2. Academic Validation
  3. Autophagy inhibitors 3-MA and BAF may attenuate hippocampal neuronal necroptosis after global cerebral ischemia-reperfusion injury in male rats by inhibiting the interaction of the RIP3/AIF/CypA complex

Autophagy inhibitors 3-MA and BAF may attenuate hippocampal neuronal necroptosis after global cerebral ischemia-reperfusion injury in male rats by inhibiting the interaction of the RIP3/AIF/CypA complex

  • J Neurosci Res. 2024 Feb;102(2):e25301. doi: 10.1002/jnr.25301.
Chen Zhang 1,  Renhui Liu 1,  Mengmeng Chen 1,  Yang Xu 2 3,  Xiaoqin Jin 4,  Bing Shen 5 6,  Jingye Wang 1
Affiliations

Affiliations

  • 1 Department of Neurology, First Affiliated Hospital of Anhui Medical University, Hefei, China.
  • 2 Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institutes, First Affiliated Hospital of Wannan Medical College, Wuhu, China.
  • 3 Department of Neurology, First Affiliated Hospital of Wannan Medical College, Wuhu, China.
  • 4 Academy of Chinese Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, China.
  • 5 School of Basic Medical Sciences, Anhui Medical University, Hefei, China.
  • 6 Dr. Neher's Biophysics Laboratory for Innovative Drug Discovery, State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China.
PMID: 38361405 DOI: 10.1002/jnr.25301
Abstract

Our previous study found that receptor interacting protein 3 (RIP3) and apoptosis-inducing factor (AIF) were involved in neuronal programmed necrosis during global cerebral ischemia-reperfusion (I/R) injury. Here, we further studied its downstream mechanisms and the role of the Autophagy inhibitors 3-methyladenine (3-MA) and bafilomycin A1 (BAF). A 20-min global cerebral I/R injury model was constructed using the 4-vessel occlusion (4-VO) method in male rats. 3-MA and BAF were injected into the lateral ventricle 1 h before ischemia. Spatial and activation changes of proteins were detected by immunofluorescence (IF), and protein interaction was determined by immunoprecipitation (IP). The phosphorylation of H2AX (γ-H2AX) and activation of Mixed Lineage Kinase domain-like protein (p-MLKL) occurred as early as 6 h after reperfusion. RIP3, AIF, and cyclophilin A (CypA) in the neurons after I/R injury were spatially overlapped around and within the nucleus and combined with each other after reperfusion. The survival rate of CA1 neurons in the 3-MA and BAF groups was significantly higher than that in the I/R group. Autophagy was activated significantly after I/R injury, which was partially inhibited by 3-MA and BAF. Pretreatment with both 3-MA and BAF almost completely inhibited nuclear translocation, spatial overlap, and combination of RIP3, AIF, and CypA proteins. These findings suggest that after global cerebral I/R injury, RIP3, AIF, and CypA translocated into the nuclei and formed the DNA degradation complex RIP3/AIF/CypA in hippocampal CA1 neurons. Pretreatment with Autophagy inhibitors could reduce neuronal Necroptosis by preventing the formation of the RIP3/AIF/CypA complex and its nuclear translocation.

Keywords

apoptosis inducing factor; autophagy; cyclophilin A; ischemia-reperfusion injury; necroptosis; receptor-interacting protein 3.

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