Genistein Improves Skin Flap Viability in Rats: A Preliminary In Vivo and In Vitro Investigation
Abstract
:1. Introduction
2. Results
2.1. Macroscopic Analysis of Flap Survival
2.2. Flap Histology
2.3. Proliferation and Migration of Endothelial Cells
2.4. Gene Expression Profile of Endothelial Cells
2.5. Bcl-2 Quantification after Genistein Treatment
2.6. CAT and SOD Activity in Endothelial Cells
2.7. Genistein and ROS
3. Discussion
4. Materials and Methods
4.1. Animal Model
4.2. Genistein Treatment
4.3. Measurement of Survived Area on the Skin Flap
4.4. Basic Histology
4.5. Endothelial Cells
4.6. Cell Viability Assay
4.7. In Vitro Migration (Wound Healing) Assay
4.8. RNA Isolation and cDNA Synthesis
4.9. Gene Expression Profiling
4.10. PCR Data Analysis and Statistics
4.11. Flow Cytometric (FCM) Analysis of Genistein Effect on Pro-/Anti-Apoptotic Gene Expressions
4.12. CAT and SOD Activity
4.13. Detection of Intracellular Oxidative Stress
4.14. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Not available. |
Group/Subgroup | No. of Rats | Ovariectomy (O)/ Sham Surgery (S) | Treatment Started | Dose mg/kg | |
---|---|---|---|---|---|
NOVX | -C | 8 | S—3M prior flap surgery | from the day of flap surgery | 0 |
-G0 | 8 | S—3M prior flap surgery | from the day of flap surgery | 1 | |
-G3 | 8 | S—3M prior flap surgery | 3 days prior flap surgery | 1 | |
OVX | -C | 8 | O—3M prior flap surgery | from the day of flap surgery | 0 |
-G0 | 8 | O—3M prior flap surgery | from the day of flap surgery | 1 | |
-G3 | 8 | O—3M prior flap surgery | 3 days prior flap surgery | 1 |
Gene Symbol | Gene Name | Genistein + VEGF/VEGF |
---|---|---|
COL18A1 | Collagen Type XVIII Alpha 1 Chain | 2.2 |
CTGF | Connective Tissue Growth Factor | 2.4 |
CXCL5 | C-X-C Motif Chemokine Ligand 5 | 2.8 |
IL-6 | Interleukin 6 | 2.1 |
ITGB3 | Integrin Subunit Beta 3 | 2.6 |
MMP-14 | Matrix Metallopeptidase 14 | 2.4 |
TIMP-2 | TIMP Metallopeptidase Inhibitor 2 | 2.2 |
TIMP-3 | TIMP Metallopeptidase Inhibitor 3 | 2.2 |
VEGF-A | Vascular Endothelial Growth Factor A | 2.4 |
Gene Symbol | Gene Name | Genistein/Control |
---|---|---|
CXCL10 | C-X-C Motif Chemokine Ligand 10 | −2.7 |
SERPINF1 | Serpin Family F Member 1 | −2.1 |
Analysis | Staining Solution | Manufacturer |
---|---|---|
Protein analysis | Bcl-2 (124) Mouse mAb (PE Conjugate) 1:50 | Cell Signaling Technology, Danvers, MA, USA |
Phospho-Bcl-2 (Ser70) Rabbit mAb (Alexa Fluor® 488 Conjugate) 1:50 |
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Fáber, L.; Kováč, I.; Mitrengová, P.; Novotný, M.; Varinská, L.; Vasilenko, T.; Kello, M.; Čoma, M.; Kuruc, T.; Petrová, K.; et al. Genistein Improves Skin Flap Viability in Rats: A Preliminary In Vivo and In Vitro Investigation. Molecules 2018, 23, 1637. https://doi.org/10.3390/molecules23071637
Fáber L, Kováč I, Mitrengová P, Novotný M, Varinská L, Vasilenko T, Kello M, Čoma M, Kuruc T, Petrová K, et al. Genistein Improves Skin Flap Viability in Rats: A Preliminary In Vivo and In Vitro Investigation. Molecules. 2018; 23(7):1637. https://doi.org/10.3390/molecules23071637
Chicago/Turabian StyleFáber, Lenka, Ivan Kováč, Petra Mitrengová, Martin Novotný, Lenka Varinská, Tomáš Vasilenko, Martin Kello, Matúš Čoma, Tomáš Kuruc, Klaudia Petrová, and et al. 2018. "Genistein Improves Skin Flap Viability in Rats: A Preliminary In Vivo and In Vitro Investigation" Molecules 23, no. 7: 1637. https://doi.org/10.3390/molecules23071637