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Accumulation of triacylglycerol in cultured rat hepatocytes is increased by ethanol and by insulin and dexamethasone.

ABSTRACT: Isolated hepatocytes from female rats were cultured in HI-WO/BA medium for 6 days. To the medium was added oleate, ethanol, dexamethasone and insulin. With oleate To alone, triacylglycerol accumulated; ethanol augmented the accumulation by 90%. To the best of our knowledge, this is the first demonstration that ethanol in vitro increases the content of triacylglycerol in liver cells. Further addition of dexamethasone or insulin did not alter the accumulation of triacylglycerol, indicating that these hormones did not play permissive roles for the effect of ethanol in the present system. Dexamethasone and insulin, in the absence of ethanol, increased the accumulation of triacylglycerol by 30% and 50% respectively. The concentration of glycerol 3-phosphate was increased in the presence of ethanol; however, with time the concentration of glycerol 3-phosphate declined almost to control values, while the accumulation of triacylglycerol continued linearly; this suggests that the effect of ethanol was not mediated via fluctuations in the concentration of glycerol 3-phosphate. These results are discussed in relation to earlier investigations in vivo and in vitro.

SUBMITTER: Dich J

PROVIDER: S-EPMC1153135 | biostudies-other | 1983 Jun

REPOSITORIES: biostudies-other

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Project description:DNA microarray is a powerful tool in biomedical research. However, transcriptomic profiling using DNA microarray is subject to many variations including biological variability. To evaluate the different sources of variation in mRNA gene expression profiles, gene expression profiles were monitored using the Affymetrix RatTox U34 arrays in cultured primary hepatocytes derived from six rats over a 26 hour period at 6 time points (0 h, 2h, 5h, 8h, 14 h and 26 h) with two replicate arrays at each time point for each animal. In addition, the impact of sample size on the variability of differentially expressed gene lists and the consistency of biological responses were also investigated. Excellent intra-animal reproducibility was obtained at all time points with 0 out of 370 present probe sets across all time points showing significant difference between the 2 replicate arrays (3-way ANOVA, p <or= 0.0001). However, large inter-animal biological variation in mRNA expression profiles was observed with 337 out of 370 present probe sets showing significant differences among 6 animals (3-way ANOVA, p <or= 0.05). Principal Component Analysis (PCA) revealed that time effect (PC1) in this data set accounted for 47.4% of total variance indicating the dynamics of transcriptomics. The second and third largest effects came from animal difference, which accounted for 16.9% (PC2 and PC3) of the total variance. The reproducibility of gene lists and their functional classification was declined considerably when the sample size was decreased. Overall, our results strongly support that there is significant inter-animal variability in the time-course gene expression profiles, which is a confounding factor that must be carefully evaluated to correctly interpret microarray gene expression studies. The consistency of the gene lists and their biological functional classification are also sensitive to sample size with the reproducibility decreasing considerably under small sample size.

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Accumulation of triacylglycerol in cultured rat hepatocytes is increased by ethanol and by insulin and dexamethasone.

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