Objective To study the effect of negative emotion evocation on the autonomic function activity of hearts in healthy subjects, and to initially discuss the mechanism by which negative emotion is involved in the development of coronary heart disease. Methods We selected 69 healthy university students as our subjects. A computer game was used to induce the stress among them; the skin code, heart rate and heart rate variability (HRV) were collected and compared at different phases. Results The high frequency (HF) value was significantly decreased(F =10.773,P<0.01), the low frequency (LF) value was significantly increased(F =5.620,P<0.01), and LF/HF ratio was significantly increased(F =40.327,P<0.01) at both expecting phase and coping phase, which indicates significant changes of cardiac sympathovagal activity, including the relative excitation of sympathetic nerve, relative depression of the vagus nerve, and aggravated autonomic imbalance. Conclusion Exposure of healthy individual to acute emotional stress can induce autonomic variation similar to that of coronary heart disease, indicating that long-term negative emotional stress may be one of the risk factors for vagal dysfunction.