Climate change and human activities exert significant impact on the mechanism of runoff generation and confluence. Comprehending the reasons of runoff change is crucial for the sustainable development of water resources. Taking the Upper Shule River as the research area, the M-K test and the moving t test were used to diagnose the runoff mutation time. Furthermore, the slope changing ratio of cumulative quantity method (SCRCQ), climate elasticity method, and Budyko equation were utilized to quantitatively evaluate the impacts and contribution rates of climate change and human activities. The following results were obtained: (1) The Upper Shule River experienced a significant increase in runoff from 1972 to 2021, with 1998 marking the year of abrupt change. (2) The runoff sensitivity showed a downward trend from 1972 to 2021. The main factor affecting the decrease in runoff sensitivity was the characteristic parameters of underlying surface (n), followed by precipitation (P), while the influence of potential evapotranspiration (ET0) was the weakest. (3) The response of runoff changes to runoff sensitivity and influencing factors were 90.32% and 9.68%, respectively. (4) The results of three attribution methods indicated that climate change was the primary factor causing the alteration of runoff in the Upper Shule River. The research results supplement the hydrological change mechanisms of the Upper Shule River and provide a scientific basis for future water resources management and flood control measures.