Investigation of the bortezomib effects on endothelial cellsin the inflammatory co-culture model induced by monosodium urate crystals

Gout is a metabolic disease characterised by inflammation in response to the deposition of monosodium urate (MSU) crystals in joints and surrounding structures. In this thesis, the effect of bortezomib, one of the NF-κB inhibitors, on the inflammatory response induced by MSU crystals was investigated. The effect of bortezomib on cell proliferation and migration, apoptotic effect and effect on inflammation in endothelial cells was investigated by creating an in vitro gout co-culture model using HUVEC and THP-1 cells. The effects of bortezomib and MSU on cell viability were investigated by MTT method in THP-1 and HUVEC cells and the effect in HUVEC cell proliferation in co-culture medium in real-time analysis system. The migration effect of bortezomib on endothelial cells was determined by real-time analysis system and scratch assay. In co-culture inflammation medium, the apoptotic effect of bortezomib on HUVEC cells, caspase-3 activity and mitochondrial membrane integrity, and the amounts of NLRP3 inflammasome-associated TNF-α and IL-1β, IL-6, IL-8, IL-10 cytokines were analyzed by flow cytometry. As a result, concentration-dependent anti-inflammatory effects of bortezomib were determined in MSU-induced inflammation. It was observed that HUVEC cell proliferation decreased depending on the bortezomib concentration increase and IC50 value at 24th hour was calculated as 0.127µM. Bortezomib increased apoptotic effect and caspase-3 activation in vitro gout model. In particular, TNF-α, IL-6, IL-8 levels decreased and IL-10 level increased depending on the concentration of bortezomib. In the light of the results obtained, we think that the potential use of bortezomib as a therapeutic agent in the treatment of inflammatory diseases such as gout will provide preliminary data to the literature and provide an alternative perspective in clinical drug therapy.