Impact of a Formulation Containing Chaga Extract, Coenzyme Q10, and Alpha-Lipoic Acid on Mitochondrial Dysfunction and Oxidative Stress: NMR Metabolomic Insights into Cellular Energy
Objectives: The aim of this study was to evaluate the impact of a novel antioxidant formulation (RE:PAIR, RP-25) containing CoQ10, alpha-lipoic acid, and Chaga extract on mitochondrial dysfunction and oxidative stress. To explore the activity of the formulation on neuronal cells, we explored cell me...
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2025-06-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/14/6/753 |
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| author | Maria D’Elia Carmen Marino Rita Celano Enza Napolitano Chiara Colarusso Rosalinda Sorrentino Anna Maria D’Ursi Luca Rastrelli |
| author_facet | Maria D’Elia Carmen Marino Rita Celano Enza Napolitano Chiara Colarusso Rosalinda Sorrentino Anna Maria D’Ursi Luca Rastrelli |
| author_sort | Maria D’Elia |
| collection | DOAJ |
| description | Objectives: The aim of this study was to evaluate the impact of a novel antioxidant formulation (RE:PAIR, RP-25) containing CoQ10, alpha-lipoic acid, and Chaga extract on mitochondrial dysfunction and oxidative stress. To explore the activity of the formulation on neuronal cells, we explored cell metabolism and its activity as an antioxidant, using a combination of NMR-based metabolomics and UHPLC-HRMS analytical techniques. Methods: SH-SY5Y neuroblastoma cells were treated with RP-25, and cell viability was assessed via CCK-8 assay. Metabolomic profiles of the treated and untreated cells were analyzed by 1D-NMR, providing insights into both intracellular metabolites (endometabolome) and excreted metabolites (exometabolome). Additionally, a UHPLC-HRMS method was developed for quality control and analysis of the RP-25 formulation. Multivariate statistical approaches, including PLS-DA and volcano plot analyses, were used to identify key metabolic changes. Changes in mitochondrial membrane potential were assessed by means of TMRE assay, while radical oxygen species (ROS) were measured by means of the DCHF assay. Results: RP-25 treatment did not affect cell viability but significantly increased metabolic pathways, including amino acid biosynthesis, oxidative phosphorylation, and glycolysis. Higher levels of ATP, glutamate, tyrosine, and proline were observed in treated cells than in control cells, indicating enhanced cellular energy production, as also proved by the increased stability of the mitochondrial membrane after RP-25 treatment, an index of preserved mitochondrial functions. In support, the formulation RP-25 showed antioxidant activity when cells underwent peroxide oxygen stimulation. This effect was mainly due to the combination of Chaga, CoQ10, and ALA, main components of the RP25 formulation. Moreover, the analysis of enriched pathways highlighted that RP formulation influenced mitochondrial energy and oxidative stress response. Conclusions: RP-25 demonstrated biological activity in that it mitigated mitochondrial dysfunction and oxidative stress in neuronal cells, with potential implications in neuronal diseases associated with dysfunctional mitochondria. |
| format | Article |
| id | doaj-art-c41df545ced8498e8b06b7ddd00aec63 |
| institution | Kabale University |
| issn | 2076-3921 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj-art-c41df545ced8498e8b06b7ddd00aec632025-08-20T03:32:26ZengMDPI AGAntioxidants2076-39212025-06-0114675310.3390/antiox14060753Impact of a Formulation Containing Chaga Extract, Coenzyme Q10, and Alpha-Lipoic Acid on Mitochondrial Dysfunction and Oxidative Stress: NMR Metabolomic Insights into Cellular EnergyMaria D’Elia0Carmen Marino1Rita Celano2Enza Napolitano3Chiara Colarusso4Rosalinda Sorrentino5Anna Maria D’Ursi6Luca Rastrelli7Department of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, ItalyDepartment of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, ItalyDepartment of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, ItalyDepartment of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, ItalyDepartment of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, ItalyDepartment of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, ItalyDepartment of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, ItalyDepartment of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, ItalyObjectives: The aim of this study was to evaluate the impact of a novel antioxidant formulation (RE:PAIR, RP-25) containing CoQ10, alpha-lipoic acid, and Chaga extract on mitochondrial dysfunction and oxidative stress. To explore the activity of the formulation on neuronal cells, we explored cell metabolism and its activity as an antioxidant, using a combination of NMR-based metabolomics and UHPLC-HRMS analytical techniques. Methods: SH-SY5Y neuroblastoma cells were treated with RP-25, and cell viability was assessed via CCK-8 assay. Metabolomic profiles of the treated and untreated cells were analyzed by 1D-NMR, providing insights into both intracellular metabolites (endometabolome) and excreted metabolites (exometabolome). Additionally, a UHPLC-HRMS method was developed for quality control and analysis of the RP-25 formulation. Multivariate statistical approaches, including PLS-DA and volcano plot analyses, were used to identify key metabolic changes. Changes in mitochondrial membrane potential were assessed by means of TMRE assay, while radical oxygen species (ROS) were measured by means of the DCHF assay. Results: RP-25 treatment did not affect cell viability but significantly increased metabolic pathways, including amino acid biosynthesis, oxidative phosphorylation, and glycolysis. Higher levels of ATP, glutamate, tyrosine, and proline were observed in treated cells than in control cells, indicating enhanced cellular energy production, as also proved by the increased stability of the mitochondrial membrane after RP-25 treatment, an index of preserved mitochondrial functions. In support, the formulation RP-25 showed antioxidant activity when cells underwent peroxide oxygen stimulation. This effect was mainly due to the combination of Chaga, CoQ10, and ALA, main components of the RP25 formulation. Moreover, the analysis of enriched pathways highlighted that RP formulation influenced mitochondrial energy and oxidative stress response. Conclusions: RP-25 demonstrated biological activity in that it mitigated mitochondrial dysfunction and oxidative stress in neuronal cells, with potential implications in neuronal diseases associated with dysfunctional mitochondria.https://www.mdpi.com/2076-3921/14/6/753coenzyme Q10lipoic acid<i>Inonotus obliquus</i>SH-SY5Y cellsUHPLC-HRMS/MS<sup>1</sup>H-NMR metabolomics |
| spellingShingle | Maria D’Elia Carmen Marino Rita Celano Enza Napolitano Chiara Colarusso Rosalinda Sorrentino Anna Maria D’Ursi Luca Rastrelli Impact of a Formulation Containing Chaga Extract, Coenzyme Q10, and Alpha-Lipoic Acid on Mitochondrial Dysfunction and Oxidative Stress: NMR Metabolomic Insights into Cellular Energy Antioxidants coenzyme Q10 lipoic acid <i>Inonotus obliquus</i> SH-SY5Y cells UHPLC-HRMS/MS <sup>1</sup>H-NMR metabolomics |
| title | Impact of a Formulation Containing Chaga Extract, Coenzyme Q10, and Alpha-Lipoic Acid on Mitochondrial Dysfunction and Oxidative Stress: NMR Metabolomic Insights into Cellular Energy |
| title_full | Impact of a Formulation Containing Chaga Extract, Coenzyme Q10, and Alpha-Lipoic Acid on Mitochondrial Dysfunction and Oxidative Stress: NMR Metabolomic Insights into Cellular Energy |
| title_fullStr | Impact of a Formulation Containing Chaga Extract, Coenzyme Q10, and Alpha-Lipoic Acid on Mitochondrial Dysfunction and Oxidative Stress: NMR Metabolomic Insights into Cellular Energy |
| title_full_unstemmed | Impact of a Formulation Containing Chaga Extract, Coenzyme Q10, and Alpha-Lipoic Acid on Mitochondrial Dysfunction and Oxidative Stress: NMR Metabolomic Insights into Cellular Energy |
| title_short | Impact of a Formulation Containing Chaga Extract, Coenzyme Q10, and Alpha-Lipoic Acid on Mitochondrial Dysfunction and Oxidative Stress: NMR Metabolomic Insights into Cellular Energy |
| title_sort | impact of a formulation containing chaga extract coenzyme q10 and alpha lipoic acid on mitochondrial dysfunction and oxidative stress nmr metabolomic insights into cellular energy |
| topic | coenzyme Q10 lipoic acid <i>Inonotus obliquus</i> SH-SY5Y cells UHPLC-HRMS/MS <sup>1</sup>H-NMR metabolomics |
| url | https://www.mdpi.com/2076-3921/14/6/753 |
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