The aim of this work implicates modern system pharmacological studies which is the elucidation of complex physiological and pathological cellular processes and the analysis and identification of molecules of interests. The overall purpose of this study is to observe dysfunction processes of the blood-brain barrier (BBB) which obverse leads to pathological circumstances caused by neurodegeneration. This pilot study focuses on the proteomic analysis of BBB in vitro models, the influence of

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... nt cultivation conditions on barrier tightness and the proteomic pattern. Experiments were conducted with human umbilical vein endothelial cells (HUVEC) and porcine brain microvascular endothelial cells (PBMEC/C1-2). The in vitro investigations were conducted under three different cultivation conditions to observe changes on the proteome pattern. Both cell lines were cultivated as follows (a) "co-culture system", (b) "induced system", (c)" uninduced system". The "co-culture system" comprises of the experimental cell lines (PBMEC/C1-2 or HUVEC) and astrocytic glial cells origin from rat brain. The "induced system" consists of enriced medium containing astrocytic factors (ACM) to induce BBB characteristics by changes in tight junctions (TJ). It is known that astrocytic derived soluble factors may induce the expression of BBB properties in endothelial cells in vitro (Janzer and Raff 1987) Using transendothelial electrical resistance (TEER) measurements to verify the tightness of the cellular monolayer in Transwell filter systems. Subsequently, cells were lysed for proteomic analysis with 2D DIGE and MALDI TOF MS. Statistical analyses revealed differentially expressed protein spots. The identification of "molecules of interests" which indicate significantly changes of the proteome pattern inside the different cultivation conditions was performed via mass spectrometry. Examples for identified proteins as heat shock protein 27 (HSP27), annexin A5, aldehyde dehydrogenase and catalase are target molecules for neurodegeneration oxi [...]