Mikroglial hücrelerde dietil maleat'in NLRP3 inflamazom aktivasyonu üzerine etkilerinin araştirilmasi

Microglial cells are tissue-resident macrophages of the Central Nervous System (CNS) (Dubbelaar, Kracht et al. 2018). In case of an injury in the brain, microglia act through their phagocytic function and eliminate invading microbes and other substances that harm the CNS. Also, they are responsible for secretion of cytokines assisting the immune response within the CNS (Dubbelaar, Kracht et al. 2018). Inflammasome activation initiates inflammatory responses leading to secretion of IL-1β and IL-18 cytokines and activation of caspase-1, and may also lead to pyroptotic cell death (Yang, Wang et al. 2019). For this reason, inflammasome activation should be prevented under such conditions. Chemicals with anti-inflammatory effects are appropriate candidates to inhibit inflammasome activation. Diethyl Maleate (DEM) is a Nrf2 activator and correlatively, DEM has been estimated to inhibit secretion of inflammatory cytokines and possible cell death. It is hypothesized that DEM represses LPS+ATP induced NLRP3 inflammasome activation caused cell death of N9 cells. In this thesis, effects of DEM on NLRP3 inflammasome activation in N9 microglia was investigated. NLRP3 inflammasome activation via LPS+ATP was performed. N9 cells were treated via DEM in order to determine the effect on inflammasome activation. The cytotoxicity of DEM in regarding conditions was determined via LDH (Lactate Dehydrogenase) and PI visualization. Afterwards, inflammasome markers were examined at mRNA level via PCR, at protein level via Western Blotting and ELISA. It was determined that DEM pretreatment represses inflammatory cytokine secretion and cell death arising from NLRP3 inflammasome activation. Keywords: Diethyl Maleate, microglia, N9, inflammasome, NLRP3, pyroptosis, LPS, ATP