Background and Purpose Although the receptor-interacting protein l kinase (RIPIK)-regulated necroptosis can be evoked by cerebral ischemia, its effects and mechanisms in mediating neuronal and astrocytic cell injury remain poorly understood. This study evaluates the contribution of RIPIK to ischemic stroke-induced neuronal and astrocytic cell injury,associating with autophagic-lysosomal pathway activation. Methods Oxygen and glucose deprivation (OGD) was induced in neurons or astrocytes. Permanent middle cerebral artery occlusion (pMCAO) was established in rats or mice. Lentivirally-delivered shRNA against RIPIK (shRNA RIPIK) or a necroptosis inhibitor Nec-1 was used to inhibit RIPIK. Brain infarct,behavioral test, neuronal or astrocytic cell death, RIPIK, RIPIK-RIP3K complex formation,autophagic-lysosomal related proteins and lysosomal membrane stability were assessed. Results RIPIK was much richer in astrocytes than that in neurons. pMCAO or OGD led to an increase in RIPIK and RIPIK-RIP3K complex formation. Icv administration of shRNA RIPIK or Nec-1 reduced infarct volume, improved neurological deficits, increased MAP2 and GFAP levels, and attenuated the necrotic neuronal or astrocytic cell death in the ischemic cortex. shRNA RIPIK or Nec-l treatment decreased OGD-induced LDH leakage from neurons or astrocytes.Simultaneously, inhibition of RIPIK decreased RIPIK-RIP3K complex formation, LC311 and active cathepsin B levels and lysosomal membrane permeability (LMP) in vivo and/or in vitro.Furthermore, a combination of Nec-I and an inhibitor of autophagy or cathepsin B produced synergistic protective effect on neuronal or astrocytic cell death. Conclusions RIPIK-mediated necroptosis may play a significant role in ischemia-induced neuronal and astrocytic cell death,associating with activation of the autophagic-lysosomal pathway.