Assessment of Anti-Inflammatory and Antioxidant Effects of Citrus unshiu Peel (CUP) Flavonoids on LPS-Stimulated RAW 264.7 Cells
Abstract
:1. Introduction
2. Results
2.1. Flavonoids from CUP
2.2. Cell Toxicity of CUP Flavonoids
2.3. CUP Flavonoids Inhibit NO Production and iNOS Expression in LPS-Stimulated Macrophages
2.4. CUP Flavonoids Inhibit PGE2 Production and COX2 Expression in LPS-Stimulated Macrophages
2.5. Effects of CUP Flavonoids on Pro-Inflammatory Cytokines Production in LPS-Stimulated Macrophages
2.6. CUP Flavonoids Disturb the NF-κb Activation and Induce Anti-Inflammatory Effects in LPS-Stimulated Macrophages
2.7. Inhibition of CUP Flavonoids on ROS Production in LPS-Stimulated Macrophages
3. Discussion
3.1. Flavonoids Identified from CUP
3.2. Anti-Inflammatory Potential of CUP Flavonoids
3.3. Antioxidant Potential of CUP Flavonoids
4. Materials and Methods
4.1. Plant Material, Standards, and Reagents
4.2. HPLC-MS/MS Analysis and Quantification of Flavonoids
4.3. Cell Culture
4.4. Cell Viability Assay
4.5. Nitric Oxide Determination
4.6. Measurement of PGE2, TNF-α, and IL-1β Levels
4.7. Measurement of Reactive Oxygen Species
4.8. RNA Isolation and qRT-qPCR Analysis
4.9. Protein Isolation and Western Blot Analysis
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Peak. No | Compound | Chemical Formula | Retention Time (Min) | UV Max | Ionization Mode[M-H]− | M/Z | MS/MS | Reference |
---|---|---|---|---|---|---|---|---|
1 | Rutin | C27H30O16 | 19.25 | 264, 354 | [M-H]− | 609 | 301 | 15 |
2 | Naringin | C27H32O14 | 19.66 | 282, 336 | [M-H]− | 579 | 459, 313, 271 | 16 |
3 | Hesperidin | C28H34O15 | 20.19 | 284 | [M-H]− | 609 | 343, 325, 301, 286 | 17 |
4 | Poncirin | C28H34O14 | 23.09 | 284, 326 | [M-H]− | 593 | 387, 327, 309, 285 | 18 |
5 | Un known | - | 25.75 | 345, 268 | [M-H]− | 723 | 417, 403, 360 | - |
S. No | Standards | Calibration Curve | R2 | LOD (mg/L) | LOQ (mg/L) | Quantity (Mg/Kg) |
---|---|---|---|---|---|---|
1 | Rutin | y = 11.488x − 90.932 | 0.9998 | 0.927 | 2.780 | 24.07 |
2 | Naringin | y = 32.386x + 645.38 | 0.999 | 0.105 | 0.315 | 256.87 |
3 | Hesperidin | y = 32.297x + 166.45 | 1 | 0.075 | 0.225 | 79.17 |
4 | Poncirin | y = 34.263x + 165.96 | 1 | 0.064 | 0.192 | 3.94 |
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Karthikeyan, A.; Kim, H.H.; Preethi, V.; Moniruzzaman, M.; Lee, K.H.; Kalaiselvi, S.; Kim, G.S.; Min, T. Assessment of Anti-Inflammatory and Antioxidant Effects of Citrus unshiu Peel (CUP) Flavonoids on LPS-Stimulated RAW 264.7 Cells. Plants 2021, 10, 2209. https://doi.org/10.3390/plants10102209
Karthikeyan A, Kim HH, Preethi V, Moniruzzaman M, Lee KH, Kalaiselvi S, Kim GS, Min T. Assessment of Anti-Inflammatory and Antioxidant Effects of Citrus unshiu Peel (CUP) Flavonoids on LPS-Stimulated RAW 264.7 Cells. Plants. 2021; 10(10):2209. https://doi.org/10.3390/plants10102209
Chicago/Turabian StyleKarthikeyan, Adhimoolam, Hun Hwan Kim, Vetrivel Preethi, Mohammad Moniruzzaman, Ki Ho Lee, Senthil Kalaiselvi, Gon Sup Kim, and Taesun Min. 2021. "Assessment of Anti-Inflammatory and Antioxidant Effects of Citrus unshiu Peel (CUP) Flavonoids on LPS-Stimulated RAW 264.7 Cells" Plants 10, no. 10: 2209. https://doi.org/10.3390/plants10102209