A Novel Highly Sensitive Method for Measuring Inflammatory Neural-Derived APC Activity in Glial Cell Lines, Mouse Brain and Human CSF
Abstract
:1. Background
2. Results
2.1. Assay Validation
2.2. Cell Cultures
2.3. Mice, Healthy Control, and Mild Traumatic Brain Injury (mTBI)
2.4. The Effect of Inflammation, LPS in Mice, and Human Inflammatory CSF
3. Discussion
4. Methods
4.1. Cell Culture
4.2. Animals
4.3. Mild Traumatic Brain Injury (mTBI) Induction
4.4. Mice Lipopolysaccharide (LPS) Treatment
4.5. Human Patients
4.6. APC Activity Assay
4.7. Thrombin, FXa, Plasma, and PC
4.8. Experimental Design
4.8.1. Thrombin/FXa/Plasma
4.8.2. Cells
4.8.3. Brain Tissue
4.9. Western Blot Analysis
4.9.1. Samples Preparation
4.9.2. Protein Detection
4.10. Statistics
Author Contributions
Funding
Acknowledgments
Disclosure
Conflicts of Interest
Abbreviations
Activated protein C | APC |
Thrombomodulin | TM |
Mild traumatic brain injury | mTBI |
Cerebrospinal fluid | CSF |
Lipopolysaccharide | LPS |
Endothelial protein C receptor | EPCR |
Protease activator receptor 1 | PAR1 |
Peripheral nervous system | PNS |
Central nervous system | CNS |
Normal pressure hydrocephalus | NPH |
Phenylmethylsulfonyl fluoride | PMSF |
Alpha-naphthylsulphonylglycyl-4-amidinophenylalanine piperidine | NAPAP |
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Golderman, V.; Gofrit, S.G.; Maggio, N.; Gera, O.; Gerasimov, A.; Laks, D.; Chapman, J.; Shavit-Stein, E. A Novel Highly Sensitive Method for Measuring Inflammatory Neural-Derived APC Activity in Glial Cell Lines, Mouse Brain and Human CSF. Int. J. Mol. Sci. 2020, 21, 2422. https://doi.org/10.3390/ijms21072422
Golderman V, Gofrit SG, Maggio N, Gera O, Gerasimov A, Laks D, Chapman J, Shavit-Stein E. A Novel Highly Sensitive Method for Measuring Inflammatory Neural-Derived APC Activity in Glial Cell Lines, Mouse Brain and Human CSF. International Journal of Molecular Sciences. 2020; 21(7):2422. https://doi.org/10.3390/ijms21072422
Chicago/Turabian StyleGolderman, Valery, Shany G. Gofrit, Nicola Maggio, Orna Gera, Alexandra Gerasimov, Dar Laks, Joab Chapman, and Efrat Shavit-Stein. 2020. "A Novel Highly Sensitive Method for Measuring Inflammatory Neural-Derived APC Activity in Glial Cell Lines, Mouse Brain and Human CSF" International Journal of Molecular Sciences 21, no. 7: 2422. https://doi.org/10.3390/ijms21072422