Anti-Inflammatory Effect of Meriania hexamera Sprague by Targeting Syk Kinase in NF-κB Signaling
Abstract
:1. Introduction
2. Results
2.1. Mh-ME Ameliorated NO Production without Inducing Death of RAW264.7 Cells
2.2. Analysis of the Components of Mh-ME
2.3. Effects of Mh-ME on the Transcriptional Activation of Pro-Inflammatory Proteins
2.4. Mh-ME Lowered Phosphorylation of Proteins Inducing the NF-κB Acitvation
2.5. Mh-ME Showed Therapeutic Potential on Acute Gastritis
2.6. Acute Toxicity Test
3. Discussion
4. Materials and Methods
4.1. Materials and Reagents
4.2. Mh-ME Preparation and GC-MS
4.3. Measurement of Total Phenolic and Flavonoid Contents
4.4. LC-MS/MS
4.5. Cell Culture
4.6. Nitric Oxide (NO) Assay
4.7. Cell Viability Assay
4.8. Semiquantitative RT-PCR and Quantitative Real-Time PCR
4.9. Luciferase Reproter Assay
4.10. Preparing Whole Lysates and Western Blotting
4.11. Plasmid Transfection for Exogeneous Expression of Syk and Src Kinase
4.12. Cellualr Thermal Shift Assay (CETSA)
4.13. Animal Experiments
4.14. Serum ALT and AST Activity Detection
4.15. Statistical Anaylsis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviation
PAMPs | pathogen-associated molecular patterns |
PRRs | pattern recognition receptors |
TLRs | toll-like receptors |
LPS | lipopolysaccharide |
TRIF | TIR-domain-containing adapter-inducing interferon-β |
MyD88 | myeloid differentiation response gene 88 |
DAMPs | damage-associated molecular patterns |
NF-κB | nuclear factor-kappa B |
AP-1 | activated protein-1 |
Syk | spleen tyrosinase kinase |
AKT | protein kinase B |
IκBα | inhibitor of kappa B alpha |
iNOS | nitric oxide synthase |
IL-1β | interleukin-1β |
TNF-α | tumor necrosis factor-alpha |
COX-2 | cyclooxygenase-2 |
Mh-ME | M. hexamera Sprague-Methanol Extract |
Aa-EE | A. asiatica-Ethanol Extract |
GC-MS | gas chromatography-mass spectrometry |
RPMI 1640 | Roswell Park Memorial Institute 1640 |
DMEM | Dulbecco’s modified Eagle’s medium |
MTT | 3-(4-5-dimetyhlathiaol-2-yl)-2-5-diphenyltetrazolium bromide |
DMSO | dimethyl sulfoxide |
SDS | sodium dodecyl sulfate |
PBS | phosphate-buffered saline |
CMC | carboxymethylcellulose |
GAPDH | glyceraldehyde 3-phosphate dehydrogenase |
PEI | polyethylenimine |
TBST | tris-buffered saline with Tween 20 |
CETSA | cellular thermal shift assay |
ALT | alanine aminotransferase |
AST | aspartate aminotransferase |
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Peak No. | RT (min) | Compound Name | Corrected Area | % of Total |
---|---|---|---|---|
1 | 8.780 | 5-Hydroxymethylfurfural | 5,310,668 | 1.388 |
2 | 9.806 | 1,1-diethyl-2-(1-methyl propyl)-Hydrazine | 7,381,173 | 1.929 |
3 | 10.301 | 2,7-Oxepanedione | 4,908,751 | 1.283 |
4 | 10.810 | 1,2,3-Benzenetriol | 251,749,615 | 65.794 |
5 | 11.193 | 1,2,3-Benzenetriol | 5,064,771 | 1.324 |
6 | 12.231 | 1,6-anhydro-beta-D-Glucopyranose | 28,492,675 | 7.446 |
7 | 13.946 | 4-hydroxy-Benzenepropanoic acid | 7,835,372 | 2.048 |
8 | 14.221 | 4-hydroxy-3-methoxy- Benzenepropanol | 7,587,564 | 1.983 |
9 | 16.218 | Neophytadiene | 4,036,831 | 1.055 |
10 | 16.663 | Z-8-Methyl-9-tetradecen-1-ol formate | 2,380,805 | 0.622 |
11 | 17.410 | n-Hexadecanoic acid | 20,611,816 | 5.387 |
12 | 19.114 | (Z,Z,Z)-9,12,15-Octadecatrienoic acid | 9,550,118 | 2.496 |
13 | 19.299 | Octadecanoic acid | 2,647,456 | 0.692 |
14 | 29.383 | gamma-Sitosterol | 2,5075,899 | 6.554 |
Extract | Total Phenolic Contents 1 | Total Flavonoid Contents 2 |
---|---|---|
Mh-ME | 301.33 ± 0.98 | 19.05 ± 1.19 |
RT (min) | Component Name | Formula |
---|---|---|
0.94 | Wogonoside | C22H20O11 |
0.98 | Kaempferol-7-O-α-L-rhamnoside | C21H20O10 |
2.65 | Bavachinin | C21H22O4 |
2.90 | Kaempferol-3-gentiobioside | C27H30O16 |
Gene Name | Sequence (5′-3′) | |
---|---|---|
iNOS | Forward Reverse | TGCCAGGGTCACAACTTTACA ACCCCAAGCAAGACTTGGAC |
IL-1β | Forward Reverse | CAGGATGAGGACATGAGCACC CTCTGCAGACTCAAACTCCAC |
IL-6 | Forward Reverse | GGAAATCGTGGAAATGAG GCTTAGGCATAACGCACT |
GAPDH | Forward Reverse | GAAGGTCGGTGTGAACGGAT AGTGATGGCATGGACTGTGG |
Gene Name | Sequence (5′-3′) | |
---|---|---|
IL-1β | Forward Reverse | GTGAAATGCCACCTTTTACAGTG CCTGCCTGAAGCTCTTGTTG |
TNF-α | Forward Reverse | TGCCTATGTCTCAGCCTCTT GAGGCCATTTGGGAACTTCT |
GAPDH | Forward Reverse | GGAGAGTGTTTCCTCGTCCC ATGAAGGGGTCGTTGATGGC |
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Share and Cite
Kwon, K.W.; Jang, W.Y.; Kim, J.W.; Noh, J.K.; Yi, D.-K.; Cho, J.Y. Anti-Inflammatory Effect of Meriania hexamera Sprague by Targeting Syk Kinase in NF-κB Signaling. Plants 2023, 12, 3044. https://doi.org/10.3390/plants12173044
Kwon KW, Jang WY, Kim JW, Noh JK, Yi D-K, Cho JY. Anti-Inflammatory Effect of Meriania hexamera Sprague by Targeting Syk Kinase in NF-κB Signaling. Plants. 2023; 12(17):3044. https://doi.org/10.3390/plants12173044
Chicago/Turabian StyleKwon, Ki Woong, Won Young Jang, Ji Won Kim, Jin Kyoung Noh, Dong-Keun Yi, and Jae Youl Cho. 2023. "Anti-Inflammatory Effect of Meriania hexamera Sprague by Targeting Syk Kinase in NF-κB Signaling" Plants 12, no. 17: 3044. https://doi.org/10.3390/plants12173044