RESUMO
The research aimed to investigate the suppression effect of Mai Shu which contains hawthorn, hippophae, medlar, phytosterolsï¼ß-sitosterol, stigmasterol and campesterolï¼, ß-glucan and lycopeneon formation of atherosclerotic plaque in apolipoprotein E knock-out (ApoE-/-) mice. Liquid chromatography-ultravioletmass spectrometryï¼LC-UV-MCï¼ methods were used to analyze the main chemical composition of Mai Shu. Atherosclerotic mice models were established by high-fat diet. The mice were administrated with Mai Shu (1, 2, 4 g·kg-1·d-1) or other contrast materials by intragastric route for 10 weeks continuously. At the end of administration, the blood of mice was collected for tests of the serum total cholesterolï¼TCï¼, total triglycerideï¼TGï¼ and high density lipoprotein cholesterolï¼HDL-Cï¼ level. Atherosclerotic lesions in aorta and aortic root were assessed by calculating the relative area of lesionsï¼oil red O stainedï¼. Intravital fluorescence microscopic system was used to evaluate the leukocyte-endothelial adhesion in mesenteric artery of mice by detecting the rolling velocity of white blood cellsï¼WBCï¼. Collagenous fibers and macrophages in lesions were detected by sirius red staining and immunological histological chemistry to evaluate the atherosclerotic plaque stability. Results showed that Mai Shu contains various flavonoidsï¼9.5%ï¼, phytosterolsï¼23.8%ï¼ and polysaccharidesï¼8.9%ï¼. The serum lipid level of model animals was significantly higher than the control animals. Serum TC level was decreased by Mai Shu (4 g·kg-1, P < 0.001) compared to the untreated model. Serum TG level was reduced by Mai Shu (1, 2, 4 g·kg-1) compared to modelï¼P < 0.01ï¼. Area of atherosclerotic lesions in aorta and aortic root was decreased in Mai Shu group (aorta: 1 g·kg-1, P < 0.05; 2 g·kg-1, P < 0.01; 4 g·kg-1, P < 0.001; aortic root: 2, 4 g·kg-1, P < 0.01). Rolling velocity of white blood cells of Mai Shu (4 g·kg-1, P < 0.001) group was increased over the untreated model. Collagenous fibers in lesions were observationally increased by Mai Shu (1, 2 g·kg-1) and macrophages were decreased (2, 4 g·kg-1) compared to model. These results demonstrate that Mai Shu can obviously decrease the serum lipid levels and the risk of leukocyte-endothelial adhesion in ApoE-/- mice. The effect of Mai Shu may be associated with the decrease of macrophages in plaque.
Assuntos
Aterosclerose/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Placa Aterosclerótica/tratamento farmacológico , Animais , Aorta/patologia , Colesterol/sangue , Dieta Hiperlipídica , Modelos Animais de Doenças , Macrófagos/citologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout para ApoE , Fitosteróis/farmacologia , Triglicerídeos/sangueRESUMO
OBJECTIVE: Cutaneous wound is a common health problem of humans. Loropetalum chinens, a medicinal plant, is widely used to treat wounds among the people. The research aims to observe whether L. chinens can promote the rats' wounds healing process, isolate the extracts primarily and commit the wound healing selection, which provide work basis for wound healing research of L. chinens. METHOD: First we analyzed the possible components with HC-MS/MS, then committed our wound healing experiments for L. chinens in the rat incision wound model and excision wound model, which are commonly used worldwide. After that, we carried on the preliminary isolation of the L. chinens and we screened the heal-promoting effects of the isolations in incision wound model. RESULT: L. chinens significantly accelerates the wound healing of rat's skin, shortens the healing period, enhances the healing intensity and promotes the cell proliferation and blood vessels formation around the wounds. The isolations, which are petroleum ether layer, ethyl acetate layer and n-butyl alcohol layer, exert heal-promoting effects. It indicates that the possible morphon that promotes wound healing may exist in these three components, with small polar. CONCLUSIONS: L. chinens possesses strong wound healing promoting effects, and the active constituent, with small polar, exists in petroleum ether layer, ethyl acetate layer and n-butyl alcohol layer, and we should focus on these three layers when carrying on further studies.
Assuntos
Medicamentos de Ervas Chinesas/administração & dosagem , Hamamelidaceae/química , Dermatopatias/tratamento farmacológico , Pele/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Animais , Humanos , Masculino , Fitoterapia , Ratos , Ratos Wistar , Pele/lesões , Pele/fisiopatologia , Dermatopatias/fisiopatologiaRESUMO
A chronic inflammatory response possibly mediated by Amyloid ß (Aß) is believed to be a major factor in the pathology of Alzheimer's disease (AD). Studies suggest that the mediators of the inflammatory response, which might contribute to brain damage, involve cytokines, such as IL-1ß. IL-1ß could play an important part in the development of pathologic conditions. There is also an endogenous interleukin-1 receptor antagonist (IL-1RA) in IL-1 family, which could prevent the actions of IL-1ß by competing for receptor binding without inducing any signal transduction. Therefore, the balance of IL-1ß vs IL-1RA is a critical parameter in determining not only whether excessive host inflammation will occur, but also the degree of subsequent host cell damage and associated toxicity. In our previous study, it has been determined that the anti-inflammatory action of Gossypium herbaceam L. extracts (GHE) was involved in its neuroprotection. However, the effects of GHE on IL-1ß and IL-1RA have not been clearly defined in the experimental rat model of AD induced by Aß. Therefore, the current study is performed to evaluate whether GHE could affect the disequilibrium of IL-1RA/IL-1ß ratio in the hippocampus of rats after Aß treatment. Subsequently, we further identify that GHE could efficaciously promote Akt and GSK3ß phosphorylation, and thereby contribute to IL-1ß release decrease as well as a concurrent increase in the level of IL-1RA through NF-κB and MAPK pathways. As a consequence, GHE is potentially beneficial to maintain the endogenous IL-1RA/ IL-1ß balance in the hippocampus of rats and it might be a potential agent to ameliorate inflammatory process in AD.
Assuntos
Peptídeos beta-Amiloides/toxicidade , Fármacos Neuroprotetores/farmacologia , Extratos Vegetais/farmacologia , Transdução de Sinais/efeitos dos fármacos , Doença de Alzheimer/imunologia , Doença de Alzheimer/metabolismo , Animais , Western Blotting , Ensaio de Desvio de Mobilidade Eletroforética , Ensaio de Imunoadsorção Enzimática , Gossypium , Imuno-Histoquímica , Inflamação/imunologia , Inflamação/metabolismo , Proteína Antagonista do Receptor de Interleucina 1/imunologia , Proteína Antagonista do Receptor de Interleucina 1/metabolismo , Interleucina-1beta/imunologia , Interleucina-1beta/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/imunologiaRESUMO
Elevated levels of low-density cholesterol (LDL-C) are highly correlated with increased risk of cardiovascular diseases (CVD). Thus, current guidelines have recommended progressively lower LDL-C for cholesterol treatment and CVD prevention as the primary goal of therapy. Even so, some patients in the high risk category fail to achieve recommended LDL-C targets with currently available medications. Thereby, additional pharmaceutical strategies are urgently required. In the review, we aim to provide an overview of both current and emerging LDL-C lowering drugs. As for current available LDL-C lowering agents, attentions are mainly focused on statins, niacin, bile acid sequestrants, ezetimibe, fibrates and omega-3 fatty acids. On the other hand, the emerging drugs differ from mechanisms are including: intervention of cholesterol biosynthesis downstream enzyme (squalene synthase inhibitors), inhibition of lipoprotein assembly (antisense mRNA inhibitors of apolipoprotein B and microsomal transfer protein inhibitors), enhanced lipoprotein clearance (proprotein convertase subtilisin kexin type 9, thyroid hormone analogues), inhibition of intestinal cholesterol absorption (Niemann-Pick C1-like 1 protein and acyl coenzyme A:cholesterol acyltransferase inhibitors) and interrupting enterohepatic circulation (apical sodium-dependent bile acid transporter inhibitors). Several ongoing agents are in their different stages of clinical trials, in expectation of promising antihyperlipidemic drugs. Therefore, alternative drugs monotherapy or in combination with statins will be sufficient to reduce LDL-C concentrations to optimal levels, and a new era for better LDL-C managements is plausible.
Assuntos
Anticolesterolemiantes/uso terapêutico , LDL-Colesterol/sangue , Hipercolesterolemia/tratamento farmacológico , Animais , Humanos , Hipercolesterolemia/sangueRESUMO
BACKGROUND: Asperosaponin X was isolated from the roots of Dipsacus asper. In this study, we investigated the anti-myocardial ischemia and reperfusion (I/R) injury effects of Asperosaponin X in vivo and elucidated the potential mechanism in vitro. RESULTS: Asperosaponin X significantly attenuated hypoxia-induced cytotoxicity in a concentration-dependent manner in H9c2 cells. Treatment of H9c2 cells with Asperosaponin X 5 µM or 10 µM blocked TNF-α-induced nuclear factor kappaB (NF-κB) phosphorylation by blocking HMGB1 expression. Treatment of rats with Asperosaponin X 10mg/kg, (i.v.) protected the animals from myocardial I/R injury as indicated by a decrease in infarct volume, improvement in hemodynamics and reduction of myocardial damage severity. Treatment with Asperosaponin X also lowered serum levels of pro-inflammatory factors and reduced High mobility group box-1 protein (HMGB1), phosphorylated IκB-α and NF-κB expression in ischemic myocardial tissue. Additionally, continuous i.v. of Asperosaponin X 14 days attenuated cardiac remodeling. CONCLUSIONS: These protective effects suggested that Asperosaponin X may be due to block of myocardial inflammatory cascades through an HMGB1-dependent NF-κB signaling pathway.
Assuntos
Dipsacaceae , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Preparações de Plantas/uso terapêutico , Animais , Western Blotting , Células Cultivadas , Modelos Animais de Doenças , Proteína HMGB1/biossíntese , Proteínas I-kappa B/biossíntese , Masculino , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Miocárdio/metabolismo , Miocárdio/patologia , Inibidor de NF-kappaB alfa , NF-kappa B/antagonistas & inibidores , Ratos , Ratos Sprague-DawleyRESUMO
Connective tissue growth factor (CTGF) plays a pathogenic role in diabetic nephropathy (DN). Loganin, an iridoid glucoside compound was isolated from Cornus officinalis Sieb. et Zucc. This study was conducted to investigate the efficacy of loganin on DN and to elucidate the potential mechanism. High glucose (HG) stimulated cultured human renal proximal tubular epithelial cells (HK-2) analyzed CTGF expression by Western blotting and investigated whether extracellular signal-regulated kinase (ERK) signaling pathway was involved. Streptozotocin (STZ)-induced experimental DN, randomized to receive intragastric (i.g.) of loganin. Renal tissue, blood and urine samples were collected to determine and analyze. In vitro study, loganin reduced CTGF excretion in HG-induced HK-2 cells through the ERK signaling pathway. In vivo study, I.g. of loganin 5 mg/kg or 10 mg/kg significantly ameliorated renal function and increased body weight. Meanwhile, loganin reduced renal CTGF expression by immunohistochemical staining, reduced serum levels of CTGF. Besides, there were no significant differences in blood sugar levels between the loganin groups compared to the STZ-treated group. Furthermore, loganin ameliorated renal pathology. These results suggested that loganin exerts an early renal protective role to DN. Inhibition of CTGF may be a potential target in DN therapy, which highlights the possibility of using loganin to treat DN.
Assuntos
Fator de Crescimento do Tecido Conjuntivo/análise , Nefropatias Diabéticas/induzido quimicamente , Nefropatias Diabéticas/tratamento farmacológico , Iridoides/farmacologia , Animais , Glicemia/química , Linhagem Celular , Proliferação de Células , Fator de Crescimento do Tecido Conjuntivo/sangue , Fator de Crescimento do Tecido Conjuntivo/urina , Cornus/química , Cistatina C/química , Diabetes Mellitus Experimental/fisiopatologia , Nefropatias Diabéticas/patologia , Células Epiteliais/efeitos dos fármacos , Glucose/farmacologia , Humanos , Imuno-Histoquímica , Iridoides/administração & dosagem , Iridoides/química , Rim/efeitos dos fármacos , Rim/patologia , Sistema de Sinalização das MAP Quinases , Masculino , Ratos , Ratos Sprague-Dawley , Aumento de PesoRESUMO
The present study investigated the effects of Forsythoside B on an experimental model of sepsis induced by caecal ligation and puncture (CLP) in rats and elucidated the potential mechanism in cultured RAW 264.7 cells. Results showed that Forsythoside B concentration-dependently down-regulated the levels of TNF-α, IL-6 and high-mobility group-box 1 protein (HMGB1) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells, inhibited the IκB kinase (IKK) pathway and modulated nuclear factor (NF)- κB. Intravenous injection (i.v.) of Forsythoside B alone or plus Imipenem reduced serum levels of TNF-α, IL-6, HMGB1, triggering receptor expressed on myeloid cells (TREM-1) and endotoxin, while the serum level of IL-10 was up-regulated and myeloperoxidase (MPO) in lung, liver and small intestine was reduced. Meanwhile, i.v. of Forsythoside B alone or plus Imipenem reduced CLP-induced lethality in rats. These data indicated that the antisepsis effect of Forsythoside B is mediated by decreasing local and systemic levels of a wide spectrum of inflammatory mediators. Its antisepsis mechanism may be that Forsythoside B binds to LPS and reduces the biological activity of serum LPS, and inhibits NF-κB activition. Our studies enhance the case for the use of Forsythoside B in sepsis. Forsythoside B itself has promise as a therapy for the treatment of sepsis in humans.
Assuntos
Ácidos Cafeicos/farmacologia , Glucosídeos/farmacologia , Mediadores da Inflamação/metabolismo , Sepse/tratamento farmacológico , Animais , Linhagem Celular , Proteína HMGB1/metabolismo , Quinase I-kappa B/metabolismo , Imipenem/farmacologia , Interleucina-10/sangue , Interleucina-6/metabolismo , Intestino Delgado/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Fígado/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Masculino , Camundongos , NF-kappa B/metabolismo , Peroxidase/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Imunológicos/metabolismo , Receptor Gatilho 1 Expresso em Células Mieloides , Fator de Necrose Tumoral alfa/metabolismoRESUMO
This study explores the effects of 3'-deoxyadenosine, a compound from Cordyceps militaris, on lipid metabolic disorder induced by a high-fat-diet in C57BL/6 mice. These mice had an obese body, lipid metabolic disorder and insulin resistance and were treated orally with 100 mg/kg/day 3'-deoxyadenosine (DA), 15 mg/kg/day rosiglitazone and 150 mg/kg/day fenofibrate, respectively. Compared to the model mice, the body weight gain in DA-treated mice were decreased by 66.5%, serum triglyceride and total cholesterol levels were decreased by 20.7% and 16.7%, respectively, and the triglyceride content in the skeletal muscle was reduced by 41.2%. This treatment also had a significant effect on insulin resistance. In DA-treated mice, the serum insulin levels and homeostasis model assessment of the insulin resistance index were decreased by 30% and 46%, respectively, and the areas under the glucose-time curve were depressed by 18% in the insulin tolerance test and by 21.5% in the oral glucose tolerance test. Finally, the value of glucose infusion rates and insulin induced-glucose uptake into the skeletal muscle in the hyperinsulinemic-euglycemic clamp test were increased by 18% and 41%, respectively, compared to those in the model mice. This data suggests that the effects of DA on lipid metabolic disorder induced by a high-fat-diet may be linked to its improvement on insulin resistance, especially concerning the increase of insulin sensitivity in the skeletal muscle.
Assuntos
Cordyceps/química , Desoxiadenosinas/uso terapêutico , Gorduras na Dieta/efeitos adversos , Resistência à Insulina/fisiologia , Transtornos do Metabolismo dos Lipídeos/tratamento farmacológico , Metabolismo dos Lipídeos/efeitos dos fármacos , Obesidade/tratamento farmacológico , Animais , Área Sob a Curva , Desoxiadenosinas/farmacologia , Gorduras na Dieta/metabolismo , Modelos Animais de Doenças , Fenofibrato/farmacologia , Glucose/metabolismo , Técnica Clamp de Glucose , Intolerância à Glucose/induzido quimicamente , Intolerância à Glucose/tratamento farmacológico , Intolerância à Glucose/metabolismo , Insulina/metabolismo , Transtornos do Metabolismo dos Lipídeos/induzido quimicamente , Transtornos do Metabolismo dos Lipídeos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/metabolismo , Obesidade/induzido quimicamente , Obesidade/metabolismo , Fitoterapia , Rosiglitazona , Tiazolidinedionas/farmacologia , Aumento de Peso/efeitos dos fármacosRESUMO
This study was conducted to investigate the efficacy of cornin, an iridoid glycoside, in an experimental cerebral ischemia induced by middle cerebral artery occlusion (MCAO) and reperfusion (I/R), and to elucidate the potential mechanism. Adult male Sprague-Dawley rats were subjected to MCAO for 1 h, then reperfusion for 23 h. Behavioral tests were used to evaluate the damage to central nervous system. The cerebral infarct volume and histopathological damage were assessed to evaluate the brain pathophysiological changes. Spectrophotometric assay methods were used to determine the activities of superoxide dismutase (SOD) and glutathione-peroxidase (GPx). Contents of malondialdehyde (MDA), the generation of reactive oxygen species (ROS) as well as respiratory control ratio and respiratory enzymes of the brain mitochondria were also determined. The results showed that cornin significantly decreased neurological deficit scores, and reduced cerebral infarct volume and degenerative neurons. Meanwhile, cornin significantly increased the brain ATP content, improved mitochondrial energy metabolism, inhibited the elevation of MDA content and ROS generation, and attenuated the decrease of SOD and GPx activities in brain mitochondria. These findings indicate that cornin has protective potential against cerebral ischemia injury and its protective effects may be due to amelioration of cerebral mitochondrial function and its antioxidant property.
Assuntos
Infarto da Artéria Cerebral Média/tratamento farmacológico , Iridoides/uso terapêutico , Fitoterapia , Verbena , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Cálcio/metabolismo , Respiração Celular/efeitos dos fármacos , Transporte de Elétrons/efeitos dos fármacos , Glutationa Peroxidase/metabolismo , Infarto da Artéria Cerebral Média/metabolismo , Infarto da Artéria Cerebral Média/patologia , Glicosídeos Iridoides , Masculino , Malondialdeído/metabolismo , Fluidez de Membrana/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Fármacos Neuroprotetores/uso terapêutico , Fosfolipídeos/metabolismo , Extratos Vegetais/uso terapêutico , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Traumatismo por Reperfusão/tratamento farmacológico , Superóxido Dismutase/metabolismo , Água/metabolismoRESUMO
This study was carried out to investigate whether rosmarinic acid (RA) has antifibrotic effect on experimental liver fibrosis in vitro and in vivo and its possible mechanism. Culture of hepatic stellate cells (HSCs) determine proliferation and expression of transforming growth factor-beta1 (TGF-beta1), connective transforming growth factor (CTGF) and alpha-smooth muscle actin (alpha-SMA). In carbon tetrachloride (CCL(4))-induced rat liver fibrosis model, determined biochemical indicator, liver fibrosis grade and histopathological changes, immunohistochemical detected liver TGF-beta1 and CTGF expression. The results indicated that RA could inhibit HSCs proliferation, inhibit TGF-beta1, CTGF and alpha-SMA expression in cultured HSCs. It has marked evident in reducing fibrosis grade, ameliorating biochemical indicator and histopathological morphology, reducing liver TGF-beta1 and CTGF expression in CCL(4)-induced liver fibrosis. These findings suggest that RA has potentially conferring antifibrogenic effects.
Assuntos
Antioxidantes/uso terapêutico , Intoxicação por Tetracloreto de Carbono/tratamento farmacológico , Cinamatos/uso terapêutico , Depsídeos/uso terapêutico , Células Estreladas do Fígado/efeitos dos fármacos , Cirrose Hepática/tratamento farmacológico , Fígado/efeitos dos fármacos , Extratos Vegetais/uso terapêutico , Actinas/metabolismo , Animais , Antioxidantes/farmacologia , Intoxicação por Tetracloreto de Carbono/complicações , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Cinamatos/farmacologia , Fator de Crescimento do Tecido Conjuntivo/antagonistas & inibidores , Depsídeos/farmacologia , Modelos Animais de Doenças , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/etiologia , Cirrose Hepática/metabolismo , Masculino , Fitoterapia , Extratos Vegetais/farmacologia , Ratos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta1/antagonistas & inibidores , Ácido RosmarínicoRESUMO
This study was conducted to investigate the efficacy of cornuside, a secoiridoid glucoside compound, in cultured macrophages as well as in an experimental model of sepsis induced by cecal ligation and puncture (CLP) in rats. Cornuside was added to cultured macrophages at different concentrations, and all CLP rats were randomized to receive an intravenous injection of the corresponding drug followed by observation of its antisepsis effect. Our results showed that cornuside downregulated the levels of TNF- alpha, IL-6, and NO production in a dose-dependent manner in activated macrophages, while it upregulated the level of IL-10. Intravenous injection of cornuside or imipenem alone or in combination reduced CLP-induced lethality in rats after CLP. In addition, serum levels of TNF- alpha, IL-6, triggering receptor expressed on myeloid cells, and endotoxin were downregulated. On the other hand, the serum levels of IL-10 were upregulated. Decreased bacterial counts in blood, peritoneum, spleen, liver, and mesenteric lymph nodes and decreased myeloperoxidase in lung, liver, and small intestine also were found after cornuside injection. These data indicate that the antisepsis therapeutic effect of cornuside is mediated by decreased local and systemic levels of a wide spectrum of inflammatory mediators. This work provides first evidence for the clinic use of cornuside as a new immunomodulatory drug that has the capacity to inhibit the inflammatory response in sepsis.
Assuntos
Antibacterianos/uso terapêutico , Cornus/química , Glucosídeos/uso terapêutico , Fatores Imunológicos/uso terapêutico , Mediadores da Inflamação/sangue , Extratos Vegetais/uso terapêutico , Piranos/uso terapêutico , Sepse/tratamento farmacológico , Animais , Antibacterianos/farmacologia , Contagem de Colônia Microbiana , Citocinas/sangue , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Regulação para Baixo , Quimioterapia Combinada , Endotoxinas/sangue , Frutas , Glucosídeos/isolamento & purificação , Glucosídeos/farmacologia , Imipenem/farmacologia , Imipenem/uso terapêutico , Fatores Imunológicos/farmacologia , Macrófagos/efeitos dos fármacos , Masculino , Células Mieloides/efeitos dos fármacos , Óxido Nítrico/sangue , Peroxidase/metabolismo , Fitoterapia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Piranos/isolamento & purificação , Piranos/farmacologia , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Sepse/sangue , Regulação para CimaRESUMO
Excitotoxicity is one of the most extensively studied processes of neuronal death and plays an important role in Alzheimer's disease. In the present study, the protective effects of Gossypium herbaceam extracts (GHE) on learning and memory impairment induced by excitatory neurotoxin ibotenic acid were examined in vivo using Morris water maze. Furthermore, neuroprotective effects of GHE were investigated with methods of immunohistochemistry and biochemistry. Our data showed that oral administration with GHE at the doses of 35, 70 and 140 mg/kg exerted an improved effect on the learning and memory impairment in rats induced by intracerebral injection of ibotenic acid. To confirm the precise mechanism of memory improvement by presence of GHE, we further investigated the potential protection on the hippocampus. Our findings suggest that GHE afforded a beneficial inhibition on pro-apoptosis proteins expression following ibotenic acid. Additionally, calcium pump activity and calbindin-D28k expression were dramatically increased after GHE treatment, implicating that the modulation of calcium homeostasis could be involved in the mechanism underlying neuroprotection of GHE against ibotenic acid-induced excitotoxicity. These data suggested that GHE could be a potential agent for preventing or retarding the development or progression of Alzheimer's disease.
Assuntos
Gossypium/química , Hipocampo/fisiopatologia , Síndromes Neurotóxicas/tratamento farmacológico , Fitoterapia/métodos , Preparações de Plantas/uso terapêutico , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Calbindina 1 , Calbindinas , ATPases Transportadoras de Cálcio/metabolismo , Caspase 3/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Reação de Fuga/efeitos dos fármacos , Agonistas de Aminoácidos Excitatórios/toxicidade , Hipocampo/efeitos dos fármacos , Hipocampo/patologia , Ácido Ibotênico/toxicidade , Injeções Intraventriculares/métodos , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Síndromes Neurotóxicas/etiologia , Síndromes Neurotóxicas/metabolismo , Síndromes Neurotóxicas/fisiopatologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Sprague-Dawley , Proteína G de Ligação ao Cálcio S100/metabolismo , Fatores de Tempo , Proteína X Associada a bcl-2/metabolismoRESUMO
In this study, we established a drug screening system based on transcriptional regulation of vascular endothelial growth factor (VEGF) under hypoxia-inducible factor-1alpha control. We cloned the neomycin-resistance gene into the plasmid GL (pGL)3-promoter vector to generate the pGL3-promoter-neo vector. The 3 copies of the 47-bp fragment that contained the hypoxia response element of VEGF were synthesized and inserted in front of the minimal promoter of the pGL3-promoter-neo vector to generate p3HRE-luc-neo. The recombinant reporter gene vectors were transfected into EAhy926 cells, and stable cell lines were obtained. The positive cell line was selected for its ability to express luciferase in response to hypoxia. We demonstrated that CoCl(2) significantly enhances luciferase activity in a concentration-dependent fashion. We then optimized the cell density and incubation time under hypoxia which were used to screen. The assay exhibited a low background and was an ideal model for high-throughput screening for human VEGF regulators.
Assuntos
Fator 1 Induzível por Hipóxia/fisiologia , Regiões Promotoras Genéticas/genética , Fator A de Crescimento do Endotélio Vascular/biossíntese , Fator A de Crescimento do Endotélio Vascular/genética , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cobalto/farmacologia , DNA/biossíntese , DNA/genética , Avaliação Pré-Clínica de Medicamentos , Resistência a Medicamentos , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Humanos , Luciferases/biossíntese , Luciferases/genética , Neomicina/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Plasmídeos/genética , Inibidores da Síntese de Proteínas/farmacologia , Reprodutibilidade dos Testes , Sais de Tetrazólio , Tiazóis , TransfecçãoRESUMO
Hydroxysafflor yellow A (HSYA) is a main active monomer purified from Carthamus tinctorius L. The research is to study the inhibitory effect of HSYA on the inflammatory signal transduction pathway related factors which were induced by permanent cerebral ischemia in rats. By using the successive administration at a 30 min interval of HSYA and the rats permanent focal cerebral ischemia model established by a intraluminal suture occlusion method. After cerebral artery occlusion 3, 6, 12 and 24 h, cortex was removed for the next experiments. Western blotting was used to detect the expression of p65 protein and the phospho-IkappaB-alpha (pIkappaB-alpha) in the cytoplasm and nucleus. Nuclear factor-kappaB (NF-kappaB) DNA binding activity was measured by Trans-AM transcription factor assay kits. mRNA expression of cytokines TNF-alpha, IL-1beta, IL-6 and IL-10 was measured by the RT-PCR method. The result showed that intravenous injection of HSYA (10 mg x kg(-1)) to rats after cerebral occlusion, the p65 translocation activity and the phosphorylation of IkappaB-alpha were significantly inhibited. At the same time, HSYA suppressed p65 binding activity and the transcriptional level of pro-inflammatory cytokines including TNF-alpha, IL-1beta and IL-6, and promoted the mRNA expression of anti-inflammatory cytokine IL-10. In conclusion, the anti-cerebral ischemic mechanism of HSYA may be due to its inhibition of NF-kappaB activity and the mRNA expression of cytokines in the inflammatory transduction pathway.
Assuntos
Isquemia Encefálica/metabolismo , Chalcona/análogos & derivados , Citocinas/biossíntese , Quinonas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição RelA/metabolismo , Animais , Carthamus/química , Chalcona/isolamento & purificação , Chalcona/farmacologia , Citocinas/genética , Flores/química , Proteínas I-kappa B/metabolismo , Interleucina-10/biossíntese , Interleucina-10/genética , Interleucina-1beta/biossíntese , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/biossíntese , Interleucina-6/genética , Masculino , Inibidor de NF-kappaB alfa , Fármacos Neuroprotetores/isolamento & purificação , Fármacos Neuroprotetores/farmacologia , Fosforilação/efeitos dos fármacos , Plantas Medicinais/química , Transporte Proteico , Quinonas/isolamento & purificação , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/metabolismoRESUMO
In present study, we investigated the mechanism of regulating HIF-1alpha expression by hydroxysafflor yellow A (HSYA) in Eahy 926 cell line under 1% O2 hypoxia. Eahy 926 cells were incubated with HSYA (100, 10 and 1 micromol x L(-1)) under hypoxia for the indicated time after treatment. Cell proliferation rate was detected using MTT assays. VHL and p53 location and protein expression were analyzed by immunocytochemical stain. HIF-1alpha, VHL and p53 mRNA expression were detected by RT-PCR. Protein expression of HIF-1alpha, VHL and p53 were assayed by Western blotting method. HSYA at 100 micromol x L(-1) increased Eahy 926 cells proliferation rate under hypoxia. HIF-1alpha mRNA and protein expression were up-regulated in the presence of HSYA. VHL, p53 mRNA and protein expression decreased significantly after 8 hours of treatment under hypoxia. HSYA protected Eahy 926 cells from hypoxia, and up-regulated HIF-1alpha expression partially via its inhibition of VHL and p53 expression.
Assuntos
Chalcona/análogos & derivados , Células Endoteliais/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/biossíntese , Quinonas/farmacologia , Proteína Supressora de Tumor p53/biossíntese , Proteína Supressora de Tumor Von Hippel-Lindau/biossíntese , Carthamus tinctorius/química , Hipóxia Celular , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Chalcona/isolamento & purificação , Chalcona/farmacologia , Células Endoteliais/citologia , Flores/química , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Plantas Medicinais/química , Quinonas/isolamento & purificação , RNA Mensageiro/metabolismo , Proteína Supressora de Tumor p53/genética , Veias Umbilicais/citologia , Regulação para Cima , Proteína Supressora de Tumor Von Hippel-Lindau/genéticaRESUMO
Amyloid-beta (Abeta) is considered to be responsible for the pathogenesis of Alzheimer's disease. In the present study, the protective effect of Gossypium herbaceam extracts (GHE) on learning and memory impairment induced by Abeta were examined in vivo using Morris water maze and step through task. Furthermore, the antioxidant activity and neuroprotective effect of GHE was investigated with methods of histochemistry and biochemistry. These data showed that oral administration with GHE at the doses of 35, 70 and 140 mg/kg exerted an improved effect on the learning and memory impairment in rats induced by intracerebroventricular (i.c.v.) injection of 10 microg of Abeta(25-35). Subsequently, the GHE afforded a beneficial action on promotion on the activity of glutathione peroxidase and catalase, as well as inhibition on the NF-kappaB activation in the hippocampus followed by the presence of Abeta(25-35). Meanwhile, the number of degenerating neurons with an apoptotic feature was dramatically decreased in hippocampus after treatment with GHE, implicating that its antioxidant stress and inhibition of NF-kappaB activation could be involved in the mechanism underlying neuroprotection of GHE against Abeta-induced cell death. These findings suggested that GHE might be a potential agent for treatment of Alzheimer's disease.
Assuntos
Precursor de Proteína beta-Amiloide/metabolismo , Gossypium , Hipocampo/metabolismo , Hipocampo/patologia , Transtornos da Memória/metabolismo , Transtornos da Memória/prevenção & controle , NF-kappa B/antagonistas & inibidores , Degeneração Neural/patologia , Degeneração Neural/terapia , Fitoterapia , Percepção Espacial , Animais , Cromatografia Líquida de Alta Pressão , Imuno-Histoquímica , Masculino , Aprendizagem em Labirinto , Degeneração Neural/metabolismo , Extratos Vegetais , Ratos , Ratos Sprague-DawleyRESUMO
With the development of molecular biology, genome science becomes an important subject currently. Characterized by high-throughput, high-integration, high-parallelism, miniaturization and automation, it is the integrated study of gene properties on a large scale. Stroke, an important cerebral vascular disease, is one of the threats to human health. The utilization of microarray study for the pathogenesis of stroke, not only reveals the essentials of the disease in the overall level of genes, but also contributes to the detection of therapeutic targets and the development of novel drugs for stroke. Referring to our own work, this discussion focuses on the progress of the mechanisms underlying experimental cerebral ischemia investigation in vivo as well as anti-cerebral ischemia agents by gene chip technology.
Assuntos
Isquemia Encefálica/genética , Perfilação da Expressão Gênica , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Traumatismo por Reperfusão/genética , Acidente Vascular Cerebral/genética , Animais , Encéfalo/irrigação sanguínea , Isquemia Encefálica/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Regulação da Expressão Gênica , Humanos , Fator 1 Induzível por Hipóxia/metabolismo , Interleucina-6/metabolismo , Precondicionamento Isquêmico , Neovascularização Fisiológica/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Traumatismo por Reperfusão/metabolismo , Acidente Vascular Cerebral/metabolismoRESUMO
AIM: To develop a cell-based model by stable transfection of SH-SY5Y with mutant A53T human alpha-synuclein, recapitulating neurotoxicity of alpha -synuclein overexpression. METHODS: The overexpression of mutant alpha -synuclein was analyzed by Western blotting, immunocytochemistry, and RT-PCR. Cell viability was processed when treated with different concentrations of 1-methyl-4-phenylpyridinium (MPP+) and exogenous dopamine (DA) for 24, 48, and 72 h by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Early apoptosis and late apoptosis/necrosis were analyzed by flow cytometry using Annexin V and propidium iodide double staining, respectively. DNA was isolated and applied to agarose gel for electrophoresis; the typical DNA "ladder"represented severe apoptosis. We also used this model to screen 99 compounds with therapeutic potential by MTT assay. RESULTS: One of the stably-transfected clones overexpressed exogenous genes on both the protein level and the transcriptive level. Significant differences in cytotoxicity were found between the pcDNA3.1(+) group and the pcDNA3.1(+)-hm alpha-synuclein group in the presence of the same concentration of MPP+ and DA within the same incubation time. The level of either early apoptosis or late apoptosis/necrosis was remarkably increased in transfected cells compared with the control after treatment with 100 micromol/L MPP+ for 24 h. In addition, the presence of the typical DNA "ladder" was observed in the pcDNA3.1(+)-hm alpha-synuclein group when treated with 200 micromol/L MPP+ for 48 h. After the screening experiment, 12 of the 99 compounds were found to decrease DA-induced cytotoxicity on cell viability. CONCLUSION: We established a cell-based model which is useful for studying the function of alpha-synuclein and screening compounds with therapeutic potential. In addition, it was identified that cells overexpressing A53T mutant alpha-synuclein were significantly vulnerable against MPP+ or dopamine exposures.
Assuntos
Antiparkinsonianos/química , Antiparkinsonianos/uso terapêutico , Avaliação Pré-Clínica de Medicamentos , Modelos Biológicos , Doença de Parkinson/tratamento farmacológico , alfa-Sinucleína/metabolismo , Animais , Antiparkinsonianos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Humanos , Estrutura Molecular , Neuroblastoma , Doença de Parkinson/patologia , Transcrição Gênica , alfa-Sinucleína/genéticaRESUMO
AIM: To investigate the protective effect of hydroxysafflor yellow A (HSYA), a soluble element extracted from Carthamus tinctorius L., on focal cerebral ischemia in rats. METHODS: Focal cerebral ischemia in male Wistar-Kyoto (WKY) rats were induced by permanent middle cerebral artery occlusion (MCAO). Three doses of 1.5, 3.0 and 6.0 mg x kg(-1) of HSYA were administrated to three groups of rats, separately, via sublingular vein injection 30 min after the onset of ischemia. 24 h after ischemia in rats, neurological deficit scores were evaluated and the infarction area of brain was assessed by quantitative image analysis. The in vitro neuroprotective effect of HSYA was tested in cultured fetal cortical neurons exposed to glutamate and sodium cyanide (NaCN). RESULTS: HSYA at doses of 3.0 and 6.0 mg x kg(-1) exerted significant neuroprotective effects on rats with focal cerebral ischemic injury as expressed by neurological deficit scores and reduced the infarct area as compared with saline group, and the potency of HSYA at dose of 6.0 mg x kg(-1) was similar to that of 0.2 mg x kg(-1) of nimodipine. In vitro studies, HSYA significantly inhibited neurons damage induced by exposure to glutamate and NaCN in cultured fetal cortical cells. CONCLUSION: HSYA has potential neuroprotective action against focal cerebral ischemia in rats and cultured rat fetal cortical neurons as well.
Assuntos
Isquemia Encefálica/patologia , Encéfalo/patologia , Carthamus tinctorius , Chalcona/análogos & derivados , Fármacos Neuroprotetores/farmacologia , Quinonas/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Isquemia Encefálica/etiologia , Isquemia Encefálica/fisiopatologia , Carthamus tinctorius/química , Células Cultivadas , Córtex Cerebral/citologia , Chalcona/isolamento & purificação , Chalcona/farmacologia , Ácido Glutâmico , Infarto da Artéria Cerebral Média/complicações , L-Lactato Desidrogenase/metabolismo , Masculino , Neurônios/citologia , Neurônios/metabolismo , Fármacos Neuroprotetores/isolamento & purificação , Plantas Medicinais/química , Quinonas/isolamento & purificação , Ratos , Ratos Endogâmicos WKY , Cianeto de Sódio/antagonistas & inibidoresRESUMO
A chemical investigation of the roots of Cynanchum stauntonii has resulted in the characterization of a new hydroperoxide with a 13,14 : 14,15-disecopregnane-type skeleton, named stauntonine (1), together with three related compounds, anhydrohirundigenin (2), anhydrohirundigenin monothevetoside (3), and glaucogenin-C mono- D-thevetoside (4). Their structures were established by spectroscopic methods, including X-ray crystallographic diffraction analysis of stauntonine that confirmed its relative stereochemistry. The compound 1 showed dose-dependent relaxation on aortic rings with endothelium contracted by phenylepherine or KCl.