Porcine placenta extract (PPE) is widely accepted as an ingredient in complementary and alternative medicine and previous studies have reported its availability, however, its underlying action mechanism remains unclear. In this study, we investigated the underlying mechanism of PPE-induced ceramide synthase 3 upregulation. PPE enhanced the expression of ceramide synthase 3 at both the mRNA and protein levels in HaCaT cells. Moreover, PPE-induced ceramide synthase3 upregulation was suppressed by the Akt inhibitor, suggesting the involvement of Akt in the underlying mechanism. As the PI3K inhibitor did not affect PPE-induced ceramide synthase 3 upregulation, the factors upstream of Akt were estimated. Inhibition and small interfering RNA experiments demonstrated that the peroxisome proliferator-activated receptors δ (PPARδ) and integrin-linked kinase (ILK) are involved in the phosphorylation of Akt. Next, we explored the factors downstream of Akt and found that PPE induced phosphorylation of the STAT3 while PPE-induced upregulation of ceramide synthase 3 was significantly suppressed by the inhibitor of the mTOR, suggesting that the mTOR/STAT3 pathway is involved in the downstream of Akt. These results demonstrate that PPE upregulates CerS3 expression via the PPARδ/ILK/Akt/mTOR/STAT3 pathway.
References
[1]
Natsuga, K. (2014) Epidermal Barriers. Cold Spring Harbor Perspective in Medicine, 4, a018218. https://doi.org/10.1101/cshperspect.a018218
[2]
Proksch, E., Brandner, J.M. and Jensen, J.M. (2008) The Skin: An Indispensable Barrier. Experimental Dermatology, 12, 1063-1072. https://doi.org/10.1111/j.1600-0625.2008.00786.x
[3]
Madison, K.C. (2003) Barrier Function of the Skin: “la raison d’être” of the Epidermis. Journal of Investigative Dermatology, 121, 231-241. https://doi.org/10.1046/j.1523-1747.2003.12359.x
[4]
Wood, L.C., Jackson, S.M., Elias, P.M., Grunfeld, C. and Feingold, K.R. (1992) Cutaneous Barrier Perturbation Stimulates Cytokine Production in the Epidermis of Mice. Journal of Clinical Investigation, 90, 482-487. https://doi.org/10.1172/JCI115884
[5]
Wang, X.P., Schunck, M., Kallen, K.J., Neumann, C., Trautwein, C., Rose-John, S. and Proksch, E. (2004) The Interleukin-6 Cytokine System Regulates Epidermal Permeability Barrier Homeostasis. Journal of Investigative Dermatology, 123, 124-131. https://doi.org/10.1111/j.0022-202X.2004.22736.x
[6]
Jensen, J.M., Schütze, S., Förl, M., Krönke, M. and Proksch, E. (1999) Roles for Tumor Necrosis Factor Receptor p55 and Sphingomyelinase in Repairing the Cutaneous Permeability Barrier. Journal of Clinical Investigation, 104, 1761-1770. https://doi.org/10.1172/JCI5307
[7]
Man, M.Q., Choi, E.H., Schmuth, M., Crumrine, D., Uchida, Y., Elias, P.M., Holleran, W.M. and Feingold, K.R. (2006) Basis for Improved Permeability Barrier Homeostasis Induced by PPAR and LXR Activators: Liposensors Stimulate Lipid Synthesis, Lamellar Body Secretion, and Post-Secretory Lipid Processing. Journal of Investigative Dermatology, 126, 386-392. https://doi.org/10.1038/sj.jid.5700046
[8]
Mizutani, Y., Sun, H., Ohno, Y., Sassa, T., Wakashima, T., Obara, M., Yuyama, K., Kihara, A. and Igarashi, Y. (2013) Cooperative Synthesis of Ultra Long-Chain Fatty Acid and Ceramide during Keratinocyte Differentiation. PLOS ONE, 8, e67317. https://doi.org/10.1371/journal.pone.0067317
[9]
Hong, J.W., Lee, W.J., Hahn, S.B., Kim, B.J. and Lew, D.H. (2010) The Effect of Human Placenta Extract in a Wound Healing Model. Annals of Plastic Surgery, 65, 96-100. https://doi.org/10.1097/SAP.0b013e3181b0bb67
[10]
Hong, K.B., Park, Y., Kim, J.H., Kim, J.M. and Suh, H.J. (2015) Effects of Porcine Placenta Extract Ingestion on Ultraviolet B-Induced Skin Damage in Hairless Mice. Korean Journal of Food Science of Animal Resources, 35, 413-420. https://doi.org/10.5851/kosfa.2015.35.3.413
[11]
O’Keefe, E.J., Payne, R.E. and Russell, N. (1985) Keratinocyte Growth-Promoting Activity from Human Placenta. Journal of Cellular Physiology, 124, 439-445. https://doi.org/10.1002/jcp.1041240312
[12]
Cho, H.R., Ryou, J.H., Lee, J.W. and Lee, M.H. (2008) The Effects of Placental Extract on Fibroblast Proliferation. Journal of Cosmetic Science, 59, 195-202.
[13]
Aioi, A., Muromoto, R., Mogami, S., Nishikawa, M., Ogawa, S. and Matsuda, T. (2021) Porcine Placenta Extract Reduced Wrinkle Formation by Potentiating Epidermal Hydration. Journal of Cosmetic Dermatological Science Application, 11, 101-109. https://doi.org/10.4236/jcdsa.2021.112011
[14]
Sassa, T., Hirayama, T. and Kihara, A. (2016) Enzyme Activities of the Ceramide Synthases CERS2-6 Are Regulated by Phosphorylation in the C-Terminal Region. Journal of Biological Chemistry, 291, 7477-7487. https://doi.org/10.1074/jbc.M115.695858
[15]
Mullen, T.D., Hannun, Y.A. and Obeid, L.M. (2012) Ceramide Synthases at the Centre of Sphingolipid Metabolism and Biology. Biochemistry Journal, 441, 789-802. https://doi.org/10.1042/BJ20111626
[16]
Levy, M. and Futerman, A.H. (2010) Mammalian Ceramide Synthases. IUBMB Life, 62, 347-356. https://doi.org/10.1002/iub.319
[17]
Nicholson, K.M. and Anderson, N.G. (2002) The Protein Kinase B/Akt Signaling Pathway in Human Malignancy. Cellular Signaling, 14, 381-395. https://doi.org/10.1016/S0898-6568(01)00271-6
[18]
Persad, S., Attwell, S., Gray, V., Mawji, N., Deng, J.T., Leung, D., Yan, J., Sanghera, J., Walsh, M.P. and Dedhar, S. (2001) Regulation of Protein Kinase B/Akt-Serine 473 Phosphorylation by Integrin-Linked Kinase: Critical Roles for Kinase Activity and Amino Acids Arginine 211 and Serine 343. Journal of Biological Chemistry, 276, 27462-27469. https://doi.org/10.1074/jbc.M102940200
[19]
Di-Poï, N., Tan, N.S., Michalik, L., Wahli, W. and Desvergne, B. (2002) Antiapoptotic Role of PPARβ in Keratinocytes via Transcriptional Control of the Akt1 Signaling Pathway. Molecular Cell, 10, 721-733. https://doi.org/10.1016/S1097-2765(02)00646-9
[20]
Dibble, C.C. and Cantley, L.C. (2015) Regulation of mTORC1 by PI3K Signaling. Trends in Cell Biology, 25, 545-555. https://doi.org/10.1016/j.tcb.2015.06.002
[21]
Buerger, C., Shirsath, N., Lang, V., Berard, A., Diehl, S., Kaufmann, R., Boehncke, W.H. and Wolf, P. (2017) Inflammation Dependent mTORC1 Signaling Interferes with the Switch from Keratinocyte Proliferation to Differentiation. PLOS ONE, 12, e0180853. https://doi.org/10.1371/journal.pone.0180853
[22]
Li, J., Huang, Q., Long, X., Zhang, J., Huang, X., Aa, J., Yang, H., Chen, Z. and Xing, J. (2015) CD147 Reprograms Fatty Acid Metabolism in Hepatocellular Carcinoma Cells through Akt/mTOR/SREBP1c and P38/PPARα Pathways. Journal of Hepatology, 63, 1378-1389. https://doi.org/10.1016/j.jhep.2015.07.039
[23]
Aioi, A. (2022) Royal Jelly Extract Accelerates Keratinocyte Proliferation, and Upregulates Laminin α3 and Integrin β1 mRNA Expression, via Akt/mTOR/HIF-1α Pathway. Journal of Cosmetics, Dermatological Sciences and Applications, 12, 83-94. https://doi.org/10.4236/jcdsa.2022.122007
[24]
Zhou, J., Wulfkuhle, J., Zhang, H., Gu, P., Yang, Y., Deng, J., Margolick, J.B., Liotta, L.A., Petricoin, E. and Zhang, Y. (2007) Activation of the PTEN/mTOR/STAT3 Pathway in Breast Cancer Stem-Like Cells Is Required for Viability and Maintenance. Proceedings of the National Academy of Sciences USA, 104, 16158-16163. https://doi.org/10.1073/pnas.0702596104
Bromberg, J. and Darnell, J.E. (2000) The Role of STATs in Transcriptional Control and Their Impact on Cellular Function. Oncogene, 19, 2468-2473. https://doi.org/10.1038/sj.onc.1203476
[27]
Levy, D.E. and Lee, C.K. (2002) What Does Stat3 Do? Journal of Clinical Investigation, 109, 1143-1148. https://doi.org/10.1172/JCI0215650
[28]
Sano, S., Itami, S., Takeda, K., Tarutani, M., Yamaguchi, Y., Miura, H., Yoshikawa, K., Akira, S. and Takeda, J. (1999) Keratinocyte-Specific Ablation of Stat3 Exhibits Impaired Skin Remodeling, But Does Not Affect Skin Morphogenesis. EMBO Journal, 18, 4657-4668. https://doi.org/10.1093/emboj/18.17.4657
