Similarly, in the control rabbits, 1 had intimal lesions and 3 had both intimal and medial layer lesions. We conclude that removal of the endothelium with a balloon catheter promotes arterial wall injury deeper than expected. This unexpected result could influence the effect of interventions usually employed in experimental atherosclerosis.
Assuntos
Aorta/patologia , Cateterismo/efeitos adversos , Endotélio Vascular/patologia , Animais , Arteriosclerose/etiologia , Colesterol/administração & dosagem , Dieta , Masculino , CoelhosRESUMO
1. Although the effects of therapeutic interventions upon infarct size are frequently assessed on the basis of left ventricular ejection fraction and segmental contraction, the correlation of these variables with infarct size has not been thoroughly evaluated. To explore such relationships, we occluded the left anterior descending coronary artery of 22 closed-chest dogs and performed contrast ventriculography one week later. Regional myocardial function was evaluated by a computer system in 60 radial segments. 2. Infarct size, measured by triphenyl-tetrazolium chloride staining, ranged from 1.4 to 43.6% of the left ventricle. Infarcted dogs were arbitrarily divided into 3 groups by percentage of necrotic area: Group 1 (less than or equal to 15%, N = 5), Group 2 (15 to 30%, N = 10) and Group 3 (greater than or equal to 30%, N = 7). 3. Although the ejection fraction was significantly reduced in infarcted animals as compared to preselected normal controls (38.9 +/- 11.6 [SD] vs 74.1 +/- 7.5%, P less than 0.001), it was similar within each infarct subgroup. 4. There was a linear inverse correlation between ejection fraction and percentage of abnormally contracting segments (R = -0.63, P = 0.0017). However, neither ejection fraction nor abnormally contracting segments were correlated with infarct size (R = -0.17 and R = 0.11, respectively). 5. A more detailed analysis revealed that infarcted or infarct-adjacent segments were less depressed in Group 1 than in Group 2 or 3 and extent of depression was similar between Groups 2 and 3. Conversely, the extent of shortening of non-infarcted inferior wall segments increased from Group 1 to Group 3. 6. Thus, regional myocardial contraction is significantly affected by non-necrotic infarct-adjacent segments and the ejection fraction is significantly influenced by non-ischemic myocardium. For infarcted areas up to 40% of the left ventricle, a single post-infarction determination of ejection fraction or the percentual of abnormally contracting segments seems unreliable, on a population basis, to judge the effects of infarct-sparing interventions.