Immune-Enhancing Effects of a High Molecular Weight Fraction of Cynanchum wilfordii Hemsley in Macrophages and Immunosuppressed Mice
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals and Reagents
2.2. Preparation of a High Molecular Weight Fraction of Cynanchum wilfordii
2.3. Extraction of a Crude Polysaccharide
2.4. Chemical Analyses
2.5. Monosaccharide Composition and Molecular Weight Distribution
2.6. Protease Treatment and Periodate Oxidation of HMFO
2.7. Cell Culture
2.8. Cell Viability Assay
2.9. Measurement of Nitric Oxide (NO)
2.10. Determination of TNF-α, (IL-6) and IL-1β Production
2.11. Phagocytosis Assay in RAW 264.7 Cells
2.12. Western Blot Analysis
2.13. Animals and Treatments
2.14. Preparation of Mouse Splenocytes
2.15. Splenocyte Proliferation Assay
2.16. Flow Cytometric Analysis
2.17. NK Cell Activity
2.18. Statistical Analysis
3. Results
3.1. Effects of HMF on NO Production and Cytokine and Phagocytic Activity
3.2. Effects of HMF on Decreased Immune Function in CYC-Treated Mice
3.3. Effects of a Crude Polysaccharide of HMF on the Immunostimulatory Activity in RAW 264.7 Macrophages
3.3.1. Effects of Chemical and Enzymatic Treatments of HMFO on Immunostimulatory Activity in RAW 264.7 Macrophages
3.3.2. Effects of HMFO on NO Production and iNOS Expression in RAW 264.7 Macrophages
3.3.3. Effects of HMFO on the Phosphorylation of IκB-α and IKK-α/β in RAW 264.7 Macrophages
3.3.4. Effects of HMFO on Mitogen-Activated Protein Kinase (MAPK) Phosphorylation in RAW 264.7 Macrophages
3.4. Chemical Compositions and Molecular Weight Distribution
4. Discussion
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Group | Initial Body Weight (g) | Terminal Body Weight (g) | Spleen Weight | Thymus Weight | ||
---|---|---|---|---|---|---|
Absolute (g) | Relative (%) | Absolute (g) | Relative (%) | |||
Negative control | 19.11 ± 0.75 a | 20.03 ± 0.65 c | 0.103 ± 0.009 a | 0.512 ± 0.027 ab | 0.047 ± 0.004 a | 0.233 ± 0.017 a |
CYC | 18.82 ± 0.94 a | 18.10 ± 0.57 a | 0.085 ± 0.005 b | 0.471 ± 0.022 a | 0.041 ± 0.005 a | 0.225 ± 0.023 a |
HMF (100 mg/kg) | 18.38 ± 0.52 a | 18.41 ± 0.53 ab | 0.100 ± 0.002 a | 0.544 ± 0.014 b | 0.042 ± 0.003 a | 0.230 ± 0.012 b |
HMF (200 mg/kg) | 19.11 ± 0.34 a | 19.52 ± 0.63 bc | 0.108 ± 0.007 a | 0.554 ± 0.021 b | 0.045 ± 0.003 a | 0.232 ± 0.009 b |
Positive control | 19.32 ± 0.64 a | 19.69 ± 0.85 c | 0.100 ± 0.008 a | 0.510 ± 0.037 ab | 0.047 ± 0.005 a | 0.238 ± 0.022 b |
HMFO | |
---|---|
Chemical composition (%) | |
Neutral sugar 1 | 63.8 ± 0.9 |
Uronic acid 2 | 33.1 ± 1.9 |
KDO 3 | 0.3 ± 0.1 |
Protein 4 | 2.9 ± 0.2 |
Component monosaccharide (mg/g) | |
Arabinose | 86.0 ± 1.6 |
Galactose | 76.5 ± 2.9 |
Rhamnose | 23.0 ± 0.4 |
Xylose | 2.2 ± 0.6 |
Glucose | 241.2 ± 6.5 |
Mannose | 9.0 ± 1.6 |
Fucose | 5.7 ± 0.2 |
Fructose | ND 5 |
Galacturonic acid | 178.8 ± 2.5 |
Glucuronic acid | 4.9 ± 0.2 |
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Jang, M.; Lim, T.-G.; Ahn, S.; Hong, H.-D.; Rhee, Y.K.; Kim, K.-T.; Lee, E.; Lee, J.H.; Lee, Y.J.; Jung, C.S.; et al. Immune-Enhancing Effects of a High Molecular Weight Fraction of Cynanchum wilfordii Hemsley in Macrophages and Immunosuppressed Mice. Nutrients 2016, 8, 600. https://doi.org/10.3390/nu8100600
Jang M, Lim T-G, Ahn S, Hong H-D, Rhee YK, Kim K-T, Lee E, Lee JH, Lee YJ, Jung CS, et al. Immune-Enhancing Effects of a High Molecular Weight Fraction of Cynanchum wilfordii Hemsley in Macrophages and Immunosuppressed Mice. Nutrients. 2016; 8(10):600. https://doi.org/10.3390/nu8100600
Chicago/Turabian StyleJang, Mi, Tae-Gyu Lim, Sungeun Ahn, Hee-Do Hong, Young Kyoung Rhee, Kyung-Tack Kim, Eunjung Lee, Jeong Hoon Lee, Yun Ji Lee, Chan Sik Jung, and et al. 2016. "Immune-Enhancing Effects of a High Molecular Weight Fraction of Cynanchum wilfordii Hemsley in Macrophages and Immunosuppressed Mice" Nutrients 8, no. 10: 600. https://doi.org/10.3390/nu8100600