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PMID:11578593

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Citation

Pang, Y, Cai, Z and Rhodes, PG (2001) Analysis of genes differentially expressed in astrocytes stimulated with lipopolysaccharide using cDNA arrays. Brain Res. 914:15-22

Abstract

We used cDNA arrays to investigate differentially expressed genes in astrocytes challenged with lipopolysaccharide (LPS). Astrocyte cultures were prepared from 1-day-old rat brains. Purified astrocytes were treated with LPS (1 microg/ml) for 2, 8 and 48 h. Differentially expressed genes in these astrocytes were examined with Atlas rat cDNA arrays. At all the three time points studied, three genes were found consistently up-regulated: I-kappaB alpha chain, NF-kappaB, and interferon induced protein. In addition to these three, six other genes were also up-regulated at 2 and 8 h. They were genes encoding vascular cell adhesion protein 1 (VCAM-1), interferon regulatory factor 1 (IRF-1), mitochondrial hydroxymethylglutaryl-CoA synthase (HMG-CoA synthase), aldehyde dehydrogenase 2, macrophage inflammatory protein 1 (MIP-1) and neurotensin receptor 2. At these two time points, three genes were down-regulated: copper-zinc-containing superoxide dismutase 1 (SOD-1), insulin-like growth factor binding protein 1 (IGFBP-1), and insulin-like growth factor binding protein 3 (IGFBP-3). Expression of several differentially expressed genes in cDNA array (I-kappaB, VCAM-1 and MIP-3) were further confirmed by reverse transcription polymerase chain reaction study. The prominently modulated genes could be classified into three categories: nuclear transcription factors, pro-inflammatory cytokines/chemokines and metabolic enzymes. Application of pyrrolidine dithiocarbamate, an inhibitor of nuclear factor-kB (NF-kappaB), prior to LPS stimulation not only prevented up-regulation of NF-kappaB gene expression, but also completely blocked up-regulation of pro-inflammatory cytokine genes (TNF-alpha and interleukin-1beta) and two chemokine genes: CXC chemokine LIX and CC chemokine MIP-3 alpha. These results indicate that both up-regulation of inflammatory cytokine expression and down-regulation of growth factor expression are probably involved in the response of astrocytes upon exposure to LPS.

Links

PubMed

Keywords

Animals; Animals, Newborn; Astrocytes/drug effects; Astrocytes/immunology; Astrocytes/metabolism; Bacterial Infections/complications; Bacterial Infections/immunology; Bacterial Infections/metabolism; Cells, Cultured/cytology; Cells, Cultured/drug effects; Cells, Cultured/metabolism; Cytokines/genetics; Cytokines/immunology; Cytokines/metabolism; Down-Regulation/genetics; Encephalitis/genetics; Encephalitis/immunology; Encephalitis/metabolism; Gene Expression Regulation/immunology; Growth Substances/genetics; Growth Substances/immunology; Growth Substances/metabolism; Lipopolysaccharides/immunology; Lipopolysaccharides/metabolism; Lipopolysaccharides/pharmacology; Oligonucleotide Array Sequence Analysis; Rats; Rats, Sprague-Dawley; Transcription Factors/genetics; Transcription Factors/immunology; Transcription Factors/metabolism; Up-Regulation/genetics

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

RAT:HMCS2

involved_in

GO:0071222: cellular response to lipopolysaccharide

ECO:0000270: expression pattern evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:ALDH2

involved_in

GO:0032496: response to lipopolysaccharide

ECO:0000270: expression pattern evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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