Project description:The study was performed using primary rat hepatocyte in culture from 4 adult male Sprague-Dawley rats to investigate the changes in gene expression under low dose (4μM) and short exposure (3hrs) of cadmium chloride. By comparing the gene expression profiles of control and cadmium-treated cells, the most dramatic and significant changes were for those genes associated with transcriptional regulation, antioxidant response and control of protein integrity. Changes in other genes involved in cellular physiological responses such as inflammation, growth and apoptosis were also observed. Results were further confirmed by quantitative real time polymerase chain reaction (qRT-PCR). Keywords: early protective responses
Project description:The study was performed using primary rat hepatocyte in culture from 4 adult male Sprague-Dawley rats to investigate the changes in gene expression under low dose (4μM) and short exposure (3hrs) of cadmium chloride. By comparing the gene expression profiles of control and cadmium-treated cells, the most dramatic and significant changes were for those genes associated with transcriptional regulation, antioxidant response and control of protein integrity. Changes in other genes involved in cellular physiological responses such as inflammation, growth and apoptosis were also observed. Results were further confirmed by quantitative real time polymerase chain reaction (qRT-PCR). Experiment Overall Design: Primary hepatocytes from four rats were isolated using the collagenase-hyaluronidase perfusion and digestion method. The cells of greater than 85% viability were used in this study. The cells were plated at 90% confluency on culture dishes coated with collagen and maintained in Waymouth's medium containing BSA for 4hrs prior to an overnight incubation with BSA free Waymouth's medium. Control samples and cadmium-treated samples from individual rats were hybridized to RAE 230A arrays (n=4 and 8 microarrays); data were generated and analyzed.
Project description:The experiment utilized a total of 54 five-week-old Sprague-Dawley rats, each weighing approximately 100 grams. These rats were randomly divided into 9 groups, consisting of 27 female and 27 male rats. Based on the intervention dose of cadmium (Cd), the rats were further randomly assigned into 3 main groups, each containing 18 rats: the control group received 1 milliliter of sodium chloride solution daily; the low-dose group received 1 milliliter of cadmium chloride (CdCl₂) solution at a concentration of 30 micrograms per kilogram (μg/kg) daily; and the high-dose group received 1 milliliter of cadmium chloride (CdCl₂) solution at a concentration of 1 milligram per kilogram (mg/kg) daily. Cadmium exposure was administered via oral gavage. Each dose group (control group, low-dose group, and high-dose group) was intervened at three different time points: 2 weeks, 4 weeks, and 8 weeks. At each intervention time point, 6 rats were randomly selected from each dose group for euthanasia, and their prefrontal brain tissues were collected for subsequent analysis. During the experiment, some rats died: 2 rats in the control group died at 2 weeks, and 1 rat in the high-dose group died at 4 weeks and 8 weeks, respectively. The experimental design ensured a systematic evaluation of the effects of cadmium exposure on rats under different doses and time points, while recording the mortality during the experiment for further analysis.
Project description:Gene expression profiling was performed in each of hepatocyte fraction and non-parenchymal cell fraction enriched with activated hepatic stellate cells/myofibroblasts from cirrhotic rat livers induced by repeated, low-dose diethylnitrosamine (DEN) treatment.
Project description:The molecular basis of TNF tolerance is poorly understood. In this experiment, primary human monocytes were used to examine TNF refractoriness. We detect absolute tolerance as selective, dose-dependently affecting a small group of powerful effector molecules; induction tolerance represented a more general phenomenon. Primary human monocytes were incubated for 48 h with Medium or different TNF-doses and subsequently short-term stimulated for two hours with TNF. Total RNA was prepared and analysed by use of Agilent microarrays. 6 different conditions were analyzed in total, corresponding to cells that were: 1) medium incubated (M+M), 2) medium incubated and short time TNF (400 U/ml) incubated (M+T), 3) low dose TNF (40 U/ml) preincubated (40+M), 4) low dose TNF preincubated and short time TNF incubated (40+T), 5) high dose TNF (400 U/ml) preincubated (400+M), 6) high dose TNF preincubated and short time TNF incubated (400+T). 3 biological replicates (corresponding to three different healthy donors) were analyzed regarding high-dose tolerance conditions (preincubation with 400 U TNF/ml), one of which was analyzed in duplicates (including cell culture, RNA preparation and a dye-swap microarray approach). 2 biological replicates (corresponding to two different healthy donors) were analyzed under low-dose tolerance conditions (preincubation with 40 U/ml). The different human donors were numbered and indicated in brackets (d1-d5).
Project description:This is a study to explore the transcriptional changes after cadmium treatment in adult rat testes at three time points (control--0 hour, 8 hour and 4 day). Cadmium is an environmental toxicant that is known to affect the male reproductive system. It disrupts the blood-testis barrier irreversibly, and affects the Sertoli-germ cells adhesion, causing germ cell depletion from the seminiferous epithelium. Keywords: Cadmium effect in rat testes
Project description:Environmental cadmium, with a high average dietary intake, is a severe public health risk. However, the long-term health implications of environmental exposure to cadmium in different life stages remain unclear. We used microarrays to explore the effects of early exposure to low-dose cadmium on hepatic gene expression.