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Interference with sphingosine kinase-1 reduces the hydrogen peroxide-induced oxidative stress damage in melanocytes through four and a half LIM domains 2.
- Source :
-
General physiology and biophysics [Gen Physiol Biophys] 2024 Jul; Vol. 43 (4), pp. 321-333. - Publication Year :
- 2024
-
Abstract
- Vitiligo is featured by manifestation of white maculae and primarily results from oxidative stress. Sphingosine kinase-1 (SPHK1) participates in oxidative stress. This paper was devised to explore the role of SPHK1 in vitiligo and to disclose the mechanism. PIG1 cell viability was appraised utilizing cell counting kit-8 assay while Western blot detected SPHK1 and four and a half LIM domains 2 (FHL2). The transduction efficacy of small interfering RNA (siRNA)-SPHK1, siRNA-FHL2 and pcDNA3.1 plasmid overexpressing FHL2 (Ov-FHL2) was checked using Western blot. Flow cytometry detected cell apoptotisis. Western blot detected mitochondrial cytochrome c (Mit-Cyt-c) and cytosolic cytochrome c (Cyto-Cyt-c). Dichloro-dihydro-fluorescein diacetate (DCFH-DA) detected reactive oxygen species (ROS) activity while oxidative stress markers were evaluated using corresponding assay kits. SPHK1 expression was discovered to be increased in hydrogen peroxide (H2O2)-challenged PIG1 cells and SPHK1 interference alleviated H2O2-challenged viability damage, apoptosis, oxidative stress and FHL2 expression in PIG1 cells. FHL2 depletion could suppress viability damage, apoptosis and oxidative stress in H2O2-challenged PIG1 cells. Rescue experiments demonstrated that the suppressive impacts of SPHK1 deficiency on PIG1 cell viability, apoptosis and oxidative stress induced by H2O2 were offset by FHL2 overexpression. Collectively, SPHK1 knockdown protected against vitiligo via the regulation of FHL2.
- Subjects :
- Humans
Apoptosis drug effects
Transcription Factors metabolism
Transcription Factors genetics
Muscle Proteins metabolism
Muscle Proteins genetics
Cell Line
Oxidative Stress drug effects
Phosphotransferases (Alcohol Group Acceptor) metabolism
Phosphotransferases (Alcohol Group Acceptor) genetics
Hydrogen Peroxide metabolism
LIM-Homeodomain Proteins metabolism
LIM-Homeodomain Proteins genetics
Melanocytes metabolism
Melanocytes drug effects
Cell Survival drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 0231-5882
- Volume :
- 43
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- General physiology and biophysics
- Publication Type :
- Academic Journal
- Accession number :
- 38953574
- Full Text :
- https://doi.org/10.4149/gpb_2024011