TGF-β1 Decreases Microglia-Mediated Neuroinflammation and Lipid Droplet Accumulation in an In Vitro Stroke Model
Abstract
:1. Introduction
2. Results
2.1. Accumulation of LD in Primary Microglia
2.2. TGF-β1 Regulates OGD-Induced LD Accumulation
2.3. TGF-β1 Can Regulate LPS-Induced LD Accumulation
2.4. TGF-β1 Modulates Microglia-Mediated Neuroinflammation
2.5. Inhibition of LD Formation Reduces Microglia-Mediated Neuroinflammation
2.6. TGF-β1 Plays an Important Role in Primary Neuron Viability Exposed to Hypoxia
3. Discussion
4. Materials and Methods
4.1. Isolation and Culture of Primary Microglia and Neurons
4.2. Oxygen-Glucose Deprivation (OGD)
4.3. TGF-β1 siRNA Transfection of Microglia
4.4. Cell Viability
4.5. Lipopolysaccharide (LPS) and Triacsin C Treatment
4.6. RNA Extraction and Real-Time qRT-PCR
4.7. Protein Extraction and Western Blot Analyses
4.8. Immunocytochemistry Staining
4.9. Immunostaining for Lipid Droplets in Primary Microglia
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
ANOVA | Analysis of variance |
BSA | Bovine serum albumin |
CNS | Central nervous system |
CD206 | Cluster of Differentiation 206 |
DMSO | Dimethyl sulfoxide |
DS | Donkey serum |
FBS | Fetal bovine serum |
HBSS | Hank’s Balanced Salt Solution |
iNOS | inducible nitric oxide synthase |
IL-1β | Interleukin-1β |
LD | Lipid droplet |
LPS | Lipopolysaccharide |
MTT | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
OGD | Oxygen-glucose deprivation |
PDL | Poly-d-lysine |
PBS | Phosphate-buffered saline |
PBST | PBS with Tween 20 |
PFA | Paraformaldehyde |
PLIN2 | Perilipin-2 |
qRT-PCR | quantitative real-time polymerase chain reaction |
siRNA | small interfering RNA |
TGF-β1 | Transforming growth factor-β1 |
TNF-α | Tumor necrosis factor-α |
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Xin, W.; Pan, Y.; Wei, W.; Gerner, S.T.; Huber, S.; Juenemann, M.; Butz, M.; Bähr, M.; Huttner, H.B.; Doeppner, T.R. TGF-β1 Decreases Microglia-Mediated Neuroinflammation and Lipid Droplet Accumulation in an In Vitro Stroke Model. Int. J. Mol. Sci. 2023, 24, 17329. https://doi.org/10.3390/ijms242417329
Xin W, Pan Y, Wei W, Gerner ST, Huber S, Juenemann M, Butz M, Bähr M, Huttner HB, Doeppner TR. TGF-β1 Decreases Microglia-Mediated Neuroinflammation and Lipid Droplet Accumulation in an In Vitro Stroke Model. International Journal of Molecular Sciences. 2023; 24(24):17329. https://doi.org/10.3390/ijms242417329
Chicago/Turabian StyleXin, Wenqiang, Yongli Pan, Wei Wei, Stefan T. Gerner, Sabine Huber, Martin Juenemann, Marius Butz, Mathias Bähr, Hagen B. Huttner, and Thorsten R. Doeppner. 2023. "TGF-β1 Decreases Microglia-Mediated Neuroinflammation and Lipid Droplet Accumulation in an In Vitro Stroke Model" International Journal of Molecular Sciences 24, no. 24: 17329. https://doi.org/10.3390/ijms242417329