RESUMO
Objectives of this study were to determine the physiological effects of psychological stress applied to dairy calves and to test if molasses consumption could be used to validate that a stressed condition was achieved. Twenty male calves (3 wk old) received jugular catheters and were randomly assigned to control (CTR; n = 4 pens of 1 calf per pen) or social stress treatments (STR; n = 4 pens of 4 calves per pen). The STR treatment included 5 cycles of 24-h isolation followed by regrouping with unfamiliar animals for 48 h (over 15 d). An ACTH challenge (0.1 IU/kg of body weight) was used to determine adrenal fatigue. Peak and total cortisol concentrations were greater for STR calves until the ACTH challenge. After the ACTH challenge, CTR calf cortisol increased and STR calf cortisol continued to decrease, suggesting adrenal fatigue. The number of calves that became positive for fecal shedding of Salmonella after the acute stress of being moved and the number of calves that were positive after the move decreased with each move. Fifty-six percent of STR calves changed from negative to positive for shedding after the first move compared with 18.75% of STR calves remaining negative after the third move. Difference in fecal shedding of Enterobacteriaceae from samples taken before and after moving calves on d 6 was less than that on d 2, 3, and 5. Leukocyte counts were not different, but trends for day effects were detected for neutrophil and monocyte percentages. Molasses consumption was greater for STR calves on d 2 and 11, as was total consumption. Latency to lie after eating also increased as the study progressed; STR calves required more time to lie after eating on d 12 than on d 3, and latency to lie was greater for STR than CTR on d 4, 8, 12, and 14. The STR calves also stood more than the CTR calves in the 4-h afternoon period on d 4, 5, 7, and 14. However, during the 4-h morning observations on d 14 (ACTH challenge), CTR calves stood more than STR calves. This model induced chronic stress, as characterized by adrenal fatigue, which was confirmed by molasses consumption and behavior changes. Therefore, molasses consumption could be used to confirm social stress in experimental models.
Assuntos
Doenças dos Bovinos/fisiopatologia , Aglomeração/psicologia , Estresse Psicológico/fisiopatologia , Hormônio Adrenocorticotrópico/farmacologia , Animais , Derrame de Bactérias , Peso Corporal/efeitos dos fármacos , Bovinos , Doenças dos Bovinos/etiologia , Ingestão de Alimentos/fisiologia , Enterobacteriaceae , Fezes/microbiologia , Hidrocortisona/sangue , Contagem de Leucócitos/veterinária , Masculino , Melaço , Estresse Psicológico/etiologiaRESUMO
Feed sorting is a natural behavior of dairy cows that can result in inconsistencies in the nutritive value of a total mixed ration (TMR). The objective of this study was to determine the effects of replacing high-moisture corn with glycerol on feed sorting and the feed intake pattern of transition dairy cows. Multiparous Holstein cows (n=26) were paired by expected calving date, housed in individual tie stalls, and fed diets containing either glycerol or high-moisture corn once daily from d -28 to +56 relative to calving. Glycerol was included at 11.5 and 10.8% of the ration dry matter for the pre- and postpartum diets, respectively. The feed consumption pattern was determined by measuring TMR disappearance during the intervals from 0 to 4 h, 4 to 8 h, 8 to 12 h, and 12 to 24 h relative to feed delivery. Feed sorting was determined on d -16, -9, 9, 16, and 51 relative to calving at 4, 8, 12 and 24 h after feeding. The TMR particle size profile was determined at feed delivery and at 4, 8, 12, and 24 after feed delivery by using the Penn State Particle Separator (Nasco, Fort Atkinson, WI) to yield long (>19 mm), medium (<19 mm, >8 mm), short (<8 mm, >1.18 mm), and fine (<1.18 mm) particles. Overall feed intake did not differ between diets and was 14.7±0.4 and 20.2±0.5 kg/d for the pre- and postpartum intervals, respectively. During the prepartum period, glycerol decreased the amount of feed consumed during the first 4h after feed delivery (7.22 vs. 5.59±0.35 kg; control vs. glycerol, respectively) but increased feed consumed from 12 through 24 h after feed delivery (2.22 vs. 3.82±0.35 kg; control vs. glycerol, respectively). Similar effects on the feed consumption pattern were observed after calving. During the prepartum period, cows fed the control diet sorted against long particles, whereas cows fed glycerol did not sort against long particles (77.2 vs. 101.5±3.50% of expected intake for control vs. glycerol; significant treatment effect). The data indicate that addition of glycerol to the TMR alters the feed consumption pattern to increase feed consumption late in the day at the expense of feed consumed immediately after feeding, and it reduces sorting behavior against long particles. Together, these may reduce diurnal variations in the rumen environment to promote greater rumen health in transition cows.
Assuntos
Bovinos/fisiologia , Comportamento Alimentar , Glicerol/farmacologia , Ração Animal , Animais , Bovinos/psicologia , Dieta/veterinária , Ingestão de Alimentos/fisiologia , Comportamento Alimentar/fisiologia , FemininoRESUMO
HYPOTHESIS: A lack of contraction of cerebral microarterioles to Ang II ("resilience") depends on cyclooxygenase (COX) and lipocalin type prostaglandin D sythase L-PGDS producing PGD2 that activates prostaglandin D type 1 receptors (DP1Rs) and nitric oxide synthase (NOS). MATERIALS & METHODS: Contractions were assessed in isolated, perfused vessels and NO by fluorescence microscopy. RESULTS: The mRNAs of penetrating intraparenchymal cerebral microarterioles versus renal afferent arterioles were >3000-fold greater for L-PGDS and DP1R and 5-fold for NOS III and COX 2. Larger cerebral arteries contracted with Ang II. However, cerebral microarterioles were entirely unresponsive but contracted with endothelin 1 and perfusion pressure. Ang II contractions were evoked in cerebral microarterioles from COX1 -/- mice or after blockade of COX2, L-PGDS or NOS and in deendothelialized vessels but effects of deendothelialization were lost during COX blockade. NO generation with Ang II depended on COX and also was increased by DP1R activation. CONCLUSION: The resilience of cerebral arterioles to Ang II contractions is specific for intraparenchymal microarterioles and depends on endothelial COX1 and two products that are metabolized by L-PGDS to generate PGD2 that signals via DP1Rs and NO.
Assuntos
Arteríolas/fisiologia , Células Endoteliais/metabolismo , Perfusão , Prostaglandina D2/metabolismo , Vasoconstrição , Angiotensina II , Animais , Células Cultivadas , Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/metabolismo , Células Endoteliais/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Endotélio Vascular/fisiologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microvasos/efeitos dos fármacos , Microvasos/metabolismo , Microvasos/fisiologia , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/metabolismo , Prostaglandina-Endoperóxido Sintases/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Receptores de Angiotensina/metabolismo , Receptores de Prostaglandina/metabolismo , Vasoconstrição/efeitos dos fármacosRESUMO
Thermographic imaging is a noninvasive diagnostic tool used to document the inflammatory process in many species and may be useful in the detection of subclinical bumblefoot and other inflammatory diseases. Bumblefoot is a chronic inflammation of the plantar metatarsal or digital pads of the foot (pododermatitis), or both. It is one of the major health problems in birds including chickens and is responsible for significant economic losses in commercial poultry operations. Early diagnosis of bumblefoot is essential for the prevention of economical loss and the improvement of animal well-being. The object of this study was to determine the suitability of thermography for the identification of subclinical bumblefoot in chickens. Experiment 1 was designed to validate thermography as a tool for screening avian populations for bumblefoot. The plantar surface of the feet of 150 randomly selected hens was imaged using a thermal camera. The thermal images were identified as suspect, positive, or negative for bumblefoot based on thermal patterns of the plantar surface. Visual inspection of the feet identified as suspect followed 14 d later. A visual score of clinical, mildly clinical, or negative for bumblefoot was assigned, based on gross pathological changes in the plantar surface. A correlation between initial thermal images identified as suspect for bumblefoot and a visual score of positive 14 d later was 83% (P < 0.01). In experiment 2, hens whose feet were free of lesions were inoculated in the metatarsal foot pad with Staphylococcus aureus. Thermal images and visual clinical scores were taken, prechallenge and 1, 2, 3, 4, and 7 d postchallenge. The correlation between thermal images classified as clinical and a visual score of clinical for bumblefoot was 86.7% (P < 0.001). However, the correlation between the thermal images classified as mild (subclinical) and a visual score of mild was only 26.7%, suggesting that thermography is a more sensitive indicator of subclinical infection than visual appraisal. Thermography may thus provide a useful tool for screening avian populations for signs of bumblefoot, early in the course of the disease, which will improve recovery percentages and bird well-being.
Assuntos
Galinhas , Doenças do Pé/veterinária , Doenças das Aves Domésticas/diagnóstico , Infecções Cutâneas Estafilocócicas/veterinária , Staphylococcus aureus/isolamento & purificação , Termografia/veterinária , Animais , Feminino , Doenças do Pé/diagnóstico , Doenças do Pé/microbiologia , Doenças das Aves Domésticas/microbiologia , Distribuição Aleatória , Infecções Cutâneas Estafilocócicas/diagnóstico , Infecções Cutâneas Estafilocócicas/microbiologiaRESUMO
This study was conducted to determine the effect of experimentally increased glucose demand on voluntary consumption of molasses by dairy calves. Three-week-old calves received 0.365 g of phlorizin by s.c. injection. Urinary output and molasses consumption were measured hourly, and urinary glucose concentration was screened. Molasses consumption for the 24 h after treatment was (mean +/- SE) 72.0 g (+/-7) for the control group and 142 g (+/-1) for the phlorizin-treated group. Urinary output for the 8-h test period was 1.13 kg for the control group and 1.67 kg for the phlorizin-treated calves. Mean urinary glucose peaked at 10 g/L by 4 h after treatment for calves given phlorizin, whereas the concentration for the control group remained close to 0 g/L. Phlorizin treatment increased voluntary consumption of molasses in 3-wk-old Holstein calves.
Assuntos
Doenças dos Bovinos/induzido quimicamente , Doenças dos Bovinos/fisiopatologia , Ingestão de Alimentos/efeitos dos fármacos , Glicosúria/veterinária , Melaço , Florizina/farmacologia , Animais , Bovinos , Glicosúria/induzido quimicamente , Glicosúria/fisiopatologia , Masculino , Cloreto de Sódio/farmacologia , Fatores de TempoRESUMO
Micropuncture studies have shown that glomerular filtration rate (GFR) falls in response to a rise in Na(+) or Cl(-) concentrations in the loop of Henle, whereas studies in isolated kidneys have shown that GFR falls in response to osmotic diuresis. To define the separate effects of an acute increase in plasma sodium (P(Na)), chloride (P(Cl)) or osmolality (P(osmol)), changes in renal blood flow (RBF) and GFR were measured during intrarenal infusions of hypertonic NaCl, NaHCO(3), Na acetate, dextrose, NH(4)Cl or NH(4)acetate to denervated kidneys. The infusions raised P(osmol) at the experimental kidney by 30-45 mosmol. RBF increased abruptly by 10-30% with all hypertonic infusions indicating that an acute increase in plasma tonicity causes renal vasodilatation. Renal vasodilatation persisted or increased further during infusion of dextrose, NaHCO(3) and Na acetate, but GFR was unchanged. In contrast, during infusion of the two Cl-containing solutions, vasodilatation was reversed after 1-5 min and RBF and GFR decreased (P < 0.01) below preinfusion levels. Prior salt depletion doubled the vasoconstriction seen with hypertonic NaCl infusions. Overall, changes in RBF were unrelated to changes in P(Na) or fractional Na or fluid reabsorption but correlated with changes in P(Cl) (r = -0.91) and fractional Cl(-) reabsorption (r = 0.94). The intrafemoral arterial infusion of the two Cl-containing solutions did not increase femoral vascular resistance. In conclusion, hyperchloremia produces a progressive renal vasoconstriction and fall in GFR that is independent of the renal nerves, is potentiated by prior salt depletion and is related to tubular Cl(-) reabsorption. Chloride-induced vasoconstriction appears specific for the renal vessels.
Assuntos
Cloretos/sangue , Circulação Renal , Absorção , Animais , Pressão Sanguínea , Cães , Artéria Femoral , Taxa de Filtração Glomerular/efeitos dos fármacos , Soluções Hipertônicas/farmacologia , Testes de Função Renal , Masculino , Fluxo Sanguíneo Regional/efeitos dos fármacos , Circulação Renal/efeitos dos fármacosRESUMO
Some studies have indicated that PGs can modulate the single nephron tubuloglomerular feedback (TGF) response. The aim of this study was to define the specific role of the vasoconstrictor PG, TX, by administration to rats of either vehicle (group 1; n = 20) or drugs that inhibit either cyclooxygenase (indomethacin [indo], 5 mg.kg-1, group 2, n = 17), TX synthetase (UK-38,485 [UK], 100 mg.kg-1, group 3, n = 19), or TX receptors (SQ-29,548 [SQ], 8 mg.kg-1, group 4, n = 14, or L-641,953 [L], 50 mg.kg-1, group 5, n = 8). Indo reduced excretion of the prostacyclin derivative 6-keto-PGF1 alpha and TXB2 and lowered whole kidney GFR and renal plasma flow, whereas UK lowered excretion of TXB2 only and did not change basal renal hemodynamics. The TGF response (assessed from reduction in proximal tubule stop-flow pressure (Psf, mmHg) during increases in perfusion of the loop of Henle (LH) from 0 to 40 nl.min-1) was unchanged after vehicle (9.8 +/- 0.5-10.9 +/- 1.0, NS) but blunted (P less than 0.001) by 40-65% in rats of groups 2-5 (indo, 11.1 +/- 1.0-4.4 +/- 0.7; UK, 9.0 +/- 0.8-4.8 +/- 0.7; SQ, 10.3 +/- 0.6-4.8 +/- 0.6; L, 10.7 +/- 0.5-6.7 +/- 1.3). This blunting was due to lower values for Psf at zero LH flow after indo, SQ, and L, and higher values of Psf at 40 nl.min-1 LH flow after indo and UK. The fall in single nephron GFR (SNGFR, nl.min-1) with increasing LH perfusion was unchanged after vehicle (10.9 +/- 2.8-11.2 +/- 0.8) but was blunted (P less than 0.05) by 45-55% in rats given indo (13.9 +/- 1.2-6.2 +/- 2.2) or UK (12.8 +/- 2.1-7.0 +/- 1.5). UK produced dose-dependent reductions in TXB2 excretion (IC50, 15 mg.kg-1) and inhibition of the TGF response (IC50: 30 mg.kg-1). After blockade of TX receptors by SQ, UK had no further affect on the TGF response. The fall in Psf at high LH flow was blunted (P less than 0.05) by indo and UK, whereas the rise in Psf at zero LH flow was blunted by indo, SQ, and L. In conclusion, endogenous TX generation can modulate the reductions in Psf and SNGFR during increased delivery of NaCl to the LH.
Assuntos
Glomérulos Renais/fisiologia , Túbulos Renais/fisiologia , Tromboxano B2/fisiologia , Análise de Variância , Animais , Pressão Sanguínea/efeitos dos fármacos , Compostos Bicíclicos Heterocíclicos com Pontes , Inibidores de Ciclo-Oxigenase , Relação Dose-Resposta a Droga , Ácidos Graxos Insaturados , Taxa de Filtração Glomerular , Hidrazinas/farmacologia , Pressão Hidrostática , Imidazóis/farmacologia , Indometacina/farmacologia , Glomérulos Renais/efeitos dos fármacos , Túbulos Renais/efeitos dos fármacos , Masculino , Prostaglandinas F/urina , Ratos , Ratos Endogâmicos , Receptores de Prostaglandina/efeitos dos fármacos , Receptores de Tromboxanos , Tromboxano A2/antagonistas & inibidores , Tromboxano B2/urina , Tromboxano-A Sintase/antagonistas & inibidoresRESUMO
UNLABELLED: Because endogenous thromboxane A2 (TXA2) potentiates the tubuloglomerular feedback response (TGF), we studied the mechanism of action of TXA2 by using a stable TXA2/prostaglandin (PG) H2 mimetic, U-46,619. Intravenous infusion of U-46,619 at 100 ng.kg-1.min-1 reduced the GFR and the single-nephron (SN)GFR measured from the distal tubule (macula densa function intact), whereas the SNGFR measured from the proximal tubule (macula densa function interrupted) was not changed consistently. 10-100-fold higher rates of infusion of U-46,619 were required to raise blood pressure or femoral vascular resistance. The regulation of glomerular capillary pressure (PGC) by TGF was assessed in anesthetized rats from changes in proximal stop flow pressure (PSF) and/or SNGFR during perfusion of the loop of Henle (LH) with artificial tubular fluid (ATF). Orthograde loop perfusion and retrograde perfusion of U-46,619 into the macula densa segment reduced PSF. Responses to luminal U-46,619 were blunted by a TXA2-PGH2 receptor antagonist. Orthograde loop perfusions with luminal U-46,619 increased net Cl absorption, whereas coperfusion with furosemide (10(-4) M) blunted the response to U-46,619 by 68%. These data indicated that the luminal U-46,619 might increase the signal for TGF activation by increasing Cl reabsorption in macula densa cells. However, since 80 +/- 4% of [3H]U-46,619 perfused via the LH was reabsorbed peritubular capillaries (PTC) were perfused with U-46,619 to test additional extra-luminal actions. PTC perfusion with U-46,619 again increased TGF by reducing PSF selectively only while macula densa function was intact during perfusion of the LH with ATF. CONCLUSIONS: (a) TGF is potentiated by U-46,619 given systematically, via the lumen of the LH by orthograde or retrograde perfusions or via the PTC; (b) at the lower doses tested, reduction of PGC and SNGFR by U-46,619 depends on tubular fluid delivery and reabsorption by the macula densa; (c) potentiation of TGF by U-46,619 entails preglomerular vasoconstriction which may be elicited in part by an increased signal due to increased net chloride reabsorption in the LH and presumably macula densa cells and by an increased sensitivity of the arteriole to macula densa-derived signals; (d) activation of TGF may contribute to the selective vasoconstriction of the renal vascular bed by low doses of U-46,619.
Assuntos
Glomérulos Renais/fisiologia , Endoperóxidos Sintéticos de Prostaglandinas , Tromboxano A2/fisiologia , Animais , Pressão Sanguínea , Relação Dose-Resposta a Droga , Retroalimentação , Taxa de Filtração Glomerular , Glomérulos Renais/irrigação sanguínea , Glomérulos Renais/efeitos dos fármacos , Alça do Néfron/efeitos dos fármacos , Alça do Néfron/fisiologia , Masculino , Ratos , Ratos Endogâmicos , VasoconstriçãoRESUMO
These studies tested the hypothesis that delivery and/or cellular uptake of L-arginine limits macula densa nitric oxide generation and actions on tubuloglomerular feedback (TGF) during salt restriction. Maximal TGF responses were assessed from reductions in proximal stop flow pressure during loop of Henle (LH) perfusion at 40 nl/min with artificial tubular fluid containing vehicles or drugs. Orthograde LH perfusion of L-arginine (10[-3] M) reduced maximal TGF significantly in rats adapted to low salt (LS: 7.9+/-0.4-6.3+/-0.4 mmHg; P < 0.05), but not high salt (HS: 5.8+/-0.3-5.9+/-0.3; NS). The effects were stereospecific and prevented by coperfusion with NG-methyl-L-arginine. Microperfusion of L-arginine (10[-3] M) into the peritubular capillaries reduced the maximum TGF response more in nephrons of LS than HS rats (deltaTGF: LS, 32+/-6 vs. HS, 13+/-4%; P < 0.05) and restored a TGF response to luminal perfusion of NG-methyl-L-arginine in LS rats. Coperfusion of nephrons with excess L-lysine or L-homoarginine, which compete with L-arginine for system y+ transport, blocked the fall in proximal stopflow pressure produced by orthograde LH perfusion of L-arginine in LS rats. Reabsorption of [3H]arginine by the perfused loop segment was similar in LS (93+/-2%) and HS (94+/-1%) rats. Coperfusion with excess L-arginine, L-lysine, or L-homoarginine, however, reduced [3H]arginine reabsorption significantly (P < 0.05) more in HS rats than in LS rats. In conclusion, blunting of maximal TGF responses in salt-restricted rats by nephron-derived NO is limited by L-arginine availability and cellular uptake via system y+.
Assuntos
Arginina/metabolismo , Glomérulos Renais/metabolismo , Cloreto de Sódio/metabolismo , Animais , Pressão Sanguínea , Peso Corporal , Capilares/fisiologia , Homoarginina/metabolismo , Rim/anatomia & histologia , Glomérulos Renais/irrigação sanguínea , Lisina/metabolismo , Masculino , Tamanho do Órgão , Ratos , Ratos Sprague-Dawley , Equilíbrio HidroeletrolíticoRESUMO
AIM: The glomerular filtration rate (GFR) falls progressively in chronic kidney disease (CKD) which is caused by a reduction in the number of functional nephrons. The dysfunctional nephron exhibits a lower glomerular capillary pressure that is induced by an unbalance between afferent and efferent arteriole. Therefore, we tested the hypothesis that oxidative stress induced by CKD differentially impairs the structure or function of efferent vs. afferent arterioles. METHODS: C57BL/6 mice received sham operations (sham) or 5/6 nephrectomy (RRM) and three months of normal- or high-salt diet or tempol. GFR was assessed from the plasma inulin clearance, arteriolar remodelling from media/lumen area ratio, myogenic responses from changes in luminal diameter with increases in perfusion pressure and passive wall compliance from the wall stress/strain relationships. RESULTS: Mice with RRM fed a high salt (vs. sham) had a lower GFR (553 ± 25 vs. 758 ± 36 µL min-1 g-1 kidney, P < 0.01) and a larger efferent arteriolar diameter (9.6 ± 0.8 vs. 7.4 ± 0.7 µm, P < 0.05) resulting in a lower media/lumen area ratio (1.4 ± 0.1 vs. 2.4 ± 0.2, P < 0.01). These alterations were corrected by tempol. The myogenic responses of efferent arterioles were about one-half that of afferent arterioles and were unaffected by RRM or salt. Passive wall compliance was reduced by high salt in both afferent and efferent arterioles. CONCLUSION: A reduction in renal mass with a high-salt diet induces oxidative stress that leads to an outward eutrophic remodelling in efferent arterioles and reduced wall compliance in both afferent and efferent arterioles. This may contribute to the lower GFR in this model of CKD.
Assuntos
Arteríolas/fisiopatologia , Estresse Oxidativo/fisiologia , Insuficiência Renal Crônica/fisiopatologia , Cloreto de Sódio na Dieta/efeitos adversos , Remodelação Vascular/fisiologia , Animais , Modelos Animais de Doenças , Taxa de Filtração Glomerular , Glomérulos Renais/irrigação sanguínea , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Nefrectomia , Tamanho do Órgão , Insuficiência Renal Crônica/patologiaRESUMO
We previously found that K vitamin analogues caused cell growth inhibition in Hep3B hepatoma cells in vitro, which was associated with their inhibitory effects on protein tyrosine-phosphatases. In this study, we show that Cdc25A, a protein phosphatase, was inactivated by novel arylating K vitamin analogues. The inactivation of Cdc25A correlated with their effects on cell growth inhibition. Cyclin-dependent kinase (Cdk) 4, an important regulator for G(1) progression, was found to be tyrosine-phosphorylated by the arylating analogues, and this phosphorylation was correlated with the inhibitory effects of the analogues on Cdc25A activity. Furthermore, Cdk4 dephosphorylation experiments showed that Compound (Cpd) 5, a prototype arylating analogue, inhibited Cdc25A-mediated Cdk4 dephosphorylation, whereas Cpd 26, a nonarylating vitamin K analogue, had no effect on this event. We also examined Cdk4 kinase activity using retinoblastoma protein as a substrate and found that Cpd 5 inhibited retinoblastoma protein phosphorylation in a concentration-dependent manner, indicating that Cdk4 activity was inhibited by Cpd 5 treatment. Moreover, the thiol-antioxidants glutathione and N-acetyl-L-cysteine antagonized the Cpd 5-induced Cdk4 tyrosine phosphorylation, whereas the nonthiol-antioxidants catalase and superoxide dismutase did not. These results suggest that Hep3B cell growth inhibition by these K vitamin analogues may be related in part to inactivation of Cdc25A activity and support the hypothesis that Cdc25A is an attractive target for drugs designed to inhibit cancer cell growth.
Assuntos
Carcinoma Hepatocelular/enzimologia , Inibidores do Crescimento/farmacologia , Neoplasias Hepáticas/enzimologia , Proteínas Proto-Oncogênicas , Vitamina K/análogos & derivados , Fosfatases cdc25/antagonistas & inibidores , Acetilcisteína/farmacologia , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/patologia , Catalase/farmacologia , Divisão Celular/efeitos dos fármacos , Quinase 4 Dependente de Ciclina , Quinases Ciclina-Dependentes/antagonistas & inibidores , Quinases Ciclina-Dependentes/metabolismo , Interações Medicamentosas , Ativação Enzimática/efeitos dos fármacos , Glutationa/farmacologia , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/patologia , Fosforilação/efeitos dos fármacos , Proteína do Retinoblastoma/metabolismo , Compostos de Sulfidrila/farmacologia , Vitamina K/antagonistas & inibidores , Vitamina K/farmacologia , Fosfatases cdc25/metabolismoRESUMO
AIM: Renal ischaemia-reperfusion injury (IRI) increases angiotensin II (Ang II) and reactive oxygen species (ROS) that are potent modulators of vascular function. However, the roles of individual ROS and their interaction with Ang II are not clear. Here we tested the hypothesis that IRI modulates renal afferent arteriolar responses to Ang II via increasing superoxide (O2-) or hydrogen peroxide (H2 O2 ). METHODS: Renal afferent arterioles were isolated and perfused from C57BL/6 mice 24 h after IRI or sham surgery. Responses to Ang II or noradrenaline were assessed by measuring arteriolar diameter. Production of H2 O2 and O2- was assessed in afferent arterioles and renal cortex. Activity of SOD and catalase, and mRNA expressions of Ang II receptors were assessed in pre-glomerular arterioles and renal cortex. RESULTS: Afferent arterioles from mice after IRI had a reduced maximal contraction to Ang II (-27±2 vs. -42±1%, P < 0.001), but retained a normal contraction to noradrenaline. Arterioles after IRI had a 38% increase in H2 O2 (P < 0.001) and a 45% decrease in catalase activity (P < 0.01). Contractions were reduced in normal arterioles after incubation with H2 O2 (-22±2 vs. -42±1%, P < 0.05) similar to the effects of IRI. However, the impaired contractions were normalized by incubation with PEG catalase despite a reduced AT1 R expression. CONCLUSIONS: Renal IRI in mice selectively impairs afferent arteriolar responses to Ang II because of H2 O2 accumulation that is caused by a reduced catalase activity. This could serve to buffer the effect of Ang II after IRI and may be a protective mechanism.
Assuntos
Injúria Renal Aguda/fisiopatologia , Angiotensina II/farmacologia , Arteríolas/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Circulação Renal/efeitos dos fármacos , Traumatismo por Reperfusão/fisiopatologia , Vasoconstritores/antagonistas & inibidores , Vasoconstritores/farmacologia , Animais , Catalase/biossíntese , Córtex Renal/irrigação sanguínea , Córtex Renal/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Norepinefrina/farmacologia , Receptor Tipo 1 de Angiotensina/biossíntese , Receptor Tipo 2 de Angiotensina/biossíntese , Superóxido Dismutase/biossínteseRESUMO
OBJECTIVES: This study was designed to investigate disturbances in arterial blood pressure and body fluid homeostasis in stable heart transplant recipients. BACKGROUND: Hypertension and fluid retention frequently complicate heart transplantation. METHODS: Blood pressure, renal and endocrine responses to acute volume expansion were compared in 10 heart transplant recipients (57 +/- 9 years old [mean +/- SD]) 20 +/- 5 months after transplantation, 6 liver transplant recipients receiving similar doses of cyclosporine (cyclosporine control group) and 7 normal volunteers (normal control subjects). After 3 days of a constant diet containing 87 mEq/24 h of sodium, 0.154 mol/liter saline was infused at 8 ml/kg per h for 4 h. Blood pressure and plasma vasopressin, angiotensin II, aldosterone, atrial natiuretic peptide and renin activity levels were determined before and at 30, 60, 120 and 240 min during the infusion. Urine was collected at 2 and 4 h. Blood pressure, fluid balance hormones and renal function were monitored for 48 h after the infusion. RESULTS: Blood pressure did not change in the two control groups but increased in the heart transplant recipients (+15 +/- 8/8 +/- 5 mm Hg) and remained elevated for 48 h (p < or = 0.05). Urine flow and urinary sodium excretion increased abruptly in the control groups sufficient to account for elimination of 86 +/- 9% of the sodium load by 48 h; the increases were blunted (p < or = 0.05) and delayed in the heart transplant recipients, resulting in elimination of only 51 +/- 13% of the sodium load. Saline infusion suppressed vasopressin, renin activity, angiotensin II and aldosterone in the two control groups (p < or = 0.05) but not in the heart transplant recipients. Heart transplant recipients had elevated atrial natriuretic peptide levels at baseline (p < or = 0.05), but relative increases during the infusion were similar to those in both control groups. CONCLUSIONS: Blood pressure in heart transplant recipients is salt sensitive. These patients have a blunted diuretic and natriuretic response to volume expansion that may be mediated by a failure to reflexly suppress fluid regulatory hormones. These defects in blood pressure and fluid homeostasis were not seen in liver transplant recipients receiving cyclosporine and therefore cannot be attributed to cyclosporine alone. Abnormal cardiorenal neuroendocrine reflexes, secondary to cardiac denervation, may contribute to salt-sensitive hypertension and fluid retention in heart transplant recipients.
Assuntos
Transplante de Coração/efeitos adversos , Hipertensão/etiologia , Sódio na Dieta/farmacologia , Desequilíbrio Hidroeletrolítico/etiologia , Angiotensina II/sangue , Arginina Vasopressina/sangue , Fator Natriurético Atrial/sangue , Pressão Sanguínea/fisiologia , Estudos de Casos e Controles , Ciclosporina/uso terapêutico , Feminino , Coração/inervação , Transplante de Coração/fisiologia , Humanos , Hipertensão/fisiopatologia , Imunossupressores/uso terapêutico , Transplante de Fígado/fisiologia , Masculino , Pessoa de Meia-Idade , Sistema Renina-Angiotensina/fisiologia , Cloreto de Sódio , Função Ventricular Esquerda/fisiologia , Equilíbrio Hidroeletrolítico/fisiologia , Desequilíbrio Hidroeletrolítico/fisiopatologiaRESUMO
OBJECTIVES: We sought to test the hypothesis that plasma volume (PV) expansion in heart transplant recipients (HTRs) is caused by failure to reflexively suppress the renin-angiotensin-aldosterone (RAA) axis. BACKGROUND: Extracellular fluid volume expansion occurs in clinically stable HTRs who become hypertensive. We have previously demonstrated that the RAA axis is not reflexively suppressed by a hypervolemic stimulus in HTRs. METHODS: Plasma volume and fluid regulatory hormones were measured in eight HTRs (57+/-6 years old) both before and after treatment with captopril (225 mg/day). Antihypertensive and diuretic agents were discontinued 10 days before. The HTRs were admitted to the Clinical Research Center (CRC), and, after three days of a constant diet containing 87 mEq/day of Na+, PV was measured by using the modified Evans blue dye dilution technique. After approximately four months (16+/-5 weeks), the same HTRs again discontinued all antihypertensive and diuretic agents; they were progressed to a captopril dose of 75 mg three times per day over 14 days, and the CRC protocol was repeated. RESULTS: Captopril pharmacologically suppressed (p<0.05) supine rest levels of angiotensin II (-65%) and aldosterone (-75%). The reductions in vasopressin and atrial natriuretic peptide levels after captopril did not reach statistical significance. The PV, normalized for body weight (ml/kg), was significantly reduced by 12% when the HTRs received captopril. CONCLUSIONS: Extracellular fluid volume is expanded (12%) in clinically stable HTRs who become hypertensive. Pharmacologic suppression of the RAA axis with high-dose captopril (225 mg/day) returned HTRs to a normovolemic state. These findings indicate that fluid retention is partly engendered by a failure to reflexively suppress the RAA axis when HTRs become hypervolemic.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Captopril/farmacologia , Transplante de Coração/fisiologia , Volume Plasmático/efeitos dos fármacos , Sistema Renina-Angiotensina/efeitos dos fármacos , Estudos Cross-Over , Transplante de Coração/efeitos adversos , Hemodinâmica/efeitos dos fármacos , Humanos , Rim/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Sistema Renina-Angiotensina/fisiologia , Função Ventricular Esquerda/efeitos dos fármacosRESUMO
Renovascular hypertension is potentially curable but of low prevalence. A previous retrospective study has demonstrated the use of a potentiated increase in plasma renin activity after captopril administration as a diagnostic test for renovascular hypertension; this requires two blood samples for plasma renin activity determination and three inclusive criteria for a positive test result. We applied this test prospectively to screen 100 hypertensive patients for renovascular hypertension. We evaluated 29 patients with renovascular hypertension; the remainder were diagnosed as having essential hypertension. In our patient population, a postcaptopril plasma renin activity of 5.7 ng of angiotensin per milliliter per hour (ngAl.mL-1.h-1) or greater had a 100% sensitivity and an 80% specificity for renovascular hypertension. An absolute increase in plasma renin activity with captopril of 4.7 ngAl.mL-1.h-1 or greater had a lower sensitivity of 90% and a specificity of 87%, whereas a fractional increase in plasma renin activity after captopril of 150% or higher had the lowest sensitivity of 69% and a specificity of 86%. A subgroup analysis of 38 patients who were receiving diuretic therapy demonstrated that the test sensitivity was unchanged but the specificity was reduced. In conclusion, a single postcaptopril plasma renin activity value of 5.7 ngAl.mL-1.h-1 or greater is a simplified screening test for renovascular hypertension, with excellent sensitivity and acceptable specificity. This test is well tolerated, inexpensive, and easy to perform.
Assuntos
Captopril , Hipertensão Renovascular/diagnóstico , Diuréticos/uso terapêutico , Humanos , Hipertensão/diagnóstico , Hipertensão Renovascular/epidemiologia , Valor Preditivo dos Testes , Estudos Prospectivos , Curva ROC , Renina/sangue , Sensibilidade e EspecificidadeRESUMO
Radioisotopic renal scanning after angiotensin converting enzyme inhibition (ACEI) has proven to be an exciting area for research. The biologic activity of markers such as DTPA and hippuran, when combined with the physiological effects of ACEI, may provide noninvasive methods of diagnosing both renal artery stenosis and renovascular hypertension. Recent investigators have demonstrated that the sensitivities and specificities of these tests may vary widely; these differences are probably due to variations in study design, patient population, diagnostic criteria, and outcome measurements. We have reviewed these studies and discuss these possible sources of variation and their impact on the clinical usefulness of these diagnostic tests, especially in relation to the prevalence of disease in the population. Current results suggest that the post-ACEI DTPA scan is relatively accurate in the diagnosis of renal artery stenosis, with sensitivity generally greater than 90% and specificity around 95%. However, the best results in predicting the response to angioplasty or surgery in patients with renal artery stenosis have been with the use of post-ACEI hippuran in combination with furosemide (sensitivity, 96%; specificity, 95%). With confirmation of these findings and continued investigation, it is expected that accurate noninvasive tests will be available for widespread clinical use in the near future.
Assuntos
Inibidores da Enzima Conversora de Angiotensina , Rim/diagnóstico por imagem , Inibidores da Enzima Conversora de Angiotensina/efeitos adversos , Taxa de Filtração Glomerular/efeitos dos fármacos , Humanos , Hipertensão Renovascular/diagnóstico , Hipertensão Renovascular/epidemiologia , Ácido Iodoipúrico , Rim/efeitos dos fármacos , Rim/fisiopatologia , Ácido Pentético , Prevalência , Radiografia , Obstrução da Artéria Renal/diagnóstico , Sensibilidade e EspecificidadeRESUMO
8-Iso prostaglandin F2alpha (8-ISO) is formed nonenzymatically from the attack of superoxide radical on arachidonic acid. Therefore, 8-ISO is a marker of oxidative stress in vivo. We have recently shown that short-term administration of the membrane-permeable, metal-independent superoxide dismutase mimetic tempol (4-hydroxy-2, 2, 6, 6-tetramethyl piperidinoxyl) normalizes blood pressure in spontaneously hypertensive rats (SHR). The present study was designed to test whether prolonged administration of tempol ameliorates oxidative stress and hypertension in SHR. In control SHR (n=8), mean arterial pressure and heart rate were increased and renal blood flow and glomerular filtration rate were reduced compared with control Wistar-Kyoto rats (WKY) (n=7). Twenty-four-hour renal excretion of 8-ISO was significantly increased in SHR compared with WKY. Two weeks of tempol administration in the drinking water (1 mmol/L) to SHR (n=8) decreased mean arterial pressure by 18% (162+/-8 to 134+/-6 mm Hg, P<0.05), increased glomerular filtration rate by 17% (1.6+/-0.2 to 1. 9+/-0.3 mL/min), and decreased renal excretion of 8-ISO by 39% (9. 8+/-0.7 to 6.0+/-0.7 ng/24 hours, P<0.05). In contrast, tempol administration to WKY (n=6) had no significant effect on mean arterial pressure (115+/-5 versus 118+/-8 mm Hg), glomerular filtration rate (3.0+/-0.4 versus 2.5+/-0.5 mL/min), or renal excretion of 8-ISO (7.9+/-0.4 versus 6.8+/-0.7 ng/24 hours). In conclusion, the SHR is a model of hypertension and renal vasoconstriction associated with oxidative stress. Because long-term administration of a superoxide scavenger reduces blood pressure and oxidative stress in vivo, this study suggests a role for oxygen radicals in the maintenance of hypertension in SHR.
Assuntos
Antioxidantes/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Óxidos N-Cíclicos/farmacologia , Dinoprosta/análogos & derivados , Hipertensão/fisiopatologia , Rim/efeitos dos fármacos , Administração Oral , Animais , Antioxidantes/administração & dosagem , Antioxidantes/uso terapêutico , Óxidos N-Cíclicos/administração & dosagem , Óxidos N-Cíclicos/uso terapêutico , Dinoprosta/urina , F2-Isoprostanos , Taxa de Filtração Glomerular , Hipertensão/tratamento farmacológico , Hipertensão/genética , Rim/fisiologia , Rim/fisiopatologia , Masculino , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Valores de Referência , Fluxo Sanguíneo Regional , Circulação Renal , Marcadores de Spin , Fatores de Tempo , Resistência VascularRESUMO
Infusion of L-arginine, the substrate for nitric oxide synthase, causes renal vasodilation. Since dietary salt restriction blunts the renal vasoconstrictor response to inhibition of nitric oxide synthase, we investigated the hypothesis that dietary salt intake determines the renal vascular response to L-arginine. Bolus intravenous doses of L-arginine given to anesthetized Sprague-Dawley rats caused dose-dependent increases in renal blood flow and decreases in renal vascular resistance, whereas D-arginine was not effective. The response to L-arginine was prevented by pretreatment with NG-nitro-L-arginine methyl ester. Compared with rats adapted to a high salt diet, those adapted to a low salt diet were more sensitive to the reductions in blood pressure and renal vascular resistance (threshold dose of L-arginine for renal vascular resistance: low salt, 2.9 +/- 0.9 mumol . kg-1 versus high salt, 20.0 +/- 6.2; P < .025), but the maximal changes in renal vascular resistance were similar (low salt, -43 +/- 5% versus high salt, -34 +/- 5%; P = NS). Bolus doses of L-glycine also caused dose-dependent renal vasodilation, but the renal vasodilator responses were not affected by salt intake. Preinfusion of L-arginine augmented the renal vasoconstrictor response to NG-nitro-L-arginine methyl ester in low salt but not high salt rats; after L-arginine dietary salt no longer significantly affected the renal vasoconstrictor response to NG-nitro-L-arginine methyl ester. In conclusion, renal vasodilation is more sensitive to L-arginine during salt restriction. This effect is specific for L-arginine.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Arginina/análogos & derivados , Arginina/farmacologia , Circulação Renal/efeitos dos fármacos , Sódio na Dieta/farmacologia , Vasodilatação/efeitos dos fármacos , Animais , Interações Medicamentosas , Hematócrito , Injeções Intravenosas , Masculino , NG-Nitroarginina Metil Éster , Ratos , Ratos Sprague-DawleyRESUMO
Short-term infusions of angiotensin II (Ang II) increase renal vascular resistance and thereby endothelial shear stress and nitric oxide (NO) release. Prolonged stimulation of Ang II can decrease the expression of NO synthase isoforms in the macula densa, but prolonged increases in shear stress can increase transcription of endothelial NO synthase. Therefore, we designed these studies to test the hypothesis that Ang II exerts time-dependent effects on renal NO generation as assessed from renal excretion of nitrate and nitrite, percent increases in renal vascular resistance during inhibition of NO synthase with intravenous NG -nitro-L-arginine methyl ester (L-NAME), or decreases in renal vascular resistance during stimulation of endothelial NO synthase with intravenous acetylcholine. Rats were tested during graded short-term (30 to 90 minutes intravenous) or prolonged (5 to 6 days subcutaneous) Ang II infusions that led to dose-dependent increases in blood pressure and renal vascular resistance and reductions in renal blood flow. Captopril was administered for 3 to 4 days to suppress Ang II generation. The renal excretion of nitrate and nitrite was increased during short-term Ang II infusions (from 205 +/- 22 to 331 +/- 58 pmol.min-1, P < .05) but was unchanged during prolonged Ang II infusion (control group, 197 +/- 33 versus Ang II, 245 +/- 42 pmol.min-1, P=NS). The percent increase in renal vascular resistance with L-NAME was potentiated dose dependently by short-term but not long-term Ang II infusions. The increase in renal vascular resistance with L-NAME in control rats without Ang II infusions was +150 +/- 13%. At an Ang II infusion of 200 ng.kg-1.min-1, the L-NAME-induced percent increase in renal vascular resistance was significantly (P < .01) increased compared with controls in short-term Ang II-infused rats (+369 +/- 70%) but was not significantly different in prolonged infused rats (+190 +/- 33%). Intravenous acetylcholine caused dose-dependent renal vasodilation that was not significantly changed in rats receiving short-term intravenous Ang II but was significantly (P < .005) potentiated in those receiving prolonged Ang II infusions (change in renal vascular resistance with acetylcholine at 10 micrograms.kg-1.min-1 versus control, -21.5 +/- 5.0%; with short-term Ang II, -24.9 +/- 4.5%; with long-term Ang II, -52.1 +/- 7.2%). In conclusion, short- and long-term Ang II infusions caused equivalent changes in blood pressure and renal blood flow and hence presumably equivalent increases in endothelial shear stress. However, only short-term Ang II infusions increased NO generation and the dependence of the renal circulation on NO, whereas acetylcholine-induced NO release was enhanced selectively during long-term Ang II infusions. This suggests that during long-term Ang II, renal NO release may become uncoupled from shear stress yet remains highly responsive to receptor-mediated stimulation.
Assuntos
Angiotensina II/farmacologia , Óxido Nítrico/fisiologia , Circulação Renal/efeitos dos fármacos , Acetilcolina/farmacologia , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Hemodinâmica/efeitos dos fármacos , Masculino , NG-Nitroarginina Metil Éster , Óxido Nítrico Sintase/antagonistas & inibidores , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Resistência Vascular/efeitos dos fármacos , Vasodilatação/efeitos dos fármacosRESUMO
Superoxide radical (O2-) is increased in the vessel wall of spontaneously hypertensive rats (SHR) where its blockade potentiates endothelium-dependent vasodilation. The purpose of this study was to determine the role of O2- in the hypertension and renal vasoconstriction of SHR and its interaction with nitric oxide (NO). Baseline mean arterial pressure (MAP) and renal vascular resistance were markedly elevated in SHR (n=6) compared with Wistar-Kyoto rats (WKY; n=6) (145+/-4 versus 118+/-4 mmHg, P<0.05, and 24+/-3 versus 17+/-1 mmHg x mL(-1) x min(-1), respectively; P<0.05). The stable membrane-permeable superoxide dismutase mimetic 4-hydroxy-2,2,6,6-tetramethyl piperidine-1-oxyl (tempol; 72 micromol/kg i.v.) normalized MAP (103+/-9 versus 96+/-6 mm Hg for SHR and WKY, respectively) and RVR (17+/-2 versus 15+/-1 mm Hg x mL(-1) x min(-1)) of SHR. The MAP of SHR was more sensitive and responsive to graded infusions of tempol (0, 1.8, 18, 180, and 1800 micromol x kg(-1) x h(-1) i.v.) than that of WKY. To determine whether O2- increases MAP by inactivation of NO, its synthesis was blocked in SHR with NW-nitro-L-arginine methyl ester (L-NAME, 11 micromol x kg(-1) x min(-1) i.v., n=6). Whereas tempol alone significantly reduced MAP by 32% (184+/-12 to 121 +/- 18 mm Hg, P<0.05, n=6), L-NAME infusion abolished the MAP response to tempol (187+/-8 to 186+/-4 mm Hg, n=5). In contrast, tempol did reduce MAP of SHR (188+/-7 to 161+/-7 mm Hg, P<0.05) where MAP was elevated by norepinephrine (31 nmol x kg(-1) x min(-1) i.v., n=6). Finally, to determine the longer-term effect of O2-, tempol (1.5 mmol x kg(-1) x d(-1) i.p.) was given for 7 days. Tempol had no effect on MAP in WKY (96+/-1 to 97+/-1 mmHg, n=7) but significantly decreased MAP in SHR (133+/-2 to 120+/-3 mm Hg, P<0.05, n=7). These data implicate O2- in the hypertension of SHR in vivo. The antihypertensive action of tempol depends on NO synthesis presumably because O2- inactivates NO and thus diminishes its vasodilatory actions.