Light exposure at night disrupts the circadian rhythm as predetermined by clock gene expressions which increases the risk of metabolic disorders, and changes the pursuit of the oxidant-antioxidant milieu and the magnitude of their cellular mRNA, which interferes with the repair process by generating reactive oxygen species. The exploration of shift-induced changes is necessary as it is linked to chronic circadian desynchronization, and sleep restriction, frequently exhibiting an increased stress response. The systemic levels of oxidative damage, antioxidant defense enzymes, pro-oxidant levels, neuroendocrine changes, and DNA damage were studied in 115 female shift workers (aged 24–45 years) depicting significant variations upon considering their chronotype. The outcomes indicated that pro-oxidative stress and increased neuroendocrine stress are the molecular mediators of the detrimental health risks associated with circadian misalignment and sleep loss experienced by shift workers. Finally, we addressed the potential advantages of taking preventative steps to lessen the stress that shift work brings, including taking prophylactic sleep.