CX3CL1 modulator in the inflammatory processes of Alzheimer’s disease

Alzheimer's disease (AD) is the most common form of dementia characterized by the accumulation of β-amyloid plaques, tau tangles, neuroinflammation, and synaptic/neuronal loss, the latter being the strongest correlating factor with memory and cognitive impairment. Through an in vitro study on a neuron-astrocyte-microglia (NAM) co-culture system and on Blood Brain barrier organ on chip (BBB-oC) model, we have analyzed the effects of cerebrospinal fluid (CSF) samples from non-AD and AD patients. Moreover, we tested the effect of sCX3CL1 purify protein on BBB-oC. Before to evaluate the effect of the CSF samples, they are characterized for the canonical markers of AD and for the presence of sCX3CL1, the results showed an increase of presence of sCX3CL1 in AD CSF compared to non-AD CSF. After the treatment with the CSF samples the NAM co-cultures were investigate through optical microscopy, and for expression of proteins and enzyme through immunofluorescence analyses, Zymography analyses and Q-PCR analyses. The results showed the activation and up-regulation of factors involved in inflammation pathways including CX3CL1, MMPs and involved in the process of hyperphosphorylation of tau and in the formation of amyloid fragment. Morphology analyses, immunofluorescence analyses and Real Time PCR analyses showed the damage of the blood brain barrier in the BBB-oC model treated with sCX3CL1 purify protein and AD CSF. After carrying out the preliminary in vitro studies, Alzheimer was induced in rats through intracerebroventricular microinjection of Streptozotocin (STZ). The induction of AD was confirmed by dot blot and western blot analyses that showed the presence of the canonical markers in animal model. Other elements, including CX3CL1, were evaluated by western blot analysis and Real Time PCR analyses to identify elements that may be up and downregulated in the metabolic pathways after the induction of Alzheimer’s disease.