RESUMO
AIM: Argirein (rhein-arginine) is a derivative of rhein isolated from Chinese rhubarb (Rheum Officinale Baill.) that exhibits antioxidant and anti-inflammatory activities. In the present study we investigated the effects of argirein on stress-induced (hypergonadotrophic) and diabetic (hypogonadotrophic) hypogonadism in male rats. METHODS: Stress-induced and diabetic hypogonadism was induced in male rats via injection of isoproterenol (ISO) or streptozotocin (STZ). ISO-injected rats were treated with argirein (30 mg·kg(-1)·d(-1), po) or testosterone replacement (0.5 mg·kg(-1)·d(-1), sc) for 5 days, and STZ-injected rats were treated with argirein (40-120 mg·kg(-1)·d(-1), po) or aminoguanidine (100 mg·kg(-1)·d(-1), po) for 4 weeks. After the rats were euthanized, blood samples and testes were collected. Serum hormone levels were measured, and the expression of endothelin receptor A (ETA), connexin 43 (Cx43) and other proteins in testes was detected. For in vitro experiments, testis homogenate was prepared from normal male rats, and incubated with ISO (1 µmol/L) or high glucose (27 mmol/L). RESULTS: ISO injection induced hyper-gonadotrophic hypogonadism characterized by low testosterone and high FSH and LH levels in the serum, whereas STZ injection induced hypogonadotrophic hypogonadism as evidenced by low testosterone and low FSH and LH levels in the serum. In the testes of ISO- and STZ-injected rats, the expression of ETA, MMP-9, NADPH oxidase and pPKCε was significantly increased, and the expression of Cx43 was decreased. Administration of argirein attenuated both the abnormal serum hormone levels and the testis changes in ISO- and STZ-injected rats, and aminoguanidine produced similar actions in STZ-injected rats; testosterone replacement reversed the abnormal serum hormone levels, but did not affect the testis changes in ISO-injected rats. Argirein (0.3-3 µmol/L) exerted similar effects in testis homogenate incubated with ISO or high glucose in vitro. CONCLUSION: Two types of hypogonadism of male rats exhibit increased expression of ETA and depressed expression of Cx43 in testes, despite different patterns of serum FSH and LH. Argirein alleviates the two types of male hypogonadism via normalizing ETA and Cx43 in testes.
Assuntos
Antraquinonas/uso terapêutico , Arginina/uso terapêutico , Conexina 43/metabolismo , Diabetes Mellitus Experimental/complicações , Medicamentos de Ervas Chinesas/uso terapêutico , Hipogonadismo/tratamento farmacológico , Hipogonadismo/etiologia , Receptor de Endotelina A/metabolismo , Animais , Antraquinonas/química , Arginina/química , Conexina 43/análise , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/metabolismo , Combinação de Medicamentos , Medicamentos de Ervas Chinesas/química , Hipogonadismo/sangue , Hipogonadismo/metabolismo , Isoproterenol , Masculino , Ratos , Ratos Sprague-Dawley , Receptor de Endotelina A/análise , Rheum/química , Estreptozocina , Testículo/efeitos dos fármacos , Testículo/metabolismo , Testosterona/sangueRESUMO
Inflammatory changes in the cerebral network are present in early mechanisms involved in neurodegenerative disease, Alzheimer disease (AD), and aging brain. We intended to verify that these are likely due to an activation of NADPH oxidase (NOX) and endoplasmic reticulum (ER) stress. Apocynin (APO) an inhibitor of NOX is potential to ameliorate these changes. Rehmannia complex (Reh) a famous prescription in China and the triterpene acids (TTA) isolated from Reh may relieve the isoproterenol (ISO) induced chronic inflammation in the brain, compared with APO. Rats were administered with ISO for 10 days and astrocytes were incubated with ISO for 24 h. Changes in neural MMP (matrix metalloproteinase), Cx43, AQP4 (aquaporin 4), NFκB, IκBß, and p-PERK (PKB like kinase) were conducted and intervened with APO, Reh and TTA, in vivo and in vitro, respectively. An increased MDA and upregulated NOX subunit p47phox, ETA, PERK in association with abnormal MMP-2/9 and Cx40/43 were found in cerebral tissue of ISO-injected rats. Astrocytes incubated with ISO exhibited upregulated APQ4, IκBß, NFκB and p-PERK/PERK and downregulated Cx43. These were significantly abrogated by APO and Reh, in vivo, and APO and TTA in vitro. In conclusion, neural damages induced by ISO were characterized by inflammatory changes in cerebral tissue and astrocytes, which were blunted significantly by APO, Reh and TTA, respectively. Reh and TTA are potential in alleviating the early pathogenesis in neurodegenerative changes in AD in the clinical settings through suppressing NOX and ER stress in the brain.
Assuntos
Acetofenonas/farmacologia , Encéfalo/metabolismo , Estresse do Retículo Endoplasmático/fisiologia , Isoproterenol/toxicidade , NADPH Oxidases/metabolismo , Rehmannia , Triterpenos/farmacologia , Acetofenonas/uso terapêutico , Animais , Encéfalo/efeitos dos fármacos , Células Cultivadas , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Inflamação/induzido quimicamente , Inflamação/metabolismo , Inflamação/prevenção & controle , Masculino , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Triterpenos/uso terapêuticoRESUMO
AIM: Hypoxia/oxidative stress can alter the pharmacokinetics (PK) of CPU86017-RS, a novel antiarrhythmic agent. The aim of this study was to investigate the mechanisms underlying the alteration of PK of CPU86017-RS by hypoxia/oxidative stress. METHODS: Male SD rats exposed to normal or intermittent hypoxia (10% O2) were administered CPU86017-RS (20, 40 or 80 mg/kg, ig) for 8 consecutive days. The PK parameters of CPU86017-RS were examined on d 8. In a separate set of experiments, female SD rats were injected with isoproterenol (ISO) for 5 consecutive days to induce a stress-related status, then CPU86017-RS (80 mg/kg, ig) was administered, and the tissue distributions were examined. The levels of Mn-SOD (manganese containing superoxide dismutase), endoplasmic reticulum (ER) stress sensor proteins (ATF-6, activating transcription factor 6 and PERK, PRK-like ER kinase) and activation of NADPH oxidase (NOX) were detected with Western blotting. Rat liver microsomes were incubated under N2 for in vitro study. RESULTS: The Cmax, t1/2, MRT (mean residence time) and AUC (area under the curve) of CPU86017-RS were significantly increased in the hypoxic rats receiving the 3 different doses of CPU86017-RS. The hypoxia-induced alteration of PK was associated with significantly reduced Mn-SOD level, and increased ATF-6, PERK and NOX levels. In ISO-treated rats, the distributions of CPU86017-RS in plasma, heart, kidney, and liver were markedly increased, and NOX levels in heart, kidney, and liver were significantly upregulated. Co-administration of the NOX blocker apocynin eliminated the abnormalities in the PK and tissue distributions of CPU86017-RS induced by hypoxia/oxidative stress. The metabolism of CPU86017-RS in the N2-treated liver microsomes was significantly reduced, addition of N-acetylcysteine (NAC), but not vitamin C, effectively reversed this change. CONCLUSION: The altered PK and metabolism of CPU86017-RS induced by hypoxia/oxidative stress are produced by mitochondrial abnormalities, NOX activation and ER stress; these abnormalities are significantly alleviated by apocynin or NAC.
Assuntos
Antiarrítmicos/farmacocinética , Compostos Heterocíclicos de 4 ou mais Anéis/farmacocinética , NADPH Oxidases/metabolismo , Estresse Oxidativo , Animais , Área Sob a Curva , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Ativação Enzimática , Meia-Vida , Masculino , Microssomos Hepáticos/enzimologia , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: Hypoxia exposure initiates low serum testosterone levels that could be attributed to downregulated androgen biosynthesizing genes such as StAR (steroidogenic acute regulatory protein) and 3-beta-HSD (3-beta-hydroxysteroid dehydrogenase) in the testis. It was hypothesized that these abnormalities in the testis by hypoxia are associated with oxidative stress and an increase in chaperones of endoplasmic reticulum stress (ER stress) and ER stress could be modulated by a reduction in calcium influx. Therefore, we verify that if an application of CPU86017-RS (simplified as RS, a derivative to berberine) could alleviate the ER stress and depressed gene expressions of StAR and 3-beta-HSD, and low plasma testosterone in hypoxic rats, these were compared with those of nifedipine. METHODS: Adult male Sprague-Dawley rats were randomly divided into control, hypoxia for 28 days, and hypoxia treated (mg/kg, p.o.) during the last 14 days with nifedipine (Nif, 10) and three doses of RS (20, 40, 80), and normal rats treated with RS isomer (80). Serum testosterone (T) and luteinizing hormone (LH) were measured. The testicular expressions of biomarkers including StAR, 3-beta-HSD, immunoglobulin heavy chain binding protein (Bip), double-strand RNA-activated protein kinase-like ER kinase (PERK) and pro-apoptotic transcription factor C/EBP homologous protein (CHOP) were measured. RESULTS: In hypoxic rats, serum testosterone levels decreased and mRNA and protein expressions of the testosterone biosynthesis related genes, StAR and 3-beta-HSD were downregulated. These changes were linked to an increase in oxidants and upregulated ER stress chaperones: Bip, PERK, CHOP and distorted histological structure of the seminiferous tubules in the testis. These abnormalities were attenuated significantly by CPU86017-RS and nifedipine. CONCLUSION: Downregulated StAR and 3-beta-HSD significantly contribute to low testosterone in hypoxic rats and is associated with ER stress which mediates testis damage caused by oxygen deprivation. CPU86017-RS is potential in ameliorating hypoxia-induced testicular injuries, possibly by its calcium antagonist effects on the testis.
Assuntos
3-Hidroxiesteroide Desidrogenases/metabolismo , Berberina/análogos & derivados , Bloqueadores dos Canais de Cálcio/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Hipogonadismo/tratamento farmacológico , Nifedipino/farmacologia , Fosfoproteínas/metabolismo , Testosterona/biossíntese , Animais , Berberina/farmacologia , Cálcio/metabolismo , Regulação para Baixo , Glutationa Peroxidase/sangue , Hipogonadismo/metabolismo , Hipóxia/metabolismo , Hipóxia/veterinária , L-Lactato Desidrogenase/sangue , Hormônio Luteinizante/sangue , Masculino , Malondialdeído/sangue , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Testículo/lesões , Testículo/metabolismo , Testículo/patologia , Testosterona/sangue , Regulação para CimaRESUMO
AIM: Downregulation of androgen biosynthesis genes StAR (steroidogenic acute regulatory) and 3ß-HSD (3ß-hydroxysteroid dehydrogenase) contributes to low testosterone levels in hypoxic mice and is possibly related to increased expression of pro-inflammatory cytokines in the testis. The aim of this study is to investigate the effects of CPU86017-RS that block Ca(2+) influx on hypoxia-induced testis insult in mice. METHODS: Male ICR mice were divided into 5 groups: control group, hypoxia group, hypoxia group treated with nifedipine (10 mg/kg), hypoxia groups treated with CPU86017-RS (60 or 80 mg/kg). Hypoxia was induced by placing the mice in a chamber under 10%±0.5% O2 for 28 d (8 h per day). The mice were orally administered with drug in the last 14 d. At the end of experiment the testes of the mice were harvested. The mRNA and protein levels of StAR, 3ß-HSD, connexin 43 (Cx43), matrix metalloprotease 9 (MMP9), endothelin receptor A (ET(A)R) and leptin receptor (OBRb) were analyzed using RT-PCR and Western blotting, respectively. The malondialdehyde (MDA), lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and acid phosphatase (ACP) levels were measured using biochemical kits. Serum testosterone concentration was measured with radioimmunoassay. RESULTS: Hypoxia significantly increased the MDA level, and decreased the LDH, ACP and SDH activities in testes. Meanwhile, hypoxia induced significant downregulation of StAR and 3ß-HSD in testes responsible for reduced testosterone biosynthesis. It decreased the expression of Cx43, and increased the expression of MMP9, ETAR and OBRb, leading to abnormal testis function and structure. These changes were effectively diminished by CPU86017-RS (80 mg/kg) or nifedipine (10 mg/kg). CONCLUSION: Low plasma testosterone level caused by hypoxia was due to downregulation of StAR and 3ß-HSD genes, in association with an increased expression of pro-inflammatory cytokines. These changes can be alleviated by CPU86017-RS or nifedipine.
Assuntos
Androgênios/metabolismo , Berberina/análogos & derivados , Bloqueadores dos Canais de Cálcio/uso terapêutico , Hipóxia/complicações , Doenças Testiculares/tratamento farmacológico , Testículo/efeitos dos fármacos , 3-Hidroxiesteroide Desidrogenases/metabolismo , Androgênios/genética , Animais , Berberina/farmacologia , Berberina/uso terapêutico , Bloqueadores dos Canais de Cálcio/farmacologia , Conexina 43/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Metaloproteinase 9 da Matriz/genética , Camundongos , Nifedipino/farmacologia , Nifedipino/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Fosfoproteínas/sangue , Receptores para Leptina/genética , Doenças Testiculares/etiologia , Doenças Testiculares/metabolismo , Doenças Testiculares/patologia , Testículo/metabolismo , Testículo/patologia , Testosterona/sangueRESUMO
AIM: To investigate the protection of pulmonary arterial rings and cardiac myocytes of rats by raisanberine (RS), a derivative of berberine, against hypoxia injury and to elucidate the action mechanisms. METHODS: Adult SD rats were exposed to intermittent hypoxia for 17 d or 28 d. The pulmonary arterial rings were isolated and vascular activity was measured using a transducer and computer-aided system. The difference in the tension produced by phenylephrine in the presence and absence of L-nitroarginine (10 µmol/L) was referred to as the NO bioavailability; the maximum release of NO was assessed by the ratio of the maximal dilatation caused by ACh to those caused by sodium nitroprusside. After the lungs were fixed, the internal and the external diameters of the pulmonary arterioles were measured using a graphic analysis system. Cultured cardiac myocytes from neonatal rats were exposed to H(2)O(2) (10 µmol/L) to mimic hypoxia injury. ROS generation and [Ca(2+)](i) level in the myocytes were measured using DHE and Fluo-3 fluorescence, respectively. RESULTS: Oral administration of RS (80 mg/kg), the NADPH oxidase inhibitor apocynin (APO, 80 mg/kg) or Ca(2+) channel blocker nifedipine (Nif, 10 mg/kg,) significantly alleviated the abnormal increase in the vasoconstriction force and endothelium-related vasodilatation induced by the intermittent hypoxia. The intermittent hypoxia markedly decreased the NO bioavailability and maximal NO release from pulmonary arterial rings, which were reversed by APO or RS administration. However, RS administration did not affect the NO bioavailability and maximal NO release from pulmonary arterial rings of normal rats. RS, Nif or APO administration significantly attenuated the pulmonary arteriole remodeling. Treatment of cultured cardiac myocytes with RS (10 µmol/L) suppressed the ROS generation and [Ca(2+)](i) increase induced by H(2)O(2), which were comparable to those caused by APO (10 µmol/L) or Nif (0.1 µmol/L). CONCLUSION: Raisanberine relieved hypoxic/oxidant insults to the pulmonary artery and cardiac myocytes of rats by suppressing activated NADPH oxidase and increased calcium influx.
Assuntos
Antioxidantes/farmacologia , Berberina/análogos & derivados , Berberina/farmacologia , Sinalização do Cálcio/efeitos dos fármacos , Hipóxia/tratamento farmacológico , Miócitos Cardíacos/efeitos dos fármacos , NADPH Oxidases/antagonistas & inibidores , Artéria Pulmonar/efeitos dos fármacos , Administração Oral , Animais , Antioxidantes/administração & dosagem , Arteríolas/efeitos dos fármacos , Arteríolas/enzimologia , Berberina/administração & dosagem , Bloqueadores dos Canais de Cálcio/farmacologia , Canais de Cálcio/efeitos dos fármacos , Canais de Cálcio/metabolismo , Células Cultivadas , Citoproteção , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Regulação para Baixo , Inibidores Enzimáticos/farmacologia , Hipóxia/enzimologia , Hipóxia/patologia , Hipóxia/fisiopatologia , Masculino , Miócitos Cardíacos/enzimologia , NADPH Oxidases/metabolismo , Óxido Nítrico/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Artéria Pulmonar/enzimologia , Artéria Pulmonar/patologia , Artéria Pulmonar/fisiopatologia , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Fatores de Tempo , Vasoconstrição/efeitos dos fármacos , Vasoconstritores/farmacologia , Vasodilatadores/farmacologiaRESUMO
AIM: To investigate which endothelin receptors mediated isoproterenol (ISO)-induced downregulation of FKBP12.6/12 in cardiomyocytes and study whether argirhein, a novel compound containing rhein and L-arginine that has anti-inflammatory activity, could reverse the downregulation of FKBP12.6/12 induced by ISO. METHODS: Neonatal rat cardiomyocytes were incubated with ISO to downregulate FKBP12.6/12. Then the cells were treated with a selective ET(A) blocker (PD156707) and a ET(B) blocker (IRL1038), a dual ET(A)/ET(B) antagonist (CPU0213), and argirhein, respectively. FKBP12.6/12 expression was assayed by RT-PCR, Western blot, and immunocytochemistry. RESULTS: The expression of FKBP12.6 mRNA was reduced by 37.7% (P<0.01) and 28.9% (P<0.05) relative to the control by ISO 1 and 0.1 µmol/L, respectively, but no response to ISO 0.01 µmol/L was observed in vitro. FKBP12.6/12 protein expression was reduced by 47.2% (P<0.01) and 37.8% (P<0.05) by ISO 1 and 0.1 µmol/L, respectively. This decrease was reversed significantly by PD156707, or IRL1038, and CPU0213. CPU0213 was more potent than either PD156707 or IRL-1038. Argirhein 10 µmol/L blunted the downregulation of FKBP12.6/12 by ISO, as demonstrated by the rising mRNA and protein levels and by the fluorescent density of the ISO-incubated cardiomyocytes. CONCLUSION: In cardiomyocytes, the ISO induced downregulation of FKBP12.6/12 is modulated by both ET(A) and ET(B). A new compound, argirein, reversed the down-regulation of FKBP12.6/12 expression in myocardial cells stimulated with ISO.
Assuntos
Antraquinonas/farmacologia , Anti-Inflamatórios/farmacologia , Proteína 1A de Ligação a Tacrolimo/efeitos dos fármacos , Proteínas de Ligação a Tacrolimo/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Western Blotting , Células Cultivadas , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Isoproterenol/administração & dosagem , Isoproterenol/toxicidade , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/patologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor de Endotelina A/metabolismo , Receptor de Endotelina B/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína 1A de Ligação a Tacrolimo/genética , Proteínas de Ligação a Tacrolimo/genéticaRESUMO
AIM: To study whether calcium-modulating proteins CASQ2, FKBP12.6 and SERCA2a participate in diabetic cardiomyopathy, and whether the beneficial actions of testosterone, sildenafil or fructose diphosphate Sr (FDP-Sr) in the treatment of diabetic cardiomyopathy result from suppressing these molecules. METHODS: Fifty male Sprague-Dawley (SD) rats were divided into five groups. Except for the normal group (non-diabetic), the other four groups were injected with streptozotocin (STZ, 60 mg/kg, ip) to induce diabetes. Four weeks after STZ injection, the four groups received sildenafil (12 mg·kg(-1)·d(-1), ig, for 4 week), FDP-Sr (200 mg/kg, ig, for 4 week), testosterone propionate (4 mg·kg(-1)·d(-1), sc, for 4 week), or no treatment, respectively. RESULTS: In the diabetic rats, blood glucose, free fatty acids, triglycerides, total cholesterol, and low-density lipoprotein cholesterol (LDL-C) were significantly increased, while high-density lipoprotein cholesterol (HDL-C) was significantly reduced, as compared to the non-diabetic rats. Cardiac dysfunction and myocardial hypertrophy of the diabetic rats were associated with increased mRNA and protein expression of iNOS, OBRb, and PKCÉ, while expression of CASQ2, SERCA2a, and FKBP12.6 was significantly down-regulated. Sildenafil and FDP-Sr, but not testosterone, significantly attenuated the biomarker abnormalities, without changing the metabolic abnormalities. CONCLUSION: CASQ2, FKBP12.6 and SERCA2a were down-regulated in diabetic cardiomyopathy. Sildenafil and FDP-Sr, but not testosterone, attenuated the cardiac dysfunction in diabetic cardiomyopathy, without changing the metabolic abnormalities, which may results from inhibiting oxidative and inflammatory cytokines and improving calcium homeostasis.
Assuntos
Proteínas de Ligação ao Cálcio/genética , Cardiomiopatias Diabéticas/prevenção & controle , Frutose/farmacologia , Miocárdio/metabolismo , Inibidores da Fosfodiesterase 4/farmacologia , Piperazinas/farmacologia , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/genética , Sulfonas/farmacologia , Proteínas de Ligação a Tacrolimo/genética , Androgênios/metabolismo , Animais , Glicemia/metabolismo , Western Blotting , Cardiomiopatias Diabéticas/metabolismo , Homeostase , Metabolismo dos Lipídeos , Masculino , Estresse Oxidativo , Purinas/farmacologia , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Citrato de SildenafilaRESUMO
AIM: To investigate whether CPU86017, a berberine derivative, attenuates heart failure by blocking calcium influx and exerting its antioxidant activity. METHODS: Myocardial infarction was induced in male Sprague-Dawley rats for 17 d followed by isoproterenol (ISO) (5 mg/kg, sc) treatment for 5 d to reduce cardiac function. The rats were divided into 5 groups: sham operation, myocardial infarction (MI), MI plus ISO, and co-treated (in mg/kg, po) with either propranolol (PRO, 10) or CPU86017 (80). Hemodynamic measurements were conducted, and measurements of the redox system, calcium handling proteins and endothelin (ET) system in vivo were done. Furthermore, calcium flux studies and PLB immunocytochemistry were conducted in vitro. RESULTS: Compared to sham operation, HF was evident following MI and further worsened by ISO treatment. This occurred in parallel with downregulated mRNA and protein production of SERCA2a, PLB, and FKBP12.6, and was associated with upregulation of preproET-1, endothelin converting enzyme, and PKA mRNA production in the myocardium in vivo. Calcium leakage was induced by ISO treatment of isolated beating myocytes in vitro. These changes were attenuated by treatment with either PRO or CPU86017. PLB fluorescence in myocytes was downregulated by ISO treatment, and was relieved significantly by treatment with antioxidant aminoguanidine, ascorbic acid or CPU86017 in vitro. CONCLUSION: HF, calcium leakage, downregulated PLB, FKBP12.6, SERCA2a production, and upregulated PKA were caused by ISO treatment, and were abolished by CPU86017 treatment. The beneficial effects of CPU86017 are attributable to its antioxidant and calcium influx blocking effects.
Assuntos
Antioxidantes/farmacologia , Berberina/análogos & derivados , Proteínas de Ligação ao Cálcio/metabolismo , Cálcio/metabolismo , Regulação para Baixo , Insuficiência Cardíaca/prevenção & controle , Animais , Sequência de Bases , Berberina/farmacologia , Primers do DNA , Imuno-Histoquímica , Masculino , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
1. Activation of beta-receptors may modulate potassium channels and calcium handling proteins and serve as a basis for arrhythmogenesis. We determined whether an endothelin (ET) receptor antagonist CPU0213 could relieve the isoprenaline-(ISO) induced changes in I(Kr) and I(Ks) and calsequestrin 2 (CASQ2) in the heart. 2. In isolated ventricular myocytes, the I(Kr) and I(Ks) currents and expression for CASQ2, FKBP12.6, SERCA2a and ET(A)R were measured in the presence of ISO and either propranolol or CPU0213. 3. In the presence of ISO, I(Kr) and I(Ks) currents were significantly exaggerated and FKBP12.6, SERCA2a and CASQ2 were downregulated together with upregulation of ET(A)R in the myocardium. Interestingly, endothelin-1 was also effective in downregulating the expression of CASQ2. These changes were partially relieved by either CPU0213 or propranolol. 4. I(Kr) and I(Ks) currents can be separated into exaggerated/induced and basic components in the presence of ISO. The former, induced by ISO, is pathological and sensitive to either CPU0213 or propranolol. 5. Exaggerated I(Kr) and I(Ks) and downregulated CASQ2 by ISO are relevant to stress-related events in which the ET pathway is actively involved. By suppressing the ISO-exaggerated I(Kr) and I(Ks) and normalizing the expression of CASQ2, endothelin receptor antagonism is likely promising in dealing with stress-related cardiac arrhythmias.
Assuntos
Proteínas de Ligação ao Cálcio/biossíntese , Endotelinas/fisiologia , Isoproterenol/farmacologia , Miócitos Cardíacos/efeitos dos fármacos , Canais de Potássio/metabolismo , Receptores Adrenérgicos beta/metabolismo , Animais , Arritmias Cardíacas/etiologia , Arritmias Cardíacas/metabolismo , Western Blotting , Células Cultivadas , Regulação para Baixo , Antagonistas dos Receptores de Endotelina , Endotelinas/metabolismo , Cobaias , Masculino , Potenciais da Membrana/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Técnicas de Patch-Clamp , Pirazóis/farmacologia , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estresse Psicológico/complicaçõesRESUMO
BACKGROUND/AIMS: NADPH oxidase(NOX) is the main source of reactive oxygen species (ROS) in diabetic nephropathy (DN). Activation of NOX could be mediated via endothelin A (ET(A)R) and B receptors (ET(B)R) of the endothelin (ET) system. Thus, CPU0213, a dual ET receptor antagonist, was expected to attenuate DN by suppressing NOX. METHODS: Diabetes was produced in male Sprague-Dawley rats 8 weeks after a single injection of streptozotocin (STZ), and treatment with CPU0213 was initiated in the last 4 weeks. Rat mesangial cells (MCs) were incubated with 30 mM glucose for 48 h supplemented with CPU0213 or the NOX inhibitors apocynin and diphenyleneiodonium. RESULTS: After 8 weeks of hyperglycemia, abnormal renal function was associated with oxidative stress and an increased renal weight index in STZ-treated rats. Additionally, upregulation of NOX subunits and the ET system was found in diabetic rats and MCs treated with 30 mM glucose and suppressed by CPU0213 or NOX inhibitors. Except for blood glucose, CPU0213 markedly suppressed these abnormalities in DN. CONCLUSION: Upregulation of NOX is associated with upregulation of the ET pathway in the pathology of DN. The dual ET receptor antagonist (ET(A)R and ET(B)R) CPU0213 effectively normalized renal function in DN by suppressing NOX.
Assuntos
Nefropatias Diabéticas/enzimologia , NADPH Oxidases/metabolismo , Pirazóis/farmacologia , Receptores de Endotelina/metabolismo , Animais , Células Cultivadas , Diabetes Mellitus Experimental/enzimologia , Antagonistas dos Receptores de Endotelina , Masculino , Estresse Oxidativo , Ratos , Ratos Sprague-DawleyRESUMO
AIM: Spatial dispersion of bioactive substances in the myocardium could serve as pathological basis for arrhythmogenesis and cardiac impairment by beta-adrenoceptor stimulation. We hypothesized that dispersed NADPH oxidase, protein kinase Cepsilon (PKCepsilon), early response gene (ERG), and matrix metalloproteinase 9(MMP-9) across the heart by isoproterenol (ISO) medication might be mediated by the endothelin (ET) - ROS pathway. We aimed to verify if ISO induced spatially heterogeneous distribution of pPKCepsilon, NAPDH oxidase, MMP-9 and ERG could be mitigated by either an ET receptor antagonist CPU0213 or iNOS inhibitor aminoguanidine. METHODS: Rats were treated with ISO (1 mg/kg sc) for 10 days, and drug interventions (mg/kg) either CPU0213 (30 sc) or aminoguanidine (100 ip) were administered on days 8-10. Expression of NADPH oxidase, MMP-9, ERG, and PKCepsilon in the left and right ventricle (LV, RV) and septum (S) were measured separately. RESULTS: Ventricular hypertrophy was found in the LV, S, and RV, in association with dispersed QTc and oxidative stress in ISO-treated rats. mRNA and protein expression of MMP-9, PKCepsilon, NADPH oxidase and ERG in the LV, S, and RV were obviously dispersed, with augmented expression mainly in the LV and S. Dispersed parameters were re-harmonized by either CPU0213, or aminoguanidine. CONCLUSION: We found at the first time that ISO-induced dispersed distribution of pPKCepsilon, NADPH oxidase, MMP-9, and ERG in the LV, S, and RV of the heart, which were suppressed by either CPU0213 or aminoguanidine. It indicates that the ET-ROS pathway plays a role in the dispersed distribution of bioactive substances following sustained beta-receptor stimulation.
Assuntos
Cardiotônicos/farmacologia , Antagonistas dos Receptores de Endotelina , Coração/efeitos dos fármacos , Isoproterenol/farmacologia , Metaloproteinase 9 da Matriz/metabolismo , NADPH Oxidases/metabolismo , Pirazóis/farmacologia , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Guanidinas/farmacologia , Hipertrofia/induzido quimicamente , Masculino , Metaloproteinase 9 da Matriz/genética , Miocárdio/enzimologia , Miocárdio/patologia , NADPH Oxidases/genética , Óxido Nítrico Sintase/antagonistas & inibidores , Proteína Quinase C-épsilon/genética , Proteína Quinase C-épsilon/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
1. The aim of the present study was to explore the effects of CPU0213, a dual endothelin ET(A)/ET(B) receptor antagonist, and nifedipine, a calcium antagonist, in relieving pulmonary hypertension (PH). Both endothelin receptor and calcium antagonists have been reported to be effective in alleviating the remodelling of pulmonary arteries induced by monocrotaline (MCT) in rats. 2. After an initial single dose of 60 mg/kg, s.c., MCT, CPU0213 was administered to rats at doses of 25, 50 or 100 mg/kg, p.o., for 28 days. In addition, nifedipine was administered to another group of rats at a dose of 10 mg/kg, p.o., for 28 days. The haemodynamics of the right ventricle, pulmonary vascular activity, remodelling of the pulmonary arterioles (< 150 microm) and biochemical changes were evaluated. 3. Right ventricular systolic pressure (RVSP), central venous pressure (CVP), the maximum rate of uprising pressure (dP/dT(max)) and the weight index of the right ventricle were significantly elevated in MCT-treated rats. In addition, increases in pulmonary endothelin-1, malonyldialdehyde (MDA) and hydroxyproline content and a reduction in superoxide dismutase activity was found after MCT treatment. The thickness and area of the pulmonary arterial wall were significantly increased in MCT-treated rats compared with control rats. At all three doses tested, CPU0213 ameliorated these changes in a dose-dependent manner and the effects were associated with a greater reduction in the remodelling of pulmonary arterioles. However, nifedipine was only partially effective in amelerioating biochemical and haemodynamic changes induced by MCT, significantly reducing RVSP, CVP, +dp/dt(max), tissue MDA, inducible nitric oxide synthase and hydroxyproline content, increasing -dp/dt(min) and having no effect on the other parameters investigated. In addition, nifedipine had no effect on remodelling of the arterial wall. 4. In conclusion, CPU0213 is more effective than nifedipine in suppressing the remodelling of pulmonary arterioles in PH induced by MCT treatment of rats. Furthermore, CPU0213 may have promise in treating PH secondary to connective tissue disease.
Assuntos
Antagonistas do Receptor de Endotelina A , Antagonistas do Receptor de Endotelina B , Hipertensão Pulmonar/tratamento farmacológico , Pulmão/irrigação sanguínea , Pirazóis/uso terapêutico , Animais , Arteríolas/efeitos dos fármacos , Bloqueadores dos Canais de Cálcio/administração & dosagem , Bloqueadores dos Canais de Cálcio/farmacologia , Bloqueadores dos Canais de Cálcio/uso terapêutico , Modelos Animais de Doenças , Feminino , Hipertensão Pulmonar/induzido quimicamente , Hipertensão Pulmonar/metabolismo , Hipertensão Pulmonar/fisiopatologia , Pulmão/metabolismo , Masculino , Monocrotalina , Nifedipino/administração & dosagem , Nifedipino/farmacologia , Nifedipino/uso terapêutico , Pirazóis/administração & dosagem , Pirazóis/farmacologia , Ratos , Ratos Sprague-Dawley , Receptor de Endotelina A/biossíntese , Receptor de Endotelina B/biossínteseRESUMO
OBJECTIVES: We aimed to investigate hypercholesterolaemia-induced early renal lesions which result in abnormal expression of endothelin A receptor (ET(A)R), induced nitric oxide synthase (iNOS) and matrix metalloproteinase 9 (MMP-9). We hypothesized that this is due to an upregulated endothelin (ET) pathway consequent to hypercholesterolaemia and that CPU0213, a dual ET antagonist, could mitigate these changes. METHODS: Rats were randomly divided into four groups: (1), control; (2), high-fat diet for 60 days (HFD); HFD rats medicated in the last 15 days with either (3) CPU0213 (30 mg/kg daily, s.c.) or (4) simvastatin (4 mg/kg daily, p.o.). KEY FINDINGS: Body weight, serum triglycerides, total cholesterol and low-density-lipoprotein cholesterol were significantly increased, whereas high-density lipoprotein cholesterol decreased in the HFD group, relative to normal. Meanwhile, these changes were associated with upregulation of mRNA and protein of ET(A)R, iNOS and MMP-9 in the kidney. The lipid-lowering effect of simvastatin was predominant, lessening abnormal expression of these molecules in the kidney dramatically. Interestingly, CPU0213 significantly normalized expression of mRNA and protein of ET(A)R, iNOS and MMP-9, comparable with simvastatin, leaving no changes in hyperlipidaemia. CONCLUSIONS: CPU0213 relieves renal lesions by blunting hypercholesterolaemia caused by the upregulated ET system, iNOS and MMP-9 in the kidney. This indicates that CPU0213 is promising in treating patients with end stage renal disease.
Assuntos
Hipercolesterolemia/tratamento farmacológico , Nefropatias/tratamento farmacológico , Metaloproteinase 9 da Matriz/biossíntese , Óxido Nítrico Sintase Tipo II/biossíntese , Pirazóis/farmacologia , Receptor de Endotelina A/biossíntese , Sinvastatina/farmacologia , Animais , Peso Corporal/efeitos dos fármacos , Colesterol/sangue , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Antagonistas do Receptor de Endotelina A , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Hipercolesterolemia/complicações , Rim/metabolismo , Nefropatias/etiologia , Nefropatias/metabolismo , Masculino , Metaloproteinase 9 da Matriz/genética , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II/genética , Pirazóis/uso terapêutico , RNA Mensageiro/biossíntese , Ratos , Ratos Sprague-Dawley , Receptor de Endotelina A/genética , Sinvastatina/uso terapêutico , Triglicerídeos/sangue , Regulação para CimaRESUMO
OBJECTIVES: Cardiac injury induced by isoprenaline produces stress. This stress can be mediated by the activated endothelin and leptin pathway; thus, the endothelin receptor antagonist CPU0213 may reverse these changes. CPU0213 is metabolized mainly by cytochrome P450 (CYP)3A, thus, erythromycin, an inhibitor of CYP3A, could affect its effects by raising its plasma levels. METHODS: Forty rats were divided into five groups. Group 1 rats were normal. Group 2 rats were administered isoprenaline (1 mg/kg, s.c.) for 10 days. Groups 3, 4 and 5 were administered isoprenaline, but group 3 was given erythromycin (100 mg/kg, p.o.) alone on days six to ten, group 4 was given CPU0213 20 mg/kg (s.c.) on days six to ten, whilst group 5 received erythromycin plus CPU0213 on days six to ten. Measurements were conducted to observe changes in the haemodynamics, cardiac weight index, serum lactate dehydrogenase and creatine kinase levels, and expression of endothelin receptor A (ETA), leptin and its OBRb receptor. KEY FINDINGS: In isoprenaline-treated rats, cardiac hypertrophy and dysfunction were found. This was associated with upregulated myocardial leptin protein and OBRb receptor mRNA. Immunohistochemical assay of ETA was upregulated, accompanied with downregulation of FKBP12.6 (calstabin 2) in isoprenaline-treated rats. These effects were significantly reversed by CPU0213. HPLC assay presented an increased plasma level of CPU0213 by erythromycin, but no change in its effects. CONCLUSIONS: CPU0213 improved isoprenaline-induced cardiomyopathy by modulating ETA, leptin and FKBP12.6. However, erythromycin increased plasma levels but did not change its effects.
Assuntos
Cardiomegalia/tratamento farmacológico , Antagonistas dos Receptores de Endotelina , Eritromicina/farmacologia , Insuficiência Cardíaca/tratamento farmacológico , Leptina/metabolismo , Pirazóis/farmacologia , Proteínas de Ligação a Tacrolimo/metabolismo , Animais , Cardiomegalia/induzido quimicamente , Cardiomegalia/fisiopatologia , Cromatografia Líquida de Alta Pressão , Citocromo P-450 CYP3A/metabolismo , Inibidores do Citocromo P-450 CYP3A , Regulação para Baixo , Insuficiência Cardíaca/induzido quimicamente , Insuficiência Cardíaca/fisiopatologia , Hemodinâmica/efeitos dos fármacos , Isoproterenol , Masculino , Pirazóis/sangue , Pirazóis/uso terapêutico , Ratos , Ratos Sprague-Dawley , Receptores de Endotelina/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para CimaRESUMO
OBJECTIVES: Male hypogonadism is frequently associated with testopathy in patients with type 2 diabetes and in middle-aged males. We hypothesized that abnormal matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in testis have large roles to play in male hypogonadism. It has been found in diabetic rats that a novel compound, strontium fructose 1,6-diphosphate (FDP-Sr), with extra high energy supply, could reverse male hypogonadism by normalizing MMP-9 and TIMPs in the testis. We investigated whether FDP-Sr could be promising in treating diabetic testopathy. METHODS: Adult male Sprague-Dawley rats were administered a single dose of streptozocin (65 mg/kg, i.p.) to induce diabetes. The diabetic rats were treated with FDP-Sr in three doses or testosterone propionate in the final four weeks during the eight-week study. KEY FINDINGS: Serum testosterone, activity of marker enzymes, and mRNA of MMPs and TIMPs and protein of MMP-9 in the testis were detected. After eight weeks, the activity of acid phosphatase, lactate dehydrogenase, succinate dehydrogenase and g-glutamyl transpeptidase in testis were significantly decreased (P < 0.01), accompanied by down-regulated mRNA and activity of MMP-2 and MMP-9 (P < 0.01) and upregulated mRNA of TIMP-1 and TIMP-2. Downregulated MMP-9 protein and degenerative changes in histology were predominant in diabetic testis. CONCLUSIONS: FDP-Sr or testosterone propionate significantly normalized expression and activity of the MMPs-TIMPs system to attenuate changes in serum testosterone, marker enzymes and histology in testis. Effects of FDP-Sr were dose-dependent and comparable with those of testosterone propionate. By supplying extra energy, FDP-Sr could be promising in treating diabetic testopathy by normalizing abnormal MMP-9 and its endogenous inhibitors in testes.
Assuntos
Frutosedifosfatos/farmacologia , Hipogonadismo/tratamento farmacológico , Inibidores de Metaloproteinases de Matriz , Metaloproteinases da Matriz/efeitos dos fármacos , Estreptozocina/toxicidade , Fosfatase Ácida/antagonistas & inibidores , Fosfatase Ácida/sangue , Animais , Complicações do Diabetes/tratamento farmacológico , Diabetes Mellitus Tipo 2/complicações , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Expressão Gênica , Hiperglicemia/induzido quimicamente , Hipogonadismo/complicações , L-Lactato Desidrogenase/antagonistas & inibidores , L-Lactato Desidrogenase/sangue , Masculino , Metaloproteinases da Matriz/classificação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Estreptozocina/administração & dosagem , Succinato Desidrogenase/antagonistas & inibidores , Succinato Desidrogenase/sangue , Testículo/efeitos dos fármacos , Testículo/enzimologia , Testículo/ultraestrutura , Testosterona/antagonistas & inibidores , Testosterona/sangue , Inibidores Teciduais de Metaloproteinases/metabolismo , gama-Glutamiltransferase/antagonistas & inibidores , gama-Glutamiltransferase/sangueRESUMO
The development of novel anti-arrhythmic drugs is necessary, specifically agents that do not cause torsades de pointes (Tdp). Ion channelopathy that is involved in mechanisms underlying sudden cardiac death (SCD) includes both ion channels in the membrane, and the calcium-releasing channels and the calcium uptake process in the sarcoplasmic reticulum. Advances in the understanding of abnormalities of ion channels in the myocardium caused by congenital defects or by a failing heart and cardiomyopathy offer further insights into the relationship between channelopathy and SCD. Enhanced L-type Ca2+ current (ICa.L) activity has been detected in the hearts of patients with a mutation of the Cav1.2 gene; these patients exhibit a high risk of SCD. Rats with thyroxin-induced cardiomyopathy demonstrate an increase in ICa.L activity that is responsible for exacerbated ventricular fibrillation (VF). This is suppressed by propranolol or CPU-86017, a class III anti-arrhythmic agent with potent antioxidant activity. Interestingly, an increase in rapidly (IKr) and slowly (IKs) activating delayed rectifying K+ currents is caused by gain-of-function mutations of the KCNH2 and KCNQ1 genes, respectively, in patients with short QT syndrome (SQT). Increased IKr and IKs, which are associated with exacerbated VF, are also found in models of thyroxin-induced cardiomyopathy and are suppressed by CPU-86017. ICa.L, IKr and IKs can also be induced in cardiomyocytes when incubated with isoproterenol. A reversal of upstream lesions by an endothelin receptor antagonist CPU-0213 provides suppression of ventricular tachyarrhythmias and upregulates FK506 binding protein 12.6. CPU-86017 and its chiral isomer SR-CPU-86017 relieve upstream lesions, with mild suppression of IKr and moderate suppression of IKs and ICa.L. These agents may be promising as anti-arrhythmic agents that produce less Tdp tachyarrhythmias.
Assuntos
Antiarrítmicos/farmacologia , Endotelinas/fisiologia , Canais Iônicos/efeitos dos fármacos , Transdução de Sinais/fisiologia , Animais , Arritmias Cardíacas/fisiopatologia , Berberina/análogos & derivados , Berberina/farmacologia , Berberina/uso terapêutico , Morte Súbita Cardíaca , Humanos , Canais de Potássio/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Fibrilação Ventricular/fisiopatologiaRESUMO
Downregulation of FKBP12.6 and sarcoplasmic reticulum Ca(2+) ATPase (SERCA2a) contributes to sudden cardiac death and heart failure. We aimed to test the hypothesis that (i) downregulation of FKBP12.6 and SERCA2a can be taken as molecular markers for drug interventions and (ii) such downregulation is produced by crosstalk between endothelin-reactive oxygen species and beta-adrenoceptors stimulation, mediated by hyperphosphorylation of protein kinase Cvarepsilon (PKCvarepsilon). Rat cardiomyocytes were incubated with isoproterenol (1 microM), endothelin-1 (0.1 microM) or hydrogen peroxide (10 microM) for 18 h, resulting in downregulation of mRNA and protein of FKBP12.6 and SERCA2a, as well as upregulation of PKCvarepsilon mRNA and phosphorylated PKCvarepsilon protein. These changes were reversed by an application of either propranolol (1 microM), endothelin receptor antagonist CPU0213 (1 microM) or vitamin E (1 microM). As indicated by the fluorescent dye Fluo3, diastolic [Ca(2+)](i) in rat ventricular myocytes was increased after incubation with isoproterenol (0.1 microM). The increased [Ca(2+)](i) in diastole was dramatically decreased by CPU0213. Thus, the downregulation of FKBP12.6 and SERCA2a, and hyperphosphorylation of PKCvarepsilon, appear to be related to crosstalk between over-activated endothelin-reactive oxygen species and a beta-adrenoceptor pathway. CPU0213 is beneficial in treating cardiac insufficiency and preventing cardiac arrhythmias possibly by normalizing hyperphosphorylation of PKCvarepsilon and abnormal FKBP12.6 and SERCA2a. The antioxidant activity of vitamin E was sufficient to normalize the levels of FKBP12.6 and SERCA2a and phosphorylation of PKCvarepsilon. Thus by testing with biomarkers FKBP12.6 and SERCA2a, we have shown that the endothelin receptor antagonist CPU0213 and the antioxidant vitamin E may relieve risk of lethal arrhythmias and heart failure by suppressing PKCvarepsilon.
Assuntos
Antagonistas dos Receptores de Endotelina , Regulação da Expressão Gênica/efeitos dos fármacos , Pirazóis/farmacologia , Proteínas de Ligação a Tacrolimo/efeitos dos fármacos , Vitamina E/farmacologia , Animais , Antioxidantes/farmacologia , Biomarcadores Farmacológicos/metabolismo , Cálcio/metabolismo , Morte Súbita Cardíaca/prevenção & controle , Regulação para Baixo/efeitos dos fármacos , Insuficiência Cardíaca/tratamento farmacológico , Insuficiência Cardíaca/fisiopatologia , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Fosforilação , Proteína Quinase C-épsilon/efeitos dos fármacos , Proteína Quinase C-épsilon/metabolismo , Ratos , Ratos Sprague-Dawley , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/efeitos dos fármacos , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/metabolismo , Proteínas de Ligação a Tacrolimo/metabolismoRESUMO
Remodeling of the pulmonary artery is a major feature of pulmonary artery hypertension, and CPU86017, a derivative of berberine, is known to effectively alleviate hypoxic pulmonary hypertension (HPH). CPU86017 is a racemate, possessing two chiral centers: 7N and 13aC. We have compared the effects of four CPU86017 isomers, SS [(+)-7S, 13aS-CPU86017], SR [(-)-7S, 13aR-CPU86017], RR [(-)-7R, 13aR-CPU86017] and RS [(+)-7R, 13aS-CPU86017], on HPH. Sprague-Dawley rats were exposed to hypoxic conditions (10 +/- 0.5% O2 for 8 h per day) for 4 weeks and treated with CPU86017, SS, SR, RR or RS (4 mg/kg, subcutaneously) from day 15 to 28. After 4 weeks of exposure to hypoxia, remodeling of the right ventricle and the small pulmonary arteries (<150 microm) was very pronounced, and extra-cellular matrix (ECM) had been excessively produced in association with abnormal mRNA and protein expression of matrix metalloproteinase 9 (MMP9) and mRNA of tissue inhibitor of matrix metalloproteinase 1 and 2 (TIMP1, TIMP2). Expression of endothelin receptor A was upregulated, while that connexin 40 was downregulated. The administration of CPU86017 and its four isomers attenuated the changes, with the isomer RS exhibiting the most favorable effect on HPH rats. We propose that an activated endothelin pathway associated with an unbalanced MMP-TIMP system may contribute to the over-accumulation of ECM and the remodeling of the pulmonary arterioles in HPH. CPU86017 and its four isomers attenuate ECM accumulation and vascular remodeling by normalizing both the MMP-TIMP system and the ET system. The RS isomer is superior to the racemate CPU86017 in attenuating HPH.
Assuntos
Berberina/análogos & derivados , Regulação da Expressão Gênica/efeitos dos fármacos , Hipertensão Pulmonar/tratamento farmacológico , Receptor de Endotelina A/efeitos dos fármacos , Animais , Berberina/farmacologia , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Hipóxia/complicações , Masculino , Metaloproteinase 9 da Matriz/efeitos dos fármacos , Metaloproteinase 9 da Matriz/metabolismo , Artéria Pulmonar/efeitos dos fármacos , Artéria Pulmonar/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor de Endotelina A/metabolismo , Estereoisomerismo , Inibidor Tecidual de Metaloproteinase-1/efeitos dos fármacos , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Inibidor Tecidual de Metaloproteinase-2/efeitos dos fármacos , Inibidor Tecidual de Metaloproteinase-2/metabolismo , Remodelação Ventricular/efeitos dos fármacosRESUMO
An increase in reactive oxygen species (ROS) through NADPH oxidase activation frequently follows stress that activates beta-adrenoreceptors, leading to deterioration of cardiovascular disease. We hypothesized that upregulation of NADPH oxidase in the vasculature causes mild vascular spasm subsequent to chronic isoproterenol (ISO) administration, correlating significantly with activation of both ETA and ETB receptors. We tested whether the dual endothelin receptor antagonist CPU0213 is effective in reversing ISO-induced vascular abnormality by suppressing activated NADPH oxidase in the vasculature. Rats were injected with ISO (1 mg/kg, SC) for 10 days to induce vascular dysfunction and treated with CPU0213 (30 mg/kg, SC) or aminoguanidine (AMG, an inhibitor of iNOS, 100 mg/kg, PO) from day 7 to day 10. On day 11, we found an increase in vascular response to phenylephrine (Phe) and reductions in NO availability and acetylcholine (ACh)-induced relaxation in ISO-treated rats along with upregulated mRNA of ETA, ETB, iNOS, NADPH oxidase-Phox22 and Phox47, and matrix metalloproteinase 9 (MMP9). These abnormalities were attenuated by 3 days of intervention with CPU0213 but less with AMG. CPU0213 was more effective in relieving enhanced vascular constriction and reversal of ET receptor and MMP9 expression in the vasculature than was AMG. In conclusion, an upregulation of NADPH oxidase phox 22 and phox 47, ETA and ETB, and MMP9 correlates with vascular abnormality and the endothelin receptor antagonist CPU0213 was more effective than AMG in reversing ISO-induced enhanced vascular constriction by normalizing the above abnormal expression.