OX-2 expression (Fig. 3D).Inflammatory agents promoted amyloidogenesis in vitro It

OX-2 expression (Fig. 3D).Inflammatory agents promoted amyloidogenesis in vitro It is known that microglia and astrocytes are major sources of neuro-inflammation. Moreover, recent data showed that neuronal cells also have cytokine receptors such as LPS receptor (toll like receptor) as well as TNF receptor [23,24]. Neurons may be directly involved in neuro-inflammation or indirectly via the interaction with microglia and astrocytes. In order to analyze the effect of LPS induced inflammation on amyloidogenesis in vitro, cultured astrocytes from rat pups and neuronal cells from rat embryos were used. Astrocytes lend both mechanical and metabolic support for neurons, regulating the environment in which they function. Interferron-gamma (IFN-) and tumor necrosis factor-alpha (TNF-) as well as LPS were treated to induce an inflammatory reaction.Figure LPS on secretase activities and amyloidogenic proteins expression Effect of3 Effect of LPS on secretase activities and amyloidogenic proteins expression. The activities of -, -secretase (A, B) and -secretase (C) were assessed by using commercially available assay kits.Protocatechuate 3,4-dioxygenase Data represent mean S.E. (n = 5). *Significant different from control group (p 0.05). The expression of APP, BACE and C99 (D) were detected by Western blotting using specific antibodies. Each blot is representative for five experiments. -actin protein was used here as an internal control.Page 7 of(page number not for citation purposes)Journal of Neuroinflammation 2008, 5:http://www.jneuroinflammation/content/5/1/Similiar to the in vivo results, inflammatory stimuli concomitantly increased expression of amyloidogenic proteins (such as APP, BACE and C99) accompanied with the increase of expression of inflammatory proteins (such as COX-2 and iNOS) in both astrocytes (Fig.Icatibant 4A) and neuronal cells (Fig.PMID:23916866 4B). These results further indicate amyloidogenic pathway could be promoted by neuroinflammatory stimulation in in vitro and in vivo.Anti-inflammatory drug inhibited LPS-induced amyloidogenesis and memory impairment The effect of sulindac sulfide, a COX-1, 2 non-selective drugs, in vivo and in vitro system was assessed. Sulindac sulfide has been known to decrease the A secretion in N2a neuroblastoma cells stably transfected with human APP695 bearing the Swedish mutation [25]. As shown in Fig. 4C, the cells expressed low levels of APP, -site APP cleavage enzyme (BACE) and C99 protein in an unstimulated condition, whereas, expression of BACE, APP and C99 proteins increased in response to LPS (1 g/ml) after 24 hrs. Treatment with sulindac sulfide (12.5, 25, 50 M)Figure anti-inflammatory agents on expression of amyloidogenic proteins (A, B) Effect of4 Effect of anti-inflammatory agents on expression of amyloidogenic proteins (A, B). The expression of APP, BACE and C99 were detected by Western blotting using specific antibodies in astrocytes (A) and neuronal cells (B). -Actin protein was used as an internal control. Each blot is representative for five experiments. Sulindac sulfide inhibits expression of amyloidogenic proteins (C) and A12 secretion (D) induced by LPS in cultured neuronal cells. Combined Sulindac sulfide (12.5, 25, 50 M) and LPS treatment for 24 hr were used. (C), The expression of APP, BACE and C99 in neuronal cells was detected by Western blotting using specific antibodies. -Actin protein was used as an internal control. (D), Media were collected to determine an A12 secretion by ELISA. Data represent mean S.E. of three exper.