Protein expression profiling of some osteoclastogenic markers in Rheumatoid arthritis: an in-vitro and in-silico study

Abstract

Rheumatoid Arthritis is a chronic polyarthritic autoimmune condition associated with joint erosion and synovial hyperplasia. Bone resorption in RA is a characteristic of disease pathogenesis. RANKL signaling pathway is the main pathway involved in bone resorption via osteoclastogenesis. Many factors activated downstream RANKL signaling pathway exhibit elevated expression levels in RA and have been targeted for therapeutic purposes. In this study, protein expression profiling of human carbonic anhydrase II, m Calpain, NFATc3 and TNF-alpha have been elucidated by using the technique of western blotting. The results showeded their increased expression in the serum of RA patients. Furthermore, since these proteins are inflammatory markers and inflammation is tightly associated with oxidative stress, the DNA damage status due to this oxidative stress is assessed in this study using comet assay. The results showeded that lymphocytes from RA patients demonstrate detectable DNA damage. Moreover, in-silico virtual screening has been used to screen the DrugBank for the identification of new hit compounds exhibiting the potential to target human carbonic anhydrase II and inhibit its activity. For this, docking and pharmacophore modeling approaches were used. Three new hit compounds were elucidated. First one, Supiride, belongs to a class of Benzenesulphonamides which is the mostwell-studied class of compounds known till now. The other two compounds, alfuzosin and Prazosin are quinzolamines, which is a novel prediction since compounds of this class have never been reported as CAIs nor used for the treatment of RA