Hyperhalophilic Diatom Extract Protects against Lead-Induced Oxidative Stress in Rats and Human HepG2 and HEK293 Cells
Abstract
:1. Introduction
2. Results
2.1. Biochemical Composition of Diatom Halamphora Extract
2.2. Cytotoxicity Assay
2.3. Effect of Lead Acetate on Cell Viability
2.4. Cytoprotective Effect of the Extract against Apoptotic Cell Death Due to Lead Acetate
2.5. Effects of Extract Treatment on Lipid Peroxidation Levels in the Liver and Kidney
2.6. Effects of Extract Treatment on Antioxidant Enzyme Activities in the Liver and Kidney
2.7. Effects of Extract Treatment on the Serum Biochemical Parameters in Rats
2.8. Histological Study
2.8.1. Hepatocyte Histology
2.8.2. Kidney Effect
3. Discussion
4. Materials and Methods
4.1. Chemicals
4.2. Microalgal Isolation and Culture Conditions
4.3. Microalgal Extract, Preparation, and Characterization
4.3.1. Extraction of Biomolecules
4.3.2. Determination of the Total Phenol Content in the Extract
4.3.3. Determination of the Total Flavonoid Content in the Extract
4.3.4. Determination of the Content of Pigments in the Extract: Chlorophyll (a) and Carotenoids
4.3.5. Estimation of the Fatty Acid Profile in the Extract via GC–MS
4.4. In Vitro Cytotoxicity Assay
4.4.1. Cell Lines and Culture
4.4.2. In Vitro Cytotoxicity Assay
4.4.3. Cell Treatment
4.5. Animals and In Vivo Treatment
4.5.1. Experimental Design
4.5.2. Determination of Stress Biomarkers
4.5.3. Antioxidant Enzyme Status
4.5.4. Biochemical Markers in Serum
4.5.5. Histological Analysis
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Biochemical Components | Halamphora Extract |
---|---|
Yield (%DW) | 18.20 ± 0.21 |
Phenolic compounds | |
Polyphenols (mg GAE g−1 extract) | 38.27 ± 2.21 |
Flavonoids (mg QE g−1 extract) | 17.69 ± 0.70 |
Pigments | |
Carotenoids (mg g−1 extract) | 2.19 ± 0.05 |
Chlorophyll (a) (mg g−1 extract) | 6.11 ± 0.10 |
Fatty acids | |
C14:0 | 0.175 ± 0.4 |
C15:0 | 0.851 ± 0.3 |
C16:0 | 29.464 ± 0.3 |
C17:0 | 1.655 ± 0.5 |
C18:0 | 2.317 ± 0.3 |
C20:0 | 0.463 ± 0.2 |
C24:0 | 3.752 ± 0.2 |
Total SFA | 38.677 ± 0.6 |
C14:1(n-5) | 1.072 ± 0.2 |
C16:1(n-7) | 42.066 ± 0.7 |
C17:1(n-8) | 0.446 ± 0.1 |
C18:1(n-9) | 4.993 ± 0.4 |
C19:1(n-9) | 1.316 ± 0.2 |
Total MUFA | 49.893 ± 1.3 |
C16:2(n-7) | 1.459 ± 0.3 |
C18:2(n-6) | 0.567 ± 0.1 |
C20:4(n-6) (ArA) | 2.867 ± 0.2 |
C20:5(n-3) (EPA) | 6.067 ± 0.3 |
Total PUFA | 10.960 ± 0.5 |
Parameters and Treatments | MDA (nmol of MDA mg−1 Protein) | SOD (U SOD mg−1 Protein) | CAT (µmoles H2O2 Destroyed min−1 mg−1 Protein) | GPx (nmoles of GSH min−1 mg−1 Protein) |
---|---|---|---|---|
Liver | ||||
Group I | 1.014 ± 0.10 | 64.736 ± 5.08 | 272.156 ± 39.59 | 0.027 ± 0.001 |
Group II | 1.064 ± 0.09 | 65.145 ± 4.47 | 269.520 ± 42.98 | 0.027 ± 0.002 |
Group III | 1.397 ± 0.05 # | 45.305 ± 4.14 # | 132.054 ± 30.97 # | 0.020 ± 0.001 ## |
Group IV | 1.110 ± 0.02 ** | 66.323 ± 2.98 * | 259.020 ± 18.75 ** | 0.028 ± 0.002 * |
Kidney | ||||
Group I | 0.960 ± 0.05 | 42.897 ± 2.23 | 89.344 ± 10.27 | 0.022 ± 0.001 |
Group II | 0.902 ± 0.07 | 41.857 ± 4.70 | 90.501 ± 9.37 | 0.020 ± 0.001 |
Group III | 1.233 ± 0.07 # | 24.613 ± 4.41 # | 61.078 ± 4.09 # | 0.016 ± 0.0005 ### |
Group IV | 1.057 ± 0.07 * | 50.946 ± 4.00 * | 98.424 ± 8.10 ** | 0.023 ± 0.0004 *** |
Groups | Group I | Group II | Group III | Group IV |
---|---|---|---|---|
ASAT (IU/L) | 79 ± 12.8 | 80.2 ± 14.6 | 195.8 ± 38.9 ## | 123.2 ± 9.8 * |
ALAT(IU/L) | 14.8 ± 2.3 | 19.7 ± 2.7 | 32.6 ± 5.3 ## | 22.2 ± 1.4 * |
ALP (U/L) | 103 ± 22.1 | 117.5 ± 19.7 | 212.8 ± 16.1 ## | 132 ± 26.6 * |
LDH (U/L) | 780.7± 114.2 | 802.2 ± 133.8 | 1489.8 ± 168.8 ## | 1004.3 ± 75.4 * |
Urea (mmol/L) | 3.0 ± 0.44 | 4.1 ± 0.23 | 6.1 ± 0.58 ## | 4.23 ± 0.38 * |
Creatinine (µmol/L) | 20 ± 1.9 | 19.3 ± 1.8 | 30.6 ± 2.97 ## | 22.8 ± 2.5 * |
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Guermazi, W.; Boukhris, S.; Annabi-Trabelsi, N.; Rebai, T.; Sellami-Kamoun, A.; Aldahmash, W.; Plavan, G.I.; Harrath, A.H.; Ayadi, H. Hyperhalophilic Diatom Extract Protects against Lead-Induced Oxidative Stress in Rats and Human HepG2 and HEK293 Cells. Pharmaceuticals 2023, 16, 875. https://doi.org/10.3390/ph16060875
Guermazi W, Boukhris S, Annabi-Trabelsi N, Rebai T, Sellami-Kamoun A, Aldahmash W, Plavan GI, Harrath AH, Ayadi H. Hyperhalophilic Diatom Extract Protects against Lead-Induced Oxidative Stress in Rats and Human HepG2 and HEK293 Cells. Pharmaceuticals. 2023; 16(6):875. https://doi.org/10.3390/ph16060875
Chicago/Turabian StyleGuermazi, Wassim, Saoussan Boukhris, Neila Annabi-Trabelsi, Tarek Rebai, Alya Sellami-Kamoun, Waleed Aldahmash, Gabriel Ionut Plavan, Abdel Halim Harrath, and Habib Ayadi. 2023. "Hyperhalophilic Diatom Extract Protects against Lead-Induced Oxidative Stress in Rats and Human HepG2 and HEK293 Cells" Pharmaceuticals 16, no. 6: 875. https://doi.org/10.3390/ph16060875