Duality of n-3 Polyunsaturated Fatty Acids on Mcp-1 Expression in Vascular Smooth Muscle: A Potential Role of 4-Hydroxy Hexenal
Abstract
:1. Introduction
2. Methods
2.1. Reagents
2.2. Animals and Experimental Procedures
2.3. Cell Culture
2.4. Fatty Acid Treatment
2.5. Messenger RNA (mRNA) Extraction and Real-Time RT-qPCR Analysis
Forward Primer | Reverse Primer | Accession Number | |
---|---|---|---|
Mcp-1 | GCTGCTACTCATTCACTGGCAA | TGCTGCTGGTGATTCTCTTGTA | NM_031530.1 |
Hmox-1 | TCTATCGTGCTCGCATGAAC | AAGGCGGTCTTAGCCTCTTC | NM_012580.2 |
Nrf2 | GGAGCAATTCAACGAAGCTC | ACAGTTCTGAGCGGCAACTT | NM_031789.2 |
18S | TTCCGATAACGAACGAGACTCT | TGGCTGAACGCCACTTGTC | NR_046237.1 |
2.6. Quantitative Analysis of 4-HHE in Biological Samples
2.7. MTT Assay for Cell Viability
2.8. Reactive Oxygen Species (ROS) Measurement Assay
2.9. Western Blot Analysis
2.10. Statistical Analysis
3. Results
3.1. Docosahexaenoic Acid (DHA)—Though Not Eicosapentaenoic Acid (EPA)—Inhibits Mcp-1 mRNA Expression in Rat Aorta
3.2. Paradoxical Increase in Mcp-1 by DHA, EPA and 4-HHE in VSMCs
3.3. 4-HHE Derived from DHA Induces Mcp-1 Expression through the Nrf2 Pathway in Human Vascular Smooth Muscle Cells (VSMCs)
3.4. DHA Induces Apoptosis of VSMCs through 4-HHE
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
References
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Nagayama, K.; Morino, K.; Sekine, O.; Nakagawa, F.; Ishikado, A.; Iwasaki, H.; Okada, T.; Tawa, M.; Sato, D.; Imamura, T.; et al. Duality of n-3 Polyunsaturated Fatty Acids on Mcp-1 Expression in Vascular Smooth Muscle: A Potential Role of 4-Hydroxy Hexenal. Nutrients 2015, 7, 8112-8126. https://doi.org/10.3390/nu7095381
Nagayama K, Morino K, Sekine O, Nakagawa F, Ishikado A, Iwasaki H, Okada T, Tawa M, Sato D, Imamura T, et al. Duality of n-3 Polyunsaturated Fatty Acids on Mcp-1 Expression in Vascular Smooth Muscle: A Potential Role of 4-Hydroxy Hexenal. Nutrients. 2015; 7(9):8112-8126. https://doi.org/10.3390/nu7095381
Chicago/Turabian StyleNagayama, Kohji, Katsutaro Morino, Osamu Sekine, Fumiyuki Nakagawa, Atsushi Ishikado, Hirotaka Iwasaki, Takashi Okada, Masashi Tawa, Daisuke Sato, Takeshi Imamura, and et al. 2015. "Duality of n-3 Polyunsaturated Fatty Acids on Mcp-1 Expression in Vascular Smooth Muscle: A Potential Role of 4-Hydroxy Hexenal" Nutrients 7, no. 9: 8112-8126. https://doi.org/10.3390/nu7095381