Extraction, Characterization and Immunological Activity of Polysaccharides from Rhizoma gastrodiae
Abstract
:1. Introduction
2. Results and Discussion
2.1. Single-Factor Experimental Analysis
2.1.1. Effect of Extraction Time on the Yield of Rhizoma gastrodiae Polysaccharide (RGP)
2.1.2. Effect of Extraction Temperature on the Yield of RGP
2.1.3. Effect of the Liquid-to-Solid Ratio on the Yield of RGP
2.1.4. Effect of the Number of Extractions on the Yield of RGP
2.1.5. Optimum Conditions for the RSM Experiments
2.2. Optimization of the RGP Extraction
2.2.1. Predicted Model and Statistical Analysis
2.2.2. Analysis of the Response Surface
2.2.3. Verification of the Model
2.3. Purification and Preliminary Characterization of RGP
2.3.1. Purification, Homogeneity, and Molecular Weights of RGP-1a and RGP-1b
2.3.2. Preliminary Characterization of RGP-1a and RGP-1b
2.4. Effects of RGP-1a and RGP-1b on Macrophages in Vitro
2.4.1. Effects of RGP-1a and RGP-1b on Macrophage Viability
2.4.2. Effects of RGP-1a and RGP-1b on Macrophage Activation
2.4.3. Effects of RGP-1a and RGP-1b Macrophages Phagocytosis
3. Materials and Methods
3.1. Materials
3.2. Extraction and Determination of Polysaccharide
3.3. Experimental Design
3.4. Purification of RGP
3.5. Characterization of RGP
3.5.1. Carbohydrate, Protein and Uronic Acid Determination
3.5.2. Homogeneity and Molecular Weight Determination
3.5.3. Analysis of Monosaccharide Composition
3.5.4. FTIR Spectrometric Analysis
3.6. Effect of Polysaccharides on Macrophage Cell
3.6.1. RAW 264.7 Cell Viability Assays
3.6.2. Nitric Oxide (NO) Production Assay
3.6.3. Assay of Macrophages Phagocytosis
3.7. Statistical Analysis
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Run | X1 (Liquid-to-Solid Ratio, mL/g) | X2 (Extraction Temperature, °C) | X3 (Extraction Time, min) | Yield of RGP (%) |
---|---|---|---|---|
1 | 1 (60) | −1 (50) | 0 (60) | 5.78 |
2 | −1 (30) | 0 (65) | 1 (80) | 4.91 |
3 | 0 (45) | 0 (65) | 0 (60) | 5.84 |
4 | −1 (30) | 0 (65) | −1 (40) | 4.59 |
5 | 0 (45) | 0 (65) | 0 (60) | 6.03 |
6 | 1 (60) | 0 (65) | 1 (80) | 5.97 |
7 | −1 (30) | 1 (80) | 0 (60) | 5.47 |
8 | 0 (45) | 0 (65) | 0 (60) | 5.91 |
9 | 0 (45) | 0 (65) | 0 (60) | 6.10 |
10 | 1 (60) | 1 (80) | 0 (60) | 6.01 |
11 | 0 (45) | −1 (50) | −1 (40) | 5.01 |
12 | 0 (45) | 1 (80) | 1 (80) | 5.73 |
13 | 0 (45) | 0 (65) | 0 (60) | 5.89 |
14 | 0 (45) | −1 (50) | 1 (80) | 5.66 |
15 | 0 (45) | 1 (80) | −1 (40) | 5.38 |
16 | 1 (60) | 0 (65) | −1 (40) | 5.27 |
17 | −1 (30) | −1 (50) | 0 (60) | 4.92 |
Source | Sum of Squares | Df | Mean Square | F-Value | p-Value | Significance |
---|---|---|---|---|---|---|
Model | 3.41 | 9 | 0.38 | 35.28 | <0.0001 | ** |
X1 | 1.23 | 1 | 1.23 | 114.92 | <0.0001 | ** |
X2 | 0.19 | 1 | 0.19 | 17.35 | 0.0042 | ** |
X3 | 0.51 | 1 | 0.51 | 47.56 | 0.0002 | ** |
X1X2 | 0.026 | 1 | 0.026 | 2.39 | 0.1663 | – |
X1X3 | 0.036 | 1 | 0.036 | 3.37 | 0.1092 | – |
X2X3 | 0.022 | 1 | 0.022 | 2.10 | 0.1908 | – |
0.47 | 1 | 0.47 | 43.93 | 0.0003 | ** | |
0.023 | 1 | 0.023 | 2.18 | 0.1834 | – | |
0.79 | 1 | 0.79 | 74.12 | <0.0001 | ** | |
Residual | 0.075 | 7 | 0.011 | |||
Lack of fit | 0.029 | 3 | 0.010 | 0.84 | 0.5398 | – |
Pure error | 0.046 | 4 | 0.012 | |||
Cor. total | 3.48 | 16 | ||||
C.V.% = 1.86% | ||||||
R2 = 0.9784 | ||||||
R2adj = 0.9507 |
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Chen, J.; Tian, S.; Shu, X.; Du, H.; Li, N.; Wang, J. Extraction, Characterization and Immunological Activity of Polysaccharides from Rhizoma gastrodiae. Int. J. Mol. Sci. 2016, 17, 1011. https://doi.org/10.3390/ijms17071011
Chen J, Tian S, Shu X, Du H, Li N, Wang J. Extraction, Characterization and Immunological Activity of Polysaccharides from Rhizoma gastrodiae. International Journal of Molecular Sciences. 2016; 17(7):1011. https://doi.org/10.3390/ijms17071011
Chicago/Turabian StyleChen, Juncheng, Shan Tian, Xiaoying Shu, Hongtao Du, Na Li, and Junru Wang. 2016. "Extraction, Characterization and Immunological Activity of Polysaccharides from Rhizoma gastrodiae" International Journal of Molecular Sciences 17, no. 7: 1011. https://doi.org/10.3390/ijms17071011