NLRP3 Inflammasome Activation Is Involved in LPA1-Mediated Brain Injury after Transient Focal Cerebral Ischemia
Abstract
:1. Introduction
2. Results
2.1. Suppressing LPA1 Activity Attenuates NLRP3 Inflammasome Activation in an Injured Brain after tMCAO Challenge
2.2. LPA1 Contributes to NLRP3 Upregulation and NLRP3 Inflammasome Activation in LPS-Primed Macrophages Followed by LPA Exposure
2.3. LPA1 Regulates NLRP3 Upregulation in LPS-Primed Macrophages Followed by LPA Exposure by Enhancing Nuclear Factor-κB (NF-κB) Translocation and Activating Extracellular Signal-Regulated Kinase 1/2 (ERK1/2) and p38 Mitogen-Activated Protein Kinase (MAPK)
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. Transient Focal Cerebral Ischemia Challenge and AM095 Administration
4.3. Determination of Functional Neurological Deficit Score
4.4. Quantitative Real-Time PCR (qPCR) Analysis
4.5. Immunohistochemistry for NLRP3 and Double Immunofluorescence for NLRP3/Iba1 or ASC/NLRP3
4.6. BMDMs Primary Culture, Treatment, and LPA1 Knockdown
4.7. Western Blot Analysis
4.8. Measurement of IL-1β in Conditioned Medium
4.9. Determination of NF-κB Translocation
4.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ASC | Apoptosis-associated speck-like protein containing a caspase recruitment domain |
BMDMs | Bone marrow-derived macrophages |
CCA | Common carotid artery |
DAPI | 4′,6-Diamidino-2-phenylindole |
ELISA | Enzyme-linked immunosorbent assay |
ERK1/2 | Extracellular signal-regulated kinase 1/2 |
HRP | Horseradish peroxidase |
Iba1 | Ionized calcium-binding adapter molecule 1 |
IL-1β | Interleukin 1β |
IL-6 | Interleukin 6 |
JNK | c-Jun N-terminal kinase |
LPA1 | Lysophosphatidic acid receptor 1 |
LPS | Lipopolysaccharide |
MAPK | Mitogen-activated protein kinase |
α-MEM | Minimum Essential Medium alpha |
mNSS | Modified neurological severity score |
NF-κB | Nuclear factor-κB |
NLRP3 | Nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 |
PFA | Paraformaldehyde |
PVDF | Polyvinylidene difluoride |
siRNA | Small interfering RNA |
tMCAO | Transient middle cerebral artery occlusion |
TNF-α | Tumor necrosis factor-α |
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Lee, C.-H.; Sapkota, A.; Gaire, B.P.; Choi, J.W. NLRP3 Inflammasome Activation Is Involved in LPA1-Mediated Brain Injury after Transient Focal Cerebral Ischemia. Int. J. Mol. Sci. 2020, 21, 8595. https://doi.org/10.3390/ijms21228595
Lee C-H, Sapkota A, Gaire BP, Choi JW. NLRP3 Inflammasome Activation Is Involved in LPA1-Mediated Brain Injury after Transient Focal Cerebral Ischemia. International Journal of Molecular Sciences. 2020; 21(22):8595. https://doi.org/10.3390/ijms21228595
Chicago/Turabian StyleLee, Chi-Ho, Arjun Sapkota, Bhakta Prasad Gaire, and Ji Woong Choi. 2020. "NLRP3 Inflammasome Activation Is Involved in LPA1-Mediated Brain Injury after Transient Focal Cerebral Ischemia" International Journal of Molecular Sciences 21, no. 22: 8595. https://doi.org/10.3390/ijms21228595