Oligonucleotide microarray analysis of apoptosis induced by 15-methoxypinusolidic acid in microglial BV2 cells.

Abstract

BACKGROUND AND PURPOSE:

We conducted a genome wide gene expression analysis to explore the biological aspects of 15-methoxypinusolidic acid (15-MPA) isolated from Biota orientalis and tried to confirm the suitability of 15-MPA as a therapeutic candidate for CNS injuries focusing on microglia.

EXPERIMENTAL APPROACH:

Murine microglial BV2 cells were treated with 15-MPA, and their transcriptome was analysed by using oligonucleotide microarrays. Genes differentially expressed upon 15-MPA treatment were selected for RT-PCR (reverse transcription-polymerase chain reaction) analysis to confirm the gene expression. Inhibition of cell proliferation and induction of apoptosis by 15-MPA were examined by bromodeoxyuridine assay, Western blot analysis of poly-ADP-ribose polymerase and flow cytometry.

KEY RESULTS:

A total of 514 genes were differentially expressed by 15-MPA treatment. Biological pathway analysis revealed that 15-MPA induced significant changes in expression of genes in the cell cycle pathway. Genes involved in growth arrest and DNA damage [gadd45alpha, gadd45gamma and ddit3 (DNA damage-inducible transcript 3)] and cyclin-dependent kinase inhibitor (cdkn2b) were up-regulated, whereas genes involved in cell cycle progression (ccnd1, ccnd3 and ccne1), DNA replication (mcm4, orc1l and cdc6) and cell proliferation (fos and jun) were down-regulated. RT-PCR analysis for representative genes confirmed the expression levels. 15-MPA significantly reduced bromodeoxyuridine incorporation, increased poly-ADP-ribose polymerase cleavage and the number of apoptotic cells, indicating that 15-MPA induces apoptosis in BV2 cells.

CONCLUSION AND IMPLICATIONS:

15-MPA induced apoptosis in murine microglial cells, presumably via inhibition of the cell cycle progression. As microglial activation is detrimental in CNS injuries, these data suggest a strong therapeutic potential of 15-MPA.

• Authors: Y Choi, S Y Lim, H S Jeong, K A Koo, S H Sung, Y C Kim
• Journal: British journal of pharmacology (Br J Pharmacol) Vol. 157 Issue 6 Pg. 1053-64 (Jul 2009) ISSN: 1476-5381 [Electronic] England
• PMID: 19466985 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 15-methoxypinusolidic acid
• Diterpenes
• Plant Extracts

Topics

• Animals
• Apoptosis (drug effects, physiology)
• Cell Line
• Diterpenes (isolation & purification, toxicity)
• Mice
• Microglia (drug effects, pathology)
• Oligonucleotide Array Sequence Analysis (methods)
• Plant Extracts (isolation & purification, toxicity)
• Thuja