Culturing the neonatal rat myocardial cells’ hypoxia/reoxygenation modle in vitro and observe the effects of Yiqi Zhuyu Formula(Shenyuandan, SYD) on the survival rate and the degree of myocardium cells injury biomarkers, and thus to further discuss the its relieving mechanism on PMI. Methods: Neonatal rat myocardial cells were cultured in vitro to establish the hypoxia/reoxygenation model. Then use CCK-8 assay to detect the survival rate and enzymatic assay to detect the levels of CPK and LDH on each cell culture supernatant. Results: Compared with the normal group, the survival rate of the model group is lower(P<0.05) and the CPK and LDH release quantity is higher(P<0.05). The survival rate of SYD drug-containing serum group, PDTC + SYD and PDTC + blank serum group were higher than those of model group and blank group(P<0.05); serum cell enzyme(CPK, LDH)release quantity are lower than those of the model group and blank serum group(P<0.05), while there was no statistical difference among the three groups. Conclusion: Shen Yuan Dan can improve the cell survival rate of myocardial cell hypoxia/reoxygenation injury model and relieve cell injury. This may because it can reduce cell damage through inhibiting NF-κB and its mediated oxidative stress and inflammatory response. It indicates that Shen Yuan Dan could protect myocardium cells.