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OBJECTIVE: To investigate the mechanisms of ATR-induced dopaminergic toxicity by microglia activation and the response of the Keap1/ Nrf2- ARE signaling pathway. METHODS: Wistar rats were treated with 50, 100 and 200 mg/kg ATR and BV-2 microglia cells were treated with 50, 100 µM ATR or 100 ng/mL LPS, respectively. Rats behavioral responses and histopathological changes were monitored. Immunohistochemical and immunofluorescence analysis detected Iba-1 and TH+ cells in rats. Keap1/Nrf2-ARE signaling-related proteins and inflammatory factors from BV-2 cells and rats were detected using ELISA, Western blot and Real-time PCR. RESULTS: After ATR treatment, the grip strength of Wistar rats was significantly decreased, and anxiety were clearly observed. TH+ neurons were reduced, however, the number of microglia cells and Iba-1 levels were increased clearly in SN. The release of ROS, TNF-α and IL-Iß were increased, and levels of SOD and GSH-Px were significantly decreased. Keap1 mRNA expression and protein levels were decreased, while nuclear Nrf2 mRNA expression and protein levels were both increased in vivo and in vitro. CONCLUSION: ATR could significantly activate microglia and exacerbate neurotoxicity and neuroinflammation, leading to accelerate dopaminergic neuron cell death by inhibiting Keap1/Nrf2-ARE signaling pathway.
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Atrazina , Fator 2 Relacionado a NF-E2 , Animais , Neurônios Dopaminérgicos/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch/genética , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Microglia , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Ratos , Ratos Wistar , Transdução de SinaisRESUMO
Atrazine (ATR) is a widely used herbicide that can induce the degeneration of dopaminergic (DAergic) neurons in the substantia nigra, resulting in a Parkinson's disease-like syndrome. Despite the high risk of environmental exposure, few studies have investigated strategies for the prevention of ATR neurotoxicity. Our previous studies demonstrated that ATR can impair mitochondrial function, leading to metabolic failure. Cells maintain mitochondrial quality through selective autophagic elimination, termed mitophagy. Soybean isoflavones (SI) possess multiple beneficial bioactivities, including preservation of mitochondria function, so it was hypothesized that SI can protect neurons against ATR toxicity by promoting mitophagy. Pretreatment of SH-SY5Y neurons with SI prevented ATR-induced metabolic failure and cytotoxicity as assessed by intracellular ATP, Na+-K+-ATPase activity, mitochondrial membrane potential, and cell viability assays. The neuroprotective efficacy of SI was superior to the major individual components genistein, daidzein, and glycitein. Ultrastructural analyses revealed that ATR induced mitochondrial damage, while SI promoted the sequestration of damaged mitochondria into autophagic vesicles. Soybean isoflavones also induced mitophagy as evidenced by upregulated expression of BNIP3/NIX, BEX2, and LC3-II, while co-treatment with the mitophagy inhibitor Mdivi-1 blocked SI-mediated neuroprotection and prevented SI from reversing ATR-induced BEX2 downregulation. Furthermore, BEX2 knockdown inhibited SI-induced activation of the BNIP3/NIX pathway, mitophagy, and neuroprotection. These findings suggest that SI protects against ATR-induced mitochondrial dysfunction and neurotoxicity by activating the BEX2/BNIP3/NIX pathway.
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Atrazina , Isoflavonas , Mitofagia , Atrazina/toxicidade , Neurônios Dopaminérgicos , Humanos , Isoflavonas/farmacologia , Proteínas de Membrana , Proteínas do Tecido Nervoso , Proteínas Proto-Oncogênicas , Glycine max/química , Proteínas Supressoras de TumorRESUMO
BACKGROUND/AIMS: Atrazine (ATR) is a broad-spectrum herbicide in wide use around the world. However, ATR is neurotoxic and can cause cell death in dopaminergic neurons, leading to neurodegenerative disorders. Autophagy is the basic cellular catabolic process involving the degradation of proteins and damaged organelles. Studies have shown that certain plant compounds can induce autophagy and prevent neuronal cell death. This prompted us to investigate plant compounds that might reduce the neurotoxic effects of ATR. METHODS: By CCK-8 and flow cytometry, we tested the ability of five candidate compounds-isoflavones, resveratrol, quercetin, curcumin, and green tea polyphenols-to protect cells from ATR. Changes in the expression of tyrosine hydroxylase (TH) and brain-expressed X-linked 2 (BEX2), autophagy-related proteins and key factors in mTOR signaling, were detected by Western blotting. RESULTS: Isoflavones had the strongest activity against ATR-induced neuronal apoptosis. ATR reduced the expression of TH and BEX2, whereas isoflavones increased TH and BEX2 expression. In addition, ATR inhibited autophagy, whereas isoflavones induced autophagy through the accumulation of LC3-II and decreased expression of p62; this effect was abolished by 3-methyladenine (3-MA). Furthermore, BEX2 siRNA abolished isoflavone-mediated autophagy and neuroprotection in vitro. CONCLUSION: Isoflavones activate BEX2-dependent autophagy, protecting against ATR-induced neuronal apoptosis.
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Autofagia/efeitos dos fármacos , Isoflavonas/farmacologia , Proteínas do Tecido Nervoso/metabolismo , Apoptose/efeitos dos fármacos , Atrazina/farmacologia , Western Blotting , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Curcumina/farmacologia , Citometria de Fluxo , Humanos , Polifenóis/farmacologia , Quercetina/farmacologia , Reação em Cadeia da Polimerase em Tempo Real , Resveratrol , Sincalida/metabolismo , Estilbenos/farmacologia , Serina-Treonina Quinases TOR/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Dibromoacetic acid (DBAA), a haloacetic acid found in drinking water as a disinfection by-product, can cause many adverse effects, including immunotoxicity. In a previous study, we confirmed that DBAA can induce obvious immunotoxicity in mice but that the underlying mechanisms are not clearly understood. In our current study, we confirmed that DBAA induced cytotoxicity and apoptosis in thymocytes isolated from mice by a range of DBAA concentrations (0, 5, 10, 20, or 40 µM). The data showed that DBAA exposure led to a significant decrease in proliferative responses to T-cell mitogens and obvious inhibition in the production of cytokines interleukin-2 and interleukin-4. We found obvious morphological changes of apoptosis in thymocytes and observed the percentage of apoptotic thymocytes to increase significantly as the DBAA concentration increased. Further investigation showed that DBAA can cause G0/G1 arrest in cell cycle analysis, increase intracellular calcium ([Ca(2+)]i) levels, increase the expression of Fas/FasL proteins, and decrease the expression of Bcl-2 protein. It is concluded that in vitro exposure to DBAA can lead to marked cytotoxicity and apoptosis among thymocytes, and the mechanism involved is strongly related to blocking cell cycle progression, increasing intracellular calcium, and increasing Fas/FasL expressions.
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Acetatos/toxicidade , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Ciclo Celular/efeitos dos fármacos , Proteína Ligante Fas/metabolismo , Timócitos/efeitos dos fármacos , Animais , Cálcio/análise , Espaço Intracelular/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Transdução de Sinais/efeitos dos fármacosRESUMO
Atrazine (ATR, 2-chloro-4-ethylamino-6-isopropylamino-s-triazine) is used worldwide as a herbicide, and its presence in the environment has resulted in documented human exposure. A lack of strong evidence for genetic heritability of idiopathic Parkinson's disease has focused attention on environmental toxicants in the disease etiology, particularly agrichemicals. Parkinson's disease is associated with advanced age and is characterized by the degeneration of dopaminergic neurons, but it is unclear whether specific neuronal damage could result from insults during development. The juvenile period is particularly vulnerable to environmental agent, therefore, we evaluated the effects of a 28-day exposure to ATR on the dopaminergic system in pubertal rats. Sprague-Dawley rats were treated orally with ATR at 50, 100, and 200 mg/kg bw, daily from postnatal days 27 to 54. In this study, we examined the hypothesis that pubertal exposure to ATR would disrupt the development of the nigrostriatal dopamine (DA) system. The content of DA and levodopa (L-DA) were examined in striatum samples by HPLC-FL, and the mRNA and protein expression of tyrosine hydroxylase, orphan nuclear hormone receptor (Nurr1), Nurr1 interacting protein (NuIP), and cyclin-dependent kinase inhibitors of the Cip̲Kip family (p57kip2) were examined in samples of the nigrostriatum by use of fluorescence Real-Time quantitative polymerase chain reaction (PCR). Exposure of juvenile rats to the high dose of ATR led to reduced levels of DA and L-DA, genes expression of NuIP, Nurr1, and p57kip2 in animals.
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Atrazina/toxicidade , Dopamina/metabolismo , Exposição Ambiental , Maturidade Sexual/efeitos dos fármacos , Animais , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Levodopa/metabolismo , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Aprendizagem Espacial/efeitos dos fármacos , NataçãoRESUMO
Atrazine (2-chloro-4-ethytlamino-6-isopropylamine-1,3,5-triazine; ATR) is widely used as a broad-spectrum herbicide. Animal studies have demonstrated that ATR exposure can cause cell death in dopaminergic neurons. The molecular mechanisms underlying ATR-induced neuronal cell death, however, are unknown. In this study, we investigated the autophagy and apoptosis induced by ATR in dopaminergic neurons in vivo. Wistar rats were administered with ATR at doses of 10, 50 and 100 mg/kg body weight by oral gavage for three months. In terms of histopathology, the expression of autophagy- and apoptosis-related genes as well as proteins related to the Beclin-1/B-cell lymphoma 2 (Bcl-2) autophagy and apoptosis pathways were examined in the rat nigrostriatal dopaminergic system. We observed degenerative micromorphology indicative of neuronal apoptosis and mitochondrial autophagy by electron microscopy in ATR-exposed rat striatum. The rat ventral mesencephalon in the ATR-exposed groups also showed increased expression of Beclin-1, LC3-II, Bax and Caspase-9, and decreased expression of tyrosine hydroxylase (TH), Bcl-xl and Bcl-2. These findings indicate that ATR may induce autophagy- and apoptosis-related changes in doparminergic neurons. Furthermore, this induction may be regulated by the Beclin-1 and Bcl-2 autophagy and apoptosis pathways, and this may help to better understand the mechanism underlying the neurotoxicity of ATR.
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Apoptose , Atrazina/toxicidade , Autofagia , Neurônios Dopaminérgicos/efeitos dos fármacos , Herbicidas/toxicidade , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Atrazina/efeitos adversos , Proteína Beclina-1 , Caspase 9/metabolismo , Neurônios Dopaminérgicos/metabolismo , Herbicidas/efeitos adversos , Masculino , Mesencéfalo/citologia , Mesencéfalo/efeitos dos fármacos , Mesencéfalo/metabolismo , Mesencéfalo/ultraestrutura , Proteínas Associadas aos Microtúbulos/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Wistar , Proteína X Associada a bcl-2/metabolismoRESUMO
High atrazine (2-chloro-4-ethytlamino-6-isopropylamine-1,3,5-triazine; ATR) contents in the environment threaten the health conditions of organisms. We examined the effects of ATR exposure on Sprague-Dawley rats during gestation and on the dopaminergic neurons of offspring during lactation. Pregnant dams were orally treated with 0 mg/kg/day to 50 mg/kg/day of ATR from gestational day 5 to postnatal day 22. Afterward, neither offspring nor dams received ATR. Dopamine (DA) content was examined in striatum samples by HPLC-FL; the mRNA expressions of tyrosine hydroxylase (TH), orphan nuclear hormone (Nurr1), dopamine transporter (DAT), and vesicular monoamine transporter 2 (VMAT2) in the ventral midbrain samples were examined by fluorescence PCR when the offspring reached one year of age. After the pregnant rats were exposed to ATR, the DA concentrations and mRNA levels of Nurr1 were decreased in their offspring. Decreased Nurr1 levels were also accompanied by changes in the mRNA levels of VMAT2, which controls the transport and reuptake of DA.
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Atrazina/toxicidade , Neurônios Dopaminérgicos/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Herbicidas/toxicidade , Membro 2 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Proteínas Vesiculares de Transporte de Monoamina/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Corpo Estriado/metabolismo , Dopamina/análise , Dopamina/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Neurônios Dopaminérgicos/metabolismo , Feminino , Lactação , Masculino , Exposição Materna/efeitos adversos , Membro 2 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Gravidez , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Tirosina 3-Mono-Oxigenase/genética , Tirosina 3-Mono-Oxigenase/metabolismo , Proteínas Vesiculares de Transporte de Monoamina/genéticaRESUMO
The objective of the study is to determine the causal relationship and potential mechanisms between Parkinson's disease (PD) and neuroinflammatory and neurotoxic mediators. We conducted two-sample Mendelian randomization (2SMR) study and multivariable Mendelian randomization (MVMR) analysis to investigate the causality between PD and neuroinflammatory and neurotoxic mediators. The mediation analysis with MR was also conducted to determine the potential mediating effect of neuroinflammatory and neurotoxic mediators between asthma and PD. Genetically predicted levels of nine neuroinflammation were associated with changes in PD risk. The associations of PD with CCL24, galectin-3 levels, haptoglobin, and Holo-Transcobalamin-2 remained significant in multivariable analyses. The mediation analysis with MR revealed that asthma affects PD through CCL24 and galectin-3. The results showed neuroinflammation could affect the pathogenesis of PD. In the combined analysis of these nine variables, CCL24, galectin-3 levels, HP, and Holo-Transcobalamin-2 alone were found to be significant. Asthma plays an intermediary role through CCL24 and galectin-3 levels.
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The objective of the study was to explore the relationship and potential mechanism between Parkinson's disease (PD) and diabetic retinopathy (DR) using bioinformatics methods. We first examined the causal relationship between PD and DR by Mendelian randomization (MR) analysis. The datasets of PD and DR patients from the Gene Expression Omnibus database were used to identify differentially expressed genes (DEGs). Then, we performed the Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and immune infiltration analysis. We also constructed a protein-protein interaction network and receiver operating characteristic (ROC) curve. Finally, an online website was used for drug prediction. The MR analysis demonstrated a causal relationship between DR and PD (odds ratio [OR] = 0.86; 95% confidence interval [CI] 0.79-0.93; p = 3.24E - 04), in which DR acted as a protective factor against PD. There were 81 DEGs identified from the PD and DR datasets, of which 29 genes had protein interaction relationships, and enrichment analysis showed that these genes were mainly related to immune pathways. As indicated by immune cell infiltration analysis, the expression of immune cells between PD and the control group was significantly different. ROC curve results showed five genes had diagnostic value, and several potential chemical compounds were predicted to target the genes. Our findings demonstrate a reduced risk of PD in patients with DR. We also found that PD and DR are closely related in terms of inflammation, which provides clues for further exploring the common mechanisms and interaction of these two diseases.
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Two dimensional (2D) 1H-13C heteronuclear single-quantum correlation (HSQC) spectroscopy has recently been proposed for quantitative determination of typical linear low density polyethylene (LLDPE) with high accuracy. It requires highly precise measurement to achieve further reliable quantification. In this context, this paper aims at determining conditions that allow the achievement of high precision. On the basis of the optimized parameters, two time-saving strategies, nonuniform sampling (NUS) and band-selective HSQC are evaluated on model polyolefins in terms of repeatability. Precision better than 0.3% and 5% for ethylene content (E mol%) and 1-hexene content (H mol%) of the model poly(ethylene-co-1-hexene)s are obtained with 50% NUS or band-selective HSQC. Moreover, dramatic precision enhancements can be achieved with the combination of band-selective HSQC and 50% NUS, in which repeatabilities better than 0.15% and 2.5% for E mol% and H mol% are observed. The experiment times are reduced to about 0.5 h. These methods open important perspectives for rapid, precise and accurate quantitative analysis of complex polymers.
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OBJECTIVE: To assess HCH and DDT exposure levels and associated risk factors among 262 children aged 6-10 years in a northeastern rural area of China between April and May of 2008. METHODS: Eight HCH and DDT metabolites in serum samples were monitored by gas chromatography. A questionnaire was administered to identify the sources of pesticides in children' serum samples. RESULTS: At least one pesticide metabolite was detected in 81.7% of the tested children. Higher amounts of pp'DDD were detected in 50% of them. Children's age and their father's occupation as farmers, together with not changing work clothes after work, were the main risk factors for HCH and DDT exposure among them. CONCLUSION: Children living in rural areas are experiencing multiple sources of organochlorine pesticide exposure. These pesticides may have been retained in the environment for a long period of time.
Assuntos
Exposição Ambiental , Hexaclorocicloexano/toxicidade , População Rural , Criança , China , HumanosRESUMO
Wingless (Wnt) signaling regulates the proliferation and differentiation of midbrain dopamine (DA) neurons. Paraquat (PQ) and maneb (MB) are environmental pollutants that can be used to model Parkinson's disease (PD) in rodents. A previous study demonstrated that developmental exposure to PQ and MB affects the expression of Wnt1, Wnt5a, nuclear receptor-related factor 1 (NURR1) and tyrosine hydroxylase (TH). However, how Wnt signaling regulates these developmental factors in vitro is yet to be determined. To explore this, SH-SY5Y cells were exposed to PQ and MB. The results of the current study indicated that exposure to PQ and MB decreased Wnt1, ß-catenin, NURR1 and TH levels and increased Wnt5a levels. Furthermore, Wnt1 silencing has the same effect as exposure to PQ and MB. Additionally, the neurotoxicity induced by PQ and MB is more severe in siWnt1-SH-SY5Y cells compared with normal SH-SY5Y cells. Therefore, Wnt1 may serve an important role in regulating developmental DA factors, and may be a candidate gene for PD diagnosis or gene therapy.
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A growing number of reports have demonstrated that the widely-used herbicide Atrazine (ATR) can cause injury to dopamine (DA) neurons, but the exact mechanism remains unclear. In this study, we examined the effects of lactational ATR exposure in Sprague-Dawley rats on dopaminergic neuron health later in life. Compared with control rats, rats exposed to ATR during a critical period of neural development showed decreased striatal DA content and increased rates of DA turnover. The expression of Monoamine oxidase (MAO), which is associated with DA degradation, was up-regulated, and the expression of Vesicular Monoamine Transporter 2 (VMAT2), which is associated with DA transport, was down-regulated. The expression of transcription factor Nuclear Receptor Related Factor 1 (Nurr1), which is associated with DA neuron development, was down-regulated. Increased age (6-12 months old) increased the statistical significance of the differences of the above indicators in the ATR-treated rats compared to the control rats (P<0.05). Taken together, our results indicate that ATR exposure during the critical neural development period causes a down-regulation of Nurr1, which in turn affects Nurr1 target genes, including MAO, VMAT2 and DAT, which are involved in DA degradation and transport. Reduced expression of these genes impairs the capacity for vesicular storage or reuptake of DA, causing decreased levels of striatal DA, which can ultimately lead to DA neuron injury. DA neuron injuries become more severe over time, which suggests that aging can synergistically promote the ATR-associated DA neuron injuries.
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Atrazina/toxicidade , Neurônios Dopaminérgicos/efeitos dos fármacos , Herbicidas/toxicidade , Lactação/metabolismo , Ácido 3,4-Di-Hidroxifenilacético/metabolismo , Fatores Etários , Animais , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Dopamina/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Feminino , Ácido Homovanílico/metabolismo , Masculino , Mesencéfalo/efeitos dos fármacos , Mesencéfalo/metabolismo , Monoaminoxidase/genética , Monoaminoxidase/metabolismo , Membro 2 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Gravidez , Ratos Sprague-Dawley , Proteínas Vesiculares de Transporte de Monoamina/genética , Proteínas Vesiculares de Transporte de Monoamina/metabolismoRESUMO
Paraquat (PQ) and maneb (MB) are widely used herbicides. Wingless (Wnt) proteins serve a role in the development and differentiation of dopaminergic neurons. Previous studies demonstrated that combined exposure to PQ and MB damages dopaminergic neurons in the midbrain. Effects of PQ and MB exposure on midbrain Wnt proteins have also been previously reported. In the present study, from the 5th day of gestation to weaning of the offspring, pregnant SpragueDawley rats were administrated saline, or PQ and MB at two different doses: high, 15 mg/kg body weight PQ + 45 mg/kg body weight MB; or low, 10 mg/kg body weight PQ + 30 mg/kg body weight MB. Dopamine content in the striatum was examined by high performance liquid chromatography with a fluorescence detector and mRNA and protein expression of Wnt1, Wnt5a, nuclear receptor related factor 1 (Nurr1) and tyrosine hydroxylase (TH) in the midbrain was examined by reverse transcriptionquantitative polymerase chain reaction and western blotting. Combined exposure to PQ and MB during development decreased mRNA and protein expression of Wnt1, TH and Nurr1 and increased expression of Wnt5a in the offspring.
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Neurônios Dopaminérgicos/efeitos dos fármacos , Membro 2 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Proteína Wnt-5a/genética , Proteína Wnt1/genética , Animais , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/crescimento & desenvolvimento , Corpo Estriado/patologia , Dopamina/genética , Dopamina/metabolismo , Neurônios Dopaminérgicos/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Lactação/efeitos dos fármacos , Lactação/genética , Maneb/toxicidade , Paraquat/toxicidade , Gravidez , Ratos , Substância Negra/efeitos dos fármacos , Substância Negra/crescimento & desenvolvimento , Substância Negra/patologia , Tirosina 3-Mono-Oxigenase/genéticaRESUMO
AIM: To investigate the effect of exogenous ceramide-induced apoptosis on human colon carcinoma HT-29 cells. METHODS: Light microscope, transmission electron microscope and fluorescence microscope were used to observe the morphology change of apoptosis in HT-29 cells. Agarose gel electrophoresis was performed to detect the DNA fragment. Mitochondrial function was detected by MTT assay. mRNA expression of Bcl-2 family gene members was determined by reverse transcription polymerase chain reaction (RT-PCR) assay. RESULTS: After C2-ceramide treatment, typical characteristics of apoptosis, such as nuclear chromatin breakage, apoptotic body and DNA ladder, could be observed. After exposure to 50 micromol/L C2-ceramide for 12 and 24 h, cell apoptosis was 64.1% and 81.3% respectively, which had a time-and dose-effect relationship. Mitochondrial function started to decrease from 6 h after exposure to ceramide. Meanwhile, ceramide up-regulated or down-regulated the mRNA expression of Bcl-2 family gene members. CONCLUSION: Ceramide induces apoptosis of human colon carcinoma HT-29 cells by affecting the expression of Bcl-2 family gene members and impacting the mitochondrial function.
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Apoptose/efeitos dos fármacos , Neoplasias do Colo/patologia , Inibidores Enzimáticos/farmacologia , Esfingosina/análogos & derivados , Linhagem Celular Tumoral , Neoplasias do Colo/genética , Neoplasias do Colo/ultraestrutura , Fragmentação do DNA , DNA de Neoplasias/efeitos dos fármacos , DNA de Neoplasias/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Genes bcl-2/genética , Humanos , Mitocôndrias/efeitos dos fármacos , RNA Mensageiro/análise , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Esfingosina/farmacologiaRESUMO
OBJECTIVE: To investigate the effect of apoptosis and mRNA expression of apoptosis-related genes of human colon cancer HT-29 cells induced by exogenous ceramide. METHODS: After HT-29 cells exposed to C2-ceramide for different time, transmission electron microscope and fluorescence microscope were used to observe the morphology change of cells. DNA agarose gel electrophoresis was applied to detect DNA fragmentation of HT-29 cells. The mRNA expression of apoptosis-related genes was determined by reverse transcription polymerase chain reaction (RT-PCR) analysis. RESULTS: Under the contribution of C2-ceramide, typical apoptosis appearance, such as nuclear chromatin breakage, apoptotic body etc and DNA ladder were observed. Exposing to 50 micromol/L C2-ceramide for 12h and 24h, the rate of apoptotic cells were 64.7% and 81.3% respectively, which appeared time-dose-relation. Meanwhile, ceramide increased the expression of Bax, Bad and Bid mRNA, and decreased the expression of Bcl-2 and Bcl-xl mRNA. CONCLUSION: Ceramide may induce apoptosis of human colon cancer HT-29 cells via up-regulating or down-regulating the expression of apoptosis-related genes.
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Apoptose/efeitos dos fármacos , Neoplasias do Colo/patologia , Inibidores Enzimáticos/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Esfingosina/análogos & derivados , Linhagem Celular Tumoral , Neoplasias do Colo/metabolismo , Regulação para Baixo , Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Esfingosina/farmacologia , Regulação para CimaRESUMO
Atrazine (2-chloro-4-ethytlamino-6-isopropylamine-1,3,5-triazine; ATR) is a broad-spectrum herbicide with a wide range of applications worldwide. However, ATR is neurotoxic; it reduces dopamine levels in the substantia nigra and corpus striatum in the midbrain, affects the absorption of synaptic vesicles and synaptic bodies, and interferes with dopamine storage and uptake in synaptic vesicles, leading to neurodegenerative disorders. Microglia are resident immunocompetent and phagocytic cells that regulate and participate in the microenvironment in the central nervous system. They demonstrate macrophage characteristics after activation by releasing inflammatory cytokines and neurotoxic substances to increase the inflammatory response, and are thus involved in neurodegeneration. The aim of this study was to investigate the neurotoxic effects of ATR-activated microglia-mediated neuronal damage in terms of human dopaminergic neuroblastoma SH-SY5Y cell death. ATR was administered to BV-2 microglial cells at 12.5, 25, and 50 µM for 1, 6, 12, 24 and 48 h, respectively. ATR increased activated-microglia-induced overexpression of reactive oxygen species, inducible nitric oxide synthase, nitric oxide, gp91(phox), p47(phox), and the inflammatory cytokines tumor necrosis factor α and interleukin-1ß, thus reducing SH-SY5Y cell viability. These results suggest that activated microglia may play a critical role in inflammation-mediated dopaminergic neuronal death, and provide the basis for further studies on the mechanisms of ATR-induced dopaminergic system toxicity.
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Atrazina/toxicidade , Dopamina/metabolismo , Microglia/patologia , Neuroblastoma/patologia , Neurotoxinas/toxicidade , Animais , Western Blotting , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Imunofluorescência , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-1beta , Camundongos , Microglia/efeitos dos fármacos , Microglia/metabolismo , NADPH Oxidases/genética , NADPH Oxidases/metabolismo , Óxido Nítrico/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Receptores Imunológicos/genética , Receptores Imunológicos/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Phosphino-phenolate neutral nickel catalysts 1-3/B(C6F5)3, without the help of any organoaluminum compound, were found to be efficient catalytic systems for norbornene polymerization and its copolymerization with norbornene derivatives. The amount of B(C6F5)3 required for achieving a high efficiency (3 equiv.) was markedly lower compared to previous reports, and high molecular weight polymers were obtained (>10(6) g mol(-1)). Efficient incorporation of polar monomers NBC, NBA, and NBM was also achieved in a controllable fashion, yielding high molecular weight copolymers. Catalysts 1-3 were highly active for ethylene polymerization as single component catalysts, with an activity of up to 10(7) g molNi(-1) h(-1), and catalyst 3 was more readily initiated at lower temperature. Catalysts 1-3 were also efficient in incorporating norbornene (up to 30%) into the polyethylene backbone. Bisligated phosphino-phenolate nickel complex 4 and salicylaldimine complex 5 were also studied for comparison, which further verified the unique performance of catalysts 1-3. Preliminary NMR analyses were conducted to explore the norbornene polymerization mechanism.
RESUMO
AIM: To determine the effect of cis -9, trans -11-conjugated linoleic acid (c9, t11-CLA) on the cell cycle of gastric cancer cells (SGC-7901) and its possible mechanism in inhibition cancer growth. METHODS: Using cell culture and immunocytochemical techniques, we examined the cell growth, DNA synthesis, expression of PCNA, cyclin A, B(1), D(1), p16(ink4a) and p21(cip/waf1) of SGC-7901 cells which were treated with various c9, t11-CLA concentrations (25, 50, 100 and 200 micromol.L(-1))of c 9, t 11-CLA for 24 and 48h, with a negative control (0.1% ethane). RESULTS: The cell growth and DNA synthesis of SGC-7901 cells were inhibited by c9, t11-CLA.SGC-7901 cells. Eight day after treatment with various concentrations of c9, t11-CLA mentioned above, the inhibition rates were 5.92%, 20.15%, 75.61% and 82.44%, respectively and inhibitory effect of c9, t11-CLA on DNA synthesis (except for 25 micromol.L, 24h) showed significantly less (3)H-TdR incorporation than that in the negative controls (P<0.05 and P<0.01). Immunocytochemical staining demonstrated that SGC-7901 cells preincubated in media supplemented with different c9, t11-CLA concentrations at various times significantly decreased the expressions of PCNA (the expression rates were 7.2-3.0%, 24h and 9.1-0.9% at 48h, respectively), Cyclin A (11.0-2.3%, 24h and 8.5-0.5%,48h), B(1) (4.8-1.8% at 24h and 5.5-0.6% at 48h)and D(1) (3.6-1.4% at 24h and 3.7%-0 at 48h) as compared with those in the negative controls(the expressions of PCNA, Cyclin A, B(1) and D(1) were 6.5% at 24h and 9.0% at 48h, 4.2% at 24h and 5.1% at 48h, 9.5% at 24h and 6.0% at 48h,respectively)(P<0.01), whereas the expressions of P16(ink4a) and P21(cip/waf1), cyclin-dependent kinases inhibitors(CDKI), were increased. CONCLUSION: The cell growth and proliferation of SGC-7901 cell is inhibited by c9, t11-CLA via blocking the cell cycle, with reduced expressions of cyclin A,B(1) and D(1) and enhanced expressions of CDKI(P16(ink4a) and p21(cip/waf1)).
Assuntos
Adenocarcinoma/patologia , Ciclo Celular/efeitos dos fármacos , Ácidos Linoleicos Conjugados , Ácidos Linoleicos/farmacologia , Neoplasias Gástricas/patologia , Animais , Divisão Celular/fisiologia , Ciclina A/metabolismo , Ciclina B/metabolismo , Ciclina B1 , Ciclina D1/metabolismo , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Inibidor de Quinase Dependente de Ciclina p21 , Ciclinas/metabolismo , Inibidores Enzimáticos/metabolismo , Humanos , Imuno-Histoquímica , Ácidos Linoleicos/química , Antígeno Nuclear de Célula em Proliferação/metabolismo , Células Tumorais CultivadasRESUMO
Thioglycolic acid (TGA) is the active ingredient of permanent-waving solution (PWS). TGA has been shown to be a chemical of high toxicity, which can be absorbed through intact skin and cause damage to organs or systems in animals. This study evaluated the effect of TGA-containing PWS on the health of a human population in 3 substudies. Firstly, 57 female hairdressers exposed to TGA-containing PWS (cases) and 64 female schoolteachers (controls) were studied. Their menstruation state was evaluated with information obtained from interviews. The results revealed that the menoxenia rate in the cases was significantly higher than that in the controls (22.81% vs 9.38%, p<0.05). Secondly, 8 female hairdressers selected from those that participated in the above survey underwent a fluctuation test for the mutagenic activity of urine. Eight female medical students were chosen as controls. Difference in the mutagenic activity of urine on TA100 between the two groups was highly significant (110.30 +/- 45.95 vs 28.43 +/- 19.33, p<0.01). Finally, a micronucleus assay was carried out on scalp hair follicle cells in healthy volunteers. Scalp hair with the follicle cell mass was sampled from 8 male and 8 female volunteers before permanent waving and at 24, 48 and 72 h after waving. One thousand hair follicle cells were examined by light microscopy. The number of cells containing a micronucleus and the number of micronuclei in each cell was determined. The permillages of micronuclei in hair follicle cells before and after permanent waving were compared. Micronuclei presence reached its peak value (12.44) 24 h after permanent waving, which was significantly higher than that before waving (3.13, p<0.001). The rate decreased progressively after 24 h. Our results suggest that the reproductive function of hairdressers may be affected by long-term exposure to PWS, probably due to the presence of TGA, and more attention should be paid to its potential carcinogenic effects.