Ultrasonic-Cellulase Synergistic Extraction of Crude Polysaccharides from Moringa oleifera Leaves and Alleviation of Insulin Resistance in HepG2 Cells
Abstract
:1. Introduction
2. Results
2.1. Ultrasound-Assisted Cellulase Extraction of Polysaccharides from MOL Single-Factor Experiment
2.2. Plackett–Burman Experiment
2.3. Response Surface Optimisation of the Extraction Process
2.4. Inhibition of α-Glucosidase and Antioxidant Activity of MOLP
2.5. MOLP Attenuates High Glucose-Induced Oxidative Stress in IR-HepG2 Cells
2.6. Effect of MOLP on the ROS Content of IR-HepG2 Cells
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Optimization of the Extraction Process of MOLP
4.3. Preparation of Crude Polysaccharides from Moringa oleifera Leaves (MOLP)
4.4. Determination of Antioxidant Activity
4.4.1. Determination of DPPH Radical Scavenging Capacity
4.4.2. Determination of Hydroxyl Radical Scavenging Capacity
4.4.3. Superoxide Anion Radical Scavenging Capacity
4.4.4. ABTS Radical Scavenging Capacity
4.5. Detection of α-Glucosidase Inhibitory Activity of MOLP
4.5.1. Determination of the Reaction Time of α-Glucosidase with the Substrate PNPG
4.5.2. α-Glucosidase Activity Assay
4.5.3. Kinetic Analysis of the Inhibitory Effect of MOLP on α-Glucosidase Activity
4.5.4. Effect of MOLP on Insulin-Resistant HepG2 Cells
MTT Cell Viability Assay
Effect of MOLP on Glucose Consumption and Glycogen Synthesis in IR-HepG2
Effect of MOLP on CAT, SOD, GSH-Px, MDA and SOD in IR-HepG2 Cells
4.6. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Informed Consent Statement
Conflicts of Interest
References
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Std | Run | A The Additive Quantity of Cellulase (%) | B Hydrolysis Temperature (°C) | C Hydrolysis Time (°C) | D pH | E Ultrasonic Power (W) | F Ultrasonic Temperature (°C) | G Ultrasound Time (min) | H Ratio of Material to Water (g/mL) | Extraction Yield (%) |
---|---|---|---|---|---|---|---|---|---|---|
11 | 1 | 0.9 | 55 | 150 | 7 | 540 | 55 | 20 | 20 | 16.39 ± 0.34 |
4 | 2 | 0.7 | 65 | 90 | 7 | 540 | 55 | 40 | 40 | 17.25 ± 0.53 |
5 | 3 | 0.7 | 55 | 150 | 6 | 540 | 65 | 20 | 40 | 15.05 ± 0.23 |
3 | 4 | 0.9 | 55 | 150 | 7 | 420 | 65 | 40 | 40 | 15.46 ± 0.54 |
8 | 5 | 0.9 | 65 | 90 | 6 | 420 | 65 | 20 | 40 | 16.71 ± 0.46 |
1 | 6 | 0.9 | 65 | 90 | 7 | 540 | 65 | 20 | 20 | 16.98 ± 0.68 |
9 | 7 | 0.9 | 65 | 150 | 6 | 420 | 55 | 40 | 20 | 16.91 ± 0.79 |
2 | 8 | 0.7 | 65 | 150 | 6 | 540 | 65 | 40 | 20 | 17.81 ± 0.65 |
12 | 9 | 0.7 | 55 | 90 | 6 | 420 | 55 | 20 | 20 | 14.21 ± 0.84 |
6 | 10 | 0.7 | 55 | 90 | 7 | 420 | 65 | 40 | 20 | 14.21 ± 0.32 |
7 | 11 | 0.9 | 55 | 90 | 6 | 540 | 55 | 40 | 40 | 17.81 ± 0.42 |
10 | 12 | 0.7 | 65 | 150 | 7 | 420 | 55 | 20 | 40 | 14.29 ± 0.36 |
Source | Sum of Square | df | Mean Square | F Value | p Value | |
---|---|---|---|---|---|---|
Model | 20.28 | 8 | 2.54 | 10.75 | 0.0382 | * |
A The additive quantity of cellulase (%) | 4.61 | 1 | 4.61 | 19.53 | 0.0215 | * |
B Hydrolysis temperature (°C) | 3.88 | 1 | 3.88 | 16.47 | 0.027 | * |
C Hydrolysis time (°C) | 0.13 | 1 | 0.13 | 0.54 | 0.5155 | |
D pH | 1.28 | 1 | 1.28 | 5.41 | 0.1024 | |
E Ultrasonic power (W) | 7.53 | 1 | 7.53 | 31.92 | 0.011 | * |
F Ultrasonic temperature (°C) | 0.036 | 1 | 0.036 | 0.15 | 0.7233 | |
G Ultrasonic time (min) | 2.83 | 1 | 2.83 | 11.98 | 0.0406 | * |
H Ratio of material to water (g/mL) | 4.38 × 10−4 | 1 | 4.38 × 10−4 | 1.86 × 10−3 | 0.9683 | |
Residual | 0.71 | 3 | 0.24 | |||
Cor Total | 20.99 | 11 |
Std | Run | A The Additive Quantity of Cellulase (%) | B Hydrolysis Temperature (°C) | C Ultrasonic Power (W) | D Ultrasonic Time (min) | Extraction Yield (%) |
---|---|---|---|---|---|---|
19 | 1 | 0.7 | 60 | 540 | 30 | 15.47 ± 0.32 |
18 | 2 | 0.9 | 60 | 420 | 30 | 15.50 ± 0.65 |
25 | 3 | 0.8 | 60 | 480 | 30 | 17.03 ± 0.75 |
28 | 4 | 0.8 | 60 | 480 | 30 | 17.71 ± 0.32 |
26 | 5 | 0.8 | 60 | 480 | 30 | 17.67 ± 0.65 |
6 | 6 | 0.8 | 60 | 540 | 20 | 15.11 ± 0.12 |
20 | 7 | 0.9 | 60 | 540 | 30 | 15.45 ± 0.35 |
17 | 8 | 0.7 | 60 | 420 | 30 | 15.41 ± 1.12 |
7 | 9 | 0.8 | 60 | 420 | 40 | 15.62 ± 0.23 |
22 | 10 | 0.8 | 65 | 480 | 20 | 14.71 ± 0.56 |
27 | 11 | 0.8 | 60 | 480 | 30 | 17.54 ± 0.31 |
3 | 12 | 0.7 | 65 | 480 | 30 | 15.17 ± 0.43 |
15 | 13 | 0.8 | 55 | 540 | 30 | 16.02 ± 0.32 |
29 | 14 | 0.8 | 60 | 480 | 30 | 17.90 ± 0.68 |
4 | 15 | 0.9 | 65 | 480 | 30 | 15.32 ± 0.78 |
10 | 16 | 0.9 | 60 | 480 | 20 | 15.41 ± 0.36 |
2 | 17 | 0.9 | 55 | 480 | 30 | 15.90 ± 1.2 |
9 | 18 | 0.7 | 60 | 480 | 20 | 13.77 ± 0.86 |
23 | 19 | 0.8 | 55 | 480 | 40 | 14.40 ± 0.35 |
11 | 20 | 0.7 | 60 | 480 | 40 | 14.67 ± 0.65 |
21 | 21 | 0.8 | 55 | 480 | 20 | 15.55 ± 0.67 |
1 | 22 | 0.7 | 55 | 480 | 30 | 15.38 ± 0.75 |
13 | 23 | 0.8 | 55 | 420 | 30 | 14.16 ± 0.36 |
12 | 24 | 0.9 | 60 | 480 | 40 | 16.09 ± 0.53 |
8 | 25 | 0.8 | 60 | 540 | 40 | 15.89 ± 0.63 |
14 | 26 | 0.8 | 65 | 420 | 30 | 15.33 ± 0.54 |
16 | 27 | 0.8 | 65 | 540 | 30 | 15.18 ± 0.63 |
5 | 28 | 0.8 | 60 | 420 | 20 | 14.17 ± 0.35 |
24 | 29 | 0.8 | 65 | 480 | 40 | 15.80 ± 0.45 |
Source | Sum of Square | df | Mean Square | F Value | p Value | |
---|---|---|---|---|---|---|
Model | 28.7 | 14 | 2.05 | 8.71 | 0.0001 | *** |
A The additive quantity of cellulase | 1.21 | 1 | 1.21 | 5.15 | 0.0396 | * |
B Hydrolysis temperature | 1.15 × 10−3 | 1 | 1.15 × 10−3 | 4.88 × 10−3 | 0.9453 | |
C Ultrasonic power | 0.72 | 1 | 0.72 | 3.04 | 0.1032 | |
D Ultrasonic time | 1.18 | 1 | 1.18 | 5.01 | 0.0419 | * |
AB | 0.034 | 1 | 0.034 | 0.14 | 0.7117 | |
AC | 3.11 × 10−3 | 1 | 3.11 × 10−3 | 0.013 | 0.9102 | |
AD | 0.013 | 1 | 0.013 | 0.054 | 0.8196 | |
BC | 1.01 | 1 | 1.01 | 4.28 | 0.0577 | |
BD | 1.27 | 1 | 1.27 | 5.38 | 0.036 | * |
CD | 0.11 | 1 | 0.11 | 0.48 | 0.5003 | |
A2 | 7.44 | 1 | 7.44 | 31.59 | <0.0001 | |
B2 | 8.58 | 1 | 8.58 | 36.43 | <0.0001 | |
C2 | 7.88 | 1 | 7.88 | 33.48 | <0.0001 | |
D2 | 12.09 | 1 | 12.09 | 51.33 | <0.0001 | |
Residual | 3.3 | 14 | 0.24 | |||
Lack of Fit | 2.86 | 10 | 0.29 | 2.66 | 0.1796 | |
Pure Error | 0.43 | 4 | 0.11 | |||
Cor Total | 32 | 28 |
Independent Variables | Levels | ||||
---|---|---|---|---|---|
The additive quantity of cellulase (%) | 0.6 | 0.7 | 0.8 | 0.9 | 1.0 |
Hydrolysis temperature (°C) | 50 | 55 | 60 | 65 | 70 |
Hydrolysis time (°C) | 30 | 60 | 90 | 120 | 150 |
pH | 5 | 5.5 | 6 | 6.5 | 7 |
Ultrasonic power (W) | 300 | 360 | 420 | 480 | 540 |
Ultrasonic temperature (°C) | 55 | 60 | 65 | 70 | 75 |
Ultrasound time (min) | 10 | 20 | 30 | 40 | 50 |
Ratio of material to water (g/mL) | 1:20 | 1:30 | 1:40 | 1:50 | 1:60 |
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Gu, F.; Tao, L.; Chen, R.; Zhang, J.; Wu, X.; Yang, M.; Sheng, J.; Tian, Y. Ultrasonic-Cellulase Synergistic Extraction of Crude Polysaccharides from Moringa oleifera Leaves and Alleviation of Insulin Resistance in HepG2 Cells. Int. J. Mol. Sci. 2022, 23, 12405. https://doi.org/10.3390/ijms232012405
Gu F, Tao L, Chen R, Zhang J, Wu X, Yang M, Sheng J, Tian Y. Ultrasonic-Cellulase Synergistic Extraction of Crude Polysaccharides from Moringa oleifera Leaves and Alleviation of Insulin Resistance in HepG2 Cells. International Journal of Molecular Sciences. 2022; 23(20):12405. https://doi.org/10.3390/ijms232012405
Chicago/Turabian StyleGu, Fan, Liang Tao, Runling Chen, Jiao Zhang, Xingzhong Wu, Min Yang, Jun Sheng, and Yang Tian. 2022. "Ultrasonic-Cellulase Synergistic Extraction of Crude Polysaccharides from Moringa oleifera Leaves and Alleviation of Insulin Resistance in HepG2 Cells" International Journal of Molecular Sciences 23, no. 20: 12405. https://doi.org/10.3390/ijms232012405