Inflammation Induced by Lipopolysaccharide and Palmitic Acid Increases Cholesterol Accumulation via Enhancing Myeloid Differentiation Factor 88 Expression in HepG2 Cells
Abstract
:1. Introduction
2. Results
2.1. LPS and PA Treatments Inhibited HepG2 Cell Viabilities
2.2. LPS and PA Treatments Increased Cholesterol Content in HepG2 Cells
2.3. LPS and PA Treatments Increased TLR4/MyD88/NF-κB Signaling Pathway Activation and SREBP-2 Expression
2.4. Inhibition of MyD88 Reduced Cholesterol Content in LPS- and PA-Treated HepG2 Cells
2.5. Inhibition of MyD88 Suppressed TLR4/MyD88/NF-κB Signaling Pathway Activation and SREBP-2 Expression Induced by LPS and PA Treatments
2.6. Role of MyD88 in SREBP-2 Mediated Cholesterol Synthesis
3. Discussion
4. Materials and Methods
4.1. Reagents and Chemicals
4.2. Cell Culture
4.3. MTT Cell Viability Assay
4.4. Transient Transfection
4.5. Measurement of Intracellular Cholesterol
4.6. Filipin Staining
4.7. Western Blotting
4.8. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Antibody | Cat. No. | Molecular Weight (kDa) | Dilutions |
---|---|---|---|
TLR4 | sc-293072 | 95 | 1:1000 |
GAPDH | G031 | 37 | 1:2000 |
TRIF | ab139281 | 76 | 1:1000 |
TRAF6 | ab137452 | 60 | 1:1000 |
SREBP-2 | 28212-1-AP | 124 (SREBP-2 precursor) and 73 (SREBP-2 mature forms) | 1:1000 |
Lamin B1 | 12987-1-AP | 66 | 1:1000 |
MyD88 | 4283S | 33 | 1:1000 |
TNF-α | 3707S | 17 | 1:1000 |
NF-κB-p65 | 4764S | 65 | 1:1000 |
phospho-NF-κB-p65 | 3031S | 65 | 1:1000 |
β-actin | 4967S | 45 | 1:1000 |
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Chen, J.; Liu, Y.; Luo, H.; Chen, G.; Zheng, Z.; Wang, T.; Hu, X.; Zhao, Y.; Tang, J.; Su, C.; et al. Inflammation Induced by Lipopolysaccharide and Palmitic Acid Increases Cholesterol Accumulation via Enhancing Myeloid Differentiation Factor 88 Expression in HepG2 Cells. Pharmaceuticals 2022, 15, 813. https://doi.org/10.3390/ph15070813
Chen J, Liu Y, Luo H, Chen G, Zheng Z, Wang T, Hu X, Zhao Y, Tang J, Su C, et al. Inflammation Induced by Lipopolysaccharide and Palmitic Acid Increases Cholesterol Accumulation via Enhancing Myeloid Differentiation Factor 88 Expression in HepG2 Cells. Pharmaceuticals. 2022; 15(7):813. https://doi.org/10.3390/ph15070813
Chicago/Turabian StyleChen, Junbin, Yuguo Liu, Huiyu Luo, Guoxun Chen, Zhongdaixi Zheng, Tiannan Wang, Xinge Hu, Yue Zhao, Jiaqi Tang, Chuhong Su, and et al. 2022. "Inflammation Induced by Lipopolysaccharide and Palmitic Acid Increases Cholesterol Accumulation via Enhancing Myeloid Differentiation Factor 88 Expression in HepG2 Cells" Pharmaceuticals 15, no. 7: 813. https://doi.org/10.3390/ph15070813