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Objective To investigate the inhibitory effect and mechanism of sophoridine(Sop) on cell injury caused by hypoxia in rat myocardial cell line H9c2. Methods H9c2 cells were exposed to 1% O2 and cultured with different concentrations (1, 4 and 16 mmol/L) of Sop for 48 h. Cell viability was measured by MTT assay. The release level of LDH and the content of MDA were determined by the kit methods. Cell apoptosis was detected by flow cytometry. The protein expression of cleaved-caspase-3, p-PI3K and p-AKT was detected by Western blot. Results Sop alleviated the cell viability, LDH release, MDA content and hypoxia-induced apoptosis of H9c2 cells in a concentration-dependent manner (P＜0.05, P＜0.01, P＜0.001). Sop decreased the cleaved-caspase-3 expression and increased the ratio of p-PI3K/PI3K and p-AKT/AKT in a concentration-dependent manner (P＜0.05, P＜0.01, P＜0.001). Conclusions Sop reduces hypoxia-induced cell injury and has a protective effect in vitro model of H9c2 cells, the potential mechanism is enhancing the stimulation activity of PI3K/AKT signal.