Recently, cognitive demands in workplaces have surged significantly. This study explored the intricate relationship among mental workload (MWL), occupational fatigue, physiological responses, and cognitive performance in office workers by using collective semi-parametric models. One hundred office workers were selected from twenty offices involved in cognitive performance. MWL was assessed through the NASA Task Load Index (NASA-TLX), and occupational fatigue was measured using the Persian version of the Swedish Occupational Fatigue Inventory. Physiological responses, including respiratory rate, the electrical conductivity of the skin (ECS), Heart Rate (HR), and other heart-related parameters, were recorded from the participants during a work shift. Selective and Divided Attention tests were chosen to evaluate workers' cognitive function based on cognitive task analysis. The mean of MWL and occupational fatigue scores were 66.28 ± 11.76 and 1.62 ± 1.07, respectively. There was a significant moderate correlation between two dimensions, mental demand (0.429) and frustration (0.409), with functional fatigue. Also, Significant and, of course, nonlinear relationships were observed between MWL and HR (R 2 = 0.44, P-value < 0.001) and ECS (R 2 = 0.45, P-value < 0.001) and reaction time in selected (R 2 = 0.34, P-value < 0.001) and divided test (R 2 = 0.48, P-value < 0.001). Similarly, nonlinear relationships were observed between physiological responses and cognitive performance with fatigue among participants who had experienced higher levels of occupational fatigue. The MWL and fatigue seem to have a significant and non-linear effect on physiological parameters such as HR and ECS and cognitive parameters such as reaction time. Moreover, MWL can influence the dimension of functional fatigue of workers.