Betulinic acid and fluvastatin exhibits synergistic effect on toll-like receptor-4 mediated anti-atherogenic mechanism in type II collagen induced arthritis

Cardiovascular disease (CVD) is a major problem during rheumatoid arthritis which leads to morbidity and mortality in arthritic patients. So the present study emphasizes combinatorial effect of Betulinic acid, a triterpenoid and fluvastatin, an HMG CoA reductase inhibitor on atherogenesis during arthritis. Arthritis was induced by bovine type II collagen dissolved in 0.01 M acetic acid at a concentration of 4 mg/mL and emulsified in equal volume of incomplete Freund's adjuvant. Betulinic acid (2 mg/kg) and fluvastatin (5 mg/kg) alone and in combination was administered orally from day 14 to 60. At the end of 60 days, tissues and blood were isolated for evaluation of biochemical parameters. Treatment with betulinic acid and fluvastatin showed significant (p < 0.05) reduction in Arthritic index, Rheumatoid factor, C-reactive protein (CRP), total lipids and anti-CCP (cyclic citrullinated peptide) antibody. Anti-inflammatory enzyme activities and oxidative stress were significantly decreased in the peripheral blood mononuclear cells by the administration of both betulinic acid and fluvastatin than alone treatments. Combination therapy was found to be a potential enhancer of the expression of anti-inflammatory cytokine interleukin-10 whereas it significantly blocked the expression of Toll-like receptors-2 and 4, inflammatory markers such as interleukin-1β, tumor necrosis factor-α, Interferon-γ, cell adhesion molecules and nuclear translocation of NF-kappa B in aorta than drug alone treated groups. So the present study summarizes a combination therapy of betulinic acid and fluvastatin that reduces the risk of both rheumatoid arthritis and CVD by modulating the expression of various inflammatory mediators through Toll-like receptors-4-NF-κB downstream signaling pathway, atherogenic index and oxidative stress in collagen induced arthritis.