Ultrasonic-Assisted Enzymolysis Extraction and Protective Effect on Injured Cardiomyocytes in Mice of Flavonoids from Prunus mume Blossom
Abstract
:1. Introduction
2. Results and Discussion
2.1. Single-Factor Test Results
2.2. Orthogonal Test Results
2.3. Identification of Main Components in FPMB
2.4. Effect of FPMB on the Cardiomyocytes’ Survival Rate
2.5. Effect of FPMB on the ROS Contents
2.6. Effect of FPMB on the Activities of Caspase-3, Caspase-8, and Caspase-9
2.7. Effect of on Cardiomyocytes’ Apoptosis
3. Materials and Methods
3.1. Raw Materials
3.2. Extraction of FPMB
3.3. Measurements and Analyses
3.3.1. Determination of Flavonoids Content
Drawing of a Standard Curve
Flavonoids Content Measurement
3.3.2. Single-Factor Test Design
3.3.3. Orthogonal Experimental Design
3.3.4. Identification of FPMB by HPLC
3.3.5. Isolation of Cardiomyocytes
3.3.6. Design of Protective Effect Test on Cardiomyocytes
3.3.7. Measurements of Cardiomyocytes’ Activity
3.3.8. Measurements of ROS Activity
3.3.9. Measurements of Caspase-3, Caspase-8, and Caspase-9 Activity in Cardiomyocytes
3.3.10. Measurements of Cardiomyocytes’ Apoptosis
3.4. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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Groups | Extraction Rate (%) | |
---|---|---|
Cellulase mass percentage (%) | 0.5 | 1.80 ± 0.07 d |
1.0 | 2.63 ± 0.12 c | |
1.5 | 3.52 ± 0.15 b | |
2.0 | 5.81 ± 0.26 a | |
2.5 | 5.83 ± 0.23 a | |
3.0 | 5.92 ± 0.19 a | |
Hydrolysis temperature (°C) | 30 | 3.75 ± 0.11 c |
35 | 4.82 ± 0.26 b | |
40 | 5.63 ± 0.28 a | |
45 | 5.65 ± 0.26 a | |
50 | 5.58 ± 0.19 a | |
55 | 5.49 ± 0.15 a |
Groups | Extraction Rate (%) | |
---|---|---|
Ultrasonic power (W) | 200 | 3.69 ± 0.11 c |
250 | 5.07 ± 0.15 b | |
300 | 6.15 ± 0.23 a | |
350 | 6.10 ± 0.18 a | |
400 | 5.91 ± 0.21 a | |
450 | 5.85 ± 0.17 a | |
Ultrasonic time (min) | 20 | 4.27 ± 0.16 c |
30 | 5.12 ± 0.26 b | |
40 | 6.06 ± 0.21 a | |
50 | 6.17 ± 0.31 a | |
60 | 6.15 ± 0.16 a | |
70 | 6.23 ± 0.22 a |
Number | Cellulase Mass Percentage A (%) | Hydrolysis Temperature B (°C) | Ultrasonic Power C (W) | Ultrasonic Time D (S) | Extraction Rate (%) |
---|---|---|---|---|---|
1 | 1.5 | 35 | 250 | 30 | 5.62 |
2 | 1.5 | 40 | 300 | 40 | 5.78 |
3 | 1.5 | 45 | 350 | 50 | 5.95 |
4 | 2.0 | 35 | 300 | 50 | 5.86 |
5 | 2.0 | 40 | 350 | 30 | 6.28 |
6 | 2.0 | 45 | 200 | 40 | 6.03 |
7 | 2.5 | 35 | 350 | 40 | 6.31 |
8 | 2.5 | 40 | 200 | 50 | 5.97 |
9 | 2.5 | 45 | 250 | 30 | 6.15 |
K1 | 17.350 | 17.790 | 17.620 | 18.050 | |
K2 | 18.170 | 18.030 | 17.790 | 18.120 | |
K3 | 18.430 | 18.130 | 18.540 | 17.780 | |
k1 | 5.783 | 5.930 | 5.873 | 6.017 | |
k2 | 6.057 | 6.010 | 5.930 | 6.040 | |
k3 | 6.143 | 6.043 | 6.180 | 5.927 | |
R | 0.360 | 0.113 | 0.250 | 0.113 |
Variance Source | Square Sum | Degrees of Freedom | Mean Square | F-Value | Significance |
---|---|---|---|---|---|
A | 0.212 | 2 | 0.106 | 10.406 | * |
B | 0.020 | 2 | 0.010 | 1.000 | |
C | 0.160 | 2 | 0.080 | 7.848 | * |
D | 0.021 | 2 | 0.011 | 1.056 | |
Error | 0.020 | 2 | 0.010 |
Groups | Apoptosis Rate (%) |
---|---|
Control | 24.20 ± 0.67 e |
H2O2 | 61.23 ± 1.81 a |
FPMB-L | 40.80 ± 1.07 b |
FPMB-M | 30.10 ± 0.88 c |
FPMB-H | 26.60 ± 0.53 d |
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Zhu, S.; Xu, J.; Chen, H.; Lv, W. Ultrasonic-Assisted Enzymolysis Extraction and Protective Effect on Injured Cardiomyocytes in Mice of Flavonoids from Prunus mume Blossom. Molecules 2021, 26, 5818. https://doi.org/10.3390/molecules26195818
Zhu S, Xu J, Chen H, Lv W. Ultrasonic-Assisted Enzymolysis Extraction and Protective Effect on Injured Cardiomyocytes in Mice of Flavonoids from Prunus mume Blossom. Molecules. 2021; 26(19):5818. https://doi.org/10.3390/molecules26195818
Chicago/Turabian StyleZhu, Shengnan, Jicheng Xu, Huizhi Chen, and Weiqiao Lv. 2021. "Ultrasonic-Assisted Enzymolysis Extraction and Protective Effect on Injured Cardiomyocytes in Mice of Flavonoids from Prunus mume Blossom" Molecules 26, no. 19: 5818. https://doi.org/10.3390/molecules26195818