Effects of IL-1 beta on receptor-mediated poly-phosphoinositide signaling pathway in immortalized astrocytes (DITNC).

Astrocytes are known to play multi-functional roles in support of many homeostatic mechanisms in the central nervous system including host defense mechanisms. Despite the ability of cytokines to alter gene expression and cellular activity, their effect on receptor-mediated poly-phosphoinositide (poly-PI) signaling pathway has not been examined in detail. In this study, an immortalized astrocyte cell line (DITNC) was used to test the effect of IL-1 beta exposure on the poly-PI signaling pathway. Similar to primary astrocytes, DITNC cells exhibit P2-purinergic receptor response to ATP and UTP leading to transient increases in inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and intracellular calcium concentration, [Ca2+]i. Upon exposure of DITNC cells to IL-1 beta (100 U/ml) for 24 hrs, an increased response to the poly-PI agonists was observed. The increase in ATP-mediated Ins(1,4,5)P3 release could not be attributed to a shift in the ATP dose or an alteration of the time profile for the release of Ins(1,4,5)P3. Since the increase in response required a lag time of 4 hr after IL-1 beta exposure, it is unlikely that this effect was due to a direct interaction of IL-1 beta with the purinergic receptor. On the other hand, an increase in ATP response could be observed in DITNC cells exposed to conditioned medium obtained after IL-1 beta treatment. It can be concluded that exposure of astrocytes to cytokines may lead to an increase in receptor-mediated poly-PI signaling activity and this may involve compounds secreted into the culture medium, e.g., the secretory phospholipase A2.