Experimental Study On The Effect Of Diclofenac Sodium On The Energy Metabolism Of Synoviocytes In The Treatment Of Rheumatoid Arthritis

Objective To further study the mechanism of diclofenac sodium in the treatment of rheumatoid arthritis,and explore its possible mechanism from glycolysis.Methods MTT method was used to detect cell proliferation inhibition;gene chip was used to detect changes in glycolysis and mitochondrial-related genes;computer-aided drug design was used to predict protein structure-based targets,so that diclofenac could be molecularly docked with protein metabolism-related protein crystal structures.,Select targets with high binding activity prediction for detailed molecular interaction analysis;lactic acid kit to detect intracellular lactic acid changes,and transfect si-COX-2 Inhibitor,si-PKM2 Inhibitor,si-GLUT1 Inhibitor with liposome Lipofectamine 2000 To human RA FLS cells,Annexin V-FITC/PI double staining method was used to detect apoptosis,RT-qPCR method and Western blot method were used to detect relative expression levels of glycolysis related genes and proteins;2-NBDG was used to detect rat joint slip Changes of glucose uptake in membrane tissues;immunohistochemistry was used to detect the relative expression levels of COX-2 glycolysis-related proteins PKM2 and GLUT1 in rat synovial tissues.Results Diclofenac had a significant inhibitory effect on the glycolysis of RA FLS cells,among which the inhibitory effects on glycolytic enzymes PKM2 and GLUT1 were more obvious.The mechanism is that diclofenac mediated the GLUT1/ PKM2 pathway in RA FLS cells through COX-2 Induced apoptosis.Immunohistochemical results also proved that diclofenac sodium had a significant inhibitory effect on the glycolytic enzymes GLUT1 and PKM2 in rat synovial tissue.Conclusion Diclofenac can inhibit the increased glycolysis of RAFLS cells,and induce apoptosis of RAFLS cells by inhibiting COX-2 mediated GLUT1/PKM2 pathway in RAFLS cells.