Methotrexate-Induced Alteration of Renal Aquaporins 1 and 2, Oxidative Stress and Tubular Apoptosis Can Be Attenuated by Omega-3 Fatty Acids Supplementation
Abstract
:1. Introduction
2. Results
2.1. Effect of LC-PUFA on MTX-Induced Alteration of Body Weight and on Renal Function Tests
2.2. Effect of LC-PUFA on Oxidative Stress Markers in MTX-Treated Rats
2.3. Effects of LC-PUFA on MTX-Induced Renal Histopathological Alteration and Histopathological Scoring
2.4. Effect of LC-PUFA on Immunohistochemical Analysis of NF-ĸB, BAX, Bcl2, AQP1, and AQP2
2.5. Effect of LC-PUFA on Morphometric Analysis of Immunohistochemical Findings of NF-ĸB, BAX, Bcl2, AQP 1 and AQP2
3. Discussion
4. Materials and Methods
4.1. Sample Size Estimation
4.2. Ethical Statement
4.3. Animals
4.4. Research Design
4.5. Blood Sampling
4.6. Evaluation of Renal Function Tests
4.7. Preparation of Tissue Homogenates
4.8. Estimation of Oxidative Stress Markers
4.9. Histopathological Examination of the Kidney
4.10. Immunohistochemistry
4.11. Morphometric Analysis
4.12. Statistical Analysis
5. Conclusions
6. Study Limitations
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Saline Control (n = 4) | LC-PUFA Control (n = 4) | MTX (n = 4) | MTX + LC-PUFA150 (n = 4) | MTX + LC-PUFA300 (n = 4) | H Value | p Value | |
---|---|---|---|---|---|---|---|
Shrinkage of glomeruli | 0(0–0) A | 0(0–0) A | 2(1–2) B | 1(1–1) BC | 1(0–1) C | 64.778 | <0.0005 |
Dilatation of Bowman’s capsule | 0(0–0) A | 0(0–0.75) A | 2(1.25–2) B | 1(1–1) BC | 1(0–1) AC | 66.029 | <0.0005 |
Tubular dilatation | 0(0–1) A | 0(0–0) A | 2(1.25–2) B | 1(1–1) BC | 1(0–1) AC | 63.082 | <0.0005 |
Tubular cast | 0(0–0.75) A | 0(0–0) A | 2(1.25–2) B | 1(1–2) BC | 1(0–1) AC | 62.767 | <0.0005 |
Inflammatory cell infiltrations | 0(0–1) A | 0(0–0.75) A | 2(1.25–2) B | 1(1–1) BC | 1(0.25–1) AC | 60.135 | <0.0005 |
Congested blood vessels | 0(0–0.75) A | 0(0–0) A | 2(1.25–2) B | 1(1–1) BC | 1(0–1) AC | 64.346 | <0.0005 |
Score | 1(1–2) A | 1(0–2) A | 10(10–11) B | 7(7–7.75) BC | 4(4–5) C | 91.765 | <0.0005 |
Name | Cat. Number | Source and Clonality | Dilution |
---|---|---|---|
NF-kB | ABclonal A19653 | Rabbit monoclonal | 1/100 |
BAX | ABclonal A12009 | Rabbit polyclonal | 1/100 |
Bcl-2 | Genemed 60-0005-7 | Mouse monoclonal | 1/100 |
AQP1 | Servicebio GB11310-1 | Rabbit polyclonal | 1/500 |
AQP2 | Servicebio GB112259 | Rabbit polyclonal | 1/400 |
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El-Agawy, M.S.E.-d.; Badawy, A.M.M.; Rabei, M.R.; Elshaer, M.M.A.; El Nashar, E.M.; Alghamdi, M.A.; Alshehri, M.A.; Elsayed, H.R.H. Methotrexate-Induced Alteration of Renal Aquaporins 1 and 2, Oxidative Stress and Tubular Apoptosis Can Be Attenuated by Omega-3 Fatty Acids Supplementation. Int. J. Mol. Sci. 2022, 23, 12794. https://doi.org/10.3390/ijms232112794
El-Agawy MSE-d, Badawy AMM, Rabei MR, Elshaer MMA, El Nashar EM, Alghamdi MA, Alshehri MA, Elsayed HRH. Methotrexate-Induced Alteration of Renal Aquaporins 1 and 2, Oxidative Stress and Tubular Apoptosis Can Be Attenuated by Omega-3 Fatty Acids Supplementation. International Journal of Molecular Sciences. 2022; 23(21):12794. https://doi.org/10.3390/ijms232112794
Chicago/Turabian StyleEl-Agawy, Mosaab Salah El-din, Alaa Mohamed Mohamed Badawy, Mohammed R. Rabei, Mohamed Mahmoud Abdelraheem Elshaer, Eman Mohamad El Nashar, Mansour A. Alghamdi, Mohammed A. Alshehri, and Hassan Reda Hassan Elsayed. 2022. "Methotrexate-Induced Alteration of Renal Aquaporins 1 and 2, Oxidative Stress and Tubular Apoptosis Can Be Attenuated by Omega-3 Fatty Acids Supplementation" International Journal of Molecular Sciences 23, no. 21: 12794. https://doi.org/10.3390/ijms232112794