Petanin Potentiated JNK Phosphorylation to Negatively Regulate the ERK/CREB/MITF Signaling Pathway for Anti-Melanogenesis in Zebrafish
"> Figure 1
<p>The comparison of the melanin (<b>A</b>) and tyrosinase (<b>B</b>) inhibition rate of petanin in zebrafish (NC: water; PC: 11 mM arbutin; PtL: 0.38 mM petanin; PtM: 0.80 mM petanin; PtH: 1.6 mM petanin; *: <span class="html-italic">p</span> < 0.05; **: <span class="html-italic">p</span> < 0.01; ***: <span class="html-italic">p</span> < 0.001; ns: not significant).</p> "> Figure 2
<p>Typical images of the petanin inhibition of melanin production in zebrafish (NC: water; PC: 11 mM arbutin; PtL: 0.38 mM petanin; PtM: 0.80 mM petanin; PtH: 1.6 mM petanin; Scale: 1×; Zebrafish larvae: incubated for 45 h; Dotted red line: region of melanin signal intensity analysis in zebrafish head).</p> "> Figure 3
<p>The effect of petanin on the distribution of melanocytes in zebrafish (HE, 20×). (<b>A</b>) NC group; (<b>B</b>) PC group; (<b>C</b>) PtL group; (<b>D</b>) PtM group; (<b>E</b>) PtH group; →: Melanocytes.</p> "> Figure 4
<p>The effect of petanin on the arrangement of melanocytes in zebrafish (TEM, minimum scale value = 2.0 μm, De: dermis; Me: melanocyte. (<b>A</b>) NC group; (<b>B</b>) PC group; (<b>C</b>) PtL group; (<b>D</b>) PtM group; (<b>E</b>) PtH group).</p> "> Figure 5
<p>The effect of petanin on the structure of melanosomes in zebrafish (TEM, minimum scale value = 50 nm. (<b>A</b>) NC group; (<b>B</b>) PC group; (<b>C</b>) PtL group; (<b>D</b>) PtM group; (<b>E</b>) PtH group).</p> "> Figure 6
<p>The effects of different treatments on oxidoreductase activity in zebrafish ((<b>A</b>) CAT (compared to NC group, ***: <span class="html-italic">p</span> < 0.001, **: <span class="html-italic">p</span> < 0.01, *: <span class="html-italic">p</span> < 0.05, ns: not significant), (<b>B</b>) POD (compared to NC group, ***: <span class="html-italic">p</span> < 0.001, *: <span class="html-italic">p</span> < 0.05, ns: not significant), (<b>C</b>) GR (compared to NC group, ***: <span class="html-italic">p</span> < 0.001, **: <span class="html-italic">p</span> < 0.01, *: <span class="html-italic">p</span> < 0.05, ns: not significant)).</p> "> Figure 7
<p>The network pharmacological analysis of the anti-melanogenesis effect of petanin ((<b>A</b>) important intersection targets; (<b>B</b>) important life processes; (<b>C</b>) key signaling pathways).</p> "> Figure 8
<p>The molecular docking of petanin with JNK ((<b>A</b>) 3D structure; (<b>B</b>) 3D diagram of hydrogen bonds; (<b>C</b>) 2D diagram of hydrogen bonds).</p> "> Figure 9
<p>The molecular dynamics’ simulation of petanin–JNK complex ((<b>A</b>) RMSD; (<b>B</b>) RMSF; (<b>C</b>) Rg; (<b>D</b>) H-bonds number; (<b>E</b>) SASA; (<b>F</b>) Gibbs energy landscape).</p> "> Figure 10
<p>The effects of petanin on the ERK/CREB/MITF signaling pathway ((<b>A</b>) protein expression level; (<b>B</b>) MITF; (<b>C</b>) CREB; (<b>D</b>) p-ERK; (<b>E</b>) p-RSK1; (<b>F</b>) p-JNK; *: <span class="html-italic">p</span> < 0.05; **: <span class="html-italic">p</span> < 0.01).</p> "> Figure 11
<p>The effect of petanin on the transcription of mRNA related to melanogenesis in zebrafish (NC: water; PC: 11 mM arbutin; PtL: 0.38 mM petanin; PtM: 0.80 mM petanin; PtH: 1.6 mM petanin; *: <span class="html-italic">p</span> < 0.05; **: <span class="html-italic">p</span> < 0.01; ***: <span class="html-italic">p</span> < 0.001).</p> ">
Abstract
:1. Introduction
2. Results and Discussion
2.1. Melanin Inhibition Rate
2.2. Tyrosinase Activity in Zebrafish
2.3. Structure and Distribution of Melanocytes and Melanosomes
2.4. Effect of Petanin on Oxidoreductase Activity in Zebrafish
2.5. Network Pharmacological Analysis of the Anti-Melanogenic Effect of Petanin
2.6. Molecular Docking and Dynamic Simulation of Petanin with Key Signaling Pathway Proteins
2.7. Effect of Petanin on the Expression of Proteins Related to Melanin Production in Zebrafish
2.8. Effect of Petanin on Transcription of mRNAs Related to Melanogenesis in Zebrafish
3. Materials and Methods
3.1. Samples and Reagents
3.2. Melanin Inhibition Rate in Zebrafish
3.3. Tyrosinase Activity Assay in Zebrafish
3.4. Detection of CAT, POD and GR Contents in Zebrafish Larvae
3.5. HE Staining
3.6. TEM
3.7. Network Pharmacological Analysis
3.8. Molecular Docking and Molecular Dynamics’ Simulation
3.9. Western Blot
3.10. Gene Expression Analysis
3.11. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sample | Melanin Inhibition Rate | Inhibition Rate of Tyrosinase | p-Value |
---|---|---|---|
NC | - | - | - |
PC | 93.52% | 70.59% | ˂0.001 |
PtL | 13.26% | 14.12% | ˂0.05 |
PtM | 18.64% | 17.65% | ˂0.01 |
PtH | 24.86% | 25.88% | ˂0.001 |
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Ouyang, J.; Hu, N.; Wang, H. Petanin Potentiated JNK Phosphorylation to Negatively Regulate the ERK/CREB/MITF Signaling Pathway for Anti-Melanogenesis in Zebrafish. Int. J. Mol. Sci. 2024, 25, 5939. https://doi.org/10.3390/ijms25115939
Ouyang J, Hu N, Wang H. Petanin Potentiated JNK Phosphorylation to Negatively Regulate the ERK/CREB/MITF Signaling Pathway for Anti-Melanogenesis in Zebrafish. International Journal of Molecular Sciences. 2024; 25(11):5939. https://doi.org/10.3390/ijms25115939
Chicago/Turabian StyleOuyang, Jian, Na Hu, and Honglun Wang. 2024. "Petanin Potentiated JNK Phosphorylation to Negatively Regulate the ERK/CREB/MITF Signaling Pathway for Anti-Melanogenesis in Zebrafish" International Journal of Molecular Sciences 25, no. 11: 5939. https://doi.org/10.3390/ijms25115939