Objective To investigate the effects of calycosin on the injury of oxygen and glucose deprivation/ reoxygenation PC12 cells. Methods PC12 cells were randomly divided into 5 groups: control group; oxygen and glucose deprivation/ reoxygenation (OGD/ R) (model group); and high, medium, and low (0. 140 μmol/ L, 0. 070 μmol/ L, 0. 035 μmol/ L) dose groups of calycosin. The CCK-8 method was used to detect cell viability, the lactate dehydrogenase leakage rate in cells was determined with the LDH method , cell morphology was observed with an inverted microscope, cell membrane integrity was detected with PI fluorescence staining, caspase-3 expression was detected by immunohistochemistry, and Bax and Bcl-2 immunofluorescence staining was used to detect apoptosis. Results The cells in the control group grew well. Compared with the control group, the number of cells in the model group decreased, the cells appeared to shrink, cell membranes were destroyed, cell activity decreased significantly ( P < 0. 05), the lactate dehydrogenase leakage rate was significantly increased ( P < 0. 05), the number of PI red-stained cells increased significantly ( P < 0. 05), the expression of caspase-3 was significantly increased ( P < 0. 05), and the ratio of Bax/ Bcl-2 increased significantly ( P < 0. 05). There were no significant differences observed between the model group and the high dose group ( P > 0. 05). However, the number of cells in the middle and low dose groups increased, the cells showed regular morphology, the membrane damage was reduced ( P < 0. 05), the cell viability was significantly increased ( P < 0. 05), the lactate dehydrogenase leakage rate was significantly decreased ( P < 0. 05), the number of PI red-stained cells decreased ( P < 0. 05), the expression of caspase-3 was significantly decreased ( P < 0. 05), and the ratio of Bax/ Bcl-2 was significantly lower ( P < 0. 05). The changes in the above indicators were more significant in the middle dose group, with the differences between the high dose group and the model group not reaching statistical significance ( P > 0. 05). Conclusions Calycosin can optimize the growth status of PC12 cells following injury through oxygen and glucose deprivation/ reoxygenation, improve cell viability, inhibit lactate dehydrogenase leakage, and inhibit cell apoptosis by reducing caspase-3 expression and the Bax/ Bcl-2 ratio, effectively alleviating cell damage and playing a protective role.