Receptor‐interacting protein 3‐phosphorylated Ca2+/calmodulin‐dependent protein kinase II and mixed lineage kinase domain‐like protein mediate intracerebral hemorrhage‐induced neuronal necroptosis

• DOI: 10.1111/jnc.15731
• Publication Date: 2022-11-25T07:02:24Z
• Authors: Guiqiang Yuan; Cheng Cao; Demao Cao; Bing Li; Xiang Li; Haiying Li; Haitao Shen; Zhong Wang; Gang Chen
• Keywords: Neurons; Hematoma; 0303 health sciences; Tumor Necrosis Factor-alpha; Apoptosis; Rats; Necrosis; 03 medical and health sciences; Receptor-Interacting Protein Serine-Threonine Kinases; Necroptosis; Animals; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Protein Kinases; Cerebral Hemorrhage

Necroptosis-mediated cell death is an important mechanism in intracerebral hemorrhage (ICH)-induced secondary brain injury (SBI). Our previous study has demonstrated that receptor-interacting protein 1 (RIP1) mediated necroptosis SBI after ICH. However, further mechanisms, such as the roles of 3 (RIP3), mixed lineage kinase domain-like (MLKL), and Ca2+ /calmodulin-dependent II (CaMK II), remain unclear. We hypothesized RIP3, MLKL, CaMK might participate ICH, including their phosphorylation. The ICH model was induced by autologous blood injection. First, we found activation tissues surrounding hematoma (propidium iodide staining). Meanwhile, phosphorylation expression were differently up-regulated (western blotting immunofluorescent specific inhibitors could suppress (GSK'872 for necrosulfonamide KN-93 II). concrete interactions RIP3-MLKL/RIP3-CaMK also both decreased intervention (co-immunoprecipitation). Then conducted short-/long-term neurobehavioral tests, rats with inhibition mostly had better performance. less blood-brain barrier (BBB) injury, neuron loss (Nissl staining) groups, which supported tests. Besides, oxidative stress inflammation alleviated intervention, significant reactive oxygen species (ROS), tumor necrosis factor (TNF)-α, lactate dehydrogenase (LDH), Iba1, GFAP hematoma. These results confirmed RIP3-phosphorylated MLKL ICH-induced provide potential targets treatment patients.