Qixuan Chen

Bisphenol A activates the Nrf1/2-antioxidant response element pathway in HEK 293 cells.

Bisphenol A (BPA) is used in the production of polycarbonate plastics and epoxy resins for baby bottles, liners of canned food, and many other consumer products. Previously, BPA has been shown to reduce the activity of several antioxidant enzymes, which may contribute to oxidative stress. However, the underlying mechanism of the BPA-mediated effect upon antioxidant enzyme activity is unknown. Antioxidant and phase II metabolizing enzymes protect cells from oxidative stress and are transcriptionally activated by Nrf1 and Nrf2 factors through their cis-regulatory antioxidant response elements (AREs). In this work, we have assessed the effect of BPA on the Nrf1/2-ARE pathway in cultured human embryonic kidney (HEK) 293 cells. Surprisingly, glutathione and reactive oxygen species (ROS) assays revealed that BPA application created a more reduced intracellular environment in cultured HEK 293 cells. Furthermore, BPA increased the transactivation activity of ectopic Nrf1 and Nrf2 and increased the expression of ARE-target genes ho-1 and nqo1 at high (100-200 μM) BPA concentrations only. Our study suggests that BPA activates the Nrf1/2-ARE pathway at high (>10 μM) micromolar concentrations.

Exposure to a Northern Contaminant Mixture (NCM) Alters Hepatic Energy and Lipid Metabolism Exacerbating Hepatic Steatosis in Obese JCR Rats

Non-alcoholic fatty liver disease (NAFLD), defined by the American Liver Society as the buildup of extra fat in liver cells that is not caused by alcohol, is the most common liver disease in North America. Obesity and type 2 diabetes are viewed as the major causes of NAFLD. Environmental contaminants have also been implicated in the development of NAFLD. Northern populations are exposed to a myriad of persistent organic pollutants including polychlorinated biphenyls, organochlorine pesticides, flame retardants, and toxic metals, while also affected by higher rates of obesity and alcohol abuse compared to the rest of Canada. In this study, we examined the impact of a mixture of 22 contaminants detected in Inuit blood on the development and progression of NAFLD in obese JCR rats with or without co-exposure to10% ethanol. Hepatosteatosis was found in obese rat liver, which was worsened by exposure to 10% ethanol. NCM treatment increased the number of macrovesicular lipid droplets, total lipid contents, portion of mono- and polyunsaturated fatty acids in the liver. This was complemented by an increase in hepatic total cholesterol and cholesterol ester levels which was associated with changes in the expression of genes and proteins involved in lipid metabolism and transport. In addition, NCM treatment increased cytochrome P450 2E1 protein expression and decreased ubiquinone pool, and mitochondrial ATP synthase subunit ATP5A and Complex IV activity. Despite the changes in mitochondrial physiology, hepatic ATP levels were maintained high in NCM-treated versus control rats. This was due to a decrease in ATP utilization and an increase in creatine kinase activity. Collectively, our results suggest that NCM treatment decreases hepatic cholesterol export, possibly also increases cholesterol uptake from circulation, and promotes lipid accumulation and alters ATP homeostasis which exacerbates the existing hepatic steatosis in genetically obese JCR rats with or without co-exposure to ethanol.

Dietary phytosterols and phytostanols decrease cholesterol levels but increase blood pressure in WKY inbred rats in the absence of salt-loading

BackgroundThere are safety concerns regarding widespread consumption of phytosterol and phytostanol supplemented food products. The aim of this study was to determine, in the absence of excess dietary salt, the individual effects of excess accumulation of dietary phytosterols and phytostanols on blood pressure in Wistar Kyoto (WKY) inbred rats that have a mutation in the Abcg5 gene and thus over absorb phytosterols and phytostanols.MethodsThirty 35-day old male WKY inbred rats (10/group) were fed a control diet or a diet containing phytosterols or phytostanols (2.0 g/kg diet) for 5 weeks. The sterol composition of the diets, plasma and tissues were analysed by gas chromatography. Blood pressure was measured by the tail cuff method. mRNA levels of several renal blood pressure regulatory genes were measured by real-time quantitative PCR.ResultsCompared to the control diet, the phytosterol diet resulted in 3- to 4-fold increases in the levels of phytosterols in plasma, red blood cells, liver, aorta and kidney of WKY inbred rats (P < 0.05). The phytostanol diet dramatically increased (> 9-fold) the levels of phytostanols in plasma, red blood cells, liver, aorta and kidney of these rats (P < 0.05). The phytosterol diet decreased cholesterol levels by 40%, 31%, and 19% in liver, aorta and kidney, respectively (P < 0.05). The phytostanol diet decreased cholesterol levels by 15%, 16%, 20% and 14% in plasma, liver, aorta and kidney, respectively (P < 0.05). The phytostanol diet also decreased phytosterol levels by 29% to 54% in plasma and tissues (P < 0.05). Both the phytosterol and phytostanol diets produced significant decreases in the ratios of cholesterol to phytosterols and phytostanols in plasma, red blood cells, liver, aorta and kidney. Rats that consumed the phytosterol or phytostanol diets displayed significant increases in systolic and diastolic blood pressure compared to rats that consumed the control diet (P < 0.05). The phytosterol diet increased renal angiotensinogen mRNA levels of these rats.ConclusionThese data suggest that excessive accumulation of dietary phytosterols and phytostanols in plasma and tissues may contribute to the increased blood pressure in WKY inbred rats in the absence of excess dietary salt. Therefore, even though phytosterols and phytostanols lower cholesterol levels, prospective clinical studies testing the net beneficial effects of dietary phytosterols and phytostanols on cardiovascular events for subgroups of individuals that have an increased incorporation of these substances are needed.

Dietary Fructooligosaccharides and Wheat Bran Elicit Specific and Dose-Dependent Gene Expression Profiles in the Proximal Colon Epithelia of Healthy Fischer 344 Rats

Proximal colon epithelial gene responses to diets containing increasing levels of dietary fermentable material (FM) from 2 different sources were measured to determine whether gene expression patterns were independent of the source of FM. Male Fischer 344 rats (10/group) were fed for 6 wk a control diet containing 10% (g/g) cellulose (0% FM); or a 2, 5, or 10% wheat bran (WB) diet (1, 2, 5% FM); or a 2, 5, or 8% fructooligosaccharides (FOS) diet (2, 5, 8% FM). WB and FOS were substituted for cellulose to give a final 10% nondigestible material content including FM. Gene responses were relative to expression in rats fed the control diet. The gene response patterns associated with feeding ∼2% FM (5% WB and 2% FOS) were similar (∼10 gene changes ≥ 1.6-fold; P ≤ 0.01) and involved genes associated with transport (Scnn1g, Mt1a), transcription (Zbtb16, Egr1), immunity (Fkbp5), a gut hormone (Retn1β), and lipid metabolism (Scd2, Insig1). These changes were also similar to those associated with 5% FM but only in rats fed the 10% WB diet. In contrast, the 5% FOS diet (~5% FM) was associated with 68 gene expression changes ≥ 1.6-fold (P ≤ 0.01). The diet with the highest level of fermentation (8% FOS, ~8% FM) was associated with 132 changes ≥ 1.6-fold (P ≤ 0.01), including genes associated with transport, cellular proliferation, oncogene and tumor metastasis, the cell cycle, apoptosis, signal transduction, transcript regulation, immunity, gut hormones, and lipid metabolic processes. These results show that both the amount and source of FM determine proximal colon epithelial gene response patterns in rats.

A Northern contaminant mixture impairs pancreas function in obese and lean JCR rats and inhibits insulin secretion in MIN6 cells.

Rates of obesity and diabetes mellitus of Arctic populations are increasing due to multiple reasons including a departure from traditional lifestyles and alcohol consumption patterns. These populations are also exposed to a variety of anthropogenic contaminants through consumption of contaminated country foods. We have previously shown that a Northern contaminant mixture (NCM), containing 22 organic and inorganic contaminants found in the blood of Canadian Arctic populations, induces endothelial cell dysfunction and exacerbates development of non-alcoholic fatty liver disease in experimental models. In order to determine if these contaminants affect pancreas function and physiology and if obesity and alcohol can influence contaminant toxicity and the development of diabetes, lean and obese JCR rats were orally treated with NCM at 0 (vehicle), 1.6 or 16mg/kg BW for four weeks in the presence or absence of 10% (v/v) alcohol. NCM treatment altered islet morphology, increased iron deposit in pancreas, and reduced circulating and pancreatic insulin levels and circulating glucagon levels as a result of direct islet injury with β and α cell loss with or without exposure to alcohol. Studies conducted with cultured mouse insulin-secreting (MIN6) β cells further demonstrated that NCM inhibited insulin release and induced cell death through oxidative stress and mitochondrial dysfunction. 2,3,4,6-Tetrabromophenol, a minor component of the NCM, alone also inhibited insulin release from MIN6 cells after 10min of exposure. These results suggest that Northern contaminants may contribute to pancreatic dysfunction, and possibly development of diabetes, in some of the highly exposed Arctic populations. The implications and relevance of these findings to Northern populations remains to be confirmed through epidemiological studies.

Bisphenol A exposure alters release of immune and developmental modulators and expression of estrogen receptors in human fetal lung fibroblasts

Bisphenol A (BPA) has been shown to exert biological effects through estrogen receptor (ER)-dependent and ER-independent mechanisms. Recent studies suggest that prenatal exposure to BPA may increase the risk of childhood asthma. To investigate the underlying mechanisms in the actions of BPA, human fetal lung fibroblasts (hFLFs) were exposed to varying doses of BPA in culture for 24hr. Effects of BPA on localization and uptake of BPA, cell viability, release of immune and developmental modulators, cellular localization and expression of ERα, ERβ and G-protein coupled estrogen receptor 30 (GPR30), and effects of ERs antagonists on BPA-induced changes in endothelin-1 (ET-1) release were examined. BPA at 0.01-100μmol/L caused no changes in cell viability after 24hr of exposure. hFLFs expresses all three ERs. BPA had no effects on either cellular distribution or protein expression of ERα, however, at 100μmol/L (or 23μmol/L intracellular BPA) increased ERβ protein levels in the cytoplasmic fractions and GPR30 protein levels in the nuclear fractions. These paralleled with increased release of growth differentiation factor-15, decreased phosphorylation of nuclear factor kappa B p65 at serine 536, and decreased release of ET-1, interleukin-6, and interferon gamma-induced protein 10. ERs antagonists had no effects on BPA-induced decrease in ET-1 release. These data suggest that BPA at 100μmol/L altered the release of immune and developmental modulators in hFLFs, which may negatively influence fetal lung development, maturation, and susceptibility to environmental stressors, although the role of BPA in childhood asthma remains to be confirmed in in vivo studies.

Excess dietary iodine differentially affects thyroid gene expression in diabetes, thyroiditis-prone versus -resistant BioBreeding (BB) rats

SCOPE To identify genes involved in the susceptibility to iodine-induced autoimmune thyroiditis. METHODS AND RESULTS Diabetes, thyroiditis-prone (BBdp) and -resistant (BBc) rats were fed either a control or a high-iodine diet for 9 wk. Excess iodine intake increased the incidence of insulitis and thyroiditis in BBdp rats. BBdp rats fed the high-iodine diet that did not develop thyroiditis had higher mRNA levels of Fabp4, Cidec, perilipin, Pparγ and Slc36a2 than BBdp rats fed the control diet and BBc rats fed either the control or the high-iodine diet. BBdp rats fed the high-iodine diet that did develop thyroiditis had higher mRNA levels of Cidec, Icam1, Ifitm1, and Slpi than BBdp rats fed the control diet and BBc rats fed either the control or the high-iodine diet. BBdp rats that did develop thyroiditis had lower mRNA levels of Fabp4, perilipin and Slc36a2 but higher mRNA levels of Icam1, Ifitm1 and Slpi than BBdp that did not develop thyroiditis. Excess dietary iodine also increased the protein levels of Fabp4, Cidec and perilipin in BBdp rats. CONCLUSION Differential expression of thyroid genes in BBdp versus BBc rats caused by excess dietary iodine may be implicated in autoimmune thyroiditis and insulitis pathogenesis.

Acknowledgement to the 2009 Reviewers

Helmut Heseker, Paderborn, Germany Manfred Hüttinger, Vienna, Austria Mohsen Janghorbani, Isfahan, Iran Miroslaw Jarosz, Warsaw, Poland Majken K. Jensen, Boston, Mass., USA Alex Johnstone, Aberdeen, UK Anthony Kafatos, Heraklion, Greece Andre Pascal Kengne, Camperdown, N.S.W., Australia Arthur Klatsky, Oakland, Calif., USA Nanne Kleefstra, Zwolle, The Netherlands Marlena Kruger, Palmerston North, New Zealand Anura Kurpad, Bangalore, India Denis Lairon, Marseille, France Wolfgang Langhans, Zürich, Switzerland Mark Lawrence, Melbourne, Vic., Australia Donald K. Layman, Urbana, Ill., USA James V. Leonard, London, UK Lars Libuda, Dortmund, Germany Xu Lin, Shanghai, China Jakob Linseisen, Munich, Germany Dieter Luetjohann, Bonn, Germany Silvia Maggini, Basel, Switzerland Philippe Marmillot, Washington, D.C., USA Franscesco Marotta, Milano, Italy Eva Martos, Budapest, Hungary Velimir Matkovic, Columbus, Ohio, USA Regis Moreau, Corvallis, Oreg., USA Luis A. Moreno, Zaragoza, Spain Yuji Naito, Kyoto, Japan Serge Nef, Geneva, Switzerland Hee Young Paik, Seoul, Korea Mihalis I. Panagiotidis, Reno, Nev., USA Sun Ming Park, Asan-si, Korea Kristina Pentieva, Ulster, UK Carmen Perez-Rodrigo, Bilbao, Spain Nancy M. Petry, Farmington, Conn., USA John M. Pettifor, Johannesburg, South Africa Spencer Proctor, Edmonton, Alta., Canada Ana Maria Proenza Arenas, Palma de Mallorca, Spain Giuliano Ramadori, Göttingen, Germany Aune Rehema, Tartu, Estonia Lajos Rethy, Budapest, Hungary Gerald Rimbach, Kiel, Germany Glorimar Rosa, Rio de Janeiro, Brazil Catherine A. Ross, University Park, Pa., USA Elaine Rush, Auckland, NZ Lydia Afman, Wageningen, The Netherlands Tasnime Akbaraly, London, UK Anthony Alberg, Baltimore, Md., USA Hooman Allayee, Los Angeles, Calif., USA James W. Anderson, Lexington, Ky., USA Bahram Arjmandi, Tallahassee, Fla., USA Georgianne L. Arnold, Rochester, N.Y., USA Benoit Arsenault, Québec, Qué., Canada Fereidoun Azizi, Tehran, Iran Matthias Baum, Kaiserslautern, Germany Giorgio Bedogni, Milano, Italy Joe Begley, Poole, UK Aloys Berg, Freiburg, Germany Renate Bergmann, Berlin, Germany Odilia I. Bermudez, Boston, Mass., USA Francisco Blanco-Vaca, Barcelona, Spain William S. Blaner, New York, N.Y., USA Pontus Boström, Göteborg, Sweden Regina Brigelius-Flohé, Nuthetal, Germany Veronika Butterweck, Gainesville, Fla., USA Anette Buyken, Dortmund, Germany Philip C. Calder, Southampton, UK Dexter Canoy, Manchester, UK Namsoo Chang, Seoul, Korea Supranee Changbumrung, Bangkok, Thailand Marie-Aline Charles, Villejuif, France Armand B. Christophe, Ghent, Belgium Nain-Feng Chu, Taipei, Taiwan Dolores Corella, Valencia, Spain Lindsey Darrow, Atlanta, Ga., USA Tamás Decsi, Pécs, Hungary Antonio Di Stefano, Chieti, Italy Marie-Claude Dop, Rome, Italy Klaus Eder, Freising, Germany Mathias Fasshauer, Leipzig, Germany Annie Ferland, Québec, Qué., Canada Jose Fernandez, Birmingham, Ala., USA Nicola Fuiano, Foggia, Italy Claudio Galli, Milano, Italy Dieter Genser, Vienna, Austria Frank Greer, Madison, Wisc., USA M.H. Hamdaoui, Tunis, Tunisia Igor Harsch, Erlangen, Germany Hans Hauner, Munich, Germany Alan S. Hazell, Montréal, Qué., Canada Erik Hemmingsson, Stockholm, Sweden