TLR4 Signaling in MPP+-Induced Activation of BV-2 Cells

Aims. This work was conducted to establish an in vitro Parkinson’s disease (PD) model by exposing BV-2 cells to 1-methyl-4-phenylpyridinium (MPP+) and exploring the roles of TLR2/TLR4/TLR9 in inflammatory responses to MPP+. Methods/Results. MTT assay showed that cell viability of BV-2 cells was 84.7...

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Main Authors: Peng Zhou, Ruihui Weng, Zhaoyu Chen, Rui Wang, Jing Zou, Xu Liu, Jinchi Liao, Yanping Wang, Ying Xia, Qing Wang
Format: Article
Language:English
Published: Wiley 2016-01-01
Series:Neural Plasticity
Online Access:http://dx.doi.org/10.1155/2016/5076740
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author Peng Zhou
Ruihui Weng
Zhaoyu Chen
Rui Wang
Jing Zou
Xu Liu
Jinchi Liao
Yanping Wang
Ying Xia
Qing Wang
author_facet Peng Zhou
Ruihui Weng
Zhaoyu Chen
Rui Wang
Jing Zou
Xu Liu
Jinchi Liao
Yanping Wang
Ying Xia
Qing Wang
author_sort Peng Zhou
collection DOAJ
description Aims. This work was conducted to establish an in vitro Parkinson’s disease (PD) model by exposing BV-2 cells to 1-methyl-4-phenylpyridinium (MPP+) and exploring the roles of TLR2/TLR4/TLR9 in inflammatory responses to MPP+. Methods/Results. MTT assay showed that cell viability of BV-2 cells was 84.78 ± 0.86% and 81.18 ± 0.99% of the control after incubation with 0.1 mM MPP+ for 12 hours and 24 hours, respectively. Viability was not significantly different from the control group. With immunofluorescence technique, we found that MPP+ incubation at 0.1 mM for 12 hours was the best condition to activate BV-2 cells. In this condition, the levels of TNF-α, IL-1β, and iNOS protein were statistically increased compared to the control according to ELISA tests. Real time RT-PCR and western blot measurements showed that TLR4 was statistically increased after 0.1 mM MPP+ incubation for 12 hours. Furthermore, after siRNA interference of TLR4 mRNA, NF-κB activation and the levels of TNF-α, IL-1β, and iNOS were all statistically decreased in this cell model. Conclusion. MPP+ incubation at the concentration of 0.1 mM for 12 hours is the best condition to activate BV-2 cells for mimicking PD inflammation in BV-2 cells. TLR4 signalling plays a critical role in the activation of BV-2 cells and the induction of inflammation in this cell model.
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spelling doaj-art-47dfd802bd344c5aae819dc9655e87052025-08-20T03:36:07ZengWileyNeural Plasticity2090-59041687-54432016-01-01201610.1155/2016/50767405076740TLR4 Signaling in MPP+-Induced Activation of BV-2 CellsPeng Zhou0Ruihui Weng1Zhaoyu Chen2Rui Wang3Jing Zou4Xu Liu5Jinchi Liao6Yanping Wang7Ying Xia8Qing Wang9Department of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University, 600 Tianhe Road, Guangzhou, Guangdong 510630, ChinaDepartment of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University, 600 Tianhe Road, Guangzhou, Guangdong 510630, ChinaDepartment of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University, 600 Tianhe Road, Guangzhou, Guangdong 510630, ChinaDepartment of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University, 600 Tianhe Road, Guangzhou, Guangdong 510630, ChinaDepartment of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University, 600 Tianhe Road, Guangzhou, Guangdong 510630, ChinaDepartment of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University, 600 Tianhe Road, Guangzhou, Guangdong 510630, ChinaDepartment of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University, 600 Tianhe Road, Guangzhou, Guangdong 510630, ChinaDepartment of Neurology, The Second Affiliated Hospital of Guangzhou Medical University, 250 Changgang Dong Road, Guangzhou 510260, ChinaDepartment of Neurosurgery, The University of Texas Medical School at Houston, Houston, TX 77030, USADepartment of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University, 600 Tianhe Road, Guangzhou, Guangdong 510630, ChinaAims. This work was conducted to establish an in vitro Parkinson’s disease (PD) model by exposing BV-2 cells to 1-methyl-4-phenylpyridinium (MPP+) and exploring the roles of TLR2/TLR4/TLR9 in inflammatory responses to MPP+. Methods/Results. MTT assay showed that cell viability of BV-2 cells was 84.78 ± 0.86% and 81.18 ± 0.99% of the control after incubation with 0.1 mM MPP+ for 12 hours and 24 hours, respectively. Viability was not significantly different from the control group. With immunofluorescence technique, we found that MPP+ incubation at 0.1 mM for 12 hours was the best condition to activate BV-2 cells. In this condition, the levels of TNF-α, IL-1β, and iNOS protein were statistically increased compared to the control according to ELISA tests. Real time RT-PCR and western blot measurements showed that TLR4 was statistically increased after 0.1 mM MPP+ incubation for 12 hours. Furthermore, after siRNA interference of TLR4 mRNA, NF-κB activation and the levels of TNF-α, IL-1β, and iNOS were all statistically decreased in this cell model. Conclusion. MPP+ incubation at the concentration of 0.1 mM for 12 hours is the best condition to activate BV-2 cells for mimicking PD inflammation in BV-2 cells. TLR4 signalling plays a critical role in the activation of BV-2 cells and the induction of inflammation in this cell model.http://dx.doi.org/10.1155/2016/5076740
spellingShingle Peng Zhou
Ruihui Weng
Zhaoyu Chen
Rui Wang
Jing Zou
Xu Liu
Jinchi Liao
Yanping Wang
Ying Xia
Qing Wang
TLR4 Signaling in MPP+-Induced Activation of BV-2 Cells
Neural Plasticity
title TLR4 Signaling in MPP+-Induced Activation of BV-2 Cells
title_full TLR4 Signaling in MPP+-Induced Activation of BV-2 Cells
title_fullStr TLR4 Signaling in MPP+-Induced Activation of BV-2 Cells
title_full_unstemmed TLR4 Signaling in MPP+-Induced Activation of BV-2 Cells
title_short TLR4 Signaling in MPP+-Induced Activation of BV-2 Cells
title_sort tlr4 signaling in mpp induced activation of bv 2 cells
url http://dx.doi.org/10.1155/2016/5076740
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