| Objective The study was designed to investigate the action of Epithelial-specific ETS-1 (ESE1) by establishing the model of cerebral ischemia injury in vivo and in vitro. We learnt the expression variations of ESE1 in the hippocampal CA1 region following cerebral ischemia/ reperfusion(I/R) to discover the underlying mechanism on neuronal apoptosis.Methods A modified Pulsinelli four vessel occlusion method was used to make model of global cerebral ischemia and reperfusion in adult rats. The 36 healthy male Sprague-Dawley rats were randomly divided into 6 groups:I/R 6h, I/R 12h, I/R 24h, I/R 48h, I/R 72h, and the control group received sham operation (without ischemia-reperfusion injury). Western blot was used to detect the expression changes of ESE1, phosphorylation of p65 (activation index of NF-kB) and active caspase3 (apoptosis markers) in the hippocampus after cerebral ischemia and reperfusion. Again, immunohistochemistry analysis was used to investigate the expression of ESE1. Next, double immunofluorescence staining was used to clear the cellular distribution of ESE1 and the relationship with active caspase3 after transient global cerebral ischemia. Furthermore, cobalt chloride (CoCl2) stimulated PC 12 cells to establish chemical hypoxia model. Western blot was applied to detect the expression variations of ESE1 and p-p65 in the process of PC 12 cells apoptosis. Finally, ESE1 was knocked down with it’s special RNA interference carrier to discover that if ESE1 has an influence on active caspase3 and p-p65, which was detected by western blot.Results (1)In vivo, western blot analysis showed that the expression of ESE1 and p-p65 was lower in the sham group, began to increase after I/R 6h (P<0.05), reached the peak value after I/R 24h (P<0.05),then decreased but still higher than the sham group (P<0.05). In the same way, the expression of active caspase3 was lower in the sham group, began to increase after I/R 6h(P<0.05), reached the peak value after I/R 48h(P<0.05),then decreased but still higher than the sham group (P<0.05). Immunohistochemistry analysis showed that the expression of ESE1 in the hippocampal CA1 region after I/R 24h was obviously increased than the sham group. Double immunofluorescence staining indicated that ESE1 and NeuN were mostly co-located. Besides, we found that ESE1 was also co-expressed with active caspase3. (2)In vitro, the expression of ESE1、p-p65 and active caspase3 began to increase after 1h of CoCl2 stimulation in PC12 cells (P<0.05), reached the peak value after 12h of stimulation (P <0.05),then decreased but still higher than the sham group (P<0.05).When interventing the expression of ESE1 in PC 12 cells, we discoerd that the expression of p-p65 and active caspase3 were decreased distinctly after CoC12 stimulation (P<0.05)Conclusions The expression of ESE1 was elevated in the hippocampal CA1 region after global cerebral ischemia and reperfusion, and it may involves in the process of neuron apoptosis via activating the NF-κB signaling pathways. By interfering with the expression of ESE1 could inhibit the neuronal apoptosis, and Play an anti-apoptotic characteristic in global cerebral ischemia and reperfusion. This study provides a new target for cerebral ischemia reperfusion injury. |
