Protein was then detected with an enhanced chemiluminescence kit

Protein was then detected with an enhanced chemiluminescence kit (PerkinElmer Inc., Waltham, MA, USA) and visualised with a FUJI Film LAS-3000 (Tokyo, Japan). Enzyme-linked immunoabsorbent assay (ELISA) was performed with respect to TNF-α using OptEIA kits (BD Biosciences, San Jose, CA) and IL-1β using Nordic Biosite, Täby, Sweden. Supernatants from purified microglial cell cultures were collected after microglia had been stimulated for 0.5–24 h. ELISA was performed on

the supernatants according to the manufacturer’s instructions. All stimulations were performed in a total volume of 1 ml MEM. Cell lysates were produced by harvesting remaining cells with a cell scraper in 1 M NaOH, and aliquots were taken for protein find more determination. Ceritinib purchase ELISA plates were analysed at 450 nm with a Molecular Devices VersaMax microplate reader and were analysed using SoftMax Pro 4.8, both from Molecular Devices (Sunnyvale, CA, USA). The protein determination assay was performed in accordance with the manufacturer’s instructions using a DC Protein Assay (Bio-Rad, Hercules, CA, USA), based with some modifications on the method used by Lowry et al. (1951). Both standard (0–4 mg/ml BSA) and samples were

mixed with the reagents, incubated for 15 min at room temperature, read at 750 nm with a Versa-max microplate reader, and analysed using SoftMax Pro 4.8. Differences between grouped mean values were identified using one way ANOVA followed by Dunnett’s multiple comparisons test. Error bars show standard error of the mean (SEM). In a microglial cell culture we observed that exposure of LPS was associated with a release of both TNF-α and IL-1β, which increased over time. Dexamethasone and corticosterone attenuated these responses. Other investigated anti-inflammatory agents in this study, which previously have been shown to counter a LPS-dependent release of TNF-α and IL-1β in astrocytes, were not associated with corresponding effects in microglial cells. After inflammation, increases of pro-inflammatory cytokines are observed. Astrocytes seem to be better target cells for anti-inflammatory substances than microglia. The physiological relevance might

be that communication within the astrocyte networks seems to be of importance. If the signalling between astrocytes is working, thereby the microglia show a normal and non-inflammatory Endonuclease state. Thus, our findings indicate that anti-inflammatory substances have a cell-type specific capacity to modulate pro-inflammatory reactions in glial tissues. This work was supported by Edit Jacobson’s Foundation, Arvid Carlsson’s Foundation, Lena and Per Sjöberg Foundation, and the Sahlgrenska University Hospital (LUA/ALF GBG-11587), Gothenburg, Sweden. “
“Physiological and theoretical studies have argued that the sensory nervous systems of animals are evolutionarily adapted to their natural stimulus environment (for review see Reinagel, 2001).

Leave a Reply

Your email address will not be published. Required fields are marked *

*

You may use these HTML tags and attributes: <a href="" title=""> <abbr title=""> <acronym title=""> <b> <blockquote cite=""> <cite> <code> <del datetime=""> <em> <i> <q cite=""> <strike> <strong>