Genomics and proteomics analysis of cultured primary rat hepatocytes
| Autor: | Peter-Juergen Kramer, Michaela Kroeger, Philip Hewitt, Juergen Beigel, Kerstin Fella |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Proteomics
Time Factors Cell Culture Techniques Biology Animal Testing Alternatives Toxicology Gene Expression Regulation Enzymologic Superoxide dismutase chemistry.chemical_compound Gene expression Toxicity Tests Animals Humans Electrophoresis Gel Two-Dimensional Cells Cultured Oligonucleotide Array Sequence Analysis chemistry.chemical_classification Glutathione peroxidase Structural gene General Medicine Glutathione Genomics Rats Biochemistry chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Toxicity biology.protein Hepatocytes Hepatocyte dedifferentiation |
| Zdroj: | Toxicology in vitro : an international journal published in association with BIBRA. 22(1) |
| ISSN: | 0887-2333 |
| Popis: | The use of animal models in pharmaceutical research is a costly and sometimes misleading method of generating toxicity data and hence predicting human safety. Therefore, in vitro test systems, such as primary rat hepatocytes, and the developing genomics and proteomics technologies, are playing an increasingly important role in toxicological research. Gene and protein expression analysis were investigated in a time series (up to 5 days) of primary rat hepatocytes cultured on collagen coated dishes. Especially after 24 h, a significant down-regulation of many important Phase I and Phase II enzymes (e.g., cytochrome P450’s, glutathione- S -transferases, sulfotransferases) involved in xenobiotic metabolism, and antioxidative enzymes (e.g., catalase, superoxide dismutase, glutathione peroxidase) was observed. Acute-phase-response enzymes were frequently up-regulated (e.g., LPS binding protein, α-2-macro-globulin, ferritin, serine proteinase inhibitor B, haptoglobin), which is likely to be a result of cellular stress caused by the cell isolation procedure (perfusion) itself. A parallel observation was the increased expression of several structural genes (e.g., β-actin, α-tubulin, vimentin), possibly caused by other proliferating cell types in the culture, such as fibroblasts or alternatively by hepatocyte dedifferentiation. In conclusion, the careful interpretation of data derived from this in vitro system indicates that primary hepatocytes can be successfully used for short-term toxicity studies up to 24 h. However, culturing conditions need to be further optimized to reduce the massive changes of gene and protein expression of long-term cultured hepatocytes to allow practical applications as a long-term toxicity test system. |
| Databáze: | OpenAIRE |
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