Microarray analysis of differentially gene expression profile in LPS-stimulated primary Kupffer cells / 中国免疫学杂志

ABSTRACT

Objective:To investigate the changes of gene expression profile in lipopolysaccharide (LPS)-stimulated primary Kupffer cells ( KCs ) . Methods: Rat KCs were isolated and purified by means of in situ perfusion and density gradient centrifugation. After being identified by ink phagocytosis and ED2 staining test,KCs were stimulated with LPS. Gene expression profile were studied using gene microarrays,and the most significant upregulated gene was verified using real-time PCR. Results:27 genes were upregulated including Ces1f, Slc17a3, Slc21a4, Hsd17b2, Sorbs2, Ccdc116, Mgam, Myo5b, Etl4, Fabp1, Kif4b, Fosl1, Cyp4a1, Penk, Tmem221,Rpl5,Nr2f1,Hoxb1,Gpr165,Fam90a13p,Kpna6,Irak1bp1,Kcnh1 and 4 unnamed genes and 4 downregulated including Oc90,Tagln,Arxes2 and Olr830 in LPS-stimulated KCs. Among the upregulated genes, Ces1f was the most significant upregulatory gene. Real-time PCR confirmed that the levels of Ces1f were 23. 88 times higher in LPS-stimulated than control cells. Conclusion:There is a significant difference between LPS-stimulated and normal control cells in gene expression profile by microarray analysis,and Ces1f is the most significantly upregulated gene.