RESUMO
This study was designed to compare the differentially expressed proteins between antibiotic-sensitive and antibiotic-resistant Salmonella Typhimurium, Klebsiella pneumonia, and Staphylococcus aureus. The susceptibilities of wild-type (WT), ciprofloxacin (CIP) and/or oxacillin (OXA)-induced, and clinically isolated resistant (CCARM) S. Typhimurium (STWT, STCIP, and STCCARM), K. pneumoniae (KPWT, KPCIP, and KPCCARM), and S. aureus (SAWT, SACIP, SAOXA, and SACCARM) to antibiotics were determined using broth microdilution assay. STCIP was highly resistant to piperacillin (MIC > 512 µg/ml), KPCIP was resistant to chloramphenicol (128 µg/ml) and norfloxacin (16 µg/ml), SACIP was resistant to fluoroquinolones (32 µg/ml), and SAOXA was resistant to ceftriaxone (32 µg/ml). The protein profiles of antibiotic-sensitive and antibiotic-resistant strains were determined using 2-DE analysis followed by LC-MS/MS. The commonly expressed proteins of STWT-STCIP, STWT-STCCARM, KPWT-KPCIP, KPWT-KPCCARM, SAWT-SACIP, SAWT-SAOXA, and SAWT-SACCARM were 763, 677, 677, 469, 261, 259, and 226, respectively. The unique protein spots were observed 57 (6.5%), 80 (11.5%), and 68 (13.9%), respectively, for STCCARM, KPCCARM, and SACCARM. The highly up-regulated protein, PrsA (10-fold), was observed in STCIP resistant to ciprofloxacin (128-fold), levofloxacin (32-fold), norfloxacin (64-fold), and piperacillin (> 16-fold). The up-regulated proteins (YadC, FimA, and RplB) in KPCIP resistant to chloramphenicol (> 32-fold), ciprofloxacin (32-fold), levofloxacin (6-fold), norfloxacin (128-fold), and sparfloxacin (64-fold). AcrB and RpoB were up-regulated in SACCARM resistant to multiple antibiotics. The differentially expressed proteins were related to the antibiotic resistance of STWT, STCIP, STCCARM, KPWT, KPCIP, KPCCARM, SAWT, SACIP, SAOXA, and SACCARM. The resistance-associated proteins could be useful biomarkers for detecting antibiotic-resistant pathogens.
Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Farmacorresistência Bacteriana Múltipla/genética , Klebsiella pneumoniae/genética , Salmonella typhimurium/genética , Staphylococcus aureus/genética , Cloranfenicol/farmacologia , Cromatografia Líquida , Ciprofloxacina/farmacologia , Fluoroquinolonas/farmacologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/metabolismo , Levofloxacino/farmacologia , Testes de Sensibilidade Microbiana , Norfloxacino/farmacologia , Piperacilina/farmacologia , Proteômica , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/metabolismo , Espectrometria de Massas em TandemRESUMO
BACKGROUND: Glutamate (an endogenous excitatory neurotransmitter) at high concentrations contributes to the development of neurodegenerative diseases. Aronia melanocarpa (A. melanocarpa) berries contain anthocyanins and have high antioxidant activities. In this study, we evaluated whether A. melanocarpa berries could protect neuronal cells against glutamate-induced oxidative stress. METHOD: A. melanocarpa berries exerted a protective effect against cytotoxicity in HT22 mouse hippocampal cells by MTT assay. We evaluated oxidative stress parameters including ROS level, intracellular Ca2+ level, glutathione level and antioxidant enzyme activity in HT22 cells to elucidate the mechanism of its neuroprotective effect. RESULTS: A. melanocarpa berries decreased glutamate-induced death of HT22 cells. In addition, A. melanocarpa berries reduced ROS and intracellular Ca2+ levels. Glutathione level, antioxidant enzymes, glutathione reductase and glutathione peroxide activities and mitochondrial membrane potential were also increased in HT22 cells. CONCLUSION: These results suggested that A. melanocarpa berries protected HT22 cells by exerting an antioxidant effect.
Assuntos
Ácido Glutâmico/efeitos adversos , Doenças Neurodegenerativas/metabolismo , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Photinia/química , Extratos Vegetais/farmacologia , Animais , Linhagem Celular Tumoral , Frutas/química , Glutationa/metabolismo , Humanos , Camundongos , Doenças Neurodegenerativas/tratamento farmacológico , Neurônios/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Gumiganghwal-tang (GT) is a traditional herbal medicine that is widely used for its anti-inflammatory, analgesic, and antipyretic actions. Fermented GT has been reported to inhibit acetylcholinesterase (AChE) activity and to exert a neuroprotective effect. In this study, we investigated the effect of fermented GT against scopolamine-induced memory impairment in mice using the Morris water maze and passive avoidance tests. The results of the Morris water maze test indicated that fermented GT significantly decreased escape latency, as compared with that observed in the scopolamine-treated group. In the prove test, fermented GT attenuated the decreased time spent in the target quadrant observed after scopolamine treatment. The results of the passive avoidance test indicated that the treatment with fermented GT increased latency time when compared with the scopolamine-treated group. Moreover, fermented GT inhibited AChE activity in the hippocampi of the treated mice. These results suggest that fermented GT reduced scopolamine-induced amnesia in mice through AChE inhibition. Therefore, we hypothesize that fermented GT may be a useful therapeutic agent for the prevention or treatment of neurodegenerative diseases.
Assuntos
Inibidores da Colinesterase/uso terapêutico , Suplementos Nutricionais , Modelos Animais de Doenças , Transtornos da Memória/prevenção & controle , Nootrópicos/uso terapêutico , Extratos Vegetais/uso terapêutico , Acetilcolinesterase/metabolismo , Animais , Aprendizagem da Esquiva/efeitos dos fármacos , Comportamento Animal/efeitos dos fármacos , Antagonistas Colinérgicos/toxicidade , Inibidores da Colinesterase/administração & dosagem , Donepezila , Fermentação , Proteínas Ligadas por GPI/antagonistas & inibidores , Proteínas Ligadas por GPI/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/enzimologia , Indanos/uso terapêutico , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Transtornos da Memória/induzido quimicamente , Transtornos da Memória/enzimologia , Camundongos Endogâmicos ICR , Antagonistas Muscarínicos/toxicidade , Proteínas do Tecido Nervoso/antagonistas & inibidores , Proteínas do Tecido Nervoso/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/enzimologia , Nootrópicos/administração & dosagem , Piperidinas/uso terapêutico , Extratos Vegetais/administração & dosagem , República da Coreia , Escopolamina/toxicidadeRESUMO
BACKGROUND: So-Cheong-Ryong-Tang (CY), a traditional herbal formula, mainly has been shown to possess allergic rhinitis and asthma for hundreds of years in Asian countries. Although this medicine has been attracted Asian scientists with investigating mechanisms of action against inflammatory-related diseases, there is a little available information on the anti-cancer effect of CY, especially on the fermented form (FCY). In this study, we explored the chemopreventive/chemotherapeutic efficacy of FCY against cancer cells and proved the efficacy of FCY through performing in vivo xenograft assay. METHODS: CY was fermented with bacteria and lyophilized. For analysis of the constituents of CY and FCY, high performance liquid chromatography (HPLC)-DAD system was performed. To detect the anti-cancer effect of FCY, cell viability assay, caspase activity assay, cell cycle analysis, and Western blot analysis were performed in AGS human gastric cancer cells. The inhibitory effects of tumor growth by CY and FCY were evaluated in athymic nude mice inoculated with HCT116 human colon cancer cells. RESULTS: As a result of analyzing the 11components present in CY and FCY, the contents of ephedrine HCl, glycyrrhizin, gingerol, schisandrin, and gomisin A were respectively increased by fermentation in FCY. The treatment of CY or FCY inhibited the viability of AGS cells, interestingly, the inhibition of cancer cell growth was enhanced by fermentation of CY. FCY induced the apoptosis through activating the caspase-3, -8, and -9. Additionally, FCY regulated the activation of mitogen-activated protein kinases (MAPKs) including extracellular signal-regulated kinase (ERK), p38, and c-Jun NH2-terminal kinase (JNK). In vivo xenografts, administration of FCY significantly inhibited the tumor formation, and improved the anti-tumor effect compared to that of CY in athymic nude mice. CONCLUSIONS: FCY indicated significant anti-cancer effects, and its efficacy against tumor formation was improved than that of CY, therefore, FCY might be used for applications of traditional medicine against cancer in modern complementary and alternative therapeutics. Graphical Abstract á .
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Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Caspases/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Animais , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Medicamentos de Ervas Chinesas/química , Feminino , Fermentação , Células HCT116/efeitos dos fármacos , Humanos , Camundongos , Camundongos Nus , Transdução de Sinais , Neoplasias Gástricas/tratamento farmacológico , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
In this study, the effect of Codonopsis lanceolata fermented by lactic acid on controlling gene expression levels related to obesity was observed in an oligonucleotide chip microarray. Among 8170 genes, 393 genes were up regulated and 760 genes were down regulated in feeding the fermented C. lanceolata (FCL). Another 374 genes were up regulated and 527 genes down regulated without feeding the sample. The genes were not affected by the FCL sample. It was interesting that among those genes, Chytochrome P450, Dmbt1, LOC76487, and thyroid hormones, etc., were mostly up or down regulated. These genes are more related to lipid synthesis. We could conclude that the FCL possibly controlled the gene expression levels related to lipid synthesis, which resulted in reducing obesity. However, more detailed protein expression experiments should be carried out.
Assuntos
Codonopsis/metabolismo , Obesidade/tratamento farmacológico , Obesidade/genética , Extratos Vegetais/farmacologia , Animais , Proteínas de Ligação ao Cálcio , Sistema Enzimático do Citocromo P-450/biossíntese , Proteínas de Ligação a DNA , Fermentação , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Ácido Láctico/química , Lipídeos/biossíntese , Fígado/enzimologia , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Mucinas/biossíntese , Análise de Sequência com Séries de Oligonucleotídeos , Plantas Medicinais/metabolismo , Hormônios Tireóideos/biossíntese , Proteínas Supressoras de TumorRESUMO
We isolated 2,3-dihydroxy-4-methoxyacetophenone, a neuroprotective compound from Cynenchum paniculatum in our previous study. The present study was conducted to investigate the possible neuroprotective effect of 2,3-dihydroxy-4-methoxyacetophenone that has been previously isolated from Cynenchum paniculatum on hippocampal neuronal cell line, HT22 cells and its possible cognitive-enhancing effect on scopolamine-induced amnesia in mice. Neuroprotective effect against glutamate-induced neurotoxicity in HT22 cells was evaluated by MTT assay. Also, cognitive enhancing effect against scopolamine (1mg/kg, ip) induced learning and memory deficit was measured by Morris water maze test. Oral administered of 2,3-dihydroxy-4-methoxyacetophenone (1, 10, 20, 40 and 50mg/kg) to amnesic mice induced by scopolamine. In Morris water maze test, 2,3-dihydroxy-4-methoxyacetophenone (50mg/kg) improved the impairment of spatial memory induced by scopolamine. 2,3-Dihydroxy-4-methoxyacetophenone protect HT22 cells on glutamate induced cell-death in a dose-dependent manner (EC50 value: 10.94µM). Furthermore, 2,3-dihydroxy-4-methoxyacetophenone was found to inhibit [Ca(2+)] accumulation in HT22 cells and had antioxidantive activity. The results showed that 2,3-dihydroxy-4-methoxyacetophenone exert neuroprotective and cognitive-enhancing activities through its antioxidant activity. We suggest that 2,3-dihydroxy-4-methoxyacetophenone improves cognitive function and may be helpful for the treatment of Alzheimer's disease.
Assuntos
Acetofenonas/farmacologia , Apoptose/efeitos dos fármacos , Aprendizagem da Esquiva/efeitos dos fármacos , Guaiacol/análogos & derivados , Transtornos da Memória/induzido quimicamente , Fármacos Neuroprotetores/farmacologia , Escopolamina , Acetofenonas/química , Acetofenonas/isolamento & purificação , Acetofenonas/uso terapêutico , Animais , Antioxidantes/química , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Apocynaceae/química , Apocynaceae/metabolismo , Cálcio/metabolismo , Linhagem Celular , Ácido Glutâmico/química , Ácido Glutâmico/toxicidade , Guaiacol/química , Guaiacol/isolamento & purificação , Guaiacol/farmacologia , Guaiacol/uso terapêutico , Transtornos da Memória/tratamento farmacológico , Camundongos , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/isolamento & purificação , Fármacos Neuroprotetores/uso terapêuticoRESUMO
Excessive glutamate can cause oxidative stress in neuronal cells and this can significantly contribute to the etiology of neurodegenerative disease. The present study mainly aims to investigate that aloe extract (AE) and fermented aloe extract (FAE) could protect against glutamate-induced cytotoxicity by modulating oxidative stress. In this study, both AE and FAE showed potent neuroprotective activity by inhibiting ROS and Ca2+ concentration, increasing mitochondria membrane potential, and activating glutathione-related enzymes against glutamate-insulted neurotoxicity in HT22 cells. In addition, the neuroprotective activity of FAE was more potent than that of AE. HPLC analysis reveals that the chemical composition of FAE is different from that of AE. Especially, the contents of aloin A, aloin B and aloenin were higher in FAE than in AE. In conclusion, this study indicates that both AE and FAE may have effective neuroprotective activity in glutamate-insulted pathological conditions such as Alzheimer's disease by managing oxidative stress.
RESUMO
The n-BuOH fraction of O. javanica significantly protected the primary cultures of rat cortical cells exposed to glutamate. Four flavonoids yielded from this fraction through bioactivity-guidance. The isolated compounds, identified as isorhamnetin (1), afzelin (2), hyperoside (3) and persicarin (4), were evaluated in vitro for their neuroprotective activity. Persicarin (4), the main constituent of O. javanica, showed significant neuroprotective activities in glutamate-injured rat cortical cells. Persicarin diminished calcium influx and inhibited the subsequent overproduction of nitric oxide and intracellular peroxide. In addition, persicarin significantly restored the reduced activities of glutathione (GSH) reductase and glutathione peroxidase, and the contents of GSH induced by glutamate. These results support a conclusion that persicarin greatly contributes to the neuroprotective activities of O. javanica.
Assuntos
Córtex Cerebral/efeitos dos fármacos , Flavonas/farmacologia , Flavonoides/farmacologia , Fármacos Neuroprotetores/farmacologia , Oenanthe/química , Extratos Vegetais/farmacologia , Animais , Cálcio/metabolismo , Células Cultivadas , Córtex Cerebral/citologia , Feminino , Flavonas/isolamento & purificação , Flavonoides/isolamento & purificação , Ácido Glutâmico/efeitos adversos , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Estrutura Molecular , Óxido Nítrico/biossíntese , Peróxidos/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
PURPOSE: Excessive glutamate amount can give oxidative stress to neuronal cells, and the accumulation of cell death can trigger the neurodegenerative disorders. In this study, we discovered the neuroprotective effect of Lysimachia christinae Hance in the mouse hippocampal HT22 cell line. METHOD: Overnight incubated HT22 cells were pretreated with L. christinae extract dose dependently (1, 10, and 100 µg/ml). Followed by then, glutamate was treated. These treated cells were incubated several times again, and cell viability, accumulation of reactive oxygen species (ROS) and Ca2+, mitochondrial membrane potential (MMP), and glutathione-related enzyme amount were measured. RESULTS: As a result, L. christinae increases the cell viability by inhibiting the ROS and Ca2+ formation, recovering the level of MMP and enhancing the activity of glutathione production compared with only vehicle-treated groups. CONCLUSION: These draw that L. christinae may remarkably decelerate the neurodegeneration by minimizing neuronal cell damage via oxidative stress.
RESUMO
The dichloromethane fraction of the bark of Machilus thunbergii Sieb. et Zucc. (Lauraceae) significantly protected primary cultures of rat cortical cells exposed to the excitotoxic amino acid, L-glutamate. Through the activity-guided isolation from the CH(2)Cl(2) fraction, (+)-9'-hydroxygalbelgin (1), isogalcatin B (2), (7S,8S,8'R)-3',4'-dimethoxy-3,4,-methylenedioxylignan-7-ol (3), 1-hydroxy-7-hydroxymethyl-6-methoxyxanthone (4), 5,7-dimethoxy-3',4'-methylenedioxyflavan-3-ol (5), (+)-(3S,4S,6R)-3,6-dihydroxypiperitone (6), protocatechuic acid methyl ester (7) and tyrosol (8) were obtained. All of them had significant neuroprotective activities against glutamate-induced neurotoxicity in primary cultures of rat cortical cells at concentrations ranging from 0.1 microM to 10.0 microM and were comparable to MK-801, a well-known inhibitor of glutamate receptor.
Assuntos
Lauraceae/química , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/farmacologia , Casca de Planta/química , Plantas Medicinais/química , Animais , Células Cultivadas , Córtex Cerebelar/citologia , Dicroísmo Circular , Feminino , Furanos/química , Furanos/classificação , Furanos/farmacologia , Ácido Glutâmico/toxicidade , Lignanas/química , Lignanas/classificação , Lignanas/farmacologia , Estrutura Molecular , Ratos , Ratos Sprague-DawleyRESUMO
AIM OF THE STUDY: The cognitive-enhancing and antioxidant activities of KD-501, a standardized extract of the roots of Scrophularia buergeriana Miquel (Scrophulariceae) were investigated. MATERIALS AND METHODS: KD-501 was orally administered to amnesic mice induced by scopolamine and we performed passive avoidance and the Morris water maze tests. To elucidate the mechanism of cognitive-enhancing activity, the effects of KD-501 on the activities of acetylcholinesterase and antioxidant enzymes within the cortex and hippocampus of mice were evaluated. RESULTS: Acute and prolonged oral administration of KD-501 significantly ameliorated scopolamine-induced amnesia in passive avoidance test. In the Morris water maze test, acute and prolonged administration of KD-501 improved the impairment of spatial memory induced by scopolamine indicated by the formation of reference and working memories. The activity of acetylcholinesterase was significantly inhibited by KD-501 within the cortex and hippocampus. Moreover, the reduced activities or contents of glutathione reductase, superoxide dismutase (SOD) and reduced GSH within the cortex and hippocampus caused by scopolamine were elevated by the treatment of KD-501. CONCLUSIONS: Taken together, it could be postulated that KD-501 may exert its potent cognitive-enhancing activity through both anti-acetylcholinesterase and antioxidative actions.
Assuntos
Antioxidantes/farmacologia , Cognição/efeitos dos fármacos , Memória/efeitos dos fármacos , Nootrópicos/farmacologia , Extratos Vegetais/farmacologia , Scrophularia , Acetilcolinesterase/metabolismo , Administração Oral , Animais , Antioxidantes/química , Aprendizagem da Esquiva/efeitos dos fármacos , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/enzimologia , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/enzimologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos ICR , Nootrópicos/química , Extratos Vegetais/química , Raízes de Plantas , Superóxido Dismutase/metabolismoRESUMO
It was reported previously that ESP-102, a combined extract of Angelica gigas, Saururus chinensis and Schizandra chinensis, significantly improved scopolamine-induced memory impairment in mice and protected primary cultured rat cortical cells against glutamate-induced toxicity. To corroborate this effect, the action patterns of ESP-102 were elucidated using the same in vitro system. ESP-102 decreased the cellular calcium concentration increased by glutamate, and inhibited the subsequent overproduction of cellular nitric oxide and reactive oxygen species to the level of control cells. It also preserved cellular activities of antioxidative enzymes such as superoxide dismutase, glutathione peroxidase and glutathione reductase reduced in the glutamate-injured neuronal cells. While a loss of mitochondrial membrane potential was observed in glutamate treated cells, the mitochondrial membrane potential was maintained by ESP-102. These results support that the actual mechanism of neuroprotective activity of ESP-102 against glutamate-induced oxidative stress might be its antioxidative activity.
Assuntos
Ácido Glutâmico/toxicidade , Neurônios/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Angelica/química , Animais , Cálcio/metabolismo , Sobrevivência Celular , Células Cultivadas , Córtex Cerebral/citologia , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Neurônios/enzimologia , Óxido Nítrico/biossíntese , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Saururaceae/química , Schisandra/química , Superóxido Dismutase/metabolismoRESUMO
When growth-phase cell suspension cultures of Capsicum annuum were treated with cellulase-elicitor preparation at 3 microg/ml, the level of capsidiol was transiently increased in the culture media rather than in the cells reaching its maximum approx 24 h after treatment. With methyl jasmonate it took 18 h. Elicitor treatment doubled phospholiphase A(2) (PLA(2)) activity but simultaneous treatment with aristolochic acid, a PLA(2) inhibitor, inhibited sesquiterpenoid accumulation as well as PLA(2) activity. Mastoparan, a G protein activator, treatment also increased PLA(2) activity and capsidiol production. Taken together, the present study shows that induction of capsidiol production in the C. annuum is mediated by PLA(2) activation.
Assuntos
Capsicum/metabolismo , Celulase/metabolismo , Fosfolipases A2/metabolismo , Sesquiterpenos/isolamento & purificação , Sesquiterpenos/metabolismo , Acetatos/metabolismo , Ácidos Aristolóquicos/metabolismo , Células Cultivadas , Cromatografia Gasosa , Ciclopentanos/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular , Oxilipinas/metabolismo , Peptídeos/metabolismo , Inibidores de Fosfolipase A2 , Fatores de Tempo , Venenos de Vespas/metabolismoRESUMO
INTRODUCTION: Spirulina maxima was used as important nutritional source in the Aztec civilization because it is rich in proteins and vitamins. It contains various antioxidants such as phycocyanin and flavonoids. Based on abundant antioxidants, S. maxima is known to possess anti-inflammatory effect, especially on neuronal cells. MATERIALS AND METHODS: S. maxima was extracted in water and contain of phycocyanin was identified by high-performance liquid chromatography. Cell viability test was performed with treatment of S. maxima extract. After, oxidative stress-related mechanisms were evaluated by detecting the accumulation of reactive oxygen species (ROS) and Ca2+ influx, and decrease of mitochondrial membrane potential (MMP) level. Then, the glutathione (GSH) related assays were conducted. RESULTS: The water extracted S. maxima exerted the neuroprotective activity by attenuating the ROS and Ca2+ formation, maintaining the MMP level, and protecting the activity of the antioxidant enzymes by increasing reduced GSH against oxidative stress compared to control. CONCLUSION: The results suggested that water extracted S. maxima showed powerful neuroprotective effect through the mechanism related to antioxidant activity, able to preventing the radical-mediated cell death. SUMMARY: Water extracted Spirulina maxima contains C-phycocyaninWater extracted Spirulina maxima exerts neuroprotective effect on HT22 cellTo investigate the protective mechanisms, reactive oxygen species, Ca2+, mitochondrial membrane potential, Glutathione-related assays were performed. Abbreviations used: ROS: Reactive oxygen species; MMP: Mitochondrial membrane potential; GSH: Glutathione; GSSG: Glutathione disulfide, oxidized glutathione; GPx: Glutathione peroxidase; GR: Glutathione reductase; DMEM: Dulbecco's modified Eagle's medium; FBS: Fetal bovine serum; DCF-DA: 2',7'-dichlorofluorescein diacetate; PBS: Phosphate buffered serum; Rho 123: Rhodamine 123; NADPH: Nicotinamide adenine dinucleotide phosphate; DTNB: 5,5'-dithiobis-2-nitrobenzoic acid, Ellman's reagent; GSSG-R: Glutathione disulfide reductase; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; DMSO: Dimethyl sulfoxide; HPLC: High-performance liquid chromatography.
RESUMO
Magnoliae Flos (MF) is a traditional medicinal herb used for managing rhinitis, sinusitis and headache. The purpose of the present study was to determine the neuroprotective effect of MF against glutamate-induced oxidative stress and to assess the underlying mechanism. Glutamate is a major endogenous excitatory neurotransmitter in the brain and contributes to the development of neurodegenerative diseases by excessive activation. MF extract was subjected to a neuroprotective effect assay in HT22 mouse hippocampal cells. The mechanism underlying the neuroprotective effect of MF extract was evaluated by assaying reactive oxygen species (ROS) levels, intracellular Ca2+ levels, mitochondrial membrane potential, glutathione level and antioxidant enzyme activity in HT22 cells. MF extract significantly decreased glutamate-induced death of HT22 cells (80.83 ± 7.34% relative neuroprotection). MF extract reduced the intracellular ROS and Ca2+ levels and increased the glutathione level and glutathione reductase and glutathione peroxide activities. Moreover, MF extract attenuated the mitochondrial membrane potential in HT22 cells. These results suggested that MF extract exerts a neuroprotective effect against oxidative stress HT22 cells, which was mediated by its antioxidant activity.
Assuntos
Hipocampo/citologia , Magnoliaceae/química , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/farmacologia , Animais , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Linhagem Celular , Glutationa/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Espécies Reativas de Oxigênio/metabolismoRESUMO
BACKGROUND: Reynoutria sachalinensis is a well-known and used herbal medicine to treatment of arthralgia, jaundice, amenorrhea, coughs, carbuncles, and sores. OBJECTIVE: We have developed high-performance liquid chromatography analysis method for simultaneous determination of isolated four compounds, campesterol, emodin8-O-ß-D-glucopyranoside, quercetin, and isoquercitrin from R. sachalinensis is. MATERIALS AND METHODS: The four compounds were separated on Shiseido C18 column (S-5 µm, 4.6 mm I.D. ×250 mm) at a column temperature of 25°C. The mobile phase composed of water and methanol with gradient elution system, and flow rate is 1.0 ml/min. The detection wavelength was set at 205 nm. RESULTS: Validation of this analytical method was evaluated by linearity, precision, and accuracy test. This established method had good linearity (R2 > 0.997). The relative standard deviation values of intra- and inter-day testing were indicated that <2%, and accuracy is 91.66%-103.31% at intraday and 91.69%-103.31% at intraday. The results of recovery test were 92.60%-108.99%. CONCLUSION: In these results, developed method was accurate and reliable to the quality evaluation of campesterol, emodin 8-O-ß-D-glucopyranoside, quercetin, and isoquercitrin isolated from R. sachalinensis. SUMMARY: We have developed high-performance liquid analysis method for simultaneous determination of 4 compounds of Reynoutria sachalinensis. Abbreviations used: HPLC: High-performance liquid chromatography, DAD: Diode array detector, LOD: Limit of detection, LOQ: Limit of quantitation, ICH: International Conference on Harmonisation.
RESUMO
Glutamate is a neurotransmitter in central nervous system. Overexpression of glutamate leads to oxidative stress, resulting in several neurodegenerative disorders that include Alzheimer's disease. The n-hexane fraction of stems and ethyl acetate (EtOAc) fraction of flowers of Reynoutria sachalinensis provide neuroprotection against glutamate-induced oxidative toxicity in HT22 cells. In this study, 1-decanol (1), ß-amyrin (2), dammaran-3ß-ol (3), campesterol (4), daucosterol (5), ergosterol peroxide (6), emodin 8-O-ß-D-glucopyranoside (7), quercetin (8) and isoquercitrin (9) were isolated from n-hexane fractions of stems and EtOAc fractions of flowers of R. sachalinensis. Their neuroprotective activity was evaluated by MTT assay. 1-Decanol, campesterol, ergosterol peroxide, quercetin and isoquercitrin exhibited neuroprotective activity. These compounds decreased reactive oxygen species level, showed anti-oxidant activity with DPPH radical and in a H2O2 scavenging assay. Therefore, the neuroprotective activity of 1-decanol, campesterol, ergosterol peroxide, quercetin and isoquercitrin are associated with antioxidant activity.
Assuntos
Antioxidantes/farmacologia , Inibidores da Colinesterase/farmacologia , Fármacos Neuroprotetores/farmacologia , Extratos Vegetais/farmacologia , Polygonaceae/química , Acetilcolinesterase/metabolismo , Antioxidantes/química , Antioxidantes/isolamento & purificação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Inibidores da Colinesterase/química , Inibidores da Colinesterase/isolamento & purificação , Relação Dose-Resposta a Droga , Flores/química , Sequestradores de Radicais Livres/metabolismo , Humanos , Estrutura Molecular , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/isolamento & purificação , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/metabolismo , Relação Estrutura-AtividadeRESUMO
The effect of meso-dihydroguaiaretic acid (MDGA) on the staurosporine-induced neuronal apoptosis and its potential mechanism were investigated using primary cultures of rat cortical cells as an assay system. Treatment of MDGA at the concentrations of 0.1, 1.0 and 10 microM significantly protected neuronal cells against Staurosporine-induced apoptosis. The neuroprotective activity of MDGA was the most potent at the concentration of 1.0 microM and was not increased at higher concentration. MDGA reduced apoptotic characteristics induced by STS; MDGA reduced the condensed nuclei in staurosporine-injured rat cortical cells. MDGA diminished the calcium influx that accompanies the staurosporine-induced apoptosis, and inhibited the subsequent overproduction of reactive oxygen species and peroxide to the level of control cells. It also preserved cellular activity of superoxide dismutase, an antioxidative enzyme reduced by staurosporine insult. In addition, MDGA significantly inhibited caspase-3/7 activation and cytochrome c release. Taken together, these results suggested that MDGA protected neuronal cells against staurosporine-induced apoptosis through the inhibition of Ca(2+) influx, cellular oxidation, cytochrome c release and caspase-3/7 activation.
Assuntos
Córtex Cerebral/citologia , Guaiacol/análogos & derivados , Lignanas/farmacologia , Neurônios/efeitos dos fármacos , Neurotoxinas/toxicidade , Estaurosporina/toxicidade , Análise de Variância , Animais , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Caspase 3 , Caspases/metabolismo , Células Cultivadas , Citocromos c/metabolismo , Compostos de Dansil/farmacologia , Relação Dose-Resposta a Droga , Interações Medicamentosas , Embrião de Mamíferos , Galactosamina/análogos & derivados , Galactosamina/farmacologia , Guaiacol/química , Guaiacol/farmacologia , Lignanas/química , Peróxidos/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: Crataegi fructus is a herbal medicine for strong stomach, sterilization, and alcohol detoxification. Chlorogenic acid and hyperoside are the major compounds in Crataegi fructus. OBJECTIVE: In this study, we established novel high-performance liquid chromatography (HPLC)-diode array detection analysis method of chlorogenic acid and hyperoside for quality control of Crataegi fructus. MATERIALS AND METHODS: HPLC analysis was achieved on a reverse-phase C18 column (5 µm, 4.6 mm × 250 mm) using water and acetonitrile as mobile phase with gradient system. The method was validated for linearity, precision, and accuracy. About 31 batches of Crataegi fructus samples collected from Korea and China were analyzed by using HPLC fingerprint of developed HPLC method. Then, the contents of chlorogenic acid and hyperoside were compared for quality evaluation of Crataegi fructus. RESULTS: The results have shown that the average contents (w/w %) of chlorogenic acid and hyperoside in Crataegi fructus collected from Korea were 0.0438% and 0.0416%, respectively, and the average contents (w/w %) of 0.0399% and 0.0325%, respectively. CONCLUSION: In conclusion, established HPLC analysis method was stable and could provide efficient quality evaluation for monitoring of commercial Crataegi fructus. SUMMARY: Quantitative analysis method of chlorogenic acid and hyperoside in Crataegi fructus is developed by high.performance liquid chromatography.(HPLC).diode array detectionEstablished HPLC analysis method is validated with linearity, precision, and accuracyThe developed method was successfully applied for quantitative analysis of Crataegi fructus sample collected from Korea and China. Abbreviations used: HPLC: High-performance liquid chromatography, GC: Gas chromatography, MS: Mass spectrometer, LOD: Limits of detection, LOQ: Limits of quantification, RSD: Relative standard deviation, RRT: Relative retention time, RPA: Relation peak area.
RESUMO
Dianthus superbus (D. superbus) is a traditional crude drug used for the treatment of urethritis, carbuncles and carcinomas. The objective of this study was to confirm the cognitive enhancing effect of D. superbus in memory impairment induced mice and to elucidate the possible potential mechanism. Effect of D. superbus on scopolamine induced memory impairment on mice was evaluated using the Morris water maze and passive avoidance tests. We also investigated acetylcholinesterase (AChE) activity and brain-derived neurotropic factor (BDNF) expression in scopolamine-induced mice. HPLC-DAD analysis was performed to identify active compounds in D. superbus. The results revealed that D. superbus attenuated the learning and memory impairment induced by scopolamine. D. superbus also inhibited AChE levels in the hippocampi of the scopolamine-injected mice. Moreover, D. superbus increased BDNF expression in the hippocampus. Eight compounds were identified using HPLC-DAD analysis. The content of 4-hydroxyphenyl acetic acid was higher than contents of other compounds. These results indicated that D. superbus improved memory functioning accompanied by inhibition of AChE and upregulation of BDNF, suggesting that D. superbus may be a useful therapeutic agent for the prevention or treatment of Alzheimer's disease.