Autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in rats
Objective To determine the effect of suppressing autophagy on the recovery of early brain injury (EBI) after subarachnoid hemorrhage (SAH) in rats, and to reveal the mechanism in which autophagy promotes neuronal ferroptosis and then affects EBI. Methods SD rats were randomly divided into control gr...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | zho |
| Published: |
Editorial Office of Journal of Third Military Medical University
2019-08-01
|
| Series: | Di-san junyi daxue xuebao |
| Subjects: | |
| Online Access: | http://aammt.tmmu.edu.cn/Upload/rhtml/201902124.htm |
| _version_ | 1830358983651622912 |
|---|---|
| author | LIANG Yidan QIN Wang HUANG Hao XIN Yuanjun ZHAO Jun |
| author_facet | LIANG Yidan QIN Wang HUANG Hao XIN Yuanjun ZHAO Jun |
| author_sort | LIANG Yidan |
| collection | DOAJ |
| description | Objective To determine the effect of suppressing autophagy on the recovery of early brain injury (EBI) after subarachnoid hemorrhage (SAH) in rats, and to reveal the mechanism in which autophagy promotes neuronal ferroptosis and then affects EBI. Methods SD rats were randomly divided into control group, SAH group, SAH+LV-ATG5 group and SAH+LV-Scramble group, with 12 animals in each group. Rat model of SAH were established by intravascular puncture. Lentiviral vector LV-ATG5 targeting autophagy-related gene 5 (ATG5) was constructed to inhibit autophagy. Immunofluorescence assay was used to detect the expression and location of ATG5, ferritin heavy polypeptide 1 (FTH1) and glutathione peroxidase 4 (GPX4) in neurons in 24 h after SAH establishment. The mRNA expression of ATG5 was detected by PCR, and the protein expression of ATG5, LC3Ⅱ/Ⅰ, FTH1 and GPX4 were detected by Western blotting. The changes of neuronal morphology and function were detected by Nissl staining, and neurological score and brain water content were also detected. Intracellular iron deposition, iron content, malonaldehyde (MDA) content and GPX4 activity were measured to evaluate the ferroptosis. Results Immunofluorescence assay showed that ATG5, FTH1 and GPX4 were widely expressed in the cytoplasm of cortical neurons after SAH. The results of PCR and Western blotting indicated that the expression of ATG5 at mRNA and protein levels, and LC3Ⅱ/Ⅰ at protein level was significantly decreased (P < 0.05), and autophagy was successfully inhibited. When autophagy was inhibited, Nissl staining showed improvements on neuronal morphology and function, neurological function score was improved (P < 0.05), and brain water content was decreased (P < 0.05). Western blot assay indicated that the expression levels of FTH1 and GPX4 were decreased after SAH (P < 0.05), however, the levels were increased when autophagy was inhibited (P < 0.05). Furthermore, autophagy inhibition also reduced iron deposition, and cellular labile iron and MDA contents (P < 0.05), and also elevated GSH content and GPX4 activity (P < 0.05). Conclusion Autophagy may promote neuronal ferroptosis by degrading ferritin, result in cellular labile iron increase, and finally early brain injury. |
| first_indexed | 2024-12-20T02:50:21Z |
| format | Article |
| id | doaj.art-28bdd61dc6ec4833b1e9d499176b9c2b |
| institution | Directory Open Access Journal |
| issn | 1000-5404 |
| language | zho |
| last_indexed | 2024-12-20T02:50:21Z |
| publishDate | 2019-08-01 |
| publisher | Editorial Office of Journal of Third Military Medical University |
| record_format | Article |
| series | Di-san junyi daxue xuebao |
| spelling | doaj.art-28bdd61dc6ec4833b1e9d499176b9c2b2022-12-21T19:56:02ZzhoEditorial Office of Journal of Third Military Medical UniversityDi-san junyi daxue xuebao1000-54042019-08-0141151407141410.16016/j.1000-5404.201902124Autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in ratsLIANG Yidan0QIN Wang1HUANG Hao2XIN Yuanjun3ZHAO Jun4Department of Neurosurgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, ChinaDepartment of Neurosurgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, ChinaDepartment of Neurosurgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, ChinaDepartment of Neurosurgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, ChinaDepartment of Neurosurgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, ChinaObjective To determine the effect of suppressing autophagy on the recovery of early brain injury (EBI) after subarachnoid hemorrhage (SAH) in rats, and to reveal the mechanism in which autophagy promotes neuronal ferroptosis and then affects EBI. Methods SD rats were randomly divided into control group, SAH group, SAH+LV-ATG5 group and SAH+LV-Scramble group, with 12 animals in each group. Rat model of SAH were established by intravascular puncture. Lentiviral vector LV-ATG5 targeting autophagy-related gene 5 (ATG5) was constructed to inhibit autophagy. Immunofluorescence assay was used to detect the expression and location of ATG5, ferritin heavy polypeptide 1 (FTH1) and glutathione peroxidase 4 (GPX4) in neurons in 24 h after SAH establishment. The mRNA expression of ATG5 was detected by PCR, and the protein expression of ATG5, LC3Ⅱ/Ⅰ, FTH1 and GPX4 were detected by Western blotting. The changes of neuronal morphology and function were detected by Nissl staining, and neurological score and brain water content were also detected. Intracellular iron deposition, iron content, malonaldehyde (MDA) content and GPX4 activity were measured to evaluate the ferroptosis. Results Immunofluorescence assay showed that ATG5, FTH1 and GPX4 were widely expressed in the cytoplasm of cortical neurons after SAH. The results of PCR and Western blotting indicated that the expression of ATG5 at mRNA and protein levels, and LC3Ⅱ/Ⅰ at protein level was significantly decreased (P < 0.05), and autophagy was successfully inhibited. When autophagy was inhibited, Nissl staining showed improvements on neuronal morphology and function, neurological function score was improved (P < 0.05), and brain water content was decreased (P < 0.05). Western blot assay indicated that the expression levels of FTH1 and GPX4 were decreased after SAH (P < 0.05), however, the levels were increased when autophagy was inhibited (P < 0.05). Furthermore, autophagy inhibition also reduced iron deposition, and cellular labile iron and MDA contents (P < 0.05), and also elevated GSH content and GPX4 activity (P < 0.05). Conclusion Autophagy may promote neuronal ferroptosis by degrading ferritin, result in cellular labile iron increase, and finally early brain injury.http://aammt.tmmu.edu.cn/Upload/rhtml/201902124.htmsubarachnoid hemorrhageearly brain injuryautophagyferroptosisferritin |
| spellingShingle | LIANG Yidan QIN Wang HUANG Hao XIN Yuanjun ZHAO Jun Autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in rats Di-san junyi daxue xuebao subarachnoid hemorrhage early brain injury autophagy ferroptosis ferritin |
| title | Autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in rats |
| title_full | Autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in rats |
| title_fullStr | Autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in rats |
| title_full_unstemmed | Autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in rats |
| title_short | Autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in rats |
| title_sort | autophagy promotes neuronal ferroptosis by degrading ferritin and induces early brain injury after subarachnoid hemorrhage in rats |
| topic | subarachnoid hemorrhage early brain injury autophagy ferroptosis ferritin |
| url | http://aammt.tmmu.edu.cn/Upload/rhtml/201902124.htm |
| work_keys_str_mv | AT liangyidan autophagypromotesneuronalferroptosisbydegradingferritinandinducesearlybraininjuryaftersubarachnoidhemorrhageinrats AT qinwang autophagypromotesneuronalferroptosisbydegradingferritinandinducesearlybraininjuryaftersubarachnoidhemorrhageinrats AT huanghao autophagypromotesneuronalferroptosisbydegradingferritinandinducesearlybraininjuryaftersubarachnoidhemorrhageinrats AT xinyuanjun autophagypromotesneuronalferroptosisbydegradingferritinandinducesearlybraininjuryaftersubarachnoidhemorrhageinrats AT zhaojun autophagypromotesneuronalferroptosisbydegradingferritinandinducesearlybraininjuryaftersubarachnoidhemorrhageinrats |