Objective To explore whether natural ceramide has effect on lactate dehydrogenase(LDH) metabolism in primary cultured mice cortical neurons. Methods Ceramide at different concentrations of 50,100,200,500,1000,2000 nmol/L was given to the culture medium of primary cultured mice cortical neurons. The LDH concentration was assayed after 0 h,1 h,4 h,8 h,12 h,16 h,24 h,36 h respectively. Its metabolism rate and leakage rate were calculated respectively. Results Ceramide changed significantly the distribution of LDH inside and outside of the neurons. Exogenous ceramide stimulated LDH elevation in supernatant in a dose-dependent and time-dependent manner, but had little impact on the total LDH metabolism of neurons. Conclusion Ceramide elevates the leakage rate of LDH and has no effect on total LDH metabolism of neurons.