Measurement of synergistic effects of a phytopharmaceutical by microarray-analysis
Gudrun Sigrid Ulrich-Merzenich, D. Jobst, H.-J. Zeitler, J Müller, Hang Vetter · Planta Medica · 2007
Phytopharmaceuticals are complex compounds. Microarray-based gene/protein expression analyses have been proposed to be capable of relating complex mixtures to complex effects, thereby revealing synergistic effects [1,2]. Phytodolor (PD) is a standardized alcoholic plant extract of Populus tremulus (S1), Solidago virgaurea (S2) and Fraxinus excelsior (S3), indicated in rheumatic pain. With the aim of revealing molecular mechanisms of PD action, we examined the effect of PD, its single extracts and acetylsalicylic acid (ASS) on gene-expression profiles in human fibroblasts. Fibroblasts were synchronized for 72 hrs and stimulated with PD (0.05 and 0.1%), S1, S2, S3 (0.1%), ASS (30µg/ml) and LPS (10µg/ml) for 6 hrs. RNA-expressions were analysed by Piquor® Skin Patho Microarray (1126 genes) and data analyzed by Piquor Analyser and microarray ImaGenesoftware. Values above 1.7- or below 0.58-fold compared to controls were judged as up- or downregulation. The application of 0.05% PD revealed a downregulation of only two genes, whereas 0.1% PD modulated additionally 36 further genes. Identified genes (eg. PGH2, TNC, LPS stimulated: MMP-1, IL-5,-6,-8, Gro-1) are involved in immunoregulation, inflammation and apoptosis. S1, S2, S3 and ASS modulated 51, 24, 31 and 44 genes respectively. Regulated genes of PD and its single extract show taken together an overlap of 57.9%. S1 has the maximum individual overlap with PD (36.5%) and greater overlap with ASS (52.9%). Initial protein expression analyses (PGH2, IL-5, -6, -8, Gro-1) support gene expression data. Microarray analyses will develop our understanding of the functional mechanisms of complex mixtures like PD and synergism. References: [1] Ulrich-Merzenich G. et al. (2007) Phytomedicine 14: 70–82. [2] Ulrich-Merzenich G, Jobst D. (2007) Genchipanalysen an humanen Fibroblasten unter Einwirkung definierter Rindenextrakte. Vortrag: Forum univ. Arbeitsgruppen f. Naturheilverfahren u. Komplementärmedizin, München.