Discovery of Phenolic Glycoside from Hyssopus cuspidatus Attenuates LPS-Induced Inflammatory Responses by Inhibition of iNOS and COX-2 Expression through Suppression of NF-κB Activation
Abstract
:1. Introduction
2. Results
2.1. Effects of HY on the Viability of Macrophages
2.2. Effects of HY on Inflammatory Mediator Production in LPS-Induced Macrophages
2.3. Effects of HY on Inflammatory Cytokine Expression and Production in LPS-Induced Macrophages
2.4. Effects of HY on Suppressing NF-κB Activation in LPS-Induced Macrophages
2.5. Effects of HY on MAPK Signaling Activation in LPS-Induced Macrophages
2.6. HY interacts with COX-2 to Activate NF-κB
3. Discussion
4. Materials and Methods
4.1. Plant Material and Prepared HY
4.2. Animals
4.3. Cell Culture and Treatment
4.4. Preparation and Culturing Primary Peritoneal Macrophages (PPMS)
4.5. Cell Viability Analysis
4.6. NO Production Assay
4.7. RNA Isolation, cDNA Synthesis, and Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR)
4.8. Enzyme-Linked Immunosorbent Assay (ELISA)
4.9. Total and Nuclear Protein Extraction and Western Blot Analysis
4.10. Immunofluorescence Assay
4.11. Molecular Modeling
4.12. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
COX-2 | Cyclooxygenase-2 |
DMEM | Dulbecco’s modified Eagle’s medium |
DAPI | 4′,6-Diamidino-2-phenylindole4′,6-diamidino-2-phenylindole |
ELISA | Enzyme-linked immunosorbent assay |
ERK1/2 | Extracellular signal-regulated kinase 1/2 |
FBS | Fetal bovine serum |
GAPDH | Glyceraldehyde 3-phosphate dehydrogenase |
HY | Hyssopuside |
IκB | Inhibitor of NF-κB |
IL-6 | Interleukin-6 |
IL-1β | Interleukin-1β |
iNOS | Inducible nitric oxide synthase |
LPS | Lipopolysaccharide |
MAPK | Mitogen-activated protein kinase |
MTT | 3-(4,5-Dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide |
NF-κB/p65 | Nuclear factor kappa-B |
NO | Nitric oxide |
RT-qPCR | Real-time quantitative polymerase chain reaction |
TNF-α | Tumor necrosis factor-α |
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Liu, X.; Su, J.; Wang, G.; Zheng, L.; Wang, G.; Sun, Y.; Bao, Y.; Wang, S.; Huang, Y. Discovery of Phenolic Glycoside from Hyssopus cuspidatus Attenuates LPS-Induced Inflammatory Responses by Inhibition of iNOS and COX-2 Expression through Suppression of NF-κB Activation. Int. J. Mol. Sci. 2021, 22, 12128. https://doi.org/10.3390/ijms222212128
Liu X, Su J, Wang G, Zheng L, Wang G, Sun Y, Bao Y, Wang S, Huang Y. Discovery of Phenolic Glycoside from Hyssopus cuspidatus Attenuates LPS-Induced Inflammatory Responses by Inhibition of iNOS and COX-2 Expression through Suppression of NF-κB Activation. International Journal of Molecular Sciences. 2021; 22(22):12128. https://doi.org/10.3390/ijms222212128
Chicago/Turabian StyleLiu, Xingyu, Jie Su, Geng Wang, Lihua Zheng, Guannan Wang, Ying Sun, Yongli Bao, Shuyue Wang, and Yanxin Huang. 2021. "Discovery of Phenolic Glycoside from Hyssopus cuspidatus Attenuates LPS-Induced Inflammatory Responses by Inhibition of iNOS and COX-2 Expression through Suppression of NF-κB Activation" International Journal of Molecular Sciences 22, no. 22: 12128. https://doi.org/10.3390/ijms222212128