RESUMO
OBJECTIVE: To investigate the effects of ibuprofen on cardiac fibrosis in a rat model of type 1 diabetes. METHODS: The diabetic model was established by injecting streptozotocin into the rats. Then, ibuprofen or pioglitazone was given by gavage for 8 weeks. The cardiac fibrosis was assessed, and the major components of the renin-angiotensin system, the transforming growth factor ß1 (TGF-ß1) and the mammalian target of rapamycin (mTOR), were evaluated by histopathological, immunohistochemical, Western blot analysis or ELISA assay. RESULTS: Obvious cardiac fibrosis was detected in the diabetic group and was alleviated by ibuprofen treatment. Angiotensin-converting enzyme (ACE), angiotensin (Ang) II and AngII type 1 receptor (AT1-R) levels were higher, and ACE2, Ang(1-7) and Mas receptor (Mas-R) were lower in the diabetic group. The ratio of ACE to ACE2 was raised in the diabetic group. All these changes were ameliorated by ibuprofen. TGF-ß1 and mTOR were raised in the hearts of the diabetic group and were attenuated by ibuprofen treatment. There was no significant difference between the ibuprofen and the pioglitazone groups. CONCLUSION: Ibuprofen could ameliorate the cardiac fibrosis in diabetic rats by reduction of the ACE/AngII/AT1-R axis and enhancement of the ACE2/Ang(1-7)/Mas-R axis, leading to a decrease in TGF-ß1 and mTOR.
Assuntos
Cardiotônicos/farmacologia , Diabetes Mellitus Tipo 1/prevenção & controle , Angiopatias Diabéticas/prevenção & controle , Ibuprofeno/farmacologia , Miocárdio/patologia , Enzima de Conversão de Angiotensina 2 , Animais , Diabetes Mellitus Experimental/prevenção & controle , Regulação para Baixo/fisiologia , Fibrose/prevenção & controle , Masculino , Miocárdio/metabolismo , Peptidil Dipeptidase A/metabolismo , Ratos Sprague-Dawley , Serina-Treonina Quinases TOR/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Regulação para Cima/fisiologiaRESUMO
Increased plasma catecholamine levels are associated with a high risk of perioperative cardiac events in aged individuals undergoing non-cardiac surgical interventions. Given the different effects of ß1-adrenoreceptor (ß1AR) and ß2-adrenoreceptor (ß2AR) stimulation by catecholamine in cardiomyocytes, this study evaluated whether simultaneous inhibition of ß1AR and activation of ß2AR is better than separate application in reducing the risk of perioperative cardiac events in aged rats undergoing non-cardiac surgery. Male aged Sprague-Dawley (SD) rats were divided into five groups. Normal group received no treatment. Surgery group received an abdominal surgery with hypoxia. ß1- group, ß2+ group, ß2+ group and ß1+ß2+ group received surgery and hypoxia with metoprolol (100 mg/kg·d), fenoterol (250 µg/kg·d) or both, respectively. The drugs were given three days before surgery with treatment continued through post-surgical day 7. The results showed that simultaneous activation of ß2AR with a ß2AR agonist and inhibition of ß1AR with a selective ß1AR blocker normalized myocardial oxygen consumption, decreased myocardial damage, augmented cardiomyocyte survival, improved cardiac function, reduced the incidence of arrhythmia, thus decreasing the occurrence of cardiac events in perioperative aged rats undergoing non-cardiac surgery. The results demonstrated that combined use of ß2AR agonist and ß1AR blocker achieved better general effects than use of either one alone. Our results provide a new insight into preventing perioperative cardiac events for elderly patients undergoing surgical stress.
Assuntos
Agonistas de Receptores Adrenérgicos beta 2/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Metoprolol/farmacologia , Miócitos Cardíacos/efeitos dos fármacos , Receptores Adrenérgicos beta 1/efeitos dos fármacos , Receptores Adrenérgicos beta 2/efeitos dos fármacos , Estresse Fisiológico , Envelhecimento , Animais , Procedimentos Cirúrgicos Cardíacos , Coração , Masculino , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacosRESUMO
The present study was designed to investigate the effect of hydrogen sulfide on cellular senescence of human umbilical vascular endothelial cells (HUVECs CC-2517) and its underlying mechanism. The premature senescence-like phenotype HUVECs (the fourth passage) was induced by treatment with nicotinamide (NAM, an inhibitor of SIRT1, 5 mmol/L, 12 h). Cells were cultured with sodium hydrosulfide (NaHS, 12.5, 25, 50 and 100 µmol/L) for 48 h in premature senescence-like phenotype HUVECs. The fourth passage of HUVECs was considered as young group. Senescence-associated (SA)-ß-galactosidase activities were detected to evaluate cell senescence, and the expression of SA heterochromatin foci (SAHF) was visualized by DAPI DNA staining. The mRNA and protein levels of SIRT1 were detected using RT-PCR and western blotting analysis, respectively. The results showed that ß-galactosidase positive cells and the formation of SAHF were markedly increased after treatment with NAM (5 mmol/L) for 12 h. We also found that NaHS (12.5 µmol/L) had no effect on the percentage of SA ß-gal positive cells and the expression of SAHF, and the hallmarks decreased at the concentration of 25 and 50 µmol/L, reaching the minimum at 50 µmol/L, while the percentage of SA ß-gal positive cells and the expression of SAHF increased at the concentration of 100 µmol/L. Furthermore, we found that both on protein and mRNA levels of SIRT1 in the Y+N+S50 group was significantly increased compared with that in Y+N group. In conclusion, NaHS delays senescence of HUVECs induced by NAM via upregulation of SIRT1 expression.
Assuntos
Envelhecimento/efeitos dos fármacos , Senescência Celular/efeitos dos fármacos , Sulfeto de Hidrogênio/administração & dosagem , Sirtuína 1/biossíntese , Envelhecimento/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana , Humanos , Peróxido de Hidrogênio/metabolismo , Niacinamida/administração & dosagem , RNA Mensageiro/biossíntese , Sirtuína 1/antagonistas & inibidores , Ativação TranscricionalRESUMO
The present study was aimed to investigate the effect of pretreatment with Danshensu (DSS) on rat aortic endothelial cells (RAECs) senescence and the underlying mechanisms. Cultured RAECs at fourth and twelfth passages were taken as young and old groups, respectively. DSS and DSS+nicotinamide (DSS+N) groups were incubated with DSS and DSS in combination with nicotinamide, an inhibitor of silent information regulator 1 (SIRT1), from the fourth to twelfth passage, respectively. The cell status of senescence was determined by the senescence-associated ß-galactosidase (SA ß-gal) staining, and 4,6-diamino-2-phenyl indole (DAPI) fluorescent dye was used to detect senescence associated heterochromatin foci (SAHF) formation; Thiobarbituric acid (TBA) and colorimetric methods were used to evaluate malondialdehyde (MDA) and H2O2contents; Western blot was employed to analysis the expressions of xanthine oxidase (XOD), SIRT1 and superoxide dismutase 2 (SOD2) in the RAECs. The results showed that, in comparison with young group, the old group exhibited higher SA ß-gal positive and SAHF formation rates, as well as higher MDA and H2O2levels (P < 0.05 or P < 0.01), whereas DSS pretreatment reduced SA ß-gal positive and SAHF formation rates, decreased MDA and H2O2 contents (P < 0.05 or P < 0.01). The protection of DSS was reversed by nicotinamide. Compared with the young group, the old group showed higher expression levels of XOD, but lower SIRT1 and SOD2expression levels (P < 0.05 or P < 0.01). With the pretreatment of DSS, the expression of XOD was declined, and the expression levels of SIRT1 and SOD2were elevated, while nicotinamide reversed the effects of DSS. These results suggest that DSS delays senescence of RAECs via up-regulation of SIRT1.
Assuntos
Senescência Celular/efeitos dos fármacos , Células Endoteliais/citologia , Lactatos/farmacologia , Sirtuína 1/metabolismo , Superóxido Dismutase/metabolismo , Animais , Aorta/citologia , Células Cultivadas , Peróxido de Hidrogênio/metabolismo , Niacinamida/farmacologia , Ratos , Regulação para CimaRESUMO
The present study was aimed to observe the protective effect of exogenous hydrogen sulfide (H2S) on vascular structural and functional changes of aorta in D-galactose-induced subacute aging rats. Adult male SD rats were randomly divided to five groups: the vehicle group, the D-galactose (D-gal) group, and the three NaHS groups treated with low (1 µmol·kg⻹·d⻹), middle (10 µmol·kg⻹·d⻹) or high (100 µmol·kg⻹·d⻹) dose of NaHS respectively. The D-gal group rats were given subcutaneously injection of 125 mg/kg D-gal per day for eight weeks to induce subacute aging model. In the NaHS group, D-gal was administered as above but with NaHS intraperitoneally injected at a dosage of 1, 10, 100 µmol·kg⻹·d⻹ respectively. Equivalent volumes of saline were administered per day for eight weeks in vehicle group. Morphological changes of aorta were observed by HE and Masson staining. The level of H2S in serum, the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA), as well as anti-superoxide anions in vascular tissue were determined by spectrophotometry. Angiotensin II (AngII) levels in plasma were measured using competitive enzyme immunoassay. The expression of angiotensin II type 1 receptor (AT1R) in aorta was determined by Western blot. The results showed that the aging aortic morphologic changes in model rats were ameliorated in NaHS groups. Decreased vascular endothelial exfoliative cells and vascular smooth muscle cell (SMC) proliferation were shown in NaHS groups by HE staining. Masson staining analysis showed reduced relative contents of collagen fibers (P < 0.05) and SMC (P < 0.05) in NaHS groups. Compared to vehicle group, serum concentration of H2S in D-gal group was decreased, while it was increased in NaHS groups after treatment with NaHS (P < 0.05). In the D-gal group, the concentration of AngII in plasma was significantly increased compared with that in vehicle group, while it was decreased in NaHS groups (P < 0.05). Moreover, levels of vascular tissue anti-superoxide anion and the activity of SOD were obviously higher, MDA was significantly lower in all NaHS treated groups than those in the D-gal group respectively (P < 0.05). Western blot analysis showed that the expression of AT1R was increased in D-gal group compared with that in vehicle group, while it was decreased after treatment with NaHS compared with that in D-gal group (P < 0.05). These results suggest that exogenous H2S can ameliorate the age-related changes of aortic morphology, decrease the concentration of AngII in plasma, down-regulate the expression of AT1R in vascular tissue, and mitigate the level of oxidative stress. These changes delay the vascular aging in aging rats ultimately.
Assuntos
Envelhecimento/efeitos dos fármacos , Aorta/patologia , Sulfeto de Hidrogênio/farmacologia , Angiotensina II/metabolismo , Animais , Proliferação de Células , Células Endoteliais/metabolismo , Galactose/efeitos adversos , Masculino , Malondialdeído/metabolismo , Miócitos de Músculo Liso/metabolismo , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Receptor Tipo 2 de Angiotensina/metabolismo , Sulfetos/farmacologia , Superóxido Dismutase/metabolismoRESUMO
CONTEXT: Heart failure (HF) is a progressive deterioration in heart function associated with overactivity of the sympathetic nervous system. Elevated sympathetic nervous system activity down regulates the ß-adrenergic signal system, suppressing ß-adrenoceptors (ß-ARs)-mediated contractile support in the failing heart. OBJECTIVE: We investigated the effects of ß(2)-AR gene transfer on shortening amplitude of isolated ventricular myocytes under chronic exposure to isoprenaline (ISO), and further determine the contributions of ß(1)-AR and ß(2)-AR to the contraction. MATERIALS AND METHODS: Cardiomyocytes were isolated from adult rat hearts and then transfected with ß(2)-AR gene using an adenovirus vector. Four hours after the infection, cardiomyocytes were treated with ISO for another 24 hours to imitate high levels of circulating catecholamines in HF. Western blotting was performed to measure myocardial protein expression of ß(2)-AR. Video-based edge-detection system was used to evaluate basal and ISO-stimulated shortening amplitudes of cardiomyocytes. RESULTS: ß(2)-AR gene transfer increased ß(2)-AR protein content. Chronic ISO stimulation produced a negative inotropic response, whereas acute ISO stimulation showed a positive inotropic response. ß(2)-AR gene transfer had no significant effects on shortening amplitude of cardiomyocytes under normal conditions, but enhanced the blunted contraction of cardiomyocytes under pathological conditions induced by chronic ISO stimulation, and the effect was inhibited by ß(1)-AR antagonist, CGP 20712A, instead of ß(2)-AR antagonist, ICI 118,551. DISCUSSION AND CONCLUSIONS: We conclude that ß(2)-AR gene transfer in isolated ventricular myocytes under chronic ISO stimulation improves cellular contraction, and the beneficial effects might be mediated by improving ß(1)-adrenoceptor responsiveness.
Assuntos
Isoproterenol/farmacologia , Contração Miocárdica/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Receptores Adrenérgicos beta 1/metabolismo , Receptores Adrenérgicos beta 2/genética , Antagonistas Adrenérgicos beta/farmacologia , Animais , Técnicas de Transferência de Genes , Terapia Genética , Insuficiência Cardíaca/patologia , Imidazóis/farmacologia , Miócitos Cardíacos/efeitos dos fármacos , Cultura Primária de Células , Propanolaminas/farmacologia , Ratos , Receptores Adrenérgicos beta 1/efeitos dos fármacos , Receptores Adrenérgicos beta 2/efeitos dos fármacosRESUMO
BACKGROUND: The potential biological significance of hydrogen sulfide (H2S) has attracted growing interests in recent years, but its role in the myogenic response of rat cerebral arterioles has not been explored. METHODS AND RESULTS: Rats were injected with NaHS (an H2S donor, 2-200 µmol·kg⻹·day⻹, i.p.) or saline for 3 weeks. MBP was measured with a tail-cuff method. Cerebral arterioles were isolated and cannulated in an organ bath system, and vessel diameters were measured with an image-shearing device. Changes in diameter in response to stepwise increases in intravascular pressure (20-120 mmHg) were investigated under no-flow conditions. After the treatments, plasma H2S increased and MBP decreased significantly. NaHS reduced the myogenic response in a dose-dependent manner. This effect was markedly attenuated by glibenclamide, a K(ATP) channel blocker. Blockade of nitric oxide (NO) production with NG-nitro-L-arginine methyl ester (L-NAME, a NO synthase inhibitor) enhanced, whereas removal of the endothelium abolished the inhibitory role of NaHS on the myogenic response. CONCLUSIONS: For the first time it has been demonstrated that H2S decreases the myogenic response of cerebral arterioles in vivo, and this effect is endothelium-dependent and partially mediated by K(ATP) channels.
Assuntos
Cérebro/irrigação sanguínea , Sulfeto de Hidrogênio/metabolismo , Vasoconstrição , Vasodilatação , Animais , Arteríolas/efeitos dos fármacos , Arteríolas/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Relação Dose-Resposta a Droga , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Inibidores Enzimáticos/farmacologia , Glibureto/farmacologia , Sulfeto de Hidrogênio/sangue , Canais KATP/antagonistas & inibidores , Canais KATP/metabolismo , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/metabolismo , Bloqueadores dos Canais de Potássio/farmacologia , Ratos , Ratos Sprague-Dawley , Sulfetos/metabolismo , Sulfetos/farmacologia , Vasoconstrição/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos , Vasodilatadores/farmacologiaRESUMO
The present study aimed to investigate the protective effect and mechanism of hydrogen sulfide donor NaHS administration against gastric mucosal injury induced by gastric ischemia-reperfusion (GI-R) in rats. GI-R injury was induced by clamping the celiac artery of adult male SD rats for 30 min and followed by reperfusion for 1 h. The rats were randomly divided into sham group, GI-R group, NaHS group, glibenclamide group and pinacidil group. Gastric mucosal damage was analyzed with macroscopic injured area, deep damage was assessed with histopathology scores, and the hydrogen sulfide concentration in plasma was determined by colorimetric method. The results showed that pretreatment of NaHS significantly reduced the injured area and deep damage of the gastric mucosa induced by GI-R. However, NaHS did not significantly alter the levels of hydrogen sulfide in plasma 14 d after NaHS administration. The gastric protective effect of NaHS during reperfusion could be attenuated by glibenclamide, an ATP-sensitive potassium channel (K(ATP)) blocker. However, K(ATP) opener pinacidil inhibited the GI-R-induced injury. These results suggest that exogenous hydrogen sulfide plays a protective role against GI-R injury in rats possibly through modulation of K(ATP) channel opening.
Assuntos
Sulfeto de Hidrogênio/metabolismo , Precondicionamento Isquêmico/métodos , Canais KATP/fisiologia , Traumatismo por Reperfusão/prevenção & controle , Estômago/irrigação sanguínea , Animais , Mucosa Gástrica/patologia , Canais KATP/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Sulfetos/farmacologiaRESUMO
The present study was aimed to investigate the effect of pretreatment with hydrogen sulfide (H2S) on human umbilical vein endothelial cells (HUVECs) senescence and the underlying mechanism. Cultured HUVECs at twelfth and fourth passages were taken as old and young groups, respectively. Sodium hydrosulfide (NaHS, donor of H2S) group was treated with NaHS from fourth to twelfth passage. The cell senescence was determined by senescence-associated ß-galactosidase (SA ß-gal) staining. DAPI fluorescent dye was used to detect cellular apoptosis. Western blot was used to analyze the expression levels of xanthine oxidase (XOD), manganese-superoxide dismutase (Mn-SOD) and the subunits p67(phox) of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in the HUVECs. Colorimetric method was used to detect SOD activity and cellular hydrogen peroxide (H2O2) level. The results showed that, compared with young group, the old group exhibited higher SA ß-gal positive rate and cellular apoptosis, while NaHS pretreatment decreased SA ß-gal positive rate and cellular apoptosis. Compared with the young group, the old group showed increased expression levels of XOD and p67(phox), as well as lower Mn-SOD expression level. With the pretreatment of NaHS, the up-regulations of XOD and p67(phox) levels and down-regulation of Mn-SOD level were inhibited. Compared with the young group, the old group showed lower SOD activity and higher H2O2 level, whereas NaHS pretreatment reversed the changes of SOD activity and H2O2 level. These results suggest that H2S delays senescence of HUVECs through lessening oxidative stress.
Assuntos
Apoptose , Senescência Celular/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/citologia , Sulfeto de Hidrogênio/farmacologia , Estresse Oxidativo , Regulação para Baixo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Peróxido de Hidrogênio/metabolismo , Fosfoproteínas/metabolismo , Superóxido Dismutase/metabolismo , Xantina Oxidase/metabolismo , beta-Galactosidase/metabolismoRESUMO
Hydrogen sulfide (H2S) is an endogenous gasotransmitter produced in mammalian cells. It is responsible for physiological functions in many organs and systems, with attention focused mainly on the cardiovascular and nervous systems. In the vascular system, H2S produces biphasic effects in regulation of vascular tone. At lower concentrations, it induces vasoconstriction predominantly via decreasing cyclic adenosine monophosphate in smooth muscle cell and inhibiting the production and bioavailability of nitric oxide. At higher concentrations, it produces vasorelaxation mainly through opening of KATP channels and induction of intracellular acidification. Scavenging reactive oxygen species and elevation of cyclic guanosine monophosphate are also implicated in the vasorelaxant response. This review presents an overview of the current knowledge of H2S in the vascular system, with special emphasis and discussion on the involvement of various signaling pathways and ion channels based on current understanding and reported literature till date.
Assuntos
Sulfeto de Hidrogênio/metabolismo , Canais Iônicos/fisiologia , Transdução de Sinais/fisiologia , Animais , Humanos , Espécies Reativas de Oxigênio/metabolismo , Vasoconstrição/fisiologia , Vasodilatação/fisiologiaRESUMO
Low-salt (LS) diet has been considered to be beneficial in the prevention and treatment of hypertension; however, it also increases plasma angiotensin (ANG) II and may cause adverse cardiovascular effects, such as endothelial dysfunction. We assessed endothelial function of coronary arterioles and vascular superoxide production, as a function of LS diet. Dogs were fed with LS (0.05% NaCl) or a normal-salt (NS, 0.65% NaCl) diet for 2 wk. There were threefold increases in plasma ANG II, associated with a 60% reduction in flow-induced dilation (FID) in coronary arterioles of LS compared with NS dogs. In vessels of NS dogs, FID was primarily mediated by nitric oxide (NO), as indicated by an eliminated FID by N(ω)-nitro-l-arginine methyl ester (l-NAME). In vessels of LS dogs, however, FID was eliminated. Administration of apocynin, a NAD(P)H oxidase inhibitor, partially restored FID and additional l-NAME eliminated FID. Generation of superoxide, measured with dihydroethidium, was significantly greater in vessels of LS than in NS dogs, which was further increased in response to ANG II or phorbol 12,13-dibutyrate, an agonist of protein kinase C (PKC). The enhanced superoxide was normalized by apocynin, losartan (a blocker of angiotensin type 1 receptor), and chelerythrine chloride (an antagonist of PKC). Western blotting indicated an upregulation of gp91(phox) and p47(phox), associated with increased expression of phosphorylated PKC in vessels of LS dogs. In separate experiments, dogs were fed simultaneously with LS and losartan (LS + Losa) for 2 wk. There was a significant increase in plasma ANG II in LS + Losa dogs, which, however, was associated with normal FID and gp91(phox) expression in coronary arterioles. In conclusion, LS led to endothelial dysfunction, as indicated by an impaired flow-induced dilation caused by decreasing NO bioavailibility, a response that involves angiotensin-induced activation of PKC that, in turn, activates vascular NAD(P)H oxidase to produce superoxide.
Assuntos
Angiotensina II/metabolismo , Vasos Coronários/fisiopatologia , NADPH Oxidases/metabolismo , Proteína Quinase C/metabolismo , Cloreto de Sódio na Dieta/farmacologia , Vasodilatação/efeitos dos fármacos , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Animais , Arteríolas/efeitos dos fármacos , Arteríolas/metabolismo , Arteríolas/fisiopatologia , Pressão Sanguínea/efeitos dos fármacos , Vasos Coronários/efeitos dos fármacos , Vasos Coronários/metabolismo , Cães , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Endotélio Vascular/fisiopatologia , Losartan/farmacologia , Masculino , Modelos Animais , Óxido Nítrico/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Superóxidos/metabolismoRESUMO
Estrogen plays a cardioprotective role in female rat hearts subjected to ischemia/reperfusion injury. The its effects are, at least partially, associated with decreased cardiomyocyte contraction and increased expression of beta(2)-adrenoceptor (beta(2)-AR). We tested whether beta(2)-AR could be involved in cardioprotection against ischemic damage and whether the roles of beta(2)-AR were dependent on estrogenic environment. We first determined the effects of hypoxia/reoxygenation (H/R) on cardiomyocyte shortening in female rats. We then determined the roles of beta(2)-AR in cardiomyocyte shortening, lactate dehydrogenase (LDH) release in culture medium, and cell death during hypoxia in isolated myocytes from female rats. We further determined the effects of estrogen on the roles of beta(2)-AR during hypoxia. H/R induced short-term hibernation and stunning at the level of ventricular myocytes from normal female rats. Inhibition of beta(2)-AR with ICI118,551 significantly elevated adrenergic contractile reserve, myocardial injury, and cell death in normal female rats during hypoxia, whereas ovariectomy (OVX) prominently enhanced myocyte contraction, myocardial injury, and cell death, and deprived the alternations in normal female rats. These changes were restored to normal by estrogen replacement (OVX+E(2)). These data suggest that beta(2)-AR may be involved in the cardioprotection against ischemic damage, and the cardioprotection may depend on estrogenic environment.
Assuntos
Citoproteção/efeitos dos fármacos , Estrogênios/farmacologia , Ventrículos do Coração/citologia , Miócitos Cardíacos/citologia , Miócitos Cardíacos/efeitos dos fármacos , Receptores Adrenérgicos beta 2/metabolismo , Animais , Hipóxia Celular/efeitos dos fármacos , Separação Celular , Sobrevivência Celular/efeitos dos fármacos , Feminino , Isoproterenol/farmacologia , Miócitos Cardíacos/metabolismo , Ovariectomia , Propanolaminas/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
Ginkgo biloba extract is an important natural product for treatment of cerebral and cardiovascular diseases, whereas ginkgolide B (GB) is a main component of it. Its effects on ischemic heart and ventricular contractile function in Sprague-Dawley male rats are unclear yet. In the present study, we investigated the function of isolated hearts subjected to ischemia-reperfusion (IR) with or without GB pretreatment by using Millar transducer instruments. We also tested the left ventricular cardiomyocyte shortening amplitude after IR with different concentrations of GB pretreatment for 0.1, 1.0, 2.0, 5.0, and 10.0 microM. The infarct size was tested by triphenyltetrazolium chloride. The release of lactate dehydrogenase (LDH) in the coronary effluent was determined with LDH kit. The expressions of Bcl-2 and Bax were assessed by Western blotting. We found that GB improved the function of left ventricle from IR injury and decreased infarct size and the release of LDH. The cardiomyocyte shortening amplitude depended on different concentrations of GB, which increased significantly at 2.0 microM GB (P < 0.01). The expression of protein Bcl-2 was upregulated by GB and the ratio of Bax to Bcl-2 was decreased by GB. Our results showed that GB can partly prevent IR injury in rat heart.
Assuntos
Ginkgolídeos/farmacologia , Lactonas/farmacologia , Contração Miocárdica/fisiologia , Miócitos Cardíacos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Traumatismo por Reperfusão/metabolismo , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Coração/efeitos dos fármacos , Coração/fisiopatologia , Ventrículos do Coração/citologia , Ventrículos do Coração/metabolismo , Ventrículos do Coração/fisiopatologia , Masculino , Miócitos Cardíacos/metabolismo , Perfusão , Ratos , Ratos Sprague-Dawley , Transdutores/efeitos adversos , Regulação para Cima/efeitos dos fármacos , Proteína X Associada a bcl-2/metabolismoRESUMO
This study investigated the mechanisms responsible for the estrogen-dependent, cytochrome P450 (CYP)-mediated dilator responses to shear stress in arterioles of NO-deficient female rats and mice. Flow-induced dilation (FID) was assessed in isolated arterioles from N(G)-nitro-L-arginine methyl ester (L-NAME)-treated male and ovariectomized female rats before and after overnight incubation with 17beta-estradiol (17beta-E2, 10(-9) mol/L). In control conditions, prostaglandins (PGs) mediated FID, because indomethacin (INDO) abolished the responses. After incubation of the vessels with 17beta-E2, the basal tone of arterioles was significantly reduced and FID was augmented. INDO did not affect the dilation of the vessels incubated with 17beta-E2. Dilations of these vessels, however, were eliminated by PPOH and miconazole, inhibitors of CYP/epoxygenase. Simultaneous incubation of the vessels with 17beta-E2 plus ICI, 182,780, an estrogen receptor antagonist, or wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3K) phosphorylation or the transcriptional inhibitor DRB, prevented the reduced arteriolar tone and the enhanced CYP-mediated FID caused by incubation of vessels with 17beta-E2. Western blot analysis indicated a significantly increased phospho-Akt level in arterioles incubated with 17beta-E2 compared with those without 17beta-E2. The enhanced phospho-Akt in response to 17beta-E2 was localized, by immunohistochemistry, to arteriolar endothelial cells. Moreover, GC-MS analysis indicated a significantly increased production of epoxyeicosatrienoic acids, vasodilator metabolites of CYP/epoxygenase, in arterioles incubated with 17beta-E2, a response that was prevented by ICI 182780 and wortmannin, respectively. Thus, estrogen, via a receptor-dependent, PI3K/Akt-mediated pathway, transcriptionally upregulates CYP activity, leading to an enhanced arteriolar response to shear stress.
Assuntos
Arteríolas/efeitos dos fármacos , Sistema Enzimático do Citocromo P-450/fisiologia , Estradiol/análogos & derivados , Estradiol/farmacologia , Hemorreologia , Óxido Nítrico/deficiência , Fosfatidilinositol 3-Quinases/fisiologia , Processamento de Proteína Pós-Traducional/fisiologia , Proteínas Serina-Treonina Quinases , Proteínas Proto-Oncogênicas/fisiologia , Vasodilatação/efeitos dos fármacos , Androstadienos/farmacologia , Animais , Arteríolas/enzimologia , Caproatos/farmacologia , Sistema Enzimático do Citocromo P-450/biossíntese , Sistema Enzimático do Citocromo P-450/genética , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Indução Enzimática/efeitos dos fármacos , Indução Enzimática/fisiologia , Inibidores Enzimáticos/farmacologia , Feminino , Fulvestranto , Indometacina/farmacologia , Masculino , Miconazol/farmacologia , NG-Nitroarginina Metil Éster/farmacologia , Inibidores de Fosfoinositídeo-3 Quinase , Fosforilação , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Estresse Mecânico , Transcrição Gênica/efeitos dos fármacos , Vasodilatação/fisiologia , WortmaninaRESUMO
INTRODUCTION: Diabetes mellitus induces microangiopathic changes that lead to endothelial dysfunction. This study investigated the effect of Xiaokening, a type of Chinese compound medicine, on the mesenteric arteriolar endothelial cell function of diabetic rats and its underlying mechanism. METHODS: Diabetes mellitus was induced in rat models via intraperitoneal injection of 60 mg/kg streptozotocin and observed over three weeks. Mesenteric arterioles, which were isolated in a cannulated and pressurised state, were incubated with intravascular injections of 1, 3 or 5 g/L Xiaokening for 24, 48 or 72 hours. The effects of Xiaokening on the release of nitric oxide (NO) on the mesenteric arterioles were detected under shear stress of 1, 10 and 20 dyn/cm(2). Biochemical methods were used to determine the activities of superoxide dismutase (SOD) and xanthine oxidase (XO). The expressions of endothelial NO synthase (eNOS), SOD and XO in the mesenteric arterioles were assessed using Western blot. RESULTS: Compared to normal rat arterioles, less NO was released in the mesenteric arterioles of diabetic rats. Xiaokening was found to have a concentration- and time-dependent effect on NO release; when the shear stress was increased, there was a gradual increase in the release of NO. Compared to normal arterioles, the expression of eNOS in the mesenteric arterioles of diabetic rats was lower. Incubation with Xiaokening increased SOD activity and expression, and decreased XO activity and expression in the mesenteric arterioles of the diabetic rats. CONCLUSION: Xiaokening was able to significantly increase NO release and improve the endothelial function of mesenteric arterioles through antioxidative mechanisms.
Assuntos
Diabetes Mellitus Experimental/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Células Endoteliais/efeitos dos fármacos , Óxido Nítrico Sintase Tipo III/metabolismo , Animais , Antioxidantes/química , Arteríolas/enzimologia , Relação Dose-Resposta a Droga , Injeções Intraperitoneais , Masculino , Óxido Nítrico/metabolismo , Oxigênio/metabolismo , Ratos , Ratos Sprague-Dawley , Estreptozocina , Superóxido Dismutase/metabolismo , Xantina Oxidase/metabolismoRESUMO
BACKGROUND: The NF-κB pathway and chemokine (C-C motif) ligand 5 (CCL5) are involved in pain modulation; however, the precise mechanisms of their interactions in chronic neuropathic pain have yet to be established. METHODS: The present study examined the roles of spinal NF-κB and CCL5 in a neuropathic pain model after chronic constriction injury (CCI) surgery. CCI-induced pain facilitation was evaluated using the Plantar and von Frey tests. The changes in NF-κB and CCL5 expression were analyzed by immunohistochemistry and Western blot analyses. RESULTS: Spinal NF-κB and CCL5 expression increased after CCI surgery. Repeated intrathecal infusions of pyrrolidine dithiocarbamate (PDTC, a NF-κB inhibitor) decreased CCL5 expression, inhibited the activation of microglia and astrocytes, and attenuated CCI-induced allodynia and hyperalgesia. Intrathecal injection of a CCL5-neutralizing antibody attenuated CCI-induced pain facilitation and also suppressed spinal glial cell activation after CCI surgery. However, the CCL5-neutralizing antibody did not affect NF-κB expression. Furthermore, selective glial inhibitors, minocycline and fluorocitrate, attenuated the hyperalgesia induced by intrathecal CCL5. CONCLUSIONS: The inhibition of spinal CCL5 expression may provide a new method to prevent and treat nerve injury-induced neuropathic pain.
Assuntos
Quimiocina CCL5/biossíntese , NF-kappa B/biossíntese , Neuralgia/genética , Animais , Astrócitos/metabolismo , Astrócitos/patologia , Quimiocina CCL5/antagonistas & inibidores , Quimiocina CCL5/genética , Hiperalgesia/metabolismo , Hiperalgesia/fisiopatologia , NF-kappa B/genética , Neuralgia/fisiopatologia , Neuroglia/metabolismo , Neuroglia/patologia , Ratos , Medula Espinal/metabolismo , Medula Espinal/fisiopatologiaRESUMO
A rat model of gastric ischemia-reperfusion injury (GI-RI) was established by clamping the celiac artery for 30 min and allowing reperfusion for 1 h, on which the regulatory effect of the paraventricular nucleus (PVN) and its neural mechanisms were investigated. The results were: 1. Electrical stimulation of the PVN obviously attenuated the GI-RI. Microinjection of L-glutamic acid into PVN produced an effect similar to that of PVN stimulation. 2. Electrolytic ablation of the PVN aggravated the GI-RI. 3. Nucleus tractus solitarius (NTS) ablation could eliminate the protective effect of electrical stimulation of PVN on GI-RI. 4. Hypophysectomy did not alter the effect of electrical stimulation of PVN. 5. Vagotomy or sympathectomy both could increase the effect of PVN stimulation on GI-RI. These results indicate that the PVN participates in the development of GI-RI as a specific area in the CNS, exerting protective effects on the GI-RI. The NTS and vagus and sympathetic nerve may be involved in the regulative mechanism of PVN on GI-RI, but the PVN mechanism here is independent of the PVN-hypophyseal pathway.
Assuntos
Núcleo Hipotalâmico Paraventricular/fisiopatologia , Traumatismo por Reperfusão/fisiopatologia , Sistema Nervoso Simpático/fisiopatologia , Animais , Modelos Animais de Doenças , Estimulação Elétrica , Ácido Glutâmico/farmacologia , Hipofisectomia , Masculino , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/lesões , Ratos , Ratos Sprague-Dawley , Núcleo Solitário/fisiopatologia , Núcleo Solitário/cirurgia , Sistema Nervoso Simpático/cirurgia , VagotomiaRESUMO
The effects of electrical and chemical stimulation and electrolytic lesion of lateral hypothalamic area (LHA) on gastric ischemia-reperfusion injury (GI-RI) were investigated in rats whose celiac arteries were clamped for 30 min and reperfused for 60 min by removal of the clamp. The results are as follows. (1) Electrical stimulation of LHA could aggravate GI-RI in an intensity-dependent manner by using 0.2, 0.4 or 0.6 mA current respectively. Microinjection of L-glutamic acid into LHA resulted in a similar effect to that of electrical stimulation of LHA on GI-RI. After electrolytic lesion of bilateral LHA, the area of gastric mucosal injury induced by gastric ischemia-reperfusion (GI-R) was smaller than that by electrical stimulation of LHA plus GI-R. (2) Dorsal vagal complex (DVC) lesion or vagotomy could eliminate the effect of electrical stimulation of LHA on GI-RI. (3) Electrical stimulation of LHA increased the content of malondialdehyde (MDA) but decreased the activity of superoxide dismutase (SOD) of ischemia-reperfusion (I-R) gastric mucosa. (4) Electrical stimulation of LHA plus gastric I-R increased gastric juice volume and total acid output, but there were no significant changes in acidity, pepsin activity and gastric barrier mucus. These results indicate that the LHA is an area in the CNS exerting aggravate effects on GI-RI. The DVC and vagus may be involved in the regulative effects of LHA on GI-RI. These effects are associated with increases in gastric mucosal MDA content, gastric juice volume, and total acid output, and a decrease in SOD activity.Acidity, pepsin activity and gastric barrier mucus do not seem to play an important role.
Assuntos
Estimulação Elétrica , Mucosa Gástrica/patologia , Traumatismo por Reperfusão/patologia , Animais , Mucosa Gástrica/irrigação sanguínea , Mucosa Gástrica/metabolismo , Região Hipotalâmica Lateral/metabolismo , Masculino , Malondialdeído/metabolismo , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/metabolismo , Superóxido Dismutase/metabolismoRESUMO
The effects of paraventricular nucleus (PVN) stimulation and vasopressin on gastric ischemia-reperfusion injury (GI-RI) were investigated in male SD rats of which the celiac artery was clamped for 30 min and reperfused for 1 h by removal of the clamp. The results were as follows. Both electrical and chemical stimulation of the PVN obviously attenuated the GI-RI. Bilateral electrolytic lesion of the nucleus tractus solitarius (NTS) or microinjection of AVP-V(1) receptor antagonist into the NTS could eliminate the protective effect of electrical stimulation of the PVN on GI-RI. Hypophysectomy did not influence the effect of electrical stimulation of the PVN. Both vagotomy and sympathectomy could increase the effect of stimulating PVN on GI-RI. Microinjection of arginine-vasopressin (AVP) into the PVN also attenuated the effect on GI-RI. These results suggest that the PVN and AVP participate in the regulation of GI-RI and play an important role in protection against GI-RI. This protective effect of PVN on GI-RI might be mediated by activation of AVP-ergic neurons in the PVN, which release AVP from the descending projection fibers and activate the AVP-V(1) receptors on the NTS neurons. The vagus and sympathetic nerves are involved in the efferent pathway exerting their effects on GI-RI. Hypophysis does not seem to be involved in the protective effect of PVN stimulation.
Assuntos
Núcleo Hipotalâmico Paraventricular/fisiologia , Traumatismo por Reperfusão/terapia , Vasopressinas/farmacologia , Vias Aferentes/fisiologia , Animais , Estimulação Elétrica , Masculino , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/fisiopatologia , Estimulação Química , Estômago/irrigação sanguínea , Sistema Nervoso Simpático/fisiologia , Nervo Vago/fisiologiaRESUMO
The present study examined the effects of intrathecal use of resveratrol on pain hypersensitivities, spinal glia activation, and CX3CR1 expression in the model of bone cancer pain (BCP). The BCP model was established through intrathecally injecting Walker 256 mammary gland carcinoma cells to Sprague-Dawley rats. We found that spinal CX3CR1 expression and glial activation aggravated after inoculation. Resveratrol (i.t.) attenuated bone cancer-induced pain hypersensitivities, decreased CX3CR1 expression and glial activation in the spine in a BCP model. Resveratrol (i.t.) also attenuated mechanical allodynia resulting from intrathecally injecting fractalkine in rats. Inhibition of spinal glial activation and CX3CR1 upregulation may involve in resveratrol's analgesic effects. These findings demonstrated that resveratrol attenuated pain facilitation through inhibiting spinal glial activation and CX3CR1 upregulation in a BCP model.