RESUMO
Fish intestine contains different types of microbiomes, and bacteria are the dominant microbiota in fishes. Studies have identified various core gut bacteria in fishes. However, little is known about the composition and their relative functions of gut microbial community along the intestine. To explore this, the current study investigated the microbial community distribution along the gut in Anguilla japonica. By 16S rRNA gene sequencing, we profiled the gut microbiota in eel along the three regions (anterior intestine (AI), the middle intestine (MI) and the posterior intestine (PI)). Results suggested that the three regions did not have significant differences on the observed species and diversities. The cluster tree analysis showed that the bacteria community in MI was closer to PI than the AI. The dominant bacteria in AI were the Proteobacteria, in which the majority was graduated replaced by Bacteroidetes along the gut to PI region. Through PICRUSt analysis, shifts in the bacterial community along the gut were found to affect the genetic information processing pathways. Higher levels of translation and transcriptional pathway activities were found in MI and PI than in AI. The dominant bacterial species were different among the regions and contributed to various biological functions along the gut.
Assuntos
Anguilla , Microbioma Gastrointestinal , Microbiota , Animais , Bactérias/genética , RNA Ribossômico 16S/genéticaRESUMO
There has been a marked increase in the prevalence of diabetes in Asia, including China, over the last few decades. While the increased prevalence of diabetes has often been attributed to the nutritional transition associated with recent economic development, emerging data suggest that early-life exposures also play a major role in shaping developmental trajectories, and contributes to alter an individual's susceptibility to diabetes and other non-communicable diseases (NCDs). Early-life exposures such as in utero exposure to undernutrition has been consistently linked with later risk of diabetes and obesity. Furthermore, in utero exposure to maternal hyperglycemia, maternal obesity and excess gestational weight gain are all linked with increased childhood obesity and later risk of diabetes. Emerging data have also highlighted the potential link between early-feeding practices, the role of one-carbon metabolism in metabolic programming and endocrine disrupting chemicals (EDCs) with later risk of diabetes. These different developmental exposures may all be highly relevant to the current epidemic of diabetes in China. For example, the prevalence of gestational diabetes has increased markedly over the last two decades, and may contribute to the diabetes epidemic by driving macrosomia, childhood obesity and later risk of diabetes. In order to address the current burden of diabetes, a lifecourse perspective, incorporating multisectoral efforts from public health policy down to the individuals, will be needed. Several major initiatives have been launched in China as part of its national plans for NCD prevention and treatment, and the experience from these efforts would be invaluable.
Assuntos
Diabetes Mellitus Tipo 2/epidemiologia , Diabetes Gestacional/epidemiologia , Desnutrição/epidemiologia , Fenômenos Fisiológicos da Nutrição Materna , Obesidade/epidemiologia , Complicações na Gravidez/epidemiologia , China/epidemiologia , Dieta , Feminino , Comportamentos Relacionados com a Saúde , Política de Saúde , Humanos , Estilo de Vida , Metanálise como Assunto , Gravidez , Efeitos Tardios da Exposição Pré-Natal , Prevalência , Saúde Pública , Fatores de RiscoRESUMO
Perfluorooctane sulfonate (PFOS), a member of the perfluorinated chemical family, has been convincingly demonstrated to affect lipid metabolism in animals and humans and readily crosses the placenta to exert its effects on the developing fetuses. While its exact mechanism is still not clear, PFOS exposure has long been suggested to exert its toxicity via oxidative stress and/or altered gene expression. Levels of PFOS and malondialdehyde in various organs and cell cultures have been widely determined as general indicators of non-specific lipid peroxidation after PFOS exposure. In this study, the oxidation of precise polyunsaturated fatty acids and their metabolites, derived from enzymatic and non-enzymatic pathways was determined following PFOS exposure in both adult and maternal/fetal mice. CD-1 mice were exposed to 3 mg/kg body weight/day of PFOS in corn oil by oral gavage until late gestation (GD17). We demonstrated that lipid peroxidation was particularly and exclusively affected in fetuses exposed to PFOS, but this was not the case in the maternal mice, where limited effects were observed in the enzymatic oxidation pathway. In this study, we demonstrated that PFOS-induced lipid peroxidation might have a greater impact in free radical generation in fetuses than in dams and could be responsible for affecting fetal development. In addition, antioxidant enzymes, such as superoxide dismutase and catalase, appeared to maintain oxidative stress homeostasis partially in adult mice exposed to PFOS. Taken together, our results might elucidate the mechanism of how PFOS induces oxidative stress in vivo.
Assuntos
Ácidos Alcanossulfônicos/toxicidade , Poluentes Ambientais/toxicidade , Fluorocarbonos/toxicidade , Peroxidação de Lipídeos , Estresse Oxidativo , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Animais , Catalase/metabolismo , Colesterol/sangue , Ácidos Graxos Insaturados/sangue , Feminino , Desenvolvimento Fetal/efeitos dos fármacos , Fígado/enzimologia , Camundongos , Especificidade de Órgãos , Gravidez , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Superóxido Dismutase/metabolismoRESUMO
The expression of the Ca(2) (+)-sensing receptor (Casr) in the endocrine gland known as the corpuscle of Stannius (CS) regulates the secretion of the hypocalcemic hormone stanniocalcin-1 (STC1) to inhibit gill Ca(2) (+) uptake. Although numerous studies have reported the branchial expression of Casr and Stc1, the functions of these proteins in gills have not been elucidated yet. On the basis of recent findings regarding the autocrine/paracrine functions of STC1 in mammalian models, we proposed the hypothesis that branchial CaSR has an in situ 'sensing' function to regulate STC1 that maintains local Ca(2) (+) homeostasis. In this study, we investigated Casr-mediated signaling and its regulation of Stc1 and cyclooxygenase-2 (Cox2) expression/function using a primary gill-cell culture model. The biochemical responses of gill cells isolated from Japanese eels to an increasing concentration of extracellular Ca(2) (+) (0.1-1âmM) were tested. This stimulation led to a transient increase in phosphatidylcholine-phospholipase C (PC-PLC) activity, followed by activation of ERK and inositol 1,4,5-trisphosphate-Ca(2) (+)/calmodulin-dependent protein kinase 2 (CaMK2) signaling pathways. Cotreatment with the calcimimetic R467 caused synergistic effects on Ca(2) (+)-stimulated PC-PLC activity, ERK signaling, and CaMK2 signaling. The activation of the CaSR-PLC-ERK pathway was associated with increased expression levels of Stc1 and Cox2 as confirmed by the inhibition of Erk using a chemical inhibitor, PD98059. Functionally, Ca(2) (+)/R-467 pretreatment was found to protect cells from thapsigargin-induced cell death. Inhibition of COX2 activity using NS398 abolished this protection, while transduction of STC1 lentiviral particles in the gill cells increased the protective effects. Collectively, our data revealed the expression of functional CaSR in gill tissues. The identification of the CaSR-STC1/COX2-mediated protective pathway in gill cells sheds light on a possible cellular protective mechanism against an increase in intracellular Ca(2) (+) levels associated with transepithelial Ca(2) (+) transport.
Assuntos
Cálcio/metabolismo , Receptores de Detecção de Cálcio/metabolismo , Transdução de Sinais , Animais , Cálcio/farmacologia , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Células Cultivadas , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Ativação Enzimática/efeitos dos fármacos , Expressão Gênica , Brânquias , Glicoproteínas/genética , Glicoproteínas/metabolismo , Lactonas/farmacologia , Lactonas/toxicidade , Sesquiterpenos/farmacologia , Sesquiterpenos/toxicidade , Transdução de Sinais/efeitos dos fármacosRESUMO
Perfluorinated compounds (PFCs) are man-made fluorinated hydrocarbons, which are very persistent in the environment. Since the early 1980s, the usage of PFCs has sharply increased for a wide array of industrial and commercial applications. Being the most important PFC, perfluorooctane sulfonate (PFOS) has received much attention. In the past decades, increasing surveys have been focused on this compound, to study its sources, fates and effects in the environment. According to the large production volume and wide usage in industrial and commercial products in the past, PFOS can be detected in various environmental media and matrix, even in human tissues. This article attempted to review the current status of PFOS contaminations in Asia, focusing on water systems, sediments, wide animals and human tissues. A special section is devoted to examine the pathways of human exposure to this compound, as well as human body loadings of PFOS and their possible association with diseases.
Assuntos
Ácidos Alcanossulfônicos/toxicidade , Exposição Ambiental , Poluentes Ambientais/toxicidade , Fluorocarbonos/toxicidade , Animais , Ásia , Aves/metabolismo , Carga Corporal (Radioterapia) , Monitoramento Ambiental , Peixes/metabolismo , Humanos , Mamíferos/metabolismoRESUMO
Bisphenol A (BPA), identified as an endocrine disruptor, is an industrially important chemical that is used as a raw material in the manufacture of many products such as engineering plastics (e.g., epoxy resins/polycarbonate plastics), food cans (i.e., lacquer coatings), and dental composites/sealants. The demand and production capacity of BPA in China have grown rapidly. This trend will lead to much more BPA contamination in the environmental media and in the general population in China. This paper reviews the current literature concerning the pollution status of BPA in China (the mainland, Hong Kong, and Taiwan) and its potential impact on human health. Due to potential human health risks from long-term exposure to BPA, body burden of the contaminant should be monitored.
Assuntos
Disruptores Endócrinos/análise , Exposição Ambiental/estatística & dados numéricos , Poluentes Ambientais/análise , Poluição Ambiental/estatística & dados numéricos , Fenóis/análise , Animais , Compostos Benzidrílicos , China , Dieta/estatística & dados numéricos , Disruptores Endócrinos/toxicidade , Política Ambiental , Poluentes Ambientais/toxicidade , Análise de Alimentos , Nível de Saúde , Humanos , Fenóis/toxicidade , Plásticos , Cimento de PolicarboxilatoRESUMO
Perfluorooctanesulfonate (PFOS) was produced and used by various industries and in consumer products. Because of its persistence, it is ubiquitous in air, water, soil, wildlife, and humans. Although the adverse effects of PFOS on male fertility have been reported, the underlying mechanisms have not yet been elucidated. Here, for the first time, the effects of PFOS on testicular signaling, such as gonadotropin, growth hormone, insulin-like growth factor, and inhibins/activins were shown to be directly related to male subfertility. Sexually mature 8-wk-old CD1 male mice were administered by gavages in corn oil daily with 0, 1, 5, or 10 mg/kg PFOS for 7, 14, or 21 days. Serum concentrations of testosterone and epididymal sperm counts were significantly lower in the mice after 21 days of the exposure to the highest dose compared with the controls. The expression levels of testicular receptors for gonadotropin, growth hormone, and insulin-like growth factor 1 were considerably reduced on Day 21 in mice exposed daily to 10 or 5 mg/kg PFOS. The transcript levels of the subunits of the testicular factors (i.e., inhibins and activins), Inha, Inhba, and Inhbb, were significantly lower on Day 21 of daily exposure to 10, 5, or 1 mg/kg PFOS. The mRNA expression levels of steroidogenic enzymes (i.e., StAR, CYP11A1, CYP17A1, 3beta-HSD, and 17beta-HSD) were notably reduced. Therefore, PFOS-elicited subfertility in male mice is manifested as progressive deterioration of testicular signaling.
Assuntos
Ácidos Alcanossulfônicos/toxicidade , Poluentes Ambientais/toxicidade , Fluorocarbonos/toxicidade , Sistema Hipotálamo-Hipofisário/fisiologia , Infertilidade Masculina/induzido quimicamente , Transdução de Sinais/efeitos dos fármacos , Testículo/fisiologia , Ativinas/genética , Ativinas/metabolismo , Ácidos Alcanossulfônicos/administração & dosagem , Animais , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Poluentes Ambientais/administração & dosagem , Epididimo/citologia , Epididimo/efeitos dos fármacos , Fluorocarbonos/administração & dosagem , Infertilidade Masculina/metabolismo , Inibinas/genética , Inibinas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos , Oligospermia/induzido quimicamente , RNA Mensageiro/metabolismo , Distribuição Aleatória , Receptor IGF Tipo 1/metabolismo , Receptores da Gonadotropina/metabolismo , Receptores da Somatotropina/metabolismo , Testosterona/sangueRESUMO
Sediments from Mai Po Ramsar site, Hong Kong were in general shown to be highly toxic based on the results of four toxicity tests (Microtox solid-phase test, Daphnia mortality test, algal [Microcystis aeruginosa] growth inhibition test and ryegrass [Lolium perenne] seed germination/root elongation test). Sediment of the mudflat (which is open to Deep Bay, i.e., the pollution source) was the most toxic while sediment of gei wai 24g (an enclosed freshwater pond) was the least toxic. Results of biomarker studies (tilapia hepatic metallothionein; glutathione (GSH) and EROD activity using H4IIE rat hepatoma cell) were also concordant with those in the toxicity tests. Significant liner relationships (p<0.01) were found between GSH contents in the rat hepatoma cells and PAHs, OCPs contents in the sediment extracts. It is recommended that the present suite of bioassays is useful and is biologically relevant for future ecotoxicological studies focusing on similar wetlands.
Assuntos
Daphnia/efeitos dos fármacos , Monitoramento Ambiental , Sedimentos Geológicos/análise , Lolium/efeitos dos fármacos , Microcystis/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Áreas Alagadas , Animais , Biomarcadores/análise , Linhagem Celular Tumoral , Citocromo P-450 CYP1A1/metabolismo , Ecotoxicologia , Glutationa/análise , Hong Kong , Hidrocarbonetos Clorados/análise , Hidrocarbonetos Clorados/toxicidade , Metalotioneína/análise , Metalotioneína/toxicidade , Hidrocarbonetos Policíclicos Aromáticos/análise , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Ratos , Poluentes Químicos da Água/análiseRESUMO
2-Methoxyestradiol (2ME2) is a normal physiological metabolite of 17beta-estradiol with anti-proliferative and anti-angiogenic activities. The purpose of this study is to elucidate the mechanism whereby 2ME2 induces endoreduplication of the well-differentiated nasopharyngeal carcinoma (NPC) cells. We report here that 2ME2 induces G2/M phase cell cycle arrest followed by endoreduplication of the well-differentiated HK-1 cells. The increase in chromosome number was confirmed by cytogenetic study. Analysis of stress signaling pathways revealed the phosphorylation activation of ERK, JNK and p38 MAPKs at various times after 2ME2 treatment. Pre-treatment of 2ME2-treated HK-1 cells with JNK inhibitor (SP600125), ERK inhibitor (PD98059) and p38 MAPK inhibitor (SB203580) resulted in the reduction of endoreduplicating cells. Furthermore, the increase in the phosphorylation of JNK was accompanied by an increase in the reactive oxygen species. In addition, endoreduplication was observed in cells after treatment with superoxide donor, 2,3-dimethoxy-1,4-naphoquinone (DMNQ). Confocal microscopic analysis also revealed the increase in mitochondrial superoxide anion in 2ME2-treated HK-1 cells. Pre-treatment of HK-1 cells with superoxide dismutase mimetic 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) or overexpressing the mitochondrial enzyme MnSOD resulted in the reduction of phosphorylation of JNK and the formation of endoreduplicating cells. Furthermore, the tubulin filaments in cytoplasm remain intact in 2ME2-treated HK-1 cells after pre-treatment of TEMPO. Our results suggest that 2ME2 induces endoreduplication through the induction of oxidative stress and the activation of MAPK signal pathways. The biological significance of drug-induced endoreduplication will also be discussed.
Assuntos
Estradiol/análogos & derivados , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Moduladores de Tubulina/farmacologia , 2-Metoxiestradiol , Linhagem Celular , Proliferação de Células , Relação Dose-Resposta a Droga , Ativação Enzimática , Estradiol/farmacologia , Humanos , Mitocôndrias/metabolismoRESUMO
Nine groups of food items (freshwater fish, marine fish, pork, chicken, chicken eggs, leafy, non-leafy vegetables, rice and flour) and three types of human samples (human milk, maternal serum and cord serum) were collected for the analysis of PCDD/Fs. Results of chemical analysis revealed PCDD/Fs concentrations (pg g(-1) fat) in the following ascending order: pork (0.289 pg g(-1) fat), grass carp (Ctenopharyngodon idellus) (freshwater fish) (0.407), golden thread (Nemipterus virgatus) (marine fish) (0.511), chicken (0.529), mandarin fish (Siniperca kneri) (marine fish) (0.535), chicken egg (0.552), and snubnose pompano (Trachinotus blochii) (marine fish) (1.219). The results of micro-EROD assay showed relatively higher PCDD/Fs levels in fish (2.65 pg g(-1) fat) when compared with pork (0.47), eggs (0.33), chicken (0.13), flour (0.07), vegetables (0.05 pg g(-1) wet wt) and rice (0.05). The estimated average daily intake of PCDD/Fs of 3.51 pg EROD-TEQ/kg bw/day was within the range of WHO Tolerable Daily Intake (1-4 pg WHO-TEQ/kg bw/day) and was higher than the Provisional Tolerable Daily Intake (PMTL) (70 pg for dioxins and dioxin-like PCBs) recommended by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) [Joint FAO/WHO Expert Committee on Food Additives (JECFA), Summary and conclusions of the fifty-seventh meeting, JECFA, 2001.]. Nevertheless, the current findings were significantly lower than the TDI (14 pg WHO-TEQ/kg/bw/day) recommended by the Scientific Committee on Food of the Europe Commission [European Scientific Committee on Food (EU SCF), Opinions on the SCF on the risk assessment of dioxins and dioxin-like PCBs in food, 2000.]. However, it should be noted that micro-EROD assay overestimates the PCDD/Fs levels by 2 to 7 folds which may also amplify the PCDD/Fs levels accordingly. Although the levels of PCDD/Fs obtained from micro-EROD assay were much higher than those obtained by chemical analysis by 2 to 7 folds, it provides a cost-effective and rapid screening of dioxin levels in food and human samples.
Assuntos
Benzofuranos/análise , Poluentes Ambientais/análise , Análise de Alimentos , Dibenzodioxinas Policloradas/análogos & derivados , Benzofuranos/sangue , Benzofuranos/metabolismo , Biomarcadores/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Dibenzofuranos Policlorados , Dioxinas/análise , Dioxinas/metabolismo , Ovos/análise , Poluentes Ambientais/sangue , Poluentes Ambientais/metabolismo , Feminino , Sangue Fetal/química , Farinha/análise , Hong Kong , Humanos , Carne/análise , Leite Humano/química , Oryza/química , Dibenzodioxinas Policloradas/análise , Dibenzodioxinas Policloradas/sangue , Dibenzodioxinas Policloradas/metabolismo , Medição de Risco , Verduras/químicaRESUMO
Chemoresistance and therapeutic selectivity are major obstacles to successful chemotherapy of ovarian cancer. Manganese superoxide disumutase (MnSOD) is an important antioxidant enzyme responsible for the elimination of superoxide radicals. We reported here that MnSOD was significantly elevated in ovarian cancer cells and its overexpression was one of the mechanisms that increased resistance to apoptosis in cancer cells. Knockdown of MnSOD by small-interfering RNA (siRNA) led to an increase in superoxide generation and sensitisation of ovarian cancer cells to the two front-line anti-cancer agents doxorubicin and paclitaxel whose action involved free-radical generation. This synergistic effect was not observed in non-transformed ovarian surface epithelial cells. Furthermore, our results revealed that this combination at the cellular level augmented activation of caspase-3 and caspase-9, but not caspase-8, suggesting involvement of an intrinsic apoptotic pathway. Evaluation of signalling pathways showed that MnSOD siRNA enhanced doxorubicin- and paclitaxel-induced phosphorylation of extracellular signal-regulated kinase 1/2. Akt activation was not affected. These results identify a novel chemoresistance mechanism in ovarian cancer, and show that combination of drugs capable of suppressing MnSOD with conventional chemotherapeutic agents may provide a novel strategy with a superior therapeutic index and advantage for the treatment of refractory ovarian cancer.
Assuntos
Caspase 9/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/enzimologia , Superóxido Dismutase/antagonistas & inibidores , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Caspase 9/genética , Inibidores de Caspase , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Feminino , Humanos , Oligopeptídeos/farmacologia , Neoplasias Ovarianas/genética , Paclitaxel/farmacologia , RNA Interferente Pequeno/genética , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , TransfecçãoRESUMO
In the present study, we aimed to clone an osmotic stress transcriptional factor (Ostf) from gill cells of Japanese eels. In addition, we measured its expression in Percoll-gradient-isolated gill chloride (CC) and pavement (PVC) cells and determined the regulation of its expression in primary gill cell culture. Using degenerative primers and RACE techniques, we cloned a cDNA of 615bp, encompassing the coding sequence of Ostf (204 amino acids). The cloned Ostf1 DNA sequence shared 84% DNA homology with the Ostf1 of tilapia. In general, the basal Ostf expression level was found to be significantly higher in CCs than in PVCs. In the direct transfer of fish from freshwater to seawater, a significant but transient induction of Ostf mRNA in CCs and PVCs was measured after 6h of acclimation. Compared with gill CCs, the level of induction measured at PVCs was lower. In the seawater-to-freshwater transfer, no significant change in Ostf transcript levels was detected in either CCs or PVCs. To decipher the regulatory mechanism of Ostf expression, we conducted experiments using primary gill cell culture to specifically address the involvement of two putative osmosensors (i.e. intracellular ion strength/macromolecular crowding and cytoskeleton) in the regulation of Ostf expression. Hypertonic treatment using impermeable solutes (i.e. NaCl, 500 mOsmol l(-1)) induced Ostf mRNA expression in 6h, but no noticeable effect was measured using permeable solute (i.e. urea, 500 mOsmol l(-1)). The induction was transcriptionally regulated and was abolished by the addition of organic osmolytes (i.e. betaine, inositol or taurine) into the culture media. Addition of colchicine (an inhibitor of microtubule polymerization) to hypertonic (with added NaCl, 500 mOsmol l(-1)) cells reduced Ostf mRNA expression, suggesting that an increase in intracellular ionic strength and the integrity of the cytoskeleton are involved in the activation of Ostf mRNA expression in the cells. Collectively, the results of this study reveal, for the first time, the differential expression of Ostf in isolated CCs and PVCs. The resulting knowledge can shed light on how Ostf participates in hyperosmotic adaptation in fish gills.
Assuntos
Enguias/genética , Regulação da Expressão Gênica/fisiologia , Brânquias/metabolismo , Fatores de Transcrição/genética , Equilíbrio Hidroeletrolítico/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Betaína , Células Cultivadas , Clonagem Molecular , Colchicina , Primers do DNA/genética , Enguias/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Dados de Sequência Molecular , Análise de Sequência de DNA , Homologia de Sequência , Cloreto de Sódio , Especificidade da Espécie , Fatores de Transcrição/metabolismo , UreiaRESUMO
Contamination from persistent organic pollutants is a pervasive global problem that urgently demands global concern and action. In the present study, concentrations of organochlorine (OC) pesticides and polychlorinated biphenyls (PCBs) were determined in 37 samples of female adipose tissue collected in Hong Kong hospitals. Among the pollutants analyzed, DDTs (2.79 ng/g fat), HCHs (0.72 ng/g fat), and PCBs (0.19 ng/g fat) were prominent compounds in most of the adipose tissue. p,p'-DDE and hexachlorinated biphenyls were found in all samples, whereas heptachlor epoxide and dieldrin were found only in some samples. An estimation of toxic equivalency concentration (TEQ) due to dioxin-like coplanar PCBs was also performed. The estimated TEQ(PCBs) was 2.01 pg/g fat. This study also compared our previous results obtained from the milk samples of the same donors. Significant correlations are obtained for DDTs and HCHs between milk and adipose tissue. Detailed review of available information concerning OC pesticides and PCBs in different ecological compartments indicated that bioconcentration and biomagnification of these contaminants are common phenomena of the Pearl River Delta region, which has undergone rapid socioeconomic change in the past 20 years. It is suggested to establish a regional organization in order to coordinate the monitoring of persistent organic pollutants in the region.
Assuntos
Tecido Adiposo/química , Poluentes Ambientais/análise , Hidrocarbonetos Clorados/análise , Leite Humano/química , Praguicidas/análise , Bifenilos Policlorados/análise , Adulto , Monitoramento Ambiental , Feminino , Hong Kong , HumanosRESUMO
Polychlorinated dibenzo-p-dioxins, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) have been recognized as highly potent developmental and reproductive toxins. We have previously demonstrated effects of TCDD in modulating the expression of rat Sertoli cell secretory products and markers for cell-cell interaction. In this study, we examined the direct biological effects of TCDD in rat Leydig cell primary cultures. Mature rat Leydig cells were purified by Percoll gradient centrifugation and the cell purity was determined by 3beta-hydroxysteroid dehydrogenase (3beta-HSD) staining and a testosterone induction assay. To examine TCDD-induced biological consequences, we measured the changes in the secretion of progesterone and testosterone, as well as transcript levels of some selected steroidogenic enzymes (i.e. StAR, P450scc, 3beta-HSD and CYP17alpha), in TCDD/human chorionic gonadotropin (hCG) co-treated cells. Our results indicated that TCDD (0.2 or 2 ng/ml) treatment significantly suppressed hCG (5 or 10 ng/ml)-induced testosterone secretion. The suppressive effect aligned with a reduction of progesterone secretion (P<0.05), as well as a decrease of P450scc mRNA and protein expression (P<0.05). The mechanistic action of TCDD was found to be via the reduction of cellular cAMP levels in the hCG-treated cells. This observation was further confirmed, as the TCDD-mediated suppressive effect could be reversed by dibutyryl cAMP co-treatment. The data indicate that TCDD can modulate cAMP signaling in rat Leydig cells to affect the process of steroidogenesis.
Assuntos
Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo , Células Intersticiais do Testículo/efeitos dos fármacos , Dibenzodioxinas Policloradas/farmacologia , Teratogênicos/farmacologia , 3-Hidroxiesteroide Desidrogenases/genética , 3-Hidroxiesteroide Desidrogenases/metabolismo , Animais , Western Blotting/métodos , Técnicas de Cultura de Células , AMP Cíclico/metabolismo , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Depressão Química , Células Intersticiais do Testículo/enzimologia , Masculino , Progesterona/genética , Progesterona/metabolismo , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Testosterona/genética , Testosterona/metabolismoRESUMO
Hong Kong and South China are the most developed regions within China. The industrialization in these areas has resulted in severe environmental problems. Sediment and biotic samples including tilapia (Oreochromis mossambicus), bighead carp (Aristichthys nobilis), grass carp (Ctenopharyngodon idellus), crucian carp (Carassius auratus) and mandarin fish/fresh water grouper (Siniperca chuatsi) were collected from different fishponds in the Pearl River Delta (Tanzhou, Sanjiao, Guangzhou, Shipai, Changan and Mai Po) for the analysis of polycyclic aromatic hydrocarbons (PAHs) and organochlorine (OC) pesticides. Mandarin fish, which belongs to the highest trophic level, accumulated the highest concentrations of PAHs and DDTs among all fish species. The levels of DDTs in fish samples ranged from 1.5 to 62ng g-1 (wet wt.), with more than 30% of the fish samples exceeding the limit of 14.4ng g-1 (wet wt.) for human consumption recommended by US EPA (2000). Levels of PAHs in fish samples ranged from 1.91 to 224.03ng g-1 (wet wt.), but the potency-weighted total concentrations of PAHs in all muscle tissues were below the guideline value of 0.67ng g-1 (wet wt.) for human consumption set by US EPA (2000). The guideline value calculated was based on a tissue consumption rate of 142.2g day-1 (4-5 meals per week), which is a more protective rate for populations with a high consumption of fish, like Chinese and Asians. The effect of lipid content in PAHs and DDTs accumulation in fish tissue was not significant in general.
Assuntos
DDT/análise , Diclorodifenil Dicloroetileno/análise , Peixes/metabolismo , Contaminação de Alimentos , Hidrocarbonetos Policíclicos Aromáticos/análise , Animais , China , DDT/metabolismo , Diclorodifenil Dicloroetileno/metabolismo , Monitoramento Ambiental , Sedimentos Geológicos/análise , Lipídeos/análise , Músculos/química , Resíduos de Praguicidas/análise , Resíduos de Praguicidas/metabolismo , Praguicidas/análise , Praguicidas/metabolismo , Medição de Risco , Rios , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/metabolismoRESUMO
The present study reported the efficacy of antisense oligonucleotides specific for N-methyl-D-aspartate receptor one (NR1) in reduction of motor symptom in a rat parkinsonian model, the unilateral 6-hydroxydopamine-lesioned rat. Significant reductions in apomorphine-induced contralateral rotation were only seen in the NR1 antisense-treated lesioned rats (after a single intraparenchymal dose of antisense to the lesioned neostriatum; 15 nmol in 3 microl of saline) at 1 or 2 days after the treatment. No motor effect was seen in the lesioned animals with control treatments (sham, treatment using NR1 sense oligonucleotides, randomized oligonucleotides or saline, respectively). In contrast, significant increases in expression of NR1 mRNA in the lesioned neostriatum were seen in rats with control treatments but not in rats with NR1 antisense treatment. Importantly, in the lesioned neostriatum that was treated with NR1 antisense, a significant reduction in NR1 protein expression was found and NR1 immunoreactivity was seen to reduce in perikarya of presumed striatal medium spiny neurons. The present data indicate that a single dose of NR1 antisense ameliorates motor symptom in the rat model. The efficacy of NR1 antisense is likely to be mediated by a selective knockdown in expression of NR1 mRNA and proteins in the presumed medium spiny neurons.
Assuntos
Regulação da Expressão Gênica/fisiologia , Inativação Gênica/fisiologia , Transtornos Parkinsonianos/metabolismo , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Receptores de N-Metil-D-Aspartato/biossíntese , Animais , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Inativação Gênica/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Neostriado/efeitos dos fármacos , Neostriado/metabolismo , Oligonucleotídeos Antissenso/farmacologia , Oxidopamina , Transtornos Parkinsonianos/induzido quimicamente , Transtornos Parkinsonianos/genética , Ratos , Ratos Sprague-DawleyRESUMO
In the present study, we attempted to address the modulation of the gene expression of N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) glutamate receptors in the neostriatum of the 6-hydroxydopamine (6-OHDA)-lesioned rat, an animal model of Parkinson's disease. After 2 weeks of lesion, reverse transcriptase-polymerase chain reactions (RT-PCRs) revealed significant reduction in GluR1 mRNA expression but a significant enhancement of NR1 mRNA expression in the striatal tissues of the lesioned side. No modulation in the mRNA expression of GluR2, GluR3, GluR4 and NR2B were found. Immunofluorescence with digital imaging analysis also demonstrated a significant reduction in GluR1 immunoreactivity in the lesioned neostriatum. Interestingly, the reduction in GluR1 immunoreactivity was primarily observed in presumed striatal medium spiny neurons but not in parvalbumin-labeled striatal GABAergic interneurons. Immunoreactivity for GluR2, GluR2/3, GluR4, NR1 and NR2B was unchanged in neurons of the neostriatum of the lesioned side. The present results indicate that there is an opposite trend in modulation in the gene expressions of GluR1 and NR1 in the neostriatum of 6-OHDA-lesioned rats after dopamine denervation. Modulation of GluR1 mRNA and immunoreactivity is likely to be limited in the striatal projection neurons. These findings have implications for the use of NMDA and AMPA receptor antagonists in the treatment of Parkinson's disease.
Assuntos
Corpo Estriado/metabolismo , Neostriado/metabolismo , Oxidopamina , Receptores de AMPA/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Animais , Modelos Animais de Doenças , Feminino , Imunofluorescência , Expressão Gênica , Doença de Parkinson Secundária/etiologia , Doença de Parkinson Secundária/metabolismo , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , RNA Mensageiro/metabolismo , Ratos , Receptores de AMPA/genética , Receptores de N-Metil-D-Aspartato/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The differential expression of cytochrome P4501A1 gene expression (CYP1A1) in liver, intestine and gill of juvenile tilapia following oral exposure to polychlorinated biphenyl (PCB) Aroclor-1254 or Aroclor-1260 spiked food were investigated. The fish was fed with 0.1 g/g of body weight each day of the spiked food with a concentration of approximately 8 ppm. Fish fed with betaNF- or acetone-spiked food were used as positive and negative controls, respectively. On day 1, 3 and 7 of post-treatment, the fish were sampled and analyzed for the total accumulated levels of PCBs, as well as the induced levels of CYP1A1 mRNA. Our results indicated that the fish fed with PCBs-spiked food showed a time-dependent bioaccumulation of PCBs. The time course studies also revealed that in the first 3 days of feeding, significant increases in CYP1A1 were found in intestine (betaNF: 320% and Aroclor-1260: 290%), followed by liver (betaNF: 144%, Aroclor-1260: 123% and Aroclor-1254: 110%). The transcript levels, then declined slowly. There was no induction of gill CYP1A1 mRNA expression in all treatment groups. Among all the tested tissues, the highest intestinal CYP1A1 transcripts induction has highlighted its prompt response to dietary PCBs exposure. Our study supported the important role of intestine in response to dietary PCBs exposure. In addition, in the present study the relative potency of Aroclor-1260 over Aroclor-1254 in stimulating CYP1A1 expression was discussed, indicating that the highly chlorinated PCBs had a higher potency of bioaccumulation and CYP1A1 mRNA induction.
Assuntos
Arocloros/farmacologia , Citocromo P-450 CYP1A1/biossíntese , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , RNA Mensageiro/biossíntese , Tilápia/metabolismo , Animais , Citocromo P-450 CYP1A1/genética , Alimentos , Brânquias/efeitos dos fármacos , Brânquias/enzimologia , Intestinos/efeitos dos fármacos , Intestinos/enzimologia , Fígado/efeitos dos fármacos , Fígado/enzimologia , Bifenilos Policlorados/análise , RNA Mensageiro/genética , Fatores de Tempo , Distribuição TecidualRESUMO
The present study was performed to investigate the patterns of gene expression of N-methyl-D-aspartate (NMDA) receptors (NRs) in the rat neostriatum during postnatal development. Reverse transcriptase-polymerase chain reactions (RT-PCR) indicated that levels of NR1, NR2A and NR2D mRNAs reached peak levels between postnatal days 7 (PND 7) and PND 14. The levels of NR2B and NR2C mRNAs were low at PND 1 and their levels increased at PND 7 and remained high in adults. Immunofluorescence combined with image analysis revealed that the levels of NR1 immunoreactivity rose to its maximum at PND 14. In contrast, NR1 immunoreactivity rose progressively in perikarya of striatal neurons. Levels of NR2A immunoreactivity in the neostriatum were highest in adults. However, levels of NR2A immunoreactivity were higher in striatal neurons at PND 1 and PND 7. Levels of NR2B immunoreactivity were highest at PND 7. In the perikarya of striatal neurons however, the highest levels of NR2B immunoreactivity were detected at PND 14 and adult striatal neurons. In addition, double immunofluorescence revealed that the levels of NR1 immunoreactivity increased but the levels of NR2A immunoreactivity were the same in parvalbumin (PV)-positive striatal interneurons of PND 14 and adult rats. NR2B immunoreactivity was not detected in PV-positive neurons of PND 14 rats, but intense NR2B labeling was seen in PV-positive neurons of adult rats. Last but not least, in choline acetyltransferase (ChAT)-positive striatal interneurons of PND 14 and adult rats, levels of NR1 and NR2A immunoreactivity was seen to increase. Level of NR2B immunoreactivity remained the same in ChAT-positive neurons of PND 14 and adult rats. The present results indicate that there are differential patterns of expression of NR mRNAs and immunoreactivity in the neostriatum during different stages of postnatal development. Different combinations of NR have been found in different subpopulations of striatal neurons at different developmental stages. NR1, NR2A and NR2B are the major NMDA receptor subunits expressed during development. The change in patterns of expression of NR may be related to the functional maturation of neurons in the neostriatum.