| Abstract | Alzheimer's Disease (AD) and Traumatic Brain Injuries (TBI) are global societal problems affecting millions of people and costing billions of dollars per year.1,6 Hallmarks of AD include memory loss, cognitive decline, depression, and confusion due to neuronal death induced by unchecked inflammation in the brain caused by the overproduction of pro-inflammatory cytokines by the immune system.1,9,11 TBI occurs when a sudden trauma damages brain cells, which activate the immune response potentially leading to chronic inflammation and a multitude of symptoms affecting cognitive, somatic, and emotional processes.3,4,12 There is currently no cure for AD, nor is there an effective treatment for chronic inflammation caused by TBI. In collaboration with a biotech company, P2D Bioscience®, we are testing a series of drugs for their ability to target inflammation in a BV-2 microglial cell culture model of LPS-induced inflammation.21 To understand the cellular mechanism of these novel drugs, we used SDS-PAGE electrophoresis and Western Blot analysis to measure levels of proteins involved in the activation of the NFkB signaling pathway, which modulates inflammation. We specifically measured the levels of the inhibitor of NFkB, IkBa, to determine whether the drugs were blocking the phosphorylation and degradation of IkBa; subsequently blocking the pro-inflammatory effects of activated NFkB. The ability to inhibit the activation of NFkB would indicate that the drug is a good candidate to reduce inflammation in brain microglial cells. Increasing our understanding of the cellular mechanism of action of these drugs is imperative for the progression of drug development and testing because it can be used to evaluate potential side effects. |
