Phytomedicine 19 (2012) 930–939 Contents lists available at SciVerse ScienceDirect Phytomedicine jou rn al hom epage: www.elsevier.de/phymed Novel neurological and immunological targets for salicylate-based phytopharmaceuticals and for the anti-depressant imipramine G. Ulrich-Merzenich a,* , O. Kelber b , A. Koptina a,h , A. Freischmidt c , J. Heilmann c , J. Müller b , H. Zeitler d , M.F. Seidel e , M. Ludwig f , E.U. Heinrich b , H. Winterhoff g,1 a Medizinische Poliklinik, Universitätsklinikum, Rheinische Friedrich-Wilhelms-Universität Bonn, Germany b Steigerwald Arzneimittelwerk GmbH, Darmstadt, Germany c Pharmazeutische Biologie, Universität Regensburg, Germany d Medizinische Klinik I, Universitätsklinikum, Rheinische Friedrich-Wilhelms-Universität Bonn, Germany e Medizinische Klinik I, Rheumatology Unit, Universitätsklinikum, Rheinische Friedrich-Wilhelms-Universität Bonn, Germany f Department of Clinical Chemistry and Clinical Pharmacology, Universitätsklinikum, Rheinische Friedrich-Wilhelms Universität Bonn, Germany g Institut für Pharmakologie und Toxikologie, Westfälische Wilhelms-Universität, Münster, Germany h Mari State Technical University, Yoshkar Ola, Russia a r t i c l e i n f o Keywords: Inflammation Neurological diseases Multitargeting Microarray Polyphenols Salicylates a b s t r a c t Inflammatory processes are increasingly recognised to contribute to neurological and neuropsychatric disorders such as depression. Thus we investigated whether a standardized willow bark preparation (WB) which contains among other constituents salicin, the forerunner of non-steroidal antiphlogistic drugs, would have an effect in a standard model of depression, the forced swimming test (FST), compared to the antidepressant imipramine. Studies were accompanied by gene expression analyses. In order to allocate potential effects to the different constituents of WB, fractions of the extract with different compositions of salicyl alcohol derivative and polyphenols were also investigated. Male Sprague Dawley rats (n = 12/group) were treated for 14 days (p.o.) with the WB preparation STW 33-I (group A) and its fractions (FR) (groups FR-B to E) in concentrations of 30 mg/kg. The FRs were characterized by a high content of flavone and chalcone glycosides (FR-B), flavonoid glycosides and salicyl alcohol derivatives (FR-C), salicin and related salicyl alcohol derivatives (FR-D) and proanthocyanidines (FR-E). The tricyclic antidepressant imipramine (20 mg/kg) (F) was used as positive control. The FST was performed on day 15. The cumulative immobility time was significantly (p < 0.05) reduced in group A (36%), group FR-D (44%) and by imipramine (16%) compared to untreated controls. RNA was isolated from peripheral blood. RNA samples (group A, group FR-D, and imipramine) were further analysed by rat whole genome microarray (Agilent) in comparison to untreated controls. Quantitative PCR for selected genes was performed. Genes (>2 fold, p < 0.01), affected by WB and/or FR-D and imipramine, included both inflammatory (e.g. IL-3, IL-10) and neurologically relevant targets. Common genes regulated by WB, FR-D and imipramine were GRIA 2 ↓, SRP54 ↓, CYP26B ↓, DNM1L ↑ and KITLG ↓. In addition, the hippocampus of rats treated (27 d) with WB (15–60 mg/kg WB) or imipramine (15 mg/kg bw) showed a slower serotonin turnover (5- hydroxyindol acetic acid/serotonin (p < 0.05)) depending on the dosage. Thus WB (30 mg/kg), its ethanolic fraction rich in salicyl alcohol derivatives (FR-D) (30 mg/kg) and imipramine, by being effective in the FST, modulated known and new targets relevant for neuro- and immunofunctions in rats. These findings contribute to our understanding of the link between inflammation and neurological functions and may also support the scope for the development of co-medications from salicylate-containing phytopharma- ceuticals as multicomponent mixtures with single component synthetic drugs. © 2012 Elsevier GmbH. All rights reserved. Abbreviations: 5-HT, serotonin; 5-HIAA, 5-hydroxyindol acetic acid; AD, Alzheimer’s disease; AMPA, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid; ASA, acetylsalicylic acid; CAT, catalase; CD, Sprague-Dawley; CNS, central nervous system; COX, cyclooxygenases; CRH, corticotrophin-releasing hormone; CSF, cerebral spinal fluid; CYP26B1, cytochrome P450 protein 26B1; DNM1L, dynamin like protein 1; EDNRB, endothelin B receptor gene; ER, endoplasmatic reticulum; EtOH-FR, ethanol fraction; FR, fraction; FST, Porsolt-Swimming Test; GR, glutathione reductase; GST, glutathione S-transferase; GTPase, guanosine triphosphatase; HGF, haematopoietic growth factor; HPA-axis, hypothalamic-pituitary-adrenocortical axis; MS, multiple sclerosis; NMDA, N-methyl-d-aspartate; RA, retinoic acid; SCF, stem cell factor; SNP, single nucleotide polymorphism; SNRIs, serotonin and noradrenalin-reuptake inhibitors; SOD, superoxidedismutase; SRP, signal recognition protein; SSRIs, selective serotonin-reuptake inhibitors; TNFRSF1A, TNF-receptor superfamily member 1 A; WB, willow bark. * Corresponding author. Tel.: +49 22828722674; fax: +49 22828722019. E-mail address: Gudrun.Ulrich-Merzenich@ukb.uni-bonn.de (G. Ulrich-Merzenich). 1 Deceased. 0944-7113/$ – see front matter © 2012 Elsevier GmbH. All rights reserved. http://dx.doi.org/10.1016/j.phymed.2012.05.004