Optimizing Se- methylselenocysteine concentration to enhance glutathione peroxidase 1 expression and mitigate oxidative stress in senescent human fibroblast.
Hazem K Ghneim, Fuad Alanazi, Abdulhadi M Abdulwahed, Raed Farzan, May Alrashed, Sara Al-Saigh, Yazeed A Al-Sheikh
{"title":"Optimizing Se- methylselenocysteine concentration to enhance glutathione peroxidase 1 expression and mitigate oxidative stress in senescent human fibroblast.","authors":"Hazem K Ghneim, Fuad Alanazi, Abdulhadi M Abdulwahed, Raed Farzan, May Alrashed, Sara Al-Saigh, Yazeed A Al-Sheikh","doi":"10.14715/cmb/2025.71.5.6","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":9802,"journal":{"name":"Cellular and molecular biology","volume":"71 5","pages":"33-42"},"PeriodicalIF":1.5000,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cellular and molecular biology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.14715/cmb/2025.71.5.6","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
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.
期刊介绍:
Cellular and Molecular Biology publishes original articles, reviews, short communications, methods, meta-analysis notes, letters to editor and comments in the interdisciplinary science of Cellular and Molecular Biology linking and integrating molecular biology, biophysics, biochemistry, enzymology, physiology and biotechnology in a dynamic cell and tissue biology environment, applied to human, animals, plants tissues as well to microbial and viral cells. The journal Cellular and Molecular Biology is therefore open to intense interdisciplinary exchanges in medical, dental, veterinary, pharmacological, botanical and biological researches for the demonstration of these multiple links.