Objectives A mounting evidence confirms the effect of caloric restriction and intermittent fasting in ameliorating body oxidative stress and inflammation. A growing body of evidence supports that chronic inflammation and increased level of oxidative stress augment the way for the development of metabolic diseases, such as diabetes and cancers. Objectives: This research was conducted to examine the effect of Ramadan diurnal intermittent fasting (RDIF) on the gene expression of cellular metabolism (SIRT1 and SIRT3) and antioxidant genes (TFAM, SOD2, and Nrf2). Methods One-hundred fourteen (75 males and 39 females) overweight and obese subjects and twelve healthy body weight controls were followed-up before and after Ramadan. Dietary, anthropometric and biochemical assessments were performed before and at the end of Ramadan fasting month. Results Results showed that the relative gene expressions in obese subjects in comparison to counterpart expressions of controls showed a significant (P < 0.001) increase in the anti-inflammatory cytokine (IL-10), along with significant (P < 0.001) reductions in the proinflammatory cytokines (IL-6 and TNF-alpha). Expression of TFAM, SOD2, and Nrf2 significantly increased at the end of Ramadan (90.5%, 54.1%, and 411.5%, respectively). However, the metabolism-controlling gene (SIRT3) showed a highly significant (P < 0.001) downregulation accompanied by a trend for reduction in the SIRT1 gene at the end of Ramadan month, with % decrements of 61.8% and 10.4%, respectively. Conclusions Results suggest that RIF ameliorates the genetic expression of antioxidant and anti-inflammatory and metabolic regulatory genes. Thus, RIF presumably may entail a protective impact against oxidative stress and its adverse metabolic-related derangements in non-diabetic obese patients. Funding Sources University of Sharjah, UAE.