To elucidate the therapeutic efficacy of leflunomide supplemented with Folic Acid on Type-II Collagen induced arthritic BALB/c mice

Abstract

Rheumatoid Arthritis (RA) is an autoimmune disease having a global importance. The role of disease modifying anti-rheumatic drug (DMARD) Leflunomide has been described in case of RA. The objective of this study was to characterize the anti oxidant property of leflunomide. The second major objective of this study was to improvise the efficacy of Leflunomide and reduce the cyto-toxicity by using Folic acid in-vivo. The anti-oxidative property was studied by performing DPPH assay. The anti-oxidative potential of leflunomide was shown by significant DPPH inhibition (p<0.05). A BALB/c collagen induced arthritic model was designed and characterized via histopathological changes and gross morphological changes. The treatment started showing improvement in joints after the second dose of the drug. The maximum improvement was shown in the group with dosage 17 mg LEF/kgBW/day along with 3mg folic acid/kgBW/day. Spleen indices showed a significant decrease in the size and inflammation of the spleen in the models taking Leflunomide alone with folic acid. No significant change was seen by increasing the dose of drug and supplement in this case. Pathological changes in the joints of paws were studied through histopathology analysis. In case of treatment groups where Leflunomide was supplemented with folic acid, it was observed that inflammation in synovium and hyperplasia was reduced. The cartilage and bone erosion was observed to be reduced. Immunohistochemistry was performed to check the levels of the mitochondrial proteins like dihyrooratate dehydrogense (DHODH) and Tyrosine kinase in the spleen cells of arthritic models. It was observed that the levels of these proteins were decreased in treated groups.