Premium
Oligonucleotide microarray analysis of apoptosis induced by 15‐methoxypinusolidic acid in microglial BV2 cells
Author(s) -
Choi Y,
Lim SY,
Jeong HS,
Koo KA,
Sung SH,
Kim YC
Publication year - 2009
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1111/j.1476-5381.2009.00247.x
Subject(s) - apoptosis , oligonucleotide , microglia , microarray analysis techniques , microbiology and biotechnology , chemistry , microarray , computational biology , biology , cancer research , biochemistry , immunology , gene , inflammation , gene expression
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 [ gadd45α , gadd45γ 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.