Type 2 diabetes mellitus (T2DM) is multifactorial disease. This cross-sectional study was aimed to investigate relationship between stress and risk for T2DM in college students. Seven-hundred participants (350 T2DM risk and 350 non-T2DM risk groups). Stress index levels and heart rate variability (HRV) were respectively measured as primary and secondary outcomes. Results showed that both T2DM-risk and non-T2DM-risk groups had temporary stress, but the T2DM-risk group had significantly higher level of psychological stress (P < .001). For the HRV, the T2DM-risk group had significantly lower levels of parasympathetic proxies (lnHF, SDNN, and RMSSD) (P < .001). Chi-square (chi2) test showed significant correlation of the stressful state with T2DM risk (chi2 = 159.372, P < .001, odds ratio (OR) = 9.326). In conclusion, psychological stress is a risk factor for T2DM in college students. Early detection, monitoring, and treatments of psychological stress should be implemented in this group of population.