In the context of global warming, the melting of glaciers in the Tien Shan Mountains as the important “solid reservoir” in the arid area of Central Asia is accelerating in recent decades, leading to profound changes in regional water resources. Based on the simulated glaciological data from the Python Glacier Evolution Model (PyGEM) and the measured glaciological data from the World Glacier Monitoring Service (WGMS), this paper analyzed the applicability of simulated data, the changes in glacier mass balance, and the responses of the glacier to climate change and its impacts on glacier runoff in the Tien Shan Mountains. The results show that (1) the PyGEM simulation dataset is in good agreement with the measurements, which can effectively reproduce the change in the glacier mass balance in the Tien Shan Mountains glaciers and is suitable for studying the regional scale glacier change. (2) From 1980 to 2016, the decadal average mass balance change rate of glaciers in the Tien Shan Mountains was −0.012 m w.e. yr−1. The regional mass balance showed an overall negative increasing trend (the area with increasingly negative accounted for 80.13% of the entire area), with a positive increase that only occurred in the West Tien Shan Mountains and western North Tien Shan Mountains (19.87%). (3) The correlation between the temperature and mass balance is much higher than that between the precipitation and mass balance. Temperature dominates the change and development of regional glaciers. The increase in negative glacier mass balance that was observed in the study area is mainly affected by the rising temperature, the decreasing solid precipitation in the accumulation period, and the rapid melting in the ablation period. (4) The glacier runoff in the six representative rivers showed an increasing trend. The contribution rate of glacier runoff to river runoff changed significantly after 2000 but differed among rivers. Overall, the larger the glacier area in the source region is, the greater the contribution rate of glacier runoff is, and the more the contribution rate continuously increases or fluctuates; otherwise, the contribution rate keeps declining, which means the runoff peak may have passed and future runoff may decrease. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.