CLEC7A Knockdown Alleviates Ischemic Stroke by Inhibiting Pyroptosis and Microglia Activation

Background: Ischemic stroke (IS) is the leading cause of mortality worldwide. Herein, we aimed to identify novel biomarkers and explore the role of C-type lectin domain family 7 member A (CLEC7A) in IS. Methods...

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Main Authors: Wei Li, Xiaoli Feng, Manyu Zhang, Kangmeng Wang, Kailai Huang, Zhenqiang Zhao, Min Xia
Format: Article
Language:English
Published: IMR Press 2024-12-01
Series:Journal of Integrative Neuroscience
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Online Access:https://www.imrpress.com/journal/JIN/23/12/10.31083/j.jin2312219
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author Wei Li
Xiaoli Feng
Manyu Zhang
Kangmeng Wang
Kailai Huang
Zhenqiang Zhao
Min Xia
author_facet Wei Li
Xiaoli Feng
Manyu Zhang
Kangmeng Wang
Kailai Huang
Zhenqiang Zhao
Min Xia
author_sort Wei Li
collection DOAJ
description Background: Ischemic stroke (IS) is the leading cause of mortality worldwide. Herein, we aimed to identify novel biomarkers and explore the role of C-type lectin domain family 7 member A (CLEC7A) in IS. Methods: Differentially expressed genes (DEGs) were screened using the GSE106680, GSE97537, and GSE61616 datasets, and hub genes were identified through construction of protein-protein interaction networks. An IS model was established by middle cerebral artery occlusion and reperfusion (MCAO/R). Neural function was assessed using triphenyl tetrazolium chloride, hematoxylin-eosin, and terminal deoxynucleotidyl transferase-mediated nick-end labeling. A cell counting kit was used to detect cell viability following oxygen-glucose deprivation/reperfusion (OGD/R). Inflammatory factors were detected using enzyme-linked immunosorbent assay. The mRNA and protein expression levels were detected using reverse transcription-quantitative polymerase chain reaction and western blotting, respectively. Results: Fc fragment of Immunoglobulin G (IgG) receptor IIIa (FCGR3A), Fc fragment of Immunoglobulin E (IgE) receptor Ig (FCER1G), Complement component 5a receptor 1 (C5AR1), CLEC7A, Plasminogen activator, urokinase (PLAU), and C-C motif chemokine ligand 6 (CCL6) were identified as important hub genes, from which CLEC7A was selected as the primary subject of this study. The activation of microglia and pyroptosis were observed in MCAO/R model with increased levels of interleukin (IL)-1β, IL-18, tumor necrosis factor-α, and lactate dehydrogenase. CLEC7A knockdown was found to promote cell viability in BV2 cells and inhibiting pyroptosis in HT22 cells. CLEC7A knockdown in microglia also decreased infarct volume and neurological deficit scores, and alleviated injury and neuronal apoptosis in IS rats. CLEC7A knockdown inhibited pyroptosis and microglial activation in the MCAO/R model. A pyroptosis activator reversed the effect of CLEC7A knockdown on the viability of OGD/R-treated HT22 cells. Conclusion: CLEC7A is a promising biomarker of IS. CLEC7A knockdown alleviates IS by inhibiting pyroptosis and microglial activation.
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spelling doaj-art-178c5f1c4a02469a9eb327cc6637f8212025-08-20T02:51:15ZengIMR PressJournal of Integrative Neuroscience0219-63522024-12-01231221910.31083/j.jin2312219S0219-6352(24)00836-2CLEC7A Knockdown Alleviates Ischemic Stroke by Inhibiting Pyroptosis and Microglia ActivationWei Li0Xiaoli Feng1Manyu Zhang2Kangmeng Wang3Kailai Huang4Zhenqiang Zhao5Min Xia6Department of Neurology, First Affiliated Hospital of Hainan Medical University, 570102 Haikou, Hainan, ChinaDepartment of Neurology, First Affiliated Hospital of Hainan Medical University, 570102 Haikou, Hainan, ChinaDepartment of Neurology, First Affiliated Hospital of Hainan Medical University, 570102 Haikou, Hainan, ChinaDepartment of Neurology, First Affiliated Hospital of Hainan Medical University, 570102 Haikou, Hainan, ChinaDepartment of Neurology, First Affiliated Hospital of Hainan Medical University, 570102 Haikou, Hainan, ChinaDepartment of Neurology, First Affiliated Hospital of Hainan Medical University, 570102 Haikou, Hainan, ChinaDepartment of Neurology, Hainan West Central Hospital, 571799 Danzhou, Hainan, ChinaBackground: Ischemic stroke (IS) is the leading cause of mortality worldwide. Herein, we aimed to identify novel biomarkers and explore the role of C-type lectin domain family 7 member A (CLEC7A) in IS. Methods: Differentially expressed genes (DEGs) were screened using the GSE106680, GSE97537, and GSE61616 datasets, and hub genes were identified through construction of protein-protein interaction networks. An IS model was established by middle cerebral artery occlusion and reperfusion (MCAO/R). Neural function was assessed using triphenyl tetrazolium chloride, hematoxylin-eosin, and terminal deoxynucleotidyl transferase-mediated nick-end labeling. A cell counting kit was used to detect cell viability following oxygen-glucose deprivation/reperfusion (OGD/R). Inflammatory factors were detected using enzyme-linked immunosorbent assay. The mRNA and protein expression levels were detected using reverse transcription-quantitative polymerase chain reaction and western blotting, respectively. Results: Fc fragment of Immunoglobulin G (IgG) receptor IIIa (FCGR3A), Fc fragment of Immunoglobulin E (IgE) receptor Ig (FCER1G), Complement component 5a receptor 1 (C5AR1), CLEC7A, Plasminogen activator, urokinase (PLAU), and C-C motif chemokine ligand 6 (CCL6) were identified as important hub genes, from which CLEC7A was selected as the primary subject of this study. The activation of microglia and pyroptosis were observed in MCAO/R model with increased levels of interleukin (IL)-1β, IL-18, tumor necrosis factor-α, and lactate dehydrogenase. CLEC7A knockdown was found to promote cell viability in BV2 cells and inhibiting pyroptosis in HT22 cells. CLEC7A knockdown in microglia also decreased infarct volume and neurological deficit scores, and alleviated injury and neuronal apoptosis in IS rats. CLEC7A knockdown inhibited pyroptosis and microglial activation in the MCAO/R model. A pyroptosis activator reversed the effect of CLEC7A knockdown on the viability of OGD/R-treated HT22 cells. Conclusion: CLEC7A is a promising biomarker of IS. CLEC7A knockdown alleviates IS by inhibiting pyroptosis and microglial activation.https://www.imrpress.com/journal/JIN/23/12/10.31083/j.jin2312219ischemic strokeclec7adifferential expression genespyroptosismicroglia
spellingShingle Wei Li
Xiaoli Feng
Manyu Zhang
Kangmeng Wang
Kailai Huang
Zhenqiang Zhao
Min Xia
CLEC7A Knockdown Alleviates Ischemic Stroke by Inhibiting Pyroptosis and Microglia Activation
Journal of Integrative Neuroscience
ischemic stroke
clec7a
differential expression genes
pyroptosis
microglia
title CLEC7A Knockdown Alleviates Ischemic Stroke by Inhibiting Pyroptosis and Microglia Activation
title_full CLEC7A Knockdown Alleviates Ischemic Stroke by Inhibiting Pyroptosis and Microglia Activation
title_fullStr CLEC7A Knockdown Alleviates Ischemic Stroke by Inhibiting Pyroptosis and Microglia Activation
title_full_unstemmed CLEC7A Knockdown Alleviates Ischemic Stroke by Inhibiting Pyroptosis and Microglia Activation
title_short CLEC7A Knockdown Alleviates Ischemic Stroke by Inhibiting Pyroptosis and Microglia Activation
title_sort clec7a knockdown alleviates ischemic stroke by inhibiting pyroptosis and microglia activation
topic ischemic stroke
clec7a
differential expression genes
pyroptosis
microglia
url https://www.imrpress.com/journal/JIN/23/12/10.31083/j.jin2312219
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