| Abstract | Multiple anti-inflammatory drugs are currently in use to treat neuroinflammation in the brain which can result from Alzheimer's disease and lead to neuronal death and cognitive decline. TNF-alpha is a proinflammatory cytokine that is produced by microglial cells in the brain and its overexpression can lead to neuroinflammation which is seen in Alzheimer's disease. In this study, we observe the in vitro effects of the drugs P2D2024 (IDT), P2D340, and P2D2244 made by P2DBioscience. Two different assays were developed to test the anti-inflammatory and TNF-alpha modulating capabilities of these drugs. BV2 cells in culture were mainly used for these assays to model how the drugs affect mice microglial cells which are resident in the brain. The first assay uses a luciferase reporter gene to establish if NF-kappaB promoter activity is disrupted when cells are treated with drug. The second assay uses RT-qPCR to measure changes in TNF-alpha mRNA levels when cells are treated with drug. Cells were also harvested during a time course followed by RT-qPCR to establish if the degradation time of TNF-alpha mRNA was affected by the treatment with drug. Through these assays we were able to demonstrate that the drugs P2D340 and P2D2244 are able to modulate TNF-alpha in vitro. Results indicated the drugs modulate steady-state levels of TNF-alpha mRNA in LPS-stimulated BV2 cells. Thus, these drugs need to be further studied to establish the mechanism of action of this TNF-alpha modulation. |
