RESUMO
Species of Leishmania and Trypanosoma genera are the causative agents of relevant parasitic diseases. Survival inside their hosts requires the existence of a potent antioxidant enzymatic machinery. Four iron superoxide dismutases have been described in trypanosomatids (FeSODA, FeSODB1, FeSODB2, and FeSODC) that hold a potential as therapeutic targets. Nonetheless, very few studies have been developed that make use of the purified enzymes. Moreover, FeSODC remains uncharacterised in Leishmania. In this work, for the first time, we describe the purification and enzymatic activity of recombinant versions of the four Leishmania FeSOD isoforms and establish an improved strategy for developing inhibitors. We propose a novel parameter [(V*cyt. c - Vcyt. c)/Vcyt. c] which, in contrast to that used in the classical cytochrome c reduction assay, correlates linearly with enzyme concentration. As a proof of concept, we determine the IC50 values of two ruthenium carbosilane metallodendrimers against these isoforms.
Assuntos
Antiprotozoários , Relação Dose-Resposta a Droga , Leishmania infantum , Testes de Sensibilidade Parasitária , Superóxido Dismutase , Leishmania infantum/enzimologia , Leishmania infantum/efeitos dos fármacos , Relação Estrutura-Atividade , Estrutura Molecular , Superóxido Dismutase/metabolismo , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/química , Antiprotozoários/farmacologia , Antiprotozoários/química , Antiprotozoários/síntese química , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/química , Inibidores Enzimáticos/síntese química , Leishmaniose/tratamento farmacológico , Leishmaniose/parasitologiaRESUMO
Although the inhibition of acid ceramidase (AC) is known to induce antitumor effects in various cancers, there are few reports in pancreatic cancer, and the underlying mechanisms remain unclear. Moreover, there is currently no safe administration method of AC inhibitor. Here the effects of gene therapy using siRNA and shRNA for AC inhibition with its mechanisms for pancreatic cancer were investigated. The inhibition of AC by siRNA and shRNA using an adeno-associated virus 8 (AAV8) vector had antiproliferative effects by inducing apoptosis in pancreatic cancer cells and xenograft mouse model. Acid ceramidase inhibition elicits mitochondrial dysfunction, reactive oxygen species accumulation, and manganese superoxide dismutase suppression, resulting in apoptosis of pancreatic cancer cells accompanied by ceramide accumulation. These results elucidated the mechanisms underlying the antitumor effect of AC inhibition in pancreatic cancer cells and suggest the potential of the AAV8 vector to inhibit AC as a therapeutic strategy.
Assuntos
Ceramidase Ácida/antagonistas & inibidores , Terapia Genética/métodos , Doenças Mitocondriais/etiologia , Estresse Oxidativo , Neoplasias Pancreáticas/terapia , RNA Interferente Pequeno/uso terapêutico , Ceramidase Ácida/metabolismo , Animais , Apoptose , Linhagem Celular Tumoral , Ceramidas/metabolismo , Dependovirus , Vetores Genéticos , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patologia , Distribuição Aleatória , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/antagonistas & inibidores , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Using natural products as antioxidant agents has been beneficial to replace synthetic products. Efforts have been made to profile the antioxidant capacities of natural resources, such as medicinal plants. The polyphenol content of Himalayan rhubarb, Rheum emodi wall, was measured and the antioxidant activity was determined using DPPH and ABTS+ assay, and the oxidative stress was assessed using SOD enzymatic assay. Five different solvent fractions, n-hexane, n-butanol, ethyl acetate, dichloromethane, and water, were used for screening the antioxidant capacity in effort to determine the optimum extraction solvent. The total phenolic contents for R. emodi fractions ranged from 27.76 to 209.21 mg of gallic acid equivalents (GAE)/g of dry weight. DPPH and ABTS+ assay results are presented into IC50 values, ranged from 21.52 to 2448.79 µg/mL and 90.25 to 1718.05 µg/mL, respectively. The ethyl acetate fraction had the highest antioxidant activity among other fractions. Also, n-butanol and water fractions showed significantly lower IC50 values than the positive control in DPPH radical scavenging activity. The IC50 values of SOD assay of fractions ranged from 2.31 to 64.78 µg/mL. A similar result was observed with ethyl acetate fraction showing the highest SOD radical scavenging activity. The study suggests that the ethyl acetate fraction of R. emodi possess the strongest antioxidant activity, thus the most efficient in extracting antioxidant contents. Moreover, a highly significant correlation was shown between total polyphenol content and antioxidant activity screening assays. The compounds related to the antioxidant activity of R. emodi were identified to myricitrin, myricetin 3-galloyl rhamnoside, and myricetin, which have not been reported in studies about R. emodi before.
Assuntos
Antioxidantes/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Rheum/química , Antioxidantes/química , Flavonoides/química , Flavonoides/farmacologia , Humanos , Fenóis/química , Fenóis/farmacologia , Extratos Vegetais/química , Polifenóis/química , Polifenóis/farmacologia , Solventes/química , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/químicaRESUMO
Relaxin, an ovarian polypeptide hormone, is found in the hypothalamic paraventricular nucleus (PVN) which is an important central integrative site for the control of blood pressure and sympathetic outflow. The aim of this study was to determine if superoxide anions modulate the effects of relaxin in the PVN. Experiments were performed in normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs). Relaxin mRNA and protein, and its receptor, relaxin family peptide receptor 1 (RXFP1) levels in PVN were 3.24, 3.17, and 3.64 times higher in SHRs than in WKY rats, respectively. Microinjection of relaxin-2 into the PVN dose-dependently increased mean arterial pressure (MAP), renal sympathetic nerve activity (RSNA) and heart rate (HR) in both WKY rats and SHRs, although the effects on MAP (16.87⯱â¯1.99 vs. 8.97⯱â¯1.48â¯mm Hg in 100â¯nmol), RSNA (22.60⯱â¯2.15 vs. 11.77⯱â¯1.43 % in 100â¯nmol) and HR (22.85⯱â¯3.13 vs. 12.62⯱â¯2.83 beats/min in 100â¯nmol) were greater in SHRs. Oxidative stress level was enhanced after relaxin-2 microinjection into the PVN. Pretreatment with superoxide anion scavengers or NADPH oxidase inhibitor blocked, and superoxide dismutase inhibitor potentiated the effects of relaxin-2 on MAP, RSNA and HR. RXFP1 knockdown significantly attenuated the blood pressure of SHRs, and inhibited the increases of atrial natriuretic peptide, brain natriuretic peptide, collagen I, collagen III and fibronectin in the heart of SHRs. These results demonstrated that relaxin is expressed in the PVN, and contributes to hypertension and sympathetic overdrive via oxidative stress. Down-regulation of RXFP1 in the PVN could attenuate hypertension and cardiac remodeling.
Assuntos
Pressão Sanguínea/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Relaxina/farmacologia , Animais , Eletrocardiografia , Sequestradores de Radicais Livres/farmacologia , Expressão Gênica , Frequência Cardíaca/efeitos dos fármacos , Masculino , NADPH Oxidases/antagonistas & inibidores , Núcleo Hipotalâmico Paraventricular/fisiologia , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Receptores Acoplados a Proteínas G/genética , Receptores de Peptídeos/genética , Relaxina/genética , Superóxido Dismutase/antagonistas & inibidoresRESUMO
BACKGROUND: We previously synthesized two DNA intercalative Pyrimido[4',5':4,5]thieno(2,3-b) quinolines (PTQ), 9-hydroxy-4-(3-diethylaminopropylamino)pyrimido[4',5':4,5]thieno(2,3-b) quinolines (Hydroxy- DPTQ) and 8-methoxy-4-(3-diethylaminopropylamino) pyrimido[4',5':4,5]thieno(2,3-b) quinolines (Methoxy-DPTQ), and reported their cytotoxicity against cancer cell lines. METHODS: In the present study, we sought to analyze the antitumor activity of Hydroxy-DPTQ and Methoxy-DPTQ on Ehrlich's ascites carcinoma in vivo models, along with other pharmacological activities and toxicity. RESULTS: In this study, both the test molecules studied possess potent in vivo antitumor activity without any hematological, biochemical or nephrotoxicity. Significant tumor regression was observed after treatment with both the test molecules, which is suggested by the decrease in the bodyweight of tumour-bearing mice. Mean survival time of mice with tumor was increased from 16 days to 25 and 29 days after 40 and 80 mg/kg Hydroxy- DPTQ treatment, respectively, with a similar result for Methoxy-DPTQ. A dose-dependent increase in lifespan up to 80-85% was also displayed by both Hydroxy-DPTQ and Methoxy-DPTQ. Reduction in the tumor volume of mice, upon treatment with molecules also confirmed their antitumor activity. These molecules also exhibited pharmacological activities such as antioxidant, anti-inflammatory and analgesic activities. Administration of Hydroxy-DPTQ and Methoxy-DPTQ not only reduced the level of lipid peroxidation in tumor bearing mice but also restored the superoxide dismutase, glutathione, and catalase levels to normal, substantiating the antioxidant property. Also, treatment of Hydroxy-DPTQ and Methoxy-DPTQ inhibited the pain to approximately 60-80% and 19-33%, respectively. Further, the treatment with Hydroxy-DPTQ and Methoxy-DPTQ reversed the abnormality in the RBC, WBC and haemoglobin levels, and gentamicin induced nephrotoxicity. CONCLUSION: Hydroxy-DPTQ and Methoxy-DPTQ are good antitumor molecules with pharmacological properties.
Assuntos
Analgésicos/farmacologia , Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Dor do Câncer/tratamento farmacológico , Carcinoma de Ehrlich/tratamento farmacológico , Pirimidinas/farmacologia , Quinolinas/farmacologia , Ácido Acético , Analgésicos/administração & dosagem , Analgésicos/química , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Antioxidantes/administração & dosagem , Antioxidantes/química , Carcinoma de Ehrlich/patologia , Catalase/antagonistas & inibidores , Catalase/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Glutationa/antagonistas & inibidores , Glutationa/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Camundongos , Manejo da Dor , Pirimidinas/administração & dosagem , Pirimidinas/química , Quinolinas/administração & dosagem , Quinolinas/química , Ratos , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismoRESUMO
Neocryptolepine (5-methyl-5H-indolo[2,3-b] quinoline) analogs were synthesized and evaluated in vitro and in vivo for their effect versus Ehrlich ascites carcinoma (EAC). The analogs showed stronger cytotoxic activity against EAC cells than the reference drug. The in vivo evaluation of the target compounds against EAC-induced solid tumor in the female albino Swiss mice revealed a remarkable decrease in the tumor volume (TV) and hepatic lipid peroxidation. A noticeable increase of both superoxide dismutase (SOD) and catalase (CAT) levels was reported (p < 0.001), which set-forth proof of their antioxidant effect. In addition, the in vitro antioxidant activity of the neocryptolepine analogs was screened out using the DPPH method and showed promising activities activity. The histopathological investigations affirmed that the tested analogs have a remarkable curative effect on solid tumors with minimal side-effect on the liver. The study also includes illustrated mechanism of the antitumor activity at the cell level by flow cytometry. The cell cycle analysis showed that the neocryptolepine analogs extensively increase the aggregation of tumor cells in three phases of the cell cycle (G0/G1, S and G2/M) with the emergence of a hypo-diploid DNA content peak (sub-G1) in the cell cycle experiments, which is a clear-cut for the apoptotic cell population. Furthermore, the immunological study manifested a significant elevation in splenic lymphocyte count (p < 0.001) with the elevation of the responsiveness of lymphocytes to phytohemagglutinin (PHA). These results indicate that these naturally-based neocryptolepine alkaloids exhibit marked antitumor activity in vivo and represent an important lead in the development of natural-based anticancer drugs.
Assuntos
Alcaloides/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Produtos Biológicos/farmacologia , Carcinoma de Ehrlich/tratamento farmacológico , Indóis/farmacologia , Quinolinas/farmacologia , Animais , Antineoplásicos Fitogênicos/química , Carcinoma de Ehrlich/enzimologia , Carcinoma de Ehrlich/patologia , Catalase/antagonistas & inibidores , Feminino , Técnicas In Vitro , Indóis/química , Camundongos , Quinolinas/química , Superóxido Dismutase/antagonistas & inibidores , Inibidores da Topoisomerase II/farmacologia , Células Tumorais CultivadasRESUMO
Reactive oxygen species (ROS) are rapidly eliminated and reproduced in organisms, and they always play important roles in various biological functions and abnormal pathological processes. Evaluated ROS have frequently been observed in various cancers to activate multiple pro-tumorigenic signaling pathways and induce the survival and proliferation of cancer cells. Hydrogen peroxide (H2O2) and superoxide anion (O2â¢-) are the most important redox signaling agents in cancer cells, the homeostasis of which is maintained by dozens of growth factors, cytokines, and antioxidant enzymes. Therefore, antioxidant enzymes tend to have higher activity levels to maintain the homeostasis of ROS in cancer cells. Effective intervention in the ROS homeostasis of cancer cells by chelating agents or metal complexes has already developed into an important anti-cancer strategy. We can inhibit the activity of antioxidant enzymes using chelators or metal complexes; on the other hand, we can also use metal complexes to directly regulate the level of ROS in cancer cells via mitochondria. In this review, metal complexes or chelators with ROS regulation capacity and with anti-cancer applications are collectively and comprehensively analyzed, which is beneficial for the development of the next generation of inorganic anti-cancer drugs based on ROS regulation. We expect that this review will provide a new perspective to develop novel inorganic reagents for killing cancer cells and, further, as candidates or clinical drugs.
Assuntos
Antineoplásicos/farmacologia , Quelantes/farmacologia , Complexos de Coordenação/farmacologia , Neoplasias/tratamento farmacológico , Animais , Antineoplásicos/química , Antineoplásicos/uso terapêutico , Quelantes/química , Quelantes/uso terapêutico , Complexos de Coordenação/química , Complexos de Coordenação/uso terapêutico , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/uso terapêutico , Humanos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismoRESUMO
Cancer chemotherapeutic drugs exert cytotoxic effects by modulating intracellular reactive oxygen species (ROS) levels. However, whether ROS modulates the efficacy of targeted therapeutics remains poorly understood. Previously, we reported that upregulation of the anti-apoptotic protein, BCL-XL, by KRAS activating mutations was a potential target for KRAS-mutant colorectal cancer (CRC) treatment. Here, we demonstrated that the BCL-XL targeting agent, ABT-263, increased intracellular ROS levels and targeting antioxidant pathways augmented the therapeutic efficacy of this BH3 mimetic. ABT-263 induced expression of genes associated with ROS response and increased intracellular ROS levels by enhancing mitochondrial superoxide generation. The superoxide dismutase inhibitor, 2-methoxyestradiol (2-ME), exhibited synergism with ABT-263 in KRAS-mutant CRC cell lines. This synergistic effect was attributed to the inhibition of mTOR-dependent translation of the anti-apoptotic MCL-1 protein via caspase 3-mediated cleavage of AKT and S6K. In addition, combination treatment of ABT-263 and 2-ME demonstrated a synergistic effect in in vivo patient-derived xenografts harboring KRAS mutations. Our data suggest a novel role for ROS in BH3 mimetic-based targeted therapy and provide a novel strategy for treatment of CRC patients with KRAS mutations.
Assuntos
Compostos de Anilina/farmacologia , Antioxidantes/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/tratamento farmacológico , Mutação , Proteínas Proto-Oncogênicas p21(ras)/genética , Sulfonamidas/farmacologia , Proteína bcl-X/antagonistas & inibidores , 2-Metoxiestradiol/farmacologia , Animais , Antineoplásicos/farmacologia , Apoptose , Biomarcadores Tumorais/genética , Proliferação de Células , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Superóxido Dismutase/antagonistas & inibidores , Tiorredoxinas/antagonistas & inibidores , Transcriptoma , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Nanoparticles that functionally mimic the activity of metal-containing enzymes (metallo-nanozymes) are of therapeutic importance for treating various diseases. However, it is still not clear whether such nanozymes can completely substitute the function of natural enzymes in living cells. In this work, we show for the first time that a cerium vanadate (CeVO4 ) nanozyme can substitute the function of superoxide dismutase 1 and 2 (SOD1 and SOD2) in the neuronal cells even when the natural enzyme is down-regulated by specific gene silencing. The nanozyme prevents the mitochondrial damage in SOD1- and SOD2-depleted cells by regulating the superoxide levels and restores the physiological levels of the anti-apoptotic Bcl-2 family proteins. Furthermore, the nanozyme effectively prevents the mitochondrial depolarization, leading to a significant improvement in the cellular levels of ATP under oxidative stress.
Assuntos
Trifosfato de Adenosina/metabolismo , Cério/química , Mitocôndrias/metabolismo , Nanoestruturas/química , Vanadatos/química , Materiais Biomiméticos/química , Materiais Biomiméticos/metabolismo , Linhagem Celular Tumoral , Humanos , Neurônios/citologia , Neurônios/metabolismo , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Superóxido Dismutase-1/antagonistas & inibidores , Superóxido Dismutase-1/genética , Superóxido Dismutase-1/metabolismo , Superóxidos/metabolismoRESUMO
Di-2-ethylhexyl phthalate (DEHP) has been widely used as a plasticizer in industry and can affect memory; however, the underlying mechanism remains unclear. In the present study, mouse HT22 cells, an immortalized hippocampal neuronal cell line, was utilized as an in vitro model. We showed that DEHP dramatically inhibited cell viability and increased lactate dehydrogenase (LDH) release from the cells in a dose-dependent manner, suggesting that DEHP could cause cytotoxicity of mouse HT22 cells. The protein levels of cleaved Caspase-8, cleaved Caspase-3, and Bax markedly increased in the DEHP-treated cells, whereas there was a significant decrease in the Bcl-2 protein level, implying that DEHP could induce apoptosis of mouse HT22 cells. DEHP exposure significantly increased the content of malondialdehyde, whereas it markedly decreased the level of glutathione and the activities of glutathione peroxidase and superoxide dismutase, suggesting that DEHP induced oxidative stress of the cells. Compared with the DEHP-treated group, the inhibition of cell viability and the release of LDH were rescued in the N-acetyl-l-cysteine plus DEHP group. Furthermore, inhibition of oxidative stress could rescue the induction of apoptosis by DEHP. Collectively, our results indicated that DEHP could induce apoptosis of mouse HT22 cells via oxidative stress.
Assuntos
Apoptose/efeitos dos fármacos , Dietilexilftalato/toxicidade , Neurônios/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Caspases/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Glutationa Peroxidase/antagonistas & inibidores , Hipocampo/efeitos dos fármacos , L-Lactato Desidrogenase/biossíntese , Camundongos , Proteínas Proto-Oncogênicas c-bcl-2 , Superóxido Dismutase/antagonistas & inibidoresRESUMO
It is well known that the disruption of the mitochondrial respiratory components prolongs lifespan in many species. The mitochondrial stress response can lead to an increased survival rate through the restoration of the cellular homeostasis. Therefore, developing pharmacological interventions that induce mitochondrial stress response may be desirable to delay the onset of age-related diseases and promote a healthy life. In this study, we present chemical compounds, revealed by systematic screening of chemical libraries, which inhibit mitochondrial ATP synthesis in mammalian cells. Our study demonstrates that these compounds alter the body length and promote the oxidative stress response which leads to an increased longevity in Caenorhabditis elegans. Thus, our study identifies chemical compounds that may have potential therapeutic applications through affecting the mitochondrial function.
Assuntos
Trifosfato de Adenosina/metabolismo , Proteínas de Caenorhabditis elegans/antagonistas & inibidores , Caenorhabditis elegans/crescimento & desenvolvimento , Mitocôndrias/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Bibliotecas de Moléculas Pequenas/farmacologia , Superóxido Dismutase/antagonistas & inibidores , Animais , Caenorhabditis elegans/efeitos dos fármacos , Caenorhabditis elegans/metabolismo , Ensaios de Triagem em Larga Escala , Longevidade , Mitocôndrias/metabolismo , Biogênese de OrganelasRESUMO
Acute myeloid leukemia (AML) is a difficult therapeutic hematological tumor. It is urgent to find a non-toxic natural drug to treat AML. Herein, the selenium nanoparticles (SeNPs) embedded in nanotubes consisted of triple helix ß-(1, 3)-d-glucan (BFP) from the black fungus that were wrapped to form stable inclusion complex BFP-Se, which was self-assembled and exhibited high stability in water. In vitro, the BFP-Se significantly inhibited the proliferation of AML cells and increased the cytotoxicity on AML cells. On single-cell levels, the U937 cells were gradually swelled and lysed with BFP-Se treatment on optofluidics chips. Further, the blood and bone marrow analysis indicated the anti-leukemia effects of BFP-Se in vivo. Moreover, BFP-Se increased the total antioxidant capacity of AML cells and decreased the expression of c-Jun activation domain-binding protein 1 and thioredoxin 1. Our results suggest that this biocompatible polysaccharide nanotube containing Se nanoparticles would provide a novel strategy for AML therapy.
Assuntos
Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Glucanos/farmacologia , Leucemia Mieloide Aguda/tratamento farmacológico , Nanopartículas/química , Selênio/farmacologia , Animais , Antineoplásicos/química , Antioxidantes/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Glucanos/química , Glutationa/antagonistas & inibidores , Humanos , Leucemia Mieloide Aguda/patologia , Camundongos , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/patologia , Imagem Óptica , Tamanho da Partícula , Selênio/química , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismo , Propriedades de Superfície , Células Tumorais CultivadasRESUMO
Pure compounds belonging to phenolic family were studied for their biological potential such as 5,8-dihydroxy-1,4-naphthoquinone (M1), rutin hydrate (M2), 2,3-dichloro-5,8-dihydroxy-1,4-naphthoquinone (M3), taxifolin (M4), myricetin (M5), plumbagin (M6), silibinin (M7), dihydromyricetin (M8), shikonin (M9), quercetin 3-ß-D-glucoside (M10), (±)-taxifolin hydrate (M11), cardamonin (M12),(-)-epicatechin (M13), 9-chloro-10-hydroxy-anthracene-1,4-dione (M14), 9-chloro-10-hydroxy-2,3-dimethyl-anthracene-1,4-dione (M15), 2-chloro-3-(2-hydroxy-5-methylanilino)-1,4-naphthoquinone (M16), 2-chloro-3-(4-hydroxy-phenylamino)-(1,4) naphthoquinone (M17), 2-chloro-3-(3,5-di-tert-butyl-4-hydroxy-phenyl)-(1,4)-naphtoquinone (M18), and myricitrin dihydrate (M19). These molecules were chosen based on two reasons; the limited or total absence of their exploitation in several studied activities and the use of other tests for the same activity. The evaluation of the in vitro anti-acetyl-cholinesterase (AChE), anti-5-lipoxygenase (5-LOX), anti-xanthine oxidase (XOD), anti-alpha glucosidase, anti-superoxide dismutase (SOD), anti-oxidant (DPPH (1, 1-diphenyl-2-picrylhydrazyl) and ABTS (2, 2- azinobis-3-ethylbenzothiazoline-6-sulphonate)), and anticancer activities of mentioned 19 molecules was explored during this work. M3, M14, M15, M16, M17, M18, M19 were exploited for the first time for such purposes. Tested compounds were shown to have interesting radical scavenging abilities against DPPH radicals, and the highest molecules among them were M19 and M5 (IC50 = 12.0 and 15.5 µM, respectively), and M4, M19 and M2 against ABTS (IC50= 1.9, 4.3 and 4.3 µM, respectively). Moreover, the majority of products showed very important cytotoxic activity since IC50 values were ranging between (IC50= 0.2 µM (M1) and 79 µM (M8)) against HCT116 cell line, and values of IC50= 0.2 µM for M1 against MCF7 cell line. All new molecules (non studied before) were shown to have great cytotoxic effect against both cancer cell lines.Furthermore, molecule M5 was shown to have anti-inflammatory potential via the inhibition of 5-LOX enzyme (65% at 100 µM). In addition, M19 showed important anti XOD activity with 47% of inhibition at 100 µM. Also, it has been found that compound M3 had the best anti alpha glucosidase activity with 43.8 % of inhibition at 100 µM, the highest anti-AChE effect (IC50= 14.5 µM), and the best effect towards SOD (IC50= 10.0 µM). A structure-activity relationship study was also performed.
Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Inibidores da Colinesterase/farmacologia , Flavonoides/farmacologia , Inibidores de Glicosídeo Hidrolases/farmacologia , Superóxido Dismutase/antagonistas & inibidores , Xantina Oxidase/antagonistas & inibidores , Anti-Inflamatórios/química , Antineoplásicos/farmacologia , Antioxidantes/química , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Flavonoides/química , Células HCT116 , Humanos , Células MCF-7 , Superóxido Dismutase/metabolismo , Xantina Oxidase/metabolismo , alfa-Glucosidases/metabolismoRESUMO
AIM AND OBJECTIVE: Humans continuously use pesticides in the field to control the pest population and weeds for considerable agricultural productivity. Side-by species like grazinganimals, insects and other species are adversely affected by or become resistant to pesticides. Insects, birds and cattle are highly abundant dwellers of the agriculture-field and represent three distinct phyla having versatile physiological features. Besides higher agricultural-productivity, protection to several species will maintain ecological/environmental balance. Studies on the effect of widely used pesticides on their DNA-stability and important enzymatic-activities are insufficient. MATERIALS AND METHODS: Antioxidant-activity (Superoxide-dismutase; SOD/Catalase- by gelzymogram- assay) and DNA-stability (fragmentation-assay) in hepatic/gut tissues were studied after in vitro exposure of Chlorpyrifos, Fenvalerate, Nimbecidine or Azadirachtin to goat/cow/poultry-hen/insect. RESULTS: In general, all pesticides were found to impair enzymatic-activities. However, lower organisms were affected more than higher vertebrates by azadirachtin-treatment. DNA fragmentation was found more in insects/poultry-birds than that of the cattle in hepatic/gut tissues. Inversely, toxicity/antioxidant marker-enzymes were more responsive in insect gut-tissues. However, mitochondrialtoxicity revealed variable effects on different species. It has been noticed that chlorpyrifos is the most toxic pesticide, followed by Fenvalerate/Nimbecidine (Azadirachtin, AZT). Nevertheless, AZT revealed its higher DNA-destabilizing effects on the field-insects as compared to the other animals. CONCLUSION: Field-insects are highly integrated into the ecosystem and the local bio-geo-chemical cycle, which may be impaired. Pesticides may have toxic effects on higher vertebrates and may sustain in the soil after being metabolized into their different derivatives. Some of the sensitive biochemical parameters of this organism may be used as a biomarker for pesticide toxicity.
Assuntos
Antioxidantes/farmacologia , Catalase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Praguicidas/farmacologia , Superóxido Dismutase/antagonistas & inibidores , Animais , Antioxidantes/toxicidade , Bovinos , Galinhas , Clorpirifos/farmacologia , Resistência a Medicamentos/efeitos dos fármacos , Ecossistema , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/toxicidade , Instabilidade Genômica/efeitos dos fármacos , Cabras , Insetos , Limoninas/farmacologia , Gado , Nitrilas/farmacologia , Noresteroides/farmacologia , Praguicidas/toxicidade , Piretrinas/farmacologiaRESUMO
Notopterol (NOT) is a major bioactive ingredient extracted from the rhizomes of either Notopterygium incisum Ting ex H. T. Chang or N. forbesii Boiss (Qianghuo in Chinese), a botanical drug that was adopted as a traditional Chinese medicine. NOT is suggested to show analgesic and anti-inflammatory effects in clinical practice. The inhibitory effects of NOT on human cytochrome P450 enzymes were investigated in the present study. Our results indicate that NOT inhibited the activity of CYP2D6 in a time-, concentration- and NADPH-dependent manner. The values of KI and kinact were 10.8 µM and 0.62 min-1, respectively. The calculated kobs at 10 µM was 0.29 min-1, above the 0.02 min-1 risk level. After incubation with NOT at 10 µM for 9 min, approximately 92% of CYP2D6 activity was inhibited. Such loss of enzyme activity was not restored through dialysis, which indicates that the observed enzyme inhibition was irreversible. Partition ratio of the inactivation was approximately 29. Quinidine, a competitive CYP2D6 inhibitor, demonstrated protection on enzymes against the NOT-induced inactivation, but such protection was not found in incubation systems fortified with glutathione or catalase/superoxide dismutase. Additionally, CYP3A4 was observed to function as an enzyme mainly involved in the biotransformation of NOT. Taken together, these findings indicate that NOT served as a mechanism-based inactivator of CYP2D6, meanwhile, those observed effects may induce the latent drug-drug interactions. The metabolic activation of NOT may be the key to trigger the inactivation of the enzyme.
Assuntos
Cumarínicos/metabolismo , Citocromo P-450 CYP2D6/metabolismo , Ativação Metabólica , Apiaceae/química , Apiaceae/metabolismo , Cumarínicos/química , Citocromo P-450 CYP2D6/química , Inibidores Enzimáticos/química , Inibidores Enzimáticos/metabolismo , Glutationa/antagonistas & inibidores , Glutationa/metabolismo , Humanos , Cinética , NADP/química , NADP/metabolismo , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismoRESUMO
OBJECTIVE: The aim of this study was to clarify the potential effect of zoledronic acid on alleviating oxidative stress and promoting bone marrow mesenchymal stem cells (BMSCs) osteogenesis through the SIRT3/SOD2 pathway, thus alleviating the progression of osteoporosis. MATERIALS AND METHODS: Relative expression levels of osteogenesis-related genes (ALP, RUNX2, and Bglap) were determined. Meanwhile, ALP activity and capacity of mineralization in BMSCs treated with different doses of zoledronic acid were measured. Subsequently, viability and ROS level in H2O2-induced BMSCs influenced by zoledronic acid treatment were assessed. The regulatory effect of zoledronic acid on the SIRT3/SOD2 pathway was detected by Western blot. Furthermore, the involvement of the SIRT3/SOD2 pathway in zoledronic acid-mediated BMSCs osteogenesis was evaluated. RESULTS: Zoledronic acid treatment significantly up-regulated the levels of ALP, RUNX2, and Bglap. Meanwhile, it improved ALP activity and capacity of mineralization in BMSCs dose-dependently. H2O2 induction markedly suppressed viability and enhanced ROS level in BMSCs, which were reversed by zoledronic acid treatment. Besides, zoledronic acid protected H2O2-induced SIRT3 down-regulation and AC-SOD2/SOD2 up-regulation in BMSCs. In addition, silence of SIRT3 reversed the protective effects of zoledronic acid on osteogenesis of BMSCs. CONCLUSIONS: Zoledronic acid alleviates the progression of osteoporosis. Meanwhile, it accelerates BMSCs osteogenesis by inhibiting oxidative stress via the SIRT3/SOD2 pathway.
Assuntos
Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Osteoporose/tratamento farmacológico , Sirtuína 3/metabolismo , Superóxido Dismutase/antagonistas & inibidores , Ácido Zoledrônico/farmacologia , Células Cultivadas , Relação Dose-Resposta a Droga , Humanos , Peróxido de Hidrogênio/antagonistas & inibidores , Peróxido de Hidrogênio/farmacologia , Células-Tronco Mesenquimais/metabolismo , Osteoporose/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Sirtuína 3/antagonistas & inibidores , Relação Estrutura-Atividade , Superóxido Dismutase/metabolismoRESUMO
Applying the single-cell gel electrophoresis (comet) assay, we show that the widely used organophosphorus pesticide malathion is cytotoxic, genotoxic, and induces oxidative stress in human lymphocytes.
Assuntos
Citotoxinas/toxicidade , Inseticidas/toxicidade , Linfócitos/efeitos dos fármacos , Malation/toxicidade , Mutagênicos/toxicidade , Oxidantes/toxicidade , Acetilcisteína/farmacologia , Antioxidantes/farmacologia , Catalase/antagonistas & inibidores , Catalase/metabolismo , Ensaio Cometa/métodos , Citotoxinas/antagonistas & inibidores , Feminino , Glutationa Transferase/antagonistas & inibidores , Glutationa Transferase/metabolismo , Humanos , Inseticidas/antagonistas & inibidores , Peroxidação de Lipídeos/efeitos dos fármacos , Linfócitos/metabolismo , Malation/antagonistas & inibidores , Masculino , Oxidantes/antagonistas & inibidores , Estresse Oxidativo/efeitos dos fármacos , Cultura Primária de Células , Espécies Reativas de Oxigênio/agonistas , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismo , Adulto JovemRESUMO
BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a chronic and fibrotic lung disease of unknown causes. Given the crucial role of oxidative stress in the progression of IPF, antioxidant therapy may be speculated to be an efficient therapeutic approach. Therefore, the present study aimed to evaluate the protective effects of Crocin as a potent, natural antioxidant against Bleomycin-induced PF in male Wistar rats. METHODS: Forty male Wistar rats were randomly divided into four groups. Group 1 received intratracheal saline on day 7 and oral gavage of saline for 28 days. Group 2 received a single dose of Bleomycin on day 7 and oral gavage of saline for 28 days. Groups 3 received a single dose of Bleomycin on day 7, accompanied with oral administration of Crocin for 28 days. Group 4 orally received Crocin for 28 days. Finally, the lungs were removed for measuring the biochemical and histopathological markers. RESULTS: The results showed that Crocin therapy remarkably decreased TNF-α, MDA and NO levels in the lungs of Bleomycin-exposed rats. Furthermore, a significant increase was seen in lung GSH content, catalase, and GPx activities in the Crocin/Bleomycin-treated group as compared with Bleomycin-treated group. However, Crocin could not markedly change the lung index and SOD activity. Histopathological changes, fibrosis and hydroxyproline content of lungs also significantly decreased by Crocin therapy in the Crocin/Bleomycin-treated group. CONCLUSION: In sum, Crocin therapy could modulate biochemical and histological changes induced by Bleomycin; therefore, it might be considered as an effective therapeutic approach against IPF.
Assuntos
Antioxidantes/uso terapêutico , Bleomicina/toxicidade , Carotenoides/uso terapêutico , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/prevenção & controle , Animais , Antibióticos Antineoplásicos/toxicidade , Antioxidantes/farmacologia , Carotenoides/farmacologia , Glutationa Peroxidase/antagonistas & inibidores , Glutationa Peroxidase/metabolismo , Masculino , Fibrose Pulmonar/metabolismo , Ratos , Ratos Wistar , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismoRESUMO
BACKGROUND: The cytotoxic activity of the pyridazin-3-one derivative LCS-1 was previously suggested to be due to the inhibition of superoxide dismutase 1 (SOD1). However, no direct evidence was provided that LCS-1 inhibits SOD1 within cells. METHODS: In this study, we investigated the cytotoxic activity of LCS-1 against bloodstream forms of Trypanosoma brucei, a protozoan parasite that does not express copper/zinc-containing SOD1, but an iron-containing superoxide dismutase (FeSOD). RESULTS: At 250 µM, LCS-1 did not inhibit the activity of FeSOD in cell lysates of bloodstream forms of T. brucei, confirming that the compound is a specific inhibitor of SOD1. However, LCS-1 displayed substantial trypanocidal activity with a minimum inhibitory concentration of 10 µM and a half-maximal effective concentration of 1.36 µM, indicating that the cytotoxic action of the compound cannot solely be due to inhibition of SOD1. CONCLUSION: The results of this study is an important finding as it shows that LCS-1 has more than one cytotoxic mode of action.
Assuntos
Antineoplásicos/farmacologia , Piridazinas/farmacologia , Superóxido Dismutase/antagonistas & inibidores , Trypanosoma brucei brucei/enzimologia , Ensaios Enzimáticos , Testes de Sensibilidade Parasitária , Superóxido Dismutase/metabolismo , Superóxido Dismutase-1/antagonistas & inibidores , Trypanosoma brucei brucei/efeitos dos fármacosRESUMO
Leishmaniasis is a widespread neglected tropical disease complex that is responsible of one million new cases per year. Current treatments are outdated and pose many problems that new drugs need to overcome. With the goal of developing new, safe, and affordable drugs, we have studied the in vitro activity of 12 different 5-nitroindazole derivatives that showed previous activity against different strains of Trypanosoma cruzi in a previous work. T. cruzi belongs to the same family as Leishmania spp., and treatments for the disease it produces also needs renewal. Among the derivatives tested, compounds 1, 2, 9, 10, 11, and 12 showed low J774.2 macrophage toxicity, while their effect against both intracellular and extracellular forms of the studied parasites was higher than the ones found for the reference drug Meglumine Antimoniate (Glucantime®). In addition, their Fe-SOD inhibitory effect, the infection rates, metabolite alteration, and mitochondrial membrane potential of the parasites treated with the selected drugs were studied in order to gain insights into the action mechanism, and the results of these tests were more promising than those found with glucantime, as the leishmanicidal effect of these new drug candidates was higher. The promising results are encouraging to test these derivatives in more complex studies, such as in vivo studies and other experiments that could find out the exact mechanism of action.