Effects of Ischemic Preconditioning and Postconditioning on Retinal Ganglion Cell Survival after Injury

Ligature of the ophthalmic vessels (LOV) was used as an animal model to study transient retinal ischemia/reperfusion in adult hamsters. Firstly, we quantified the loss of retinal ganglion cells (RGCs) and activation of microglia after 10 min, 30 min, 60 min or 120 min retinal ischemia at 7, 14 and 28 days post-ischemia. The results showed that after 10-min or 30-min retinal ischemia, the number of RGCs had no significant decrease compared to sham LOV group at 7 days. In the retinal ischemia 60 min group, there were 58% of the RGCs population remained alive at 7 days, 51% at 14 days and 44% at 28 days post-ischemia, respectively. In the retinal ischemia 120 min group, the number of RGCs was reduced to 22% at 7 days and 17% at 14 days, but cell death slowed down from 14 to 28 days. Meanwhile, the number of microglia was increased sharply at 7 days and decreased gradually from 7 to 28 days. At the same time, it was found that the loss of RGCs and activation of microglia in the ganglion cell layer at 7 days post-insult existed strong positive correlation.;Secondly, the effects of ischemic preconditioning (IPC) were proved to promote RGCs survival after axotomy or retinal ischemia 120 min. It was presented firstly that a 5 or 10 min brief IPC which performed 1 or 3 days prior to axotomy enhanced the RGCs survival at 7 days and 14 days post-axotomy. The number of HSP27-positive RGCs was significantly higher in the IPC plus axotomy subgroup compared with the sham-operated subgroup, while the percentage of HSP27-positive RGCs did not show significant difference between subgroups. For the IPC plus retinal ischemia 60 min group, both the number and the percentage of HSP27-positive RGCs had no significant difference between IPC and sham-operated subgroups. The number of HSP70-positive RGCs exhibited significant difference but not the percentage in IPC plus axotomy or retinal ischemia 60 min experimental groups. The thicknesses of the whole retina and GCL were similar to the normal value in the IPC plus ischemia 60 min subgroup, while in the sham-operated subgroup, these two values decreased significantly.;Consequently, the effect of remote ischemic postconditioning (RIPostC) was also explored to promote RGCs survival after axotomy. Four cycles of 10 min occlusion and 10 min release of the right femoral artery were initiated on animals at 10 min, 6 h or 24 h after axotomy. In the10 min group, the effect of RIPostC on promoting RGCs survival was significant at both 7 and 14 days post-injury. In the 6 h group, the survival of RGCs was more in the RIPostC treatment subgroup at 7 days, while there was no significant difference at 14 days post-axotomy. In the 24 h group, RGC survival was not significantly different at 7 days post-axotomy. Both the number and the percentage of HSP27-positive RGCs were significantly higher in the RIPostC treatment subgroup. The results of the induction of HSP70 only showed a priority in absolute number of the HSP70-positive RGCs in the RIPostC treatment subgroup.;In summary, the effect of IPC has been proved that it could protect RGCs against axotomy and retinal ischemia/reperfusion injury, in addition, the application of RIPostC also protected RGCs from axotomy. The proportion of HSP27-positive RGCs increased significantly in the process of RIPostC against axotomy, which may clue that the ability of axonal regeneration is stronger which induced by the RIPostC intervention. The upregulation of HSP27 might play a role in the neuroprotection of the RIPostC against axotomy. The expression of HSP70 maybe plays a little role in the neuroprotection of the IPC and RIPostC.