2% and 18. 9%, respectively. In the delayed down group, only 3. 4%, 3. 4%, and 24. 1% of the genes affected in the BF S L Ep, shikonin, and emodin treatments, respectively, displayed CHIR99021 IC50 the same regulation mode as cytopiloyne. We were curious about the detailed mechanism responsible for the similarity in the effects of BF S L Ep and cytopiloyne. For this purpose, we compared the expression profiles of the genes sharing common regula tion modes in both BF S L Ep and cytopiloyne treat ment. Genes that displayed early down regulation modes were subsequently classified into 2 types of expression profiles accord ing to the initial response of the gene in comparison with LPS treatment in THP 1 cells alone. Strikingly, all genes in the up regulation sub group showed sustained up regulation after both cytopiloyne and BF S L Ep treatments.

Genes that were down regulated by LPS only treatment produced the typical up regulation pattern at 4 h with the Asteraceae preparations. The same scenario was observed in analysis of those genes displaying early non response mode. Signaling molecules and associated pathways that may be involved in the modulation of LPS induced inflammatory response To find out the possible signaling pathways involved in the different gene expression patterns observed in cyto piloyne, BF S L Ep, shikonin and emodin treatments, we analyzed the microarray data using the TRANS PATH database. First, we analyzed those genes whose expression was up or down regulated more than threefold after 2, 4, and 12 h of LPS only treatment to verify the processing steps in the TRANSPATH data base.

Three key molecules and signaling pathways were observed, CKII, JNK JIP and p300 were the target molecules at 2, 4 and 12 h time points respectively. In this light, we reasoned that a possible target for Asteraceae preparations could be a common signaling molecule upstream of the genes shar ing same expression modes. We then subjected selected genes from two groups to key node ana lysis, which identified the ERK1 2 pathway as a single common denominator at no more than 4 steps of hier archical gene regulation at 4 h. The shikonin and emodin affected genes were ana lyzed by the same method, and a specific molecule and signaling pathway was observed for each treatment. The possible master regulator in the treatment with shikonin plus LPS at 0.

5 h was identified by a signaling database search as Rad23A. The possible master regu lator Dacomitinib in the emodin plus LPS treatment at 0. 5 h was the ubiquitin protein ligase E3A. For treatment with cytopiloyne or BF S L Ep, there was little or no significant change in gene expression at the early stage. However, among all four phytocompounds tested, only the BF S L Ep treatment showed a significant inhi bition of LPS stimulated gene expression increase in our focused array at 12 h. Key node analysis of genes with significant down regulation pointed to E6 AP as a possible master regulator.