RESUMEN
Heat stroke (HS) is a life-threatening illness and defined as when body temperature elevates above 40°C accompanied by the systemic inflammatory response syndrome that results in multiple organ dysfunctions. α-Lipoic acid (ALA) acts as a cofactor of mitochondrial enzymes and exerts anti-inflammatory and antioxidant properties in a variety of diseases. This study investigates the beneficial effects of ALA on myocardial injury and organ damage caused by experimental HS and further explores its underlying mechanism. Male Wistar rats were exposed to 42°C until their rectal core temperature reached 42.9°C and ALA was pretreared 40 or 80 mg/kg (i.v.) 1.5 h prior to heat exposure. Results showed that HS-induced lethality and hypothermia were significantly alleviated by ALA treatment that also improved plasma levels of CRE, LDH, and CPK and myocardial injury biomarkers myoglobin and troponin. In addition, ALA reduced cardiac superoxide anion formation and protein expression of cleaved caspase 3 caused by HS. Proinflammatory cytokine TNF-α and NF-κB pathways were significantly reduced by ALA treatment which may be associated with the upregulation of Hsp70. ALA significantly increased the Atg5-12 complex and LC3B II/LC3B I ratio, whereas the p62 and p-mTOR expression was attenuated in HS rats, indicating the activation of autophagy by ALA. In conclusion, ALA ameliorated the deleterious effects of HS by exerting antioxidative and anti-inflammatory capacities. Induction of Hsp70 and activation of autophagy contribute to the protective effects of ALA in HS-induced myocardial injury.
Asunto(s)
Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Golpe de Calor/tratamiento farmacológico , Golpe de Calor/patología , Inflamación/tratamiento farmacológico , Miocitos Cardíacos/citología , Miocitos Cardíacos/efectos de los fármacos , Ácido Tióctico/uso terapéutico , Animales , Autofagia/fisiología , Proteínas HSP70 de Choque Térmico/metabolismo , Golpe de Calor/metabolismo , Inflamación/metabolismo , Inflamación/patología , Masculino , FN-kappa B/metabolismo , Ratas , Ratas Wistar , Superóxidos/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUND: Loss of ovarian function, as in menopause or after ovariectomy (OVX), is closely associated with obesity and white adipose tissue (WAT) inflammation. Estrogen replacement protects against postmenopausal obesity but increases the risks of carcinogenesis. In the present study, we investigated the effects of long-term treatment of raloxifene (RAL), a selective estrogen receptor modulator, on the features of estrogen deficiency-induced obesity and explored the involvement of canonical and non-canonical Wnt regulation in vivo and in vitro. METHODS: Adult female rats received bilateral OVX and divided into 5 groups: (1) Sham, (2) OVX, (3) OVX + E2: OVX rats were administered with E2 (50 µg/kg, s.c., 3 times/week), (4) OVX + RAL: OVX rats were treated with RAL (gavage, 1 mg/kg/day) suspended in 0.8% carboxymethylcellulose (CMC), (5) OVX + CMC: 0.8% CMC as vehicle control. All treatments were given for 8 weeks beginning at 1 week after OVX. In 3 T3-L1 cells, the effects of RAL on adipogenesis and lipopolysaccharide (LPS)-induced inflammation were evaluated. RESULTS: Treatment with RAL significantly decreased body weight, visceral fat pad mass, adipocyte size and plasma levels of glucose but increased plasma adiponectin. RAL reduced the elevation of HIF-1α, VEGF-A and proinflammatory cytokines (MCP-1 and TNF-α) expression by inhibition of NF-κB p65 and JNK cascades in retroperitoneal WAT. This anti-inflammatory capacity of RAL may result from upregulation of secreted frizzle-related protein 5 (SFRP5), an adipokine that repressed Wnt5a signaling. Furthermore, RAL inhibited adipogenic factors such as PPAR-γ, C/EBP-α, and FABP4, and preserved canonical Wnt10b/ß-catenin protein expression. In 3 T3-L1 adipocytes, RAL (20 µM) diminished lipid accumulation and inhibited adipogenic factors accompanied with the induction of ß-catenin, which were effectively reversed by the ß-catenin inhibitor IWR-1-endo. In addition, RAL reduced LPS-induced NF-κB p65 and p-IκB expression as well as TNF-α secretion. Suppression of SFRP5 by small interfering RNA significantly abrogated the anti-inflammatory effects of RAL. CONCLUSIONS: Distinct activation of canonical ß-catenin on inhibition of adipogenesis and non-canonical SFRP5 on suppression of WAT inflammation may contribute to the beneficial effects of RAL. Therefore, this study provides a rationale for the therapeutic potential of RAL for postmenopausal obesity.
Asunto(s)
Adipogénesis/efectos de los fármacos , Tejido Adiposo/efectos de los fármacos , Inflamación/inducido químicamente , Clorhidrato de Raloxifeno/farmacología , Receptores de Estrógenos/antagonistas & inhibidores , Moduladores Selectivos de los Receptores de Estrógeno/farmacología , Proteínas Wnt/genética , Células 3T3-L1 , Tejido Adiposo/inmunología , Animales , Femenino , Regulación de la Expresión Génica , Lipopolisacáridos/farmacología , Ratones , Ovariectomía , Ratas , Ratas Wistar , Proteínas Wnt/metabolismo , Proteína Wnt1RESUMEN
Estrogen deficiency in postmenopausal women is linked to the higher prevalence of obesity, type 2 diabetes and metabolic syndromes. Development of beige adipocytes (browning of WAT) increases energy expenditure and could be a promising strategy for obesity management. This study aimed to investigate the effects of phytoestrogen genistein (GEN) on white adipose tissue (WAT) inflammation, browning and hepatic lipogenesis in ovariectomized rats with high-fat diet (HFD) and further explore the underlying mechanism. Female Wistar rats received ovariectomy (Ovx) and HFD (45% fat) and then were administered with 17ß-estradiol (E2, 3 times/week, subcutaneously) or GEN (15 mg/kg or 30 mg/kg, gavage, once daily) for 4 weeks. Administration of GEN decreased Ovx-induced body weight gain and adiposity and improved insulin sensitivity as well as increased insulin signaling p-IRS1 and p-AKT in retroperitoneal WAT. Adipocyte hypertrophy and production of proinflammatory cytokines MCP-1, TNF-α and IL-6 were reduced by GEN. It also suppressed the activation of NF-κB pathway evidenced by attenuation of p65 and phospho-IκB levels. Additionally, GEN elevated myokine irisin and promoted WAT browning by increasing UCP-1, PRDM-16, PGC-1α and CIDEA proteins and Ppargc1a, Ucp-1 and Tbx-1 mRNA in inguinal WAT which is associated with up-regulation of nuclear estrogen receptor-α. Plasma levels of triglyceride and cholesterol were reduced by GEN treatment accompanied with inhibition of lipogenic proteins (p-ACC, SREBP-1, FAS and CD36) in the liver. Long-term treatment with GEN attenuated estrogen-deficiency-induced obesity, WAT inflammation and hepatic lipogenesis and promoted the induction of WAT browning. It may provide a promising approach to prevent obesity during menopause.
Asunto(s)
Dieta Alta en Grasa/efectos adversos , Genisteína/farmacología , Lipogénesis/efectos de los fármacos , Hígado/efectos de los fármacos , Adipocitos/efectos de los fármacos , Adipocitos/patología , Adiponectina/sangre , Tejido Adiposo Pardo/efectos de los fármacos , Tejido Adiposo Blanco/efectos de los fármacos , Animales , Peso Corporal/efectos de los fármacos , Ingestión de Alimentos/efectos de los fármacos , Femenino , Fibronectinas/sangre , Insulina/sangre , Hígado/metabolismo , Ovariectomía , Paniculitis/tratamiento farmacológico , Paniculitis/etiología , Ratas Wistar , Proteína Desacopladora 1/metabolismoRESUMEN
Much evidence has indicated that matrix metalloproteinases (MMPs) participate in the progression of neuroinflammatory disorders. The present study was undertaken to investigate the inhibitory effect and mechanism of the antipsychotic haloperidol on MMP activation in the stimulated THP-1 monocytic cells. Haloperidol exerted a strong inhibition on tumor necrosis factor- (TNF-) α-induced MMP-9 gelatinolysis of THP-1 cells. A concentration-dependent inhibitory effect of haloperidol was observed in TNF-α-induced protein and mRNA expression of MMP-9. On the other hand, haloperidol slightly affected cell viability and tissue inhibition of metalloproteinase-1 levels. It significantly inhibited the degradation of inhibitor-κB-α (IκBα) in activated cells. Moreover, it suppressed activated nuclear factor-κB (NF-κB) detected by a mobility shift assay, NF-κB reporter gene, and chromatin immunoprecipitation analyses. Consistent with NF-κB inhibition, haloperidol exerted a strong inhibition of lipopolysaccharide- (LPS-) induced MMP-9 gelatinolysis but not of transforming growth factor-ß1-induced MMP-2. In in vivo studies, administration of haloperidol significantly attenuated LPS-induced intracerebral MMP-9 activation of the brain homogenate and the in situ in C57BL/6 mice. In conclusion, the selective anti-MMP-9 activation of haloperidol could possibly involve the inhibition of the NF-κB signal pathway. Hence, it was found that haloperidol treatment may represent a bystander of anti-MMP actions for its conventional psychotherapy.
Asunto(s)
Haloperidol/farmacología , Metaloproteinasa 9 de la Matriz/metabolismo , FN-kappa B/metabolismo , Animales , Supervivencia Celular/efectos de los fármacos , Inmunoprecipitación de Cromatina , Humanos , Proteínas I-kappa B/metabolismo , Lipopolisacáridos/farmacología , Ratones , Ratones Endogámicos C57BL , Transducción de Señal/efectos de los fármacosRESUMEN
Oestrogens have been reported to attenuate acute inflammation in sepsis. In this study, the effects of long-term oestrogen replacement with 17ß-oestradiol (E2 ) on endotoxaemia-induced circulatory dysfunction and multiple organ dysfunction syndrome were evaluated in ovariectomized (Ovx) rats. E2 (50 µg/kg, s.c., 3 times/week) was administered for 8 weeks, followed by the induction of endotoxaemia by intravenous infusion of lipopolysaccharides (LPS; 30 mg/kg/4 hrs). Oestrogen deficiency induced by ovariectomy for 9 weeks augmented the LPS-induced damage, including endotoxic shock, myocardial contractile dysfunction, renal dysfunction and rhabdomyolysis. Cardiac levels of NF-κB p65, iNOS and oxidized glutathione, free radical production in skeletal muscles, myoglobin deposition in renal tubules, and plasma levels of plasminogen activator inhibitor-1, TNF-α, and IL-6 were more pronounced in the Ovx + LPS group than in the Sham + LPS group. Long-term treatment of E2 prevented this amplified damage in Ovx rats. Six hours after LPS initiation, activation of the autophagic process, demonstrated by increases in Atg12 and LC3B-II/LC3B-I ratios, and induction of haem oxygenase (HO)-1 and heat-shock protein (HSP) 70 protein expression in myocardium were increased significantly in the Ovx + E2 + LPS group. These results suggest that activation of autophagy and induction of HO-1 and HSP70 contribute to the protective effect of long-term E2 replacement on multiple organ dysfunction syndrome in endotoxaemia.
Asunto(s)
Antiinflamatorios/farmacología , Autofagia/efectos de los fármacos , Trastornos Cerebrovasculares/tratamiento farmacológico , Endotoxemia/tratamiento farmacológico , Estradiol/farmacología , Insuficiencia Renal/tratamiento farmacológico , Rabdomiólisis/tratamiento farmacológico , Animales , Autofagia/genética , Proteína 12 Relacionada con la Autofagia/genética , Proteína 12 Relacionada con la Autofagia/metabolismo , Trastornos Cerebrovasculares/genética , Trastornos Cerebrovasculares/metabolismo , Trastornos Cerebrovasculares/patología , Esquema de Medicación , Endotoxemia/genética , Endotoxemia/metabolismo , Endotoxemia/patología , Femenino , Regulación de la Expresión Génica , Proteínas HSP70 de Choque Térmico/genética , Proteínas HSP70 de Choque Térmico/metabolismo , Hemo-Oxigenasa 1/genética , Hemo-Oxigenasa 1/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Lipopolisacáridos , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Ovariectomía , Inhibidor 1 de Activador Plasminogénico/genética , Inhibidor 1 de Activador Plasminogénico/metabolismo , Ratas , Ratas Sprague-Dawley , Insuficiencia Renal/genética , Insuficiencia Renal/metabolismo , Insuficiencia Renal/patología , Rabdomiólisis/genética , Rabdomiólisis/metabolismo , Rabdomiólisis/patología , Factor de Transcripción ReIA/genética , Factor de Transcripción ReIA/metabolismo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVE: Accumulating evidence demonstrates that raloxifene, a selective estrogen receptor modulator, possesses anti-inflammatory action. This study evaluates the preventive effects of long-term treatment of raloxifene on acute inflammation and multiple organ dysfunction syndrome (MODS) in ovariectomized (OVX) rats with endotoxemia and its underlying mechanism of action. METHODS: Adult female rats were OVX bilaterally to induce estrogen insufficiency. OVX rats were administered with raloxifene (1âmg/kg, gavage, once daily) for 8 weeks, beginning 1 week after surgery, followed by induction of sepsis via intravenous infusion of lipopolysaccharides (LPS; 30âmg/kg) for 4âhours. LPS-activated RAW 264.7 cells were used to investigate the mechanism of raloxifene. RESULTS: Ovariectomy amplified the endotoxemia-induced hypotensive effect, MODS, and superoxide anion production in the myocardium. The levels of inducible nitric oxide synthase, high mobility group box 1, and nuclear factor-κB p65 protein increased in OVX rats 6âhours after LPS initiation. Raloxifene mitigated MODS, together with reduced inducible nitric oxide synthase induction and fewer superoxide anions in organs. Raloxifene induced high levels of heat shock protein 70 (HSP70) and heme oxygenase 1 (HO-1), which are associated with an increase in the transcription factor heat shock factor-1 and Nrf-2, respectively. Pretreatment with quercetin, an inhibitor of HSP70, or SnPP, an inhibitor of HO-1, reversed the protective effects of raloxifene in septic OVX rats and LPS-activated macrophages. CONCLUSIONS: Long-term treatment with raloxifene reduces the severity of sepsis in OVX rats, attributed from up-regulation of HSP70 and HO-1 to exert the antioxidant and anti-inflammatory capacities. These findings provide new insights into bacterial infection during menopause and the molecular mechanism of raloxifene.
Asunto(s)
Endotoxemia/tratamiento farmacológico , Insuficiencia Multiorgánica/tratamiento farmacológico , Ovariectomía , Clorhidrato de Raloxifeno/uso terapéutico , Moduladores Selectivos de los Receptores de Estrógeno/uso terapéutico , Administración Oral , Animales , Modelos Animales de Enfermedad , Endotoxemia/metabolismo , Femenino , Proteínas HSP70 de Choque Térmico/metabolismo , Hemo-Oxigenasa 1/metabolismo , Insuficiencia Multiorgánica/metabolismo , Clorhidrato de Raloxifeno/administración & dosificación , Ratas , Ratas Wistar , Moduladores Selectivos de los Receptores de Estrógeno/administración & dosificaciónRESUMEN
Heat shock protein 70 (Hsp70) preconditioning induces thermotolerance, and adenosine monophosphate (AMP)-activated protein kinase (AMPK) plays a role in the process of autophagy. Here, we investigated whether 17-dimethylaminoethylamino-17-demethoxy-geldanamycin (17-DMAG) protected against heat stroke (HS) in rats by up-regulation of Hsp70 and phosphorylated AMPK (pAMPK). To produce HS, male Sprague-Dawley rats were placed in a chamber with an ambient temperature of 42°C. Physiological function (mean arterial pressure, heart rate and core temperature), hepatic and intestinal injury, inflammatory mediators and levels of Hsp70, pAMPK and light chain 3 (LC3B) in hepatic tissue were measured in HS rats or/and rats pre-treated with 17-DMAG. 17-DMAG pre-treatment significantly attenuated hypotension and organ dysfunction induced by HS in rats. The survival time during HS was also prolonged by 17-DMAG treatment. Hsp70 expression was increased, whereas pAMPK levels in the liver were significantly decreased in HS rats. Following pre-treatment with 17-DMAG, Hsp70 protein levels increased further, and pAMPK levels were enhanced. Treatment with an AMPK activator significantly increased the LC3BII/LC3BI ratio as a marker of autophagy in HS rats. Treatment with quercetin significantly suppressed Hsp70 and pAMPK levels and reduced the protective effects of 17-DMAG in HS rats. Both of Hsp70 and AMPK are involved in the 17-DMAG-mediated protection against HS. 17-DMAG may be a promising candidate drug in the clinical setting.
Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Benzoquinonas/uso terapéutico , Proteínas HSP70 de Choque Térmico/metabolismo , Golpe de Calor/tratamiento farmacológico , Golpe de Calor/metabolismo , Lactamas Macrocíclicas/uso terapéutico , Sustancias Protectoras/uso terapéutico , Animales , Autofagia/efectos de los fármacos , Benzoquinonas/farmacología , Proteínas de Unión al ADN/metabolismo , Factores de Transcripción del Choque Térmico , Golpe de Calor/fisiopatología , Íleon/efectos de los fármacos , Íleon/patología , Mediadores de Inflamación/metabolismo , Lactamas Macrocíclicas/farmacología , Masculino , Fosforilación/efectos de los fármacos , Sustancias Protectoras/farmacología , Ratas Sprague-Dawley , Análisis de Supervivencia , Factores de Transcripción/metabolismoRESUMEN
Sepsis is a systemic inflammatory disorder, accompanied with elevated oxidative stress, leading to multiple organ dysfunction syndrome (MODS), and disseminated intravascular coagulation. 17-Dimethylaminoethylamino- 17-demethoxygeldanamycin (17-DMAG), a heat shock protein (HSP) 90 inhibitor, has been reported to possess anti-inflammatory effects. In this study, the beneficial effects of 17-DMAG on lipopolysaccharide (LPS) induced MODS and DIC was evaluated in anesthetized rats. 17-DMAG (5 mg/kg, i.p.) was significantly increased survival rate, and prevented hypotension in LPS (30 mg/kg i.v. infused for 4 h) induced endotoxemia. The elevated levels of alanine aminotransferase (ALT), creatine phosphokinase (CPK), lactate dehydrogenase, creatinine, nitric oxide (NO) metabolites, IL-6, and TNF-α in LPS-exposed rat plasma were significantly reduced by 17-DMAG. Moreover, 17-DMAG suppressed LPS-induced superoxide anion production and caspase 3 activation in heart tissues. LPS induced the prolongation of prothrombin time, and a pronounced decrease in platelet count, which were improved by 17-DMAG. 17-DMAG markedly induced HSP70 and heme oxygenase (HO)-1, and suppressed inducible nitric oxide synthase (iNOS) and phosphorylated NF-κB p65 protein expression in organs 6 h after LPS initiation. Pretreatment with high dose of quercetin (300 mg/kg, i.p.), as an HSP70 inhibitor, reversed the beneficial effects of 17-DMAG on survival rate, plasma levels of ALT, CPK, creatinine, IL-6, and NO metabolites, iNOS induction, and caspase-3 activation in LPS-treated rats. In conclusion, 17-DMAG possesses the anti-inflammatory and antioxidant effects that were proved through LPS-induced acute inflammation, which is associated with induction of HSP70 and HO-1, leading to prevent MODS in sepsis.
Asunto(s)
Benzoquinonas/farmacología , Endotoxemia/tratamiento farmacológico , Proteínas HSP70 de Choque Térmico/biosíntesis , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Lactamas Macrocíclicas/farmacología , Insuficiencia Multiorgánica/tratamiento farmacológico , Alanina Transaminasa/sangre , Animales , Caspasa 3/metabolismo , Creatina Quinasa/sangre , Creatinina/sangre , Endotoxemia/inducido químicamente , Endotoxemia/metabolismo , Endotoxemia/patología , Interleucina-6/sangre , L-Lactato Deshidrogenasa/sangre , Lipopolisacáridos/toxicidad , Masculino , Insuficiencia Multiorgánica/inducido químicamente , Insuficiencia Multiorgánica/metabolismo , Insuficiencia Multiorgánica/patología , Óxido Nítrico/sangre , Ratas , Ratas Wistar , Factor de Transcripción ReIA/metabolismo , Factor de Necrosis Tumoral alfa/sangreRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Celastrol, a quinone methide extracted from the root of Tripterygium wilfordii Hook, possesses anti-oxidant and anti-inflammatory effects. Tripterygium wilfordii Hook is officially listed in the Chinese Pharmacopoeia and is used traditionally against rheumatoid arthritis, ankylosing spondylitis, and cancer. Furthermore, the circulatory protective effect of celastrol on an in vivo animal model of sepsis was investigated. AIM OF THE STUDY: Sepsis is a systemic inflammatory disorder that increases tissue oxidative stress and leads to multiple organ injury. We evaluated the beneficial effects of celastrol on multiple organ failure induced by lipopolysaccharide (LPS) in rats. MATERIALS AND METHODS: Celastrol (0.5 and 1.0 mg/kg, i.v.) was administered to anaesthetized rats 2 h before and 30 min after LPS challenge (10 mg/kg, i.v.). Eight hours later, cardiac and aortic protein expressions related to inflammatory responses, superoxide anion production, and reduced glutathione (GSH) level were measured. RESULTS: Treatment with celastrol prevented circulatory failure (bradycardia and hypotension) 8h after LPS challenge. The plasma levels of ALT, LDH, TNF-α, and nitric oxide metabolites increased markedly during sepsis, which significantly reduced after celastrol treatments. Celastrol attenuated iNOS, TNF-α, NF-κB phospho-p65 expression, superoxide anion production, and caspase 3 activity in the cardiovascular system, all of which were markedly elevated after LPS challenge. Furthermore, celastrol induced HO-1 and HSP70 expressions increase in nuclear levels of Nrf2 and HSF-1, respectively, and increase cardiac GSH level 8h after LPS challenge. CONCLUSION: Anti-inflammatory and anti-oxidant effects of celastrol contribute to prevent circulatory failure in sepsis. Induction of HO-1 and HSP70 by celastrol participates in these beneficial effects.
Asunto(s)
Endotoxemia/tratamiento farmacológico , Proteínas HSP70 de Choque Térmico/metabolismo , Hemo-Oxigenasa 1/metabolismo , Triterpenos/farmacología , Alanina Transaminasa/sangre , Animales , Relación Dosis-Respuesta a Droga , Endotoxemia/inducido químicamente , Endotoxemia/metabolismo , Glutatión/metabolismo , Proteínas HSP70 de Choque Térmico/genética , Hemo-Oxigenasa 1/genética , Lipopolisacáridos/toxicidad , Masculino , Óxido Nítrico/metabolismo , Triterpenos Pentacíclicos , Ratas , Ratas Endogámicas WKY , Superóxidos , Triterpenos/administración & dosificación , Factor de Necrosis Tumoral alfa/sangre , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Alpha-lipoic acid (ALA), a naturally occurring disulfide derivative of octanoic acid, serves as a strong antioxidant and has been reported to possess anti-inflammatory effects. The aim of the present study is to investigate the preventive and therapeutic effects of ALA on multiple organ dysfunction syndrome (MODS) caused by endotoxemia in rats. Male Wistar rats were intravenously infused with lipopolysaccharide (LPS) (10 mg/kg) to induce endotoxemia. Alpha-lipoic acid 10, 20, or 40 mg/kg was administered intravenously 60 min before (pretreatment) LPS challenge, and ALA 40 mg/kg was administered intravenously 30 min after (posttreatment) LPS challenge. Pretreatment and posttreatment with ALA significantly improved the deleterious hemodynamic changes 8 h after LPS challenge, including hypotension and bradycardia. Alpha-lipoic acid reduced the plasma levels of glutamic pyruvic transaminase, blood urea nitrogen, lactate dehydrogenase, tumor necrosis factor-α, nitric oxide metabolites, and thrombin-antithrombin complex, which increased markedly after LPS challenge. The induction of inducible nitric oxide synthase both in the liver and the lung and vascular superoxide anion production were also significantly suppressed by ALA. Moreover, ALA significantly attenuated LPS-induced caspase-3 activation in cardiomyocytes and improved survival rate. In conclusion, ALA effectively attenuated LPS-induced acute inflammatory response and improved MODS. The antioxidant and anti-inflammatory effects of ALA may contribute to these beneficial effects. Alpha-lipoic acid might be considered as a novel therapeutic strategy in the prevention of sepsis-induced MODS and inflammatory vascular diseases.
Asunto(s)
Endotoxemia/complicaciones , Insuficiencia Multiorgánica/prevención & control , Choque Séptico/prevención & control , Ácido Tióctico/farmacología , Animales , Antioxidantes/farmacología , Antitrombinas/química , Aorta/metabolismo , Caspasa 3/metabolismo , Disulfuros/química , Activación Enzimática , Hemodinámica , Inflamación/metabolismo , Hígado/enzimología , Pulmón/enzimología , Masculino , Insuficiencia Multiorgánica/etiología , Miocardio/enzimología , Nitratos/química , Nitritos/química , Ratas , Ratas Wistar , Sepsis/fisiopatología , Choque Séptico/etiología , Superóxidos/química , Trombina/químicaRESUMEN
Sepsis can cause myocardial dysfunction, which contributes to the high mortality of sepsis. Hypertonic saline (HS) has been reported to increase myocardial contractility in sepsis. In the present study, mechanisms of action of HS resuscitation (4 mL of 7.5% NaCl per kilogram) on cardiac function have been evaluated in septic rats. HS was administered 1 h after LPS (10 mg/kg, i.v.) challenge. The mean arterial blood pressure significantly decreased 4 h after LPS challenge, and septic shock was observed at the end of experiment (6 h). Posttreatment with HS prevented hypotension caused by LPS and significantly improved cardiac function, evidenced by increases in left ventricular developed pressure, mean +dP/dt and -dP/dt. The amplitude of electrical-stimulated intracellular Ca(2+) transient in isolated single cardiomyocytes was significantly reduced after 6 h LPS insult, which was recovered by HS. In addition, LPS resulted in significant increases in neutrophil myeloperoxidase activity, macrophage migration inhibitory factor (MIF), and NF-κB phospho-p65 protein levels in myocardium at 6 h, which were significantly attenuated by HS. In conclusion, HS improved myocardial contractility and prevented circulatory failure induced by endotoxemia, which may attribute to improvement of intracellular calcium handling process and inhibitory effects on neutrophil infiltration and MIF production in hearts.
Asunto(s)
Factores Inhibidores de la Migración de Macrófagos/antagonistas & inhibidores , Solución Salina Hipertónica/administración & dosificación , Sepsis/tratamiento farmacológico , Animales , Calcio/metabolismo , Humanos , Hipotensión/inducido químicamente , Hipotensión/tratamiento farmacológico , Hipotensión/patología , Lipopolisacáridos/toxicidad , Contracción Miocárdica/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Ratas , Sepsis/inducido químicamente , Sepsis/patologíaRESUMEN
Neurodegenerative brain disorders such as Alzheimer's disease (AD) have been well investigated. However, significant methods for the treatment of the progression of AD are unavailable currently. Heat shock protein 70 (Hsp70) plays important roles in neural protection from stress by assisting cellular protein folding. In this study, we investigated the effect and the molecular mechanism of YC-1, an activator of guanylyl cyclase (GC), on Aß25-35-induced cytotoxicity in differentiated PC12 cells. The results of this study showed that Aß25-35 (10 µM) significantly increased p25 protein production in a pattern that was consistent with the increase in µ-calpain expression. Moreover, Aß25-35 significantly increased tau hyperphosphorylation and induced differentiated PC12 cell death. YC-1 (0.5-10 µM) prevented the cell death induced by Aß25-35. In addition, YC-1 (1, 10 µM) significantly blocked Aß25-35-induced µ-calpain expression and decreased the formation of p25 and tau hyperphosphorylation. Moreover, YC-1 (5-20 µM) alone or combined with Aß25-35 (10 µM) significantly increased the expression of Hsp70 in differentiated PC12 cells. The neuroprotective effect of YC-1 was significantly attenuated by an Hsp70 inhibitor (quercetin, 50 µM) or in PC12 cells transfected with an Hsp70 small interfering RNA. However, pretreatment of cells with the GC inhibitor ODQ (10 µM) did not affect the neuroprotective effect of YC-1 against Aß25-35 in differentiated PC12 cells. These results suggest that the neuroprotective effect of YC-1 against Aß25-35-induced toxicity is mainly mediated by the induction of Hsp70. Thus, YC-1 is a potential agent against AD.
Asunto(s)
Péptidos beta-Amiloides/toxicidad , Diferenciación Celular/efectos de los fármacos , Proteínas HSP70 de Choque Térmico/metabolismo , Indazoles/farmacología , Fármacos Neuroprotectores/farmacología , Fragmentos de Péptidos/toxicidad , Animales , Calpaína/metabolismo , Guanilato Ciclasa/metabolismo , Proteínas HSP70 de Choque Térmico/antagonistas & inhibidores , Neurotoxinas/toxicidad , Células PC12 , Fosforilación/efectos de los fármacos , Ratas , Transducción de Señal/efectos de los fármacos , Proteínas tau/metabolismoRESUMEN
Heat stroke is a life-threatening illness characterized by an elevated core body temperature. Despite adequate lowering of the body temperature and support treatment of multiple organ-system function, heat stroke is often fatal. 3-(5'-Hydoxymethyl-2'-furyl)-1-benzyl-indazol (YC-1) been identified as an activator of soluble guanylate cyclase. To evaluate whether YC-1 protects multiple organ dysfunctions and improves survival during heat stroke and its mechanism. Male Sprague-Dawley rats untreated or treated with either YC-1 or quercetin (heat shock protein (Hsp) 70 inhibitor) were exposures to heat as a model of heat stroke. The mean arterial pressure (MAP), heart rate, rectal temperature (Tco), survival time, and plasma biochemical data, intracellular Hsp70 and heat shock factor-1 expression were measured. The value of MAP, heart rate and Tco of untreated heat stroke (HS) group were all significantly lower than that of normothermal (NT) group. Biochemical markers evidenced that liver and kidney injuries of HS group were significantly higher than that of NT groups. YC-1 (20mg/kg) pretreatment with heat stroke (YC-1+HS) group, the MAP and heart rate were return to normal, and the biochemical markers were all significantly recovered to normal. The survival time of HS group, NT group and YC-1+HS group were 21, 480, and 445 min, respectively. The expression of Hsp70 and HSF-1 in liver and renal of YC-1+HS group was significantly higher than that of HS group. All of the protective effects of YC-1 were all significantly suppressed when pretreated with quercetin (400mg/kg). Results indicate that YC-1 may improve survival due to induce Hsp70 overexpression.
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Activadores de Enzimas/farmacología , Proteínas HSP70 de Choque Térmico/metabolismo , Golpe de Calor/tratamiento farmacológico , Indazoles/farmacología , Animales , Presión Arterial/efectos de los fármacos , Proteínas de Unión al ADN/genética , Modelos Animales de Enfermedad , Regulación de la Expresión Génica/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Factores de Transcripción del Choque Térmico , Golpe de Calor/fisiopatología , Riñón/efectos de los fármacos , Riñón/patología , Hígado/efectos de los fármacos , Hígado/patología , Masculino , Quercetina/farmacología , Ratas , Ratas Sprague-Dawley , Tasa de Supervivencia , Factores de Transcripción/genéticaRESUMEN
OBJECTIVE: Adenosine monophosphate (AMP)-activated protein kinase (AMPK) plays an important role in regulating whole-body energy homeostasis. The aim of this study was to investigate dietary α-lipoic acid (α-LA) supplementation on the activation of AMPK in both central and peripheral tissues in obese rats induced by ovariectomy. METHODS: Ovariectomized (Ovx) rats were treated with α-LA (200 mg/kg) 3 to 10 weeks after surgery. Body weight, food intake, fat mass, phosphorylated AMPKα (pAMPKα), and phosphorylated acetyl-CoA carboxylase (ACC) protein expression in both the hypothalamus and white adipose tissue (WAT) as well as plasma leptin and adiponectin levels were measured in rats after either Ovx or sham operations. RESULTS: Compared with control rats, ovariectomy led to increased body weight, food intake, and WAT mass 2 to 10 weeks after surgery. Furthermore, plasma leptin and adiponectin levels as well as hypothalamic pAMPKα expression were significantly increased after ovariectomy, accompanied by a reduction in pAMPKα expression in WAT after ovariectomy. However, after treatment with α-LA, the elevation of leptin and adiponectin levels and the activation of hypothalamic AMPKα and ACC, as induced by ovariectomy, were significantly suppressed. Meanwhile, decreased fat mass and increased pAMPKα and phosphorylated ACC expression in the WAT were observed in Ovx rats treated with α-LA. CONCLUSIONS: α-LA significantly decreased appetite and fat accumulation, possibly through the regulation of central and peripheral AMPK activities in rats. Therefore, this study provides a rationale for the therapeutic use of α-LA for obesity in postmenopausal women.
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Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Obesidad/metabolismo , Ovario/metabolismo , Ácido Tióctico/farmacología , Complejo Vitamínico B/farmacología , Acetil-CoA Carboxilasa/metabolismo , Adiponectina/sangre , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/metabolismo , Animales , Peso Corporal/efectos de los fármacos , Ingestión de Alimentos/efectos de los fármacos , Femenino , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Leptina/sangre , Ovariectomía , Ovario/cirugía , Ratas , Ratas Sprague-Dawley , Ácido Tióctico/metabolismo , Complejo Vitamínico B/metabolismoRESUMEN
BACKGROUND: Receptor-interacting protein 140 (RIP140) is a corepressor for nuclear receptors with an important role in the inhibition of energy expenditure. Postmenopausal women have increased white adipose tissue (WAT), and excessive accumulation of adipose tissue (obesity) implies a health risk. The aim of the present work was to investigate the time course of RIP140 expression in WAT during the development of ovariectomy (OVX)-induced obesity in rats. METHODS: OVX was performed in female Sprague-Dawley (SD) rats 8 weeks old. Body weight and food intake were determined once a week. WAT of sham-operated, OVX and OVX plus 17ß-estradiol therapy (OVX/E2) female SD rats was weighed and used to analyse RIP140 and uncoupling protein 1 (UCP-1) expression by Western blot. RESULTS: Food intake and body weight were significantly increased during the 2-8 weeks after OVX. Even though body weight increased until killing, food intake progressively decreased from 9 to 16 weeks after OVX in rats. Meanwhile, increased WAT mass and decreased RIP140 expression in WAT were observed in OVX rats. In contrast, the expression of UCP-1, a key target gene of RIP140, in WAT of OVX rats was significantly higher than in sham-operated rats. All of these alterations caused by OVX were mostly reversed by the replacement of 17ß-estradiol. CONCLUSIONS: The down-regulation of RIP140 in WAT may play a compensatory role in OVX-induced obesity in rat.
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Proteínas Adaptadoras Transductoras de Señales/biosíntesis , Tejido Adiposo Blanco/metabolismo , Adiposidad/fisiología , Menopausia/metabolismo , Proteínas Nucleares/biosíntesis , Obesidad/metabolismo , Ovariectomía , Animales , Regulación del Apetito/fisiología , Western Blotting , Peso Corporal , Regulación hacia Abajo , Ingestión de Alimentos , Estradiol/administración & dosificación , Estradiol/metabolismo , Estrógenos/administración & dosificación , Estrógenos/metabolismo , Femenino , Canales Iónicos/biosíntesis , Proteínas Mitocondriales/biosíntesis , Proteína de Interacción con Receptores Nucleares 1 , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Factores de Tiempo , Proteína Desacopladora 1RESUMEN
OBJECTIVE: Adenosine monophosphate-activated protein kinase (AMPK) acts as a cellular energy sensor, being activated during states of low energy charge. Hypothalamic AMPK is altered by hormonal and metabolic signals and mediates the feeding response. The aims of this study were to examine whether the phosphorylation of AMPKα in the hypothalamus is affected by ovariectomy (Ovx) and thus would be involved in the development of obesity in rats. METHODS: Body weight, food intake, hypothalamic phosphorylated AMPKα (pAMPKα) protein expression, and plasma leptin and adiponectin levels were measured in female rats after either Ovx or sham operations. These patterns were also observed after treatment with 17ß-estradiol, compound C, and leptin in Ovx rats. RESULTS: Compared with control rats, Ovx led to increased body weight and food intake at 2 to 8 weeks after operation. Meanwhile, plasma leptin and adiponectin levels and hypothalamic pAMPKα expression were significantly increased after Ovx. Replacement of estradiol significantly reversed these effects. Treatment with compound C, an AMPKα inhibitor, for 1 week produced a reduction in food intake, body weight, and plasma leptin and adiponectin levels. Meanwhile, these effects were reversed upon withdrawal of compound C. In addition, central injection of leptin also significantly reduced body weight, food intake, plasma leptin and adiponectin levels, and hypothalamic pAMPKα expression relative to those of the Ovx group. CONCLUSIONS: Increased hypothalamic pAMPKα expression may contribute to hyperphagia during the development of Ovx-induced obesity in rats.
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Proteínas Quinasas Activadas por AMP/metabolismo , Hipotálamo/enzimología , Obesidad/etiología , Ovario/fisiología , Adiponectina/sangre , Análisis de Varianza , Animales , Peso Corporal , Ingestión de Alimentos , Estradiol/farmacología , Estradiol/fisiología , Femenino , Leptina/sangre , Leptina/fisiología , Modelos Animales , Obesidad/enzimología , Obesidad/fisiopatología , Ovariectomía , Fosforilación/efectos de los fármacos , Fosforilación/fisiología , Pirazoles/farmacología , Pirimidinas/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: The aim of this study was to investigate the beneficial effect of long-term treatment with raloxifene (RAL), a selective estrogen receptor modulator, on myocardial ischemia/reperfusion (MI/R) injury in ovariectomized (Ovx) rats. METHODS: Ovariectomy was performed in female Sprague-Dawley rats 8 weeks old. Ovx rats were treated with RAL 1 or 5 mg/kg (gavage, once daily) or 17beta-estradiol (E2; 50 microg/kg SC, three times a week) for 8 weeks. The cardioprotective effect of RAL was evaluated in an open-chest anesthetized rat model of MI/R, which was induced by 40-minute left coronary artery occlusion and 100-minute reperfusion. RESULTS: Long-term treatment with RAL 1 mg/kg significantly suppressed the duration of ventricular tachycardia elicited by MI. After MI/R, the levels of plasma creatine kinase-MB fraction and lactate dehydrogenase in Ovx rats were significantly higher than those in the sham group, which were significantly reduced by long-term treatment with RAL 1 mg/kg or E2. Neutrophil myeloperoxidase activity in ischemic myocardium markedly increased in the Ovx group, whereas long-term treatment with RAL 1 or 5 mg/kg or E2 significantly suppressed the elevation of myeloperoxidase activity. After MI/R, the protein expression of phosphorylated inhibitory kappaBalpha and caspase-3 in ischemic myocardium pronouncedly increased in the Ovx group and was attenuated by long-term treatment with RAL 1 mg/kg or E2. CONCLUSIONS: Long-term treatment with RAL can reduce the severity of MI-induced arrhythmias and attenuate MI/R-induced damages and apoptosis in Ovx rats. This cardioprotective effect of RAL may be associated with inhibition of neutrophil infiltration and suppression of nuclear factor-kappaB activation.
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Estradiol/farmacología , Terapia de Reemplazo de Estrógeno , Daño por Reperfusión Miocárdica/prevención & control , Posmenopausia , Clorhidrato de Raloxifeno/administración & dosificación , Moduladores Selectivos de los Receptores de Estrógeno/administración & dosificación , Taquicardia Ventricular/prevención & control , Animales , Apoptosis/efectos de los fármacos , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Femenino , Infarto del Miocardio , Ovariectomía , Ratas , Ratas Sprague-Dawley , TiempoRESUMEN
BACKGROUND: Matrix metalloproteinases (MMPs) activity may modulate hypertension-related accumulation of extracellular matrix (ECM) in arteries. We tested whether estrogen deficiency induces alterations of vascular collagen, MMP-2, membrane-type 1-MMP (MT1-MMP), or tissue inhibitor of metalloproteinases-2 (TIMP-2) expression in ovariectomized rats, which may be associated with postmenopausal hypertension. METHODS: Estrogen deficiency was induced by ovariectomy (Ovx) in female rats. Time-course changes of aortic MMPs protein expression were evaluated. Treatment with tempol or aminoguanidine was used to examine the role of oxidative stress and nitric oxide (NO) on these changes. RESULTS: The level of the active-form MMP-2 was markedly reduced during 1-4 weeks after Ovx, with a significant increase in collagen accumulation and increased MT1-MMP expression. Although active-form MMP-2 and collagen progressively returned to normal levels, the markedly increased collagen deposition appeared again at 8 weeks and persisted until 12 weeks, followed by induction of MMP-2 and MT1-MMP at 12 weeks. The TIMP-2 level reduced for 2 weeks after Ovx, but soon returned to normal. Treatment with 17beta-estradiol (E(2)), tempol, or aminoguanidine for 6 weeks prevented Ovx-induced blood pressure elevation and apparently reversed the MMPs changes. CONCLUSIONS: In an initial period, E(2) deficiency induces a reduction of active-form MMP-2 leading to collagen accumulation, and induction of MT1-MMP, which may be a compensatory response to degrade collagen. At a latter stage, the concurrent elevation of active-form MMP-2 and MT1-MMP expression may be adaptive responses to regulate ECM composition in the vascular wall. Oxidative stress and NO contribute to activity modulation of vascular MMPs in Ovx rats.
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Aorta Torácica/enzimología , Estrógenos/deficiencia , Hipertensión/metabolismo , Metaloproteinasa 14 de la Matriz/biosíntesis , Metaloproteinasa 2 de la Matriz/biosíntesis , Ovariectomía/efectos adversos , Inhibidor Tisular de Metaloproteinasa-2/biosíntesis , Animales , Aorta Torácica/fisiopatología , Presión Sanguínea/fisiología , Western Blotting , Modelos Animales de Enfermedad , Femenino , Estudios de Seguimiento , Hipertensión/fisiopatología , Estrés Oxidativo , Ratas , Ratas Sprague-Dawley , Resistencia VascularRESUMEN
OBJECTIVE: We investigated whether the effect of estrogen therapy on vascular endothelial function is mediated through increasing the bioavailability of tetrahydrobiopterin (BH4) and associated antioxidant capacity in ovariectomized (Ovx) rats. DESIGN: Aortas of sham-operated, Ovx, and Ovx plus estrogen therapy (Ovx + ET) female Sprague-Dawley rats were used to measure vascular reactivity. Plasma levels of nitric oxide (NO) metabolites, total antioxidant capacity, aortic superoxide anion (O2.), and BH4 contents were determined. RESULTS: Vascular reactivity, assessed on isolated aortic segments, indicated that phenylephrine-induced contraction in the Ovx group was significantly greater than that in the sham and Ovx + ET groups. The vasodilator responses to acetylcholine (10 to 10 M) and L-arginine (L-Arg; 10 M) in the sham and Ovx + ET groups were significantly greater than those in the Ovx group. Pretreatment with BH4 (10 M) enhanced the vasodilator responses to L-Arg in the Ovx group compared with the untreated Ovx group. An inhibitor of BH4 synthesis, 2,4-diamino-6-hydroxypyrimidine (2 mM), significantly attenuated the vasodilator response to L-Arg in the sham and Ovx + ET groups. In addition, Ovx significantly increased O2. production in aortic tissues and decreased plasma NO metabolites levels, whereas ET significantly prevented these effects. Pretreatment with BH4 also significantly decreased aortic O2. production in the Ovx group; both plasma total antioxidant capacity and aortic BH4 contents in the Ovx group decreased significantly compared with those in the sham group, which were also improved by ET. There were no significant differences in the protein expression of endothelial NO synthase in aortas in these groups. CONCLUSIONS: ET increases the availability of vascular BH4 to attenuate O2. production and restores total antioxidant capacity, leading to improved NO-mediated vasodilation in Ovx rats.
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Antioxidantes/farmacología , Biopterinas/análogos & derivados , Endotelio Vascular/efectos de los fármacos , Estradiol/farmacología , Terapia de Reemplazo de Estrógeno , Estrés Oxidativo/efectos de los fármacos , Vasodilatación/efectos de los fármacos , Animales , Aorta/efectos de los fármacos , Aorta/metabolismo , Disponibilidad Biológica , Biopterinas/sangre , Western Blotting , Femenino , Óxido Nítrico/metabolismo , Ovariectomía , Ratas , Ratas Sprague-Dawley , Superóxidos/metabolismoRESUMEN
Heme oxygenase-1 (HO-1), an inducible stress protein, has been implicated in cytoprotection against oxidative stress in vitro and in vivo. Estrogens also have antioxidant effects. This study investigated the time course of HO-1 and inducible nitric oxide synthase (iNOS) expression in the aortas of ovariectomized rats, and the regulatory relationship between the NO/NOS and the carbon monoxide/HO systems. HO-1 and iNOS protein expression was induced by ovariectomy (Ovx) and was extremely high 2-6 weeks after Ovx compared with the sham-operated group. Expression of the constitutive enzymes HO-2 and endothelial NOS did not differ significantly between sham-operated and Ovx rats. 17beta-Estradiol (E(2)) replacement reversed these changes in rats after Ovx. Long-term treatment with the antioxidant tempol significantly inhibited HO-1 and iNOS expression. The iNOS inhibitor aminoguanidine significantly suppressed the induction of HO-1. Oxidized glutathione in the hearts of Ovx rats increased gradually, with significant elevation at 3-6 weeks after Ovx compared with the sham-operated group, whereas plasma levels of NO metabolites were significantly reduced 4-6 weeks after Ovx. Treatment with the HO inhibitor zinc protoporphyrin IX blocked HO-1 induction, but significantly increased the plasma levels of NO metabolites. In conclusion, HO-1 is induced by oxidative stress resulting from E(2) depletion. The NO/iNOS system contributes to the induction of HO-1, which may subsequently suppress iNOS activity to modulate vasculoprotective effects after menopause.