Caffeine Inhibits NLRP3 Inflammasome Activation by Downregulating TLR4/MAPK/NF-κB Signaling Pathway in an Experimental NASH Model
Abstract
:1. Introduction
2. Results
2.1. Caffeine Prevents Liver Damage and Oxidative Stress in the NASH Experimental Model
2.2. Effect of Caffeine on Liver Steatosis in the NASH Experimental Model
2.3. Caffeine Effect on the Proinflammatory and Fibrogenic TLR4-MAPK Pathway
2.4. Caffeine Prevents NASH by Blocking the Activation of the NLRP3 Inflammasome Signaling Pathway
2.5. Caffeine Attenuates Fibrosis by Mitigating Hepatic Stellate Cell Activation
3. Discussion
4. Materials and Methods
4.1. Animal Treatments
Non-Alcoholic Steatohepatitis Induction and Caffeine Treatment
4.2. Reagents
4.3. Antibodies
4.4. Biochemical Analyses
4.5. Histology Determinations
4.6. Immunohistochemistry Assays
4.7. Transmission Electron Microscopy
4.8. Immunofluorescence
4.9. Western Blotting Analysis
4.10. Zymography
4.11. Statistical Analyses
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Protein | Brand (Location) | Catalog | WB Dilution | IHC Dilution | IF Dilution |
---|---|---|---|---|---|
MMP-13 | Merck-Millipore® (Burlington, MA, USA) | MAB13426 | 1:500 | ||
TGF-β | Merck-Millipore® (Burlington, MA, USA) | MAB1032 | 1:500 | 1:250 | |
NF-κB (p65) | Merck Millipore® (Burlington, MA, USA) | MAB3026 | 1:500 | 1:250 | |
JNK | Cell Signaling Technology® (Danvers, MA, USA) | 9252S | 1:500 | ||
pp38 | Cell Signaling Technology® (Danvers, MA, USA) | 9211S | 1:500 | ||
p38 | Abcam® (Cambridge, UK) | AB31828 | 1:500 | ||
Smad7 | Abcam® (Cambridge, UK) | AB90086 | 1:500 | ||
pJNK | Abcam® (Cambridge, UK) | AB131499 | 1:500 | ||
4-HNE | Abcam® (Cambridge, UK) | AB46545 | 1:500 | ||
IL-1β | Abcam® (Cambridge, UK) | AB18329 | 1:500 | ||
Nrf2 | Abcam® (Cambridge, UK) | AB31163 | 1:500 | ||
SREBP1C | Abcam® (Cambridge, UK) | AB28481 | 1:500 | ||
α-SMA | Sigma-Aldrich® (St. Louis, MI, USA) | A-5691 | 1:500 | 1:250 | 1:250 |
Desmin | Sigma-Aldrich® (St. Louis, MI, USA) | 243M-1 | 1:500 | ||
pERK | Santa Cruz Biotechnology® (Santa Cruz, CA, USA) | SC-136521 | 1:500 | ||
ERK | Santa Cruz Biotechnology® (Santa Cruz, CA, USA) | SC-292838 | 1:500 | ||
CTGF | Santa Cruz Biotechnology® (Santa Cruz, CA, USA) | SC-365970 | 1:500 | ||
IL-17 | Santa Cruz Biotechnology® (Dallas, TX, USA) | SC-374218 | 1:500 | ||
TNF-α | Santa Cruz Biotechnology® (Dallas, TX, USA) | SC-52746 | 1:500 | ||
ASC | Santa Cruz Biotechnology® (Dallas, TX, USA) | SC-514414 | 1:500 | ||
Caspase 1 | Santa Cruz Biotechnology® (Dallas, TX, USA) | SC-392736 | 1:500 | 1:250 | 1:250 |
PPAR-α | Santa Cruz Biotechnology® (Dallas, TX, USA) | SC-398394 | 1:500 | ||
NLRP3 | Novus biologicals® (Littleton, CO, USA) | NBP2-12446 | 1:500 | 1:250 | 1:250 |
TLR4 | Thermo Fisher® (Waltham, MA, USA) | 48-2300 | 1:500 | ||
β-actin | Thermo Fisher® (Waltham, MA, USA) | AM4302 | 1:500 | ||
Secondary Antibodies | |||||
Anti-Rabbit | Merck Millipore® (Burlington, MA, USA) | A0545 | 1:3000 | 1:1000 | |
Anti-Mouse | Thermo Fisher® (Waltham, MA, USA) | 62-6520 | 1:5000 | 1:1000 | |
Alexa fluor 594 | Thermo Fisher® (Waltham, MA, USA) | Z25007 | 1:1000 | ||
Alexa fluor 488 | Thermo Fisher® (Waltham, MA, USA) | Z25302 | 1:1000 |
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Vargas-Pozada, E.E.; Ramos-Tovar, E.; Rodriguez-Callejas, J.D.; Cardoso-Lezama, I.; Galindo-Gómez, S.; Talamás-Lara, D.; Vásquez-Garzón, V.R.; Arellanes-Robledo, J.; Tsutsumi, V.; Villa-Treviño, S.; et al. Caffeine Inhibits NLRP3 Inflammasome Activation by Downregulating TLR4/MAPK/NF-κB Signaling Pathway in an Experimental NASH Model. Int. J. Mol. Sci. 2022, 23, 9954. https://doi.org/10.3390/ijms23179954
Vargas-Pozada EE, Ramos-Tovar E, Rodriguez-Callejas JD, Cardoso-Lezama I, Galindo-Gómez S, Talamás-Lara D, Vásquez-Garzón VR, Arellanes-Robledo J, Tsutsumi V, Villa-Treviño S, et al. Caffeine Inhibits NLRP3 Inflammasome Activation by Downregulating TLR4/MAPK/NF-κB Signaling Pathway in an Experimental NASH Model. International Journal of Molecular Sciences. 2022; 23(17):9954. https://doi.org/10.3390/ijms23179954
Chicago/Turabian StyleVargas-Pozada, Eduardo E., Erika Ramos-Tovar, Juan D. Rodriguez-Callejas, Irina Cardoso-Lezama, Silvia Galindo-Gómez, Daniel Talamás-Lara, Verónica Rocío Vásquez-Garzón, Jaime Arellanes-Robledo, Víctor Tsutsumi, Saúl Villa-Treviño, and et al. 2022. "Caffeine Inhibits NLRP3 Inflammasome Activation by Downregulating TLR4/MAPK/NF-κB Signaling Pathway in an Experimental NASH Model" International Journal of Molecular Sciences 23, no. 17: 9954. https://doi.org/10.3390/ijms23179954