Cerebral cortices from these rats, as well as unoperated control rats, were processed for protein or mRNA tissue level analyses or were fixed and processed for immunofluorescent image analyses. p38 MAPK Rat brains implanted with IL 1b containing pellets had markedly elevated steady state levels of ApoE mRNA and of ApoE protein compared to those in rats Inhibitors,Modulators,Libraries implanted with sham pellets or to unoper ated controls. Neuroinflammatory conditions and models thereof often exhibit chain reactions of multiple effectors work ing sequentially, in parallel, or in feedback loops fomenting a persistent and progressive situation. In this vein, the ability of IL 1b to elevate the levels of IL a prompted an examination of gene expression indices of neuroinflammation in this chronic IL 1b delivery para digm.

The increase in IL 1a immunofluorescence noted above was found to be reflected at the mRNA level. Chronic IL 1b also elevated mRNA levels of endogenous IL 1b, as Inhibitors,Modulators,Libraries well as its cleavage enzyme ICE. Along with these changes in IL 1 related Inhibitors,Modulators,Libraries molecules, the mRNA for the proinflammatory cytokine TNF was elevated. These proinflammatory changes were accompanied by induction of bAPP mRNA, consistent with the immunofluorescence results and prior studies of IL 1 bAPP interactions. The induction of ApoE in the cortex by IL 1b pellets was also detectable by immunofluorescence, which demonstrated neuronal localization. IL 1b pellets also elevated expression of IL 1a in the CA1 of hippo campus. This IL 1a induction was localized principally to cells with astrocytic morphology.

Pyrami dal neurons of the CA1 overexpressed bAPP in response to the chronic delivery of IL 1b. Tissue culture studies reveal potential for indirect impacts Inhibitors,Modulators,Libraries of IL 1b on ApoE To examine the impact of IL 1b on ApoE expression in greater temporal and mechanistic detail, we utilized two types of neuronal cell culture, primary cultures of rat cortical neurons and the human NTera2 cell line. We previously demonstrated that glutamate elevates bAPP expression via Inhibitors,Modulators,Libraries a mechanism that requires the bio logical activity of ApoE. Moreover, IL 1b has been shown to influence the processing of bAPP. There fore, we tested whether ApoE expression was responsive to these agents and another derivative of bAPP, Ab1 42. In both culture types, expression of ApoE mRNA was elevated approximately two fold by exposure to IL 1b, Ab1 42, or glutamate for 20 h, the induction by sAPP exceeded six fold.

All of these agents were found to elevate ApoE protein levels as well. The ability of glutamate and bAPP fragments to impact ApoE was given additional relevance by demon stration of impacts of IL 1b on these agents. Levels of glutamate released into neuronal culture medium was elevated by IL 1b. Likewise, IL 1b elevated the levels of sAPP in the culture medium of primary neurons in Wortmannin supplier a dose dependent fashion.