Cargando…
RIP1 and RIP3 mediate hemin-induced cell death in HT22 hippocampal neuronal cells
BACKGROUND: Intracerebral hemorrhage (ICH) is a devastating neurological injury associated with significant mortality. Necroptosis is a newly identified type of programmed necrosis initiated by the activation of tumor necrosis factor alpha. Evidences had demonstrated the importance of necroptosis in...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Dove Medical Press
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6247969/ https://www.ncbi.nlm.nih.gov/pubmed/30532542 http://dx.doi.org/10.2147/NDT.S181074 |
_version_ | 1783372574305026048 |
---|---|
author | Su, Xingfen Wang, Handong Lin, Yuanxiang Chen, Fuxiang |
author_facet | Su, Xingfen Wang, Handong Lin, Yuanxiang Chen, Fuxiang |
author_sort | Su, Xingfen |
collection | PubMed |
description | BACKGROUND: Intracerebral hemorrhage (ICH) is a devastating neurological injury associated with significant mortality. Necroptosis is a newly identified type of programmed necrosis initiated by the activation of tumor necrosis factor alpha. Evidences had demonstrated the importance of necroptosis in neuronal cell death. Necrostatin-1 is a specific inhibitor of necroptosis. The present study was carried out to explore whether RIP1/RIP3 pathways participate in hemin induced cell death in HT-22 hippocampal neuronal cells and investigate the potential neuroprotection of necrostatin-1 in hemin induced cell death in HT-22. METHODS: First, different concentrations of hemin (0, 25, 50, 100 μmol/L) were added to HT-22 cells. Propidium iodide (PI) positive cells and cell viability were measured at 24 hours after hemin treatment. Then, necrostatin-1, pan-caspase inhibitor Benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (z-VAD-fmk) and reactive oxygen species (ROS) scavenger butylated hydroxyanisole (BHA) were applied to hemin-treated HT-22 cells. PI positive cells and cell viability were measured at 24 hours after hemin treatment. MitoSox Red was used to indicate ROS level. Last, the effect of RIP3 in hemin induced HT-22 cell death was explored through RIP3 knockdown using siRNA. PI positive cells, cell viability and ROS lever were measured at 24 h after hemin treatment. RESULTS: Hemin could induce a dose dependent cell death in HT22 neural cells. RIP1 specific inhibitor necrostatin-1 significantly inhibited cell death induced by hemin in HT-22 cells, greatly reducing PI positive cells, dramatically improving cell viability and decreasing ROS accumulation. BHA could significantly inhibit PI positive cells induced by hemin in HT-22 cells. Furthermore, silencing of RIP3 using siRNA attenuated hemin induced cell death in HT-22 cells, greatly reducing PI positive cells, dramatically improving cell viability and decreasing ROS accumulation. CONCLUSION: These data revealed that RIP1/RIP3 might mediate hemin induced cell death in HT-22 cells, and necrostatin-1 played a neuroprotection role in hemin induced cell death in HT-22. RIP1 and RIP3 might represent novel therapeutic targets for ICH. |
format | Online Article Text |
id | pubmed-6247969 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Dove Medical Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-62479692018-12-07 RIP1 and RIP3 mediate hemin-induced cell death in HT22 hippocampal neuronal cells Su, Xingfen Wang, Handong Lin, Yuanxiang Chen, Fuxiang Neuropsychiatr Dis Treat Original Research BACKGROUND: Intracerebral hemorrhage (ICH) is a devastating neurological injury associated with significant mortality. Necroptosis is a newly identified type of programmed necrosis initiated by the activation of tumor necrosis factor alpha. Evidences had demonstrated the importance of necroptosis in neuronal cell death. Necrostatin-1 is a specific inhibitor of necroptosis. The present study was carried out to explore whether RIP1/RIP3 pathways participate in hemin induced cell death in HT-22 hippocampal neuronal cells and investigate the potential neuroprotection of necrostatin-1 in hemin induced cell death in HT-22. METHODS: First, different concentrations of hemin (0, 25, 50, 100 μmol/L) were added to HT-22 cells. Propidium iodide (PI) positive cells and cell viability were measured at 24 hours after hemin treatment. Then, necrostatin-1, pan-caspase inhibitor Benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (z-VAD-fmk) and reactive oxygen species (ROS) scavenger butylated hydroxyanisole (BHA) were applied to hemin-treated HT-22 cells. PI positive cells and cell viability were measured at 24 hours after hemin treatment. MitoSox Red was used to indicate ROS level. Last, the effect of RIP3 in hemin induced HT-22 cell death was explored through RIP3 knockdown using siRNA. PI positive cells, cell viability and ROS lever were measured at 24 h after hemin treatment. RESULTS: Hemin could induce a dose dependent cell death in HT22 neural cells. RIP1 specific inhibitor necrostatin-1 significantly inhibited cell death induced by hemin in HT-22 cells, greatly reducing PI positive cells, dramatically improving cell viability and decreasing ROS accumulation. BHA could significantly inhibit PI positive cells induced by hemin in HT-22 cells. Furthermore, silencing of RIP3 using siRNA attenuated hemin induced cell death in HT-22 cells, greatly reducing PI positive cells, dramatically improving cell viability and decreasing ROS accumulation. CONCLUSION: These data revealed that RIP1/RIP3 might mediate hemin induced cell death in HT-22 cells, and necrostatin-1 played a neuroprotection role in hemin induced cell death in HT-22. RIP1 and RIP3 might represent novel therapeutic targets for ICH. Dove Medical Press 2018-11-15 /pmc/articles/PMC6247969/ /pubmed/30532542 http://dx.doi.org/10.2147/NDT.S181074 Text en © 2018 Su et al. This work is published and licensed by Dove Medical Press Limited The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. |
spellingShingle | Original Research Su, Xingfen Wang, Handong Lin, Yuanxiang Chen, Fuxiang RIP1 and RIP3 mediate hemin-induced cell death in HT22 hippocampal neuronal cells |
title | RIP1 and RIP3 mediate hemin-induced cell death in HT22 hippocampal neuronal cells |
title_full | RIP1 and RIP3 mediate hemin-induced cell death in HT22 hippocampal neuronal cells |
title_fullStr | RIP1 and RIP3 mediate hemin-induced cell death in HT22 hippocampal neuronal cells |
title_full_unstemmed | RIP1 and RIP3 mediate hemin-induced cell death in HT22 hippocampal neuronal cells |
title_short | RIP1 and RIP3 mediate hemin-induced cell death in HT22 hippocampal neuronal cells |
title_sort | rip1 and rip3 mediate hemin-induced cell death in ht22 hippocampal neuronal cells |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6247969/ https://www.ncbi.nlm.nih.gov/pubmed/30532542 http://dx.doi.org/10.2147/NDT.S181074 |
work_keys_str_mv | AT suxingfen rip1andrip3mediatehemininducedcelldeathinht22hippocampalneuronalcells AT wanghandong rip1andrip3mediatehemininducedcelldeathinht22hippocampalneuronalcells AT linyuanxiang rip1andrip3mediatehemininducedcelldeathinht22hippocampalneuronalcells AT chenfuxiang rip1andrip3mediatehemininducedcelldeathinht22hippocampalneuronalcells |