Effects Of E64D On Apoptosis, Autophagy, Necrosis In Hippocampus With Hypoxic-ischemic Brain Damage

Objective:The aim of this investigation is to study the injury of hypoxic-ischemia through hippocampal apoptosis、autophagy、necrosis, and make sure whether E64D has protective effect or not.Methods:All neonatal rats (7days after birth) were divided into3groups:sham operation group、HIBD group、E64D group.1.Through the method of western blot, to test actived caspase-3、beclin-1and spectrin α Ⅱ (C-3) degradative products of neonatal rat’s hippocampus in0h、2h、6h、12h、24h、48h.2. Measure percentages of brain water content after hypoxic-ischemia and using E64D at48h.3. The pathological changes of left brain tissue were observed under light microscope.Results:1. Actived caspase-3expressed obviously in12h after in HIBD group, and got peak in24h, then went down. The protein level is generally higher than Sham operation group at12h、24h、48h (P<0.05). The actived caspase-3in E64D group expressed lower than that in HIBD group24h (P<0.05).2. Beclin-1in HIBD group expressed obviously in6h, and got peak in12h, then went down. The protein level was generally higher than that in Sham operation group (P<0.05). Beclin-1in E64D group at6h,12h,24h,48h were lower than that in HIBD group (P<0.05).3. The spectrin α Ⅱ degradative products in HIBD group increased from2h, and go on increasing until48h. Spectrin α Ⅱ degradative products in HIBD group were obviously higher than that in sham operation group at any time (P<0.05). Spectrin α Ⅱ degradative products in E64D group were generally lower than that in HIBD group at2h,6h,12h (P<0.05).4. At48h after hypoxic-ischemia, left brain water content increased. The percentages of brain water content in HIBD group were obviously higher than that in sham operation group (P<0.01). The percentages of brain water content in E64D group were not lower than that in HIBD group (P>0.05).Conclusions:1.Hypoxic-ischemia can strengthen apoptosis、autophagy and necrosis.2. E64D as one protease inhibitor can reduce apoptosis、 autophagy and necrosis caused by hypoxic-ischemia.3. By measuring the percentages of brain water content, I find that hypoxic-ischemia can cause left brain edema obviously.4. E64D is not effective to reduce brain edema.