Objective To observe the influence of Bcl-2 inhibitor (TW-37) on the alleviation action of astragalus injection (AI) on apoptosis of hippocamal neurons of rats induced by hypoxia/hypoglycemia and reoxygenation (H/R). Methods The hippocampal neurons were isolated from fetal rat and cultured for eight days. The cells were randomly divided into six groups:normal control group, H/R group, AI group, AI solution group (sterile deionized water), TW-37 group, TW-37 with AI group. All groups were tested after 0 h, 0.5 h, 2 h, 6 h, 24 h, 72 h and 120 h since they were treated with hypoglycemia and reoxygenation after deprived of oxygen and glucose for 30 min except normal control group. The activity of hippocampal neurons were detected with MTT,The expression level of Caspase-3 was measured by western blotting, the expression of mRNA of Caspase-3 was tested by RT-PCR. Results Compared with normal control group, aside from 0 h group, the activity of neurons decreased obviously (P<0.05), moreover, the protein and mRNA expressions of Caspase-3 were enhanced significantly (P<0.05) in H/R groups. Compared with H/R group, the activity of neurons increased obviously in astragalus group (P<0.05), and the expressions of Caspase-3 protein and the expression of Caspase-3 mRNA in the AI groups decreased significantly (P<0.05) besides 0 h group; but there was no significant difference in both AI solution group、TW-37+H/R and TW-37+AI+H/R group (P>0.05). Conclusions Astragalus injection to increase the neurons activity and decrease the expression level of Caspase-3 in hippocampal neurons of rat by activating the effect of anti-apoptosis of Bcl-2. Bcl-2 is one of the targets exerting the inhibitory action of astragalus injection on apoptosis of hippocamal neurons of rats induced by hypoxia/hypoglycemia and reoxygenation.