Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins
We previously demonstrated that the silk protein sericin promotes pigmentation of retinal pigment epithelium (RPE) by activating the NF-κB pathway. Among numerous agents, NF-κB can be activated by hydrogen peroxide. In the present study, we explored possible associations between reactive oxygen spec...
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| Format: | Article |
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MDPI AG
2020-09-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/19/4395 |
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| author | Ayyad Zartasht Khan Catherine Joan Jackson Tor Paaske Utheim Sjur Reppe Dipak Sapkota Ole Kristoffer Olstad Bernd Thiede Jon Roger Eidet |
| author_facet | Ayyad Zartasht Khan Catherine Joan Jackson Tor Paaske Utheim Sjur Reppe Dipak Sapkota Ole Kristoffer Olstad Bernd Thiede Jon Roger Eidet |
| author_sort | Ayyad Zartasht Khan |
| collection | DOAJ |
| description | We previously demonstrated that the silk protein sericin promotes pigmentation of retinal pigment epithelium (RPE) by activating the NF-κB pathway. Among numerous agents, NF-κB can be activated by hydrogen peroxide. In the present study, we explored possible associations between reactive oxygen species and sericin-induced melanogenesis in RPE. The proteome of human fetal RPE cultured for seven days with or without 1% sericin was analyzed using ingenuity pathway analysis (IPA). The proteomic data was verified by immunofluorescence and immunoblotting. Light microscopy and scanning electron microscopy were used to assess morphology. Dihydroethidium (DHE) and dihydrorhodamine (DHR) assays were used to measure superoxide and hydrogen peroxide species. Expression levels of proteins related to inflammation, differentiation, cell survival and cell adhesion were higher in cells cultured in Dulbecco’s Modified Eagle Medium (DMEM) with 1% sericin, whereas cells cultured in DMEM alone showed higher expression levels of proteins associated with Bruch’s membrane and cytoskeleton. Despite upregulation of inflammatory proteins, sericin co-cultured RPE yielded significantly higher cell viability compared to cells cultured without sericin. Addition of sericin to culture media significantly increased hydrogen peroxide-levels without significantly affecting superoxide-levels. We suggest that sericin-induced melanogenesis in cultured RPE is associated with elevated levels of superoxide dismutase, hydrogen peroxide and inflammatory proteins. |
| first_indexed | 2024-03-10T16:04:56Z |
| format | Article |
| id | doaj.art-ad1142a7d2dd43878d399ef3bc818b6d |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T16:04:56Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-ad1142a7d2dd43878d399ef3bc818b6d2023-11-20T14:57:46ZengMDPI AGMolecules1420-30492020-09-012519439510.3390/molecules25194395Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory ProteinsAyyad Zartasht Khan0Catherine Joan Jackson1Tor Paaske Utheim2Sjur Reppe3Dipak Sapkota4Ole Kristoffer Olstad5Bernd Thiede6Jon Roger Eidet7Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, P.O. Box 1078, Blindern, 0316 Oslo, NorwayDepartment of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, NorwayDepartment of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, NorwayDepartment of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, NorwayDepartment of Oral Biology, Faculty of Dentistry, University of Oslo, Sognsvannsveien 10, P.O. Box 1052, 0316 Oslo, NorwayDepartment of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, NorwayDepartment of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, P.O. Box 1066, Blindern, 0316 Oslo, NorwayDepartment of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, NorwayWe previously demonstrated that the silk protein sericin promotes pigmentation of retinal pigment epithelium (RPE) by activating the NF-κB pathway. Among numerous agents, NF-κB can be activated by hydrogen peroxide. In the present study, we explored possible associations between reactive oxygen species and sericin-induced melanogenesis in RPE. The proteome of human fetal RPE cultured for seven days with or without 1% sericin was analyzed using ingenuity pathway analysis (IPA). The proteomic data was verified by immunofluorescence and immunoblotting. Light microscopy and scanning electron microscopy were used to assess morphology. Dihydroethidium (DHE) and dihydrorhodamine (DHR) assays were used to measure superoxide and hydrogen peroxide species. Expression levels of proteins related to inflammation, differentiation, cell survival and cell adhesion were higher in cells cultured in Dulbecco’s Modified Eagle Medium (DMEM) with 1% sericin, whereas cells cultured in DMEM alone showed higher expression levels of proteins associated with Bruch’s membrane and cytoskeleton. Despite upregulation of inflammatory proteins, sericin co-cultured RPE yielded significantly higher cell viability compared to cells cultured without sericin. Addition of sericin to culture media significantly increased hydrogen peroxide-levels without significantly affecting superoxide-levels. We suggest that sericin-induced melanogenesis in cultured RPE is associated with elevated levels of superoxide dismutase, hydrogen peroxide and inflammatory proteins.https://www.mdpi.com/1420-3049/25/19/4395sericinmelanogenesissuperoxide dismutaseinflammationretinal pigment epitheliumhydrogen peroxide |
| spellingShingle | Ayyad Zartasht Khan Catherine Joan Jackson Tor Paaske Utheim Sjur Reppe Dipak Sapkota Ole Kristoffer Olstad Bernd Thiede Jon Roger Eidet Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins Molecules sericin melanogenesis superoxide dismutase inflammation retinal pigment epithelium hydrogen peroxide |
| title | Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins |
| title_full | Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins |
| title_fullStr | Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins |
| title_full_unstemmed | Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins |
| title_short | Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins |
| title_sort | sericin induced melanogenesis in cultured retinal pigment epithelial cells is associated with elevated levels of hydrogen peroxide and inflammatory proteins |
| topic | sericin melanogenesis superoxide dismutase inflammation retinal pigment epithelium hydrogen peroxide |
| url | https://www.mdpi.com/1420-3049/25/19/4395 |
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