Synergistic protective effects of humanin and necrostatin-1 on hypoxia and ischemia/reperfusion injury

• DOI: 10.1016/j.brainres.2010.07.080
• Publication Date: 2010-08-03T08:42:52Z
• Authors: Xingshun Xu; Kao-Wei Chua; Chu C. Chua; Chun-Feng Liu; Ronald C. Hamdy; Balvin H.L. Chua
• Keywords: 0301 basic medicine; Indoles; humanin; necroptosis; 610; Apoptosis; Mice; 03 medical and health sciences; Internal Medicine; Animals; Cells, Cultured; apoptosis; Imidazoles; Intracellular Signaling Peptides and Proteins; Drug Synergism; 3. Good health; Disease Models, Animal; Neuroprotective Agents; Treatment Outcome; Reperfusion Injury; Hypoxia-Ischemia, Brain; neuroprotection; Drug Therapy, Combination; Necrostatin-1; Signal Transduction

Since several different pathways are involved in cerebral ischemia/reperfusion injury, combination therapy rather than monotherapy may be required for efficient neuroprotection. In this study, we examined the protective effects of an apoptosis inhibitor Gly(14)-humanin (HNG) and a necroptosis inhibitor necrostatin-1 (Nec-1) on hypoxia/ischemia/reperfusion injury. Cultured mouse primary cortical neurons were incubated with Nec-1, HNG or both in a hypoxia chamber for 60 min. Cell viability was determined by MTS assay at 24h after oxygen-glucose deprivation (OGD) treatment. Mice underwent middle cerebral artery occlusion for 75 min followed by 24h reperfusion. Mice were administered HNG and/or Nec-1 (i.c.v.) at 4h after reperfusion. Neurological deficits were evaluated and the cerebral infarct volume was determined by TTC staining. Nec-1 or HNG alone had protective effects on OGD-induced cell death. Combined treatment with Nec-1 and HNG resulted in more neuroprotection than Nec-1 or HNG alone. Treatment with HNG or Nec-1 reduced cerebral infarct volume from 59.3 ± 2.6% to 47.0 ± 2.3% and 47.1 ± 1.5%, respectively. Combined treatment with HNG and Nec-1 improved neurological scores and decreased infarct volume to 38.6 ± 1.5%. In summary, we demonstrated that the combination treatment of HNG and Nec-1 conferred synergistic neuroprotection on hypoxia/ischemia/reperfusion injury in vitro and in vivo. These findings provide a novel therapeutic strategy for the treatment of stroke by combining anti-apoptosis and anti-necroptosis therapy.