Investigation of mechanisms involved in neuroinflammation as therapeutic targets

Project no. 22123 · Status: Kapanmış (Yayın bekleniyor) · 2020

Summary

High blood pressure affects many organs. Sympathetic nervous system activity is closely related to hypertension and is an effective factor in the development of cardiovascular diseases. However, it is known that neuroinflammation accompanies sympathetic activation. With the discovery of inflammasomes in recent years, neuroinflammation studies have gained a new dimension.The aim of the thesis is to investigate the effect of Angiotensin (Ang) II, which is the mediator molecule in increasing sympathetic activity, on inflammasome activation in neuronal cells and to reveal the molecular targets that play a role in possible NLRP1 inflammasome activation. The glycogen synthase kinase 3β (GSK3β) enzyme investigated within the scope of the thesis is an enzyme associated with neuropsychiatric and neurodegenerative diseases. This enzyme is involved in regulating blood pressure through mechanisms in the brainstem.Within the scope of the thesis, Ang II-induced in vitro model and in vivo spontaneous hypertension model were used to examine NLRP1 inflammasome activation through IL-1β, caspase-1, IL-18, inflammasome-related genes and proteins using various biochemical and molecular techniques such as ELISA, real-time PCR, Western Blot. The response of NLRP1 activation induced by Ang II to GSK3β enzyme inhibition was evaluated by cell culture study. Additionally, the effect of GSK3β inhibitor on NLRP1 inflammasome activation was investigated in the brainstem of spontaneously hypertensive rats. Following Ang II treatment, a significant increase was observed in the levels of the NLRP1 inflammasome and its associated IL-1β, caspase-1, IL-18 in neuronal cells (p˂0.05). All of these were accompanied by an increase in GSK3β activation. NLRP1 inflammasome activation was evaluated following GSK3β inhibitors, tideglusib and LiCl applications. It can suggested that the decrease in IL-1β, caspase-1, and IL-18 levels with these applications may be related to the NLRP1 inflammasome activation and GSK3β enzyme activity. In in vivo studies, a decrease in blood pressure was observed with tideglusib administration in spontaneously hypertensive rats, and studies in the brainstem showed that the NLRP1 inflammasome activation and its related parameters (ASC, IL-1β, caspase-1, IL-18 and pNFκB(p65)) decreased. By confirming GSK3β inhibition in the brainstem, the GSK3β enzyme-NLRP1 relationship with neurogenic hypertension has been demonstrated.Thus, within the scope of the thesis, Ang II-mediated NLRP1 inflammasome activation was investigated for the first time in neuronal cells, and answers were sought for possible mechanisms that play a role in neuroinflammatory processes and the potential of these mechanisms to be presented as therapeutic targets.

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