Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation
Transient receptor potential melastatin 2 (TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mechanism underlying this pro...
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Wolters Kluwer Medknow Publications
2020-01-01
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doaj-4ad8c07af5914c60b8c7308537daf5992020-11-25T03:18:08ZengWolters Kluwer Medknow PublicationsNeural Regeneration Research1673-53742020-01-0115112154216110.4103/1673-5374.282271Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivationTao PanQiu-Jiao ZhuLi-Xiao XuXin DingJian-Qin LiBin SunJun HuaXing FengTransient receptor potential melastatin 2 (TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mechanism underlying this process involves. This study investigated the protective effect of reducing TRPM2 expression on pheochromocytoma (PC12) cells injured by oxygen-glucose deprivation (OGD). PC12 cells were transfected with plasmid encoding TRPM2 shRNAS, then subjected to OGD by incubation in glucose-free medium under hypoxic conditions for 8 hours, after which the cells were allowed to reoxygenate for 24 hours. Apoptotic cells, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels were detected using flow cytometry. The relative expression of C-X-C motif chemokine ligand 2 (CXCL2), NACHT, LRR, and PYD domain–containing protein 3 (NALP3), and caspase-1 were detected using fluorescence-based quantitative reverse transcription-polymerase chain reaction and western blotting. The rates of apoptosis, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels in the TRPM2-shRNA + OGD group were lower than those observed in the OGD group. Taken together, these results suggest that TRPM2 knockdown reduces OGD-induced neuronal injury, potentially by inhibiting apoptosis and reducing oxidative stress levels, mitochondrial membrane potentials, intracellular calcium concentrations, and NLRP3 inflammasome activation.http://www.nrronline.org/article.asp?issn=1673-5374;year=2020;volume=15;issue=11;spage=2154;epage=2161;aulast=Panapoptosis; calcium; caspase-1; nlrp3; mitochondrial impairment; oxidative stress; oxygen-glucose deprivation; pc12; shrna; trpm2 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Tao Pan Qiu-Jiao Zhu Li-Xiao Xu Xin Ding Jian-Qin Li Bin Sun Jun Hua Xing Feng |
spellingShingle |
Tao Pan Qiu-Jiao Zhu Li-Xiao Xu Xin Ding Jian-Qin Li Bin Sun Jun Hua Xing Feng Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation Neural Regeneration Research apoptosis; calcium; caspase-1; nlrp3; mitochondrial impairment; oxidative stress; oxygen-glucose deprivation; pc12; shrna; trpm2 |
author_facet |
Tao Pan Qiu-Jiao Zhu Li-Xiao Xu Xin Ding Jian-Qin Li Bin Sun Jun Hua Xing Feng |
author_sort |
Tao Pan |
title |
Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation |
title_short |
Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation |
title_full |
Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation |
title_fullStr |
Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation |
title_full_unstemmed |
Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation |
title_sort |
knocking down trpm2 expression reduces cell injury and nlrp3 inflammasome activation in pc12 cells subjected to oxygen-glucose deprivation |
publisher |
Wolters Kluwer Medknow Publications |
series |
Neural Regeneration Research |
issn |
1673-5374 |
publishDate |
2020-01-01 |
description |
Transient receptor potential melastatin 2 (TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mechanism underlying this process involves. This study investigated the protective effect of reducing TRPM2 expression on pheochromocytoma (PC12) cells injured by oxygen-glucose deprivation (OGD). PC12 cells were transfected with plasmid encoding TRPM2 shRNAS, then subjected to OGD by incubation in glucose-free medium under hypoxic conditions for 8 hours, after which the cells were allowed to reoxygenate for 24 hours. Apoptotic cells, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels were detected using flow cytometry. The relative expression of C-X-C motif chemokine ligand 2 (CXCL2), NACHT, LRR, and PYD domain–containing protein 3 (NALP3), and caspase-1 were detected using fluorescence-based quantitative reverse transcription-polymerase chain reaction and western blotting. The rates of apoptosis, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels in the TRPM2-shRNA + OGD group were lower than those observed in the OGD group. Taken together, these results suggest that TRPM2 knockdown reduces OGD-induced neuronal injury, potentially by inhibiting apoptosis and reducing oxidative stress levels, mitochondrial membrane potentials, intracellular calcium concentrations, and NLRP3 inflammasome activation. |
topic |
apoptosis; calcium; caspase-1; nlrp3; mitochondrial impairment; oxidative stress; oxygen-glucose deprivation; pc12; shrna; trpm2 |
url |
http://www.nrronline.org/article.asp?issn=1673-5374;year=2020;volume=15;issue=11;spage=2154;epage=2161;aulast=Pan |
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