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Objectives: Assessment of upstream levels of lipid peroxidation and DNA damage regulating the development of rheumatoid arthritis. Data Source: All the samples were collected from Jinnah Hospital Lahore. Study Design: Comparative cross sectional study. Period: Two years from 18-09-2014 to 24-10-2016. Setting: The Institute of Molecular Biology and Biotechnology (IMBB) in The University of Lahore-Pakistan. Material and Methods: Blood, saliva and synovial fluid of fifty (n=50) individuals diagnosed with Rheumatoid Arthritis and fifty (n=50) age-sex matched controls were added in current study. Levels of MDA were determined spectrophotometrically while the concentrations of isoprostanes, 8-OHdG and 4-HNE were measured by the help of commercially available Elisa Kit. Results: Levels of lipid peroxidation products including MDA (μmol/ml), isoprostanes (pg/ml), 4-HNE (μmol/ml) and DNA damage in the form of 8-OHdG (pg/ml) were significantly high (p<0.015, p<0.022, p<0.004 and p<0.036) in patients as relative to normal individuals. Levels of MDA in serum (1.95±0.094 vs. 0.95±0.019), saliva (0.012±0.0034 vs. 0.056±0.0056) and synovial fluid (3.26±0.65 vs. 0.019±0.0016) were differed significantly in each groups. Level of isoprostanes in serum (12.26±5.26 vs. 1.26±0.015), saliva (2.16±0.019 vs. 0.816±0.017) and synovial fluid (34.26±4.26 vs. 0.136±0.019) were recorded higher in RA patients as compared to control. Concentrations of 8-OHdG in serum (0.945±0.014 vs. 0.019±0.0035) saliva (0.0024±0.0003/ 0.0029±0.00017) and synovial fluid (1.33±0.451 vs. 0.055±0.0016) were recorded high in RA patients. Significantly higher concentration of 4-HNE in serum (4.265±1.25 vs. 1.99±0.016), saliva (1.26±0.15 vs. 0.191±0.0091) and synovial fluid (6.35±1.16 vs. 0.094±0.00165) were recorded in patients with RA. Conclusion: Present study concluded the role of oxidative biomarkers and their differential expression in the onset of autoimmunity in patients with RA. Increased stress is involved in the DNA damage and increased lipid peroxidation in the synovial fluid. Therefore, antioxidant therapy may have some prognostic role in the patients with RA by decreasing the intensity of oxidative stress and DNA damage. [ABSTRACT FROM AUTHOR]