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Copyright (c) 2025 Hazem K Ghneim, Fuad Alanazi, Abdulhadi Abdulwahed, Raed Farzan, May Alrashed, Sara Al-Saigh, Yazeed A. Al-Sheikh
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
The undersigned hereby assign all rights, included but not limited to copyright, for this manuscript to CMB Association upon its submission for consideration to publication on Cellular and Molecular Biology. The rights assigned include, but are not limited to, the sole and exclusive rights to license, sell, subsequently assign, derive, distribute, display and reproduce this manuscript, in whole or in part, in any format, electronic or otherwise, including those in existence at the time this agreement was signed. The authors hereby warrant that they have not granted or assigned, and shall not grant or assign, the aforementioned rights to any other person, firm, organization, or other entity. All rights are automatically restored to authors if this manuscript is not accepted for publication.Optimizing Se- methylselenocysteine concentration to enhance glutathione peroxidase 1 expression and mitigate oxidative stress in senescent human fibroblast
Corresponding Author(s) : Hazem K Ghneim
Cellular and Molecular Biology,
Vol. 71 No. 5: Issue 5
Abstract
Glutathione peroxidase 1 (GPx1) activity, gene expression, and several oxidative stress (OS) marker levels were investigated in the senescent passage (P) 20, 25, and 30 fibroblasts cultured in media supplemented with increasing Se-Methylselenocysteine (MSC) increments. While GPx1 activity slightly increased in cells grown in standard culture medium (CM1) compared to primary P5 cells, the enzyme exhibited significant MSC-dose-dependent elevations in cells cultured in MSC-supplemented media (CM3-CM6) compared to CM1 (p<0.001). GPx1 activity in CM5-incubated P30, P25, and P20 cells equaled 5.99±0.62, 4.72±0.48, and 4.06±0.36 µmoles/min/mg protein respectively (p<0.001), with percentage increases of 250% in P30 cells compared to 190% in P20 cells when cultured with CM1. Similarly, GPx1 expression was markedly upregulated in CM2, CM4, and CM6-incubated cells compared to primary P5 cells (p<0.001), with fold change values of 1.51±0.12, 1.99±0.16, and 2.31±0.19 in P20 cells. Percentage upregulations were 50.0±3.68%, 89.5±7.11%, and 126.5±9.74% in CM2, CM4, and CM6-incubated P20 cells respectively, and reached 248.0±18.6% in P30 cells at the highest MSC concentration. Concurrently, OS marker levels were substantially higher in CM1-cultured P25 and P30 senescent cells compared to primary P5 cells (p<0.001). Furthermore, hydrogen peroxide levels were significantly reduced in CM3-incubated cells compared to CM1 (p<0.01), reaching the lowest values in CM6 (p<0.001), with reductions of approximately 11.5%, 40%, 57%, and 58% in P30 CM3, CM4, CM5, and CM6-incubated cells respectively. MSC-Km values for GPx1 were 0.87, 1.13, and 1.92 µM in P20, P25, and P30 cells, respectively, with corresponding Vmax values of 4.59, 5.68, and 7.94 µmole/min/mg protein. These findings suggest that senescent cells utilize higher amounts of MSC to upregulate GPx1 expression and maximize its activity, supporting using Se supplements to combat OS.
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