Abstract
Apoptosis of hippocampal neurons is one of the mechanisms of hippocampal atrophy in posttraumatic stress disorder (PTSD), and it is also an important cause of memory impairment in PTSD patients. Endoplasmic reticulum stress (ERS) mediated by activated transcription factor 6α (ATF6α)/site 1 protease (S1P)/S2P is involved in cell apoptosis, but it is not clear whether it is involved in hippocampal neuron apoptosis caused by PTSD. A PTSD rat model was constructed by the single prolonged stress (SPS) method. The study was divided into three parts. Experiment 1 included the control group, SPS 1 d group, SPS 7 d group, and SPS 14 d group. Experiment 2 included the control group, SPS 7 d group, SPS 7 d + AEBSF group, and control + AEBSF group. (4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF) is an ATF6α pathway inhibitor). Experiment 3 included the control group, SPS 4 d group, SPS 4 d + AEBSF group, and control + AEBSF group. The protein and mRNA expression levels of ATF6α, glucose-regulated protein (GRP78), S1P, S2P, C/EBP homologous protein (CHOP), and caspase-12 in the hippocampus of PTSD rats were detected by immunohistochemistry, Western blotting and qRT-PCR. Apoptosis of hippocampal neurons was detected by TUNEL staining. In experiment 1, the protein and mRNA expression of ATF6α and GRP78 increased gradually in the SPS 1 d group and the SPS 7 d group but decreased in the SPS 14 d group (P < 0.01). In experiment 2, compared with that in the control group, the protein and mRNA expression of ATF6α, GRP78, S1P, S2P, CHOP, and caspase-12 and the apoptosis rate were significantly increased in the SPS 7 d group (P < 0.01). However, the protein and mRNA expression of ATF6α, GRP78, S1P, S2P, CHOP, and caspase-12 and the apoptosis rate were significantly decreased after AEBSF pretreatment (P < 0.01). In experiment 3, compared with that in the control group, the protein and mRNA expression of ATF6α, GRP78, S1P, S2P, CHOP, and caspase-12 and the apoptosis rate were increased in the SPS 14 d group (P < 0.05). However, the protein and mRNA expression of ATF6α, GRP78, S1P, S2P, CHOP, and caspase-12 and the apoptosis rate were decreased after AEBSF pretreatment (P < 0.05). SPS induced apoptosis of hippocampal neurons by activating ERS mediated by ATF6α, suggesting that ERS-induced apoptosis is involved in the occurrence of PTSD.
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The authors are grateful to all staff members of the China Medical University Experiment Center for their technical support.
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This work was supported by grants from the China National Natural Science Foundation (81571324) and the Science and Technology Project of Liao Ning Province, China (2017225011).
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Yanhao Xu and Yuxiu Shi contributed to the conception of the study and helped perform the analysis with constructive discussion. Liang han performed the experiment; contributed significantly to analysis and manuscript preparation; performed the data analyses, and wrote the manuscript.
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The animal experiments were performed according to protocols approved by the Ethical Committee of Animal Research at the China Medical University, and all efforts were made to minimize the number of animals used and their suffering. All study participants provided informed consent, and the study design was approved by the appropriate ethics review board.
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Han, L., Xu, Y. & Shi, Y. Molecular Mechanism of the ATF6α/S1P/S2P Signaling Pathway in Hippocampal Neuronal Apoptosis in SPS Rats. J Mol Neurosci 71, 2487–2499 (2021). https://doi.org/10.1007/s12031-021-01823-9
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DOI: https://doi.org/10.1007/s12031-021-01823-9