NADH Reductive Stress and Its Correlation with Disease Severity in Leigh Syndrome: A Pilot Study Using Patient Fibroblasts and a Mouse Model
Nicotinamide adenine dinucleotide (NAD) is a critical cofactor in mitochondrial energy production. The NADH/NAD<sup>+</sup> ratio, reflecting the balance between NADH (reduced) and NAD<sup>+ (</sup>oxidized<sup>)</sup>, is a key marker for the severity of mitochon...
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MDPI AG
2024-12-01
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| Online Access: | https://www.mdpi.com/2218-273X/15/1/38 |
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| author | Tamaki Ishima Natsuka Kimura Mizuki Kobayashi Chika Watanabe Eriko F. Jimbo Ryosuke Kobayashi Takuro Horii Izuho Hatada Kei Murayama Akira Ohtake Ryozo Nagai Hitoshi Osaka Kenichi Aizawa |
| author_facet | Tamaki Ishima Natsuka Kimura Mizuki Kobayashi Chika Watanabe Eriko F. Jimbo Ryosuke Kobayashi Takuro Horii Izuho Hatada Kei Murayama Akira Ohtake Ryozo Nagai Hitoshi Osaka Kenichi Aizawa |
| author_sort | Tamaki Ishima |
| collection | DOAJ |
| description | Nicotinamide adenine dinucleotide (NAD) is a critical cofactor in mitochondrial energy production. The NADH/NAD<sup>+</sup> ratio, reflecting the balance between NADH (reduced) and NAD<sup>+ (</sup>oxidized<sup>)</sup>, is a key marker for the severity of mitochondrial diseases. We recently developed a streamlined LC-MS/MS method for the precise measurement of NADH and NAD<sup>+</sup>. Utilizing this technique, we quantified NADH and NAD<sup>+</sup> levels in fibroblasts derived from pediatric patients and in a Leigh syndrome mouse model in which mitochondrial respiratory chain complex I subunit <i>Ndufs4</i> is knocked out (KO). In patient-derived fibroblasts, NAD<sup>+</sup> levels did not differ significantly from those of healthy controls (<i>p</i> = 0.79); however, NADH levels were significantly elevated (<i>p</i> = 0.04), indicating increased NADH reductive stress. This increase, observed despite comparable total NAD(H) levels between the groups, was attributed to elevated NADH levels. Similarly, in the mouse model, NADH levels were significantly increased in the KO group (<i>p</i> = 0.002), further suggesting that NADH elevation drives reductive stress. This precise method for NADH measurement is expected to outperform conventional assays, such as those for lactate, providing a simpler and more reliable means of assessing disease progression. |
| format | Article |
| id | doaj-art-b473edac79d843e7a8b926b6dc6242a6 |
| institution | Kabale University |
| issn | 2218-273X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj-art-b473edac79d843e7a8b926b6dc6242a62025-01-24T13:24:56ZengMDPI AGBiomolecules2218-273X2024-12-011513810.3390/biom15010038NADH Reductive Stress and Its Correlation with Disease Severity in Leigh Syndrome: A Pilot Study Using Patient Fibroblasts and a Mouse ModelTamaki Ishima0Natsuka Kimura1Mizuki Kobayashi2Chika Watanabe3Eriko F. Jimbo4Ryosuke Kobayashi5Takuro Horii6Izuho Hatada7Kei Murayama8Akira Ohtake9Ryozo Nagai10Hitoshi Osaka11Kenichi Aizawa12Division of Clinical Pharmacology, Department of Pharmacology, Jichi Medical University, Shimotsuke 329-0498, JapanDivision of Clinical Pharmacology, Department of Pharmacology, Jichi Medical University, Shimotsuke 329-0498, JapanDepartment of Pediatrics, Jichi Medical University, Shimotsuke 329-0498, JapanDepartment of Pediatrics, Jichi Medical University, Shimotsuke 329-0498, JapanDepartment of Pediatrics, Jichi Medical University, Shimotsuke 329-0498, JapanBiosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi 371-8512, JapanBiosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi 371-8512, JapanBiosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi 371-8512, JapanCenter for Medical Genetics, Chiba Children’s Hospital, Chiba 266-0007, JapanDepartment of Clinical Genomics, Faculty of Medicine, Saitama Medical University, Moroyama 350-0495, JapanJichi Medical University, Shimotsuke 329-0498, JapanDepartment of Pediatrics, Jichi Medical University, Shimotsuke 329-0498, JapanDivision of Clinical Pharmacology, Department of Pharmacology, Jichi Medical University, Shimotsuke 329-0498, JapanNicotinamide adenine dinucleotide (NAD) is a critical cofactor in mitochondrial energy production. The NADH/NAD<sup>+</sup> ratio, reflecting the balance between NADH (reduced) and NAD<sup>+ (</sup>oxidized<sup>)</sup>, is a key marker for the severity of mitochondrial diseases. We recently developed a streamlined LC-MS/MS method for the precise measurement of NADH and NAD<sup>+</sup>. Utilizing this technique, we quantified NADH and NAD<sup>+</sup> levels in fibroblasts derived from pediatric patients and in a Leigh syndrome mouse model in which mitochondrial respiratory chain complex I subunit <i>Ndufs4</i> is knocked out (KO). In patient-derived fibroblasts, NAD<sup>+</sup> levels did not differ significantly from those of healthy controls (<i>p</i> = 0.79); however, NADH levels were significantly elevated (<i>p</i> = 0.04), indicating increased NADH reductive stress. This increase, observed despite comparable total NAD(H) levels between the groups, was attributed to elevated NADH levels. Similarly, in the mouse model, NADH levels were significantly increased in the KO group (<i>p</i> = 0.002), further suggesting that NADH elevation drives reductive stress. This precise method for NADH measurement is expected to outperform conventional assays, such as those for lactate, providing a simpler and more reliable means of assessing disease progression.https://www.mdpi.com/2218-273X/15/1/38LC-MS/MSNADHreductive stressLeigh syndromemitochondrial diseases<i>Ndufs4</i>-KO mice |
| spellingShingle | Tamaki Ishima Natsuka Kimura Mizuki Kobayashi Chika Watanabe Eriko F. Jimbo Ryosuke Kobayashi Takuro Horii Izuho Hatada Kei Murayama Akira Ohtake Ryozo Nagai Hitoshi Osaka Kenichi Aizawa NADH Reductive Stress and Its Correlation with Disease Severity in Leigh Syndrome: A Pilot Study Using Patient Fibroblasts and a Mouse Model Biomolecules LC-MS/MS NADH reductive stress Leigh syndrome mitochondrial diseases <i>Ndufs4</i>-KO mice |
| title | NADH Reductive Stress and Its Correlation with Disease Severity in Leigh Syndrome: A Pilot Study Using Patient Fibroblasts and a Mouse Model |
| title_full | NADH Reductive Stress and Its Correlation with Disease Severity in Leigh Syndrome: A Pilot Study Using Patient Fibroblasts and a Mouse Model |
| title_fullStr | NADH Reductive Stress and Its Correlation with Disease Severity in Leigh Syndrome: A Pilot Study Using Patient Fibroblasts and a Mouse Model |
| title_full_unstemmed | NADH Reductive Stress and Its Correlation with Disease Severity in Leigh Syndrome: A Pilot Study Using Patient Fibroblasts and a Mouse Model |
| title_short | NADH Reductive Stress and Its Correlation with Disease Severity in Leigh Syndrome: A Pilot Study Using Patient Fibroblasts and a Mouse Model |
| title_sort | nadh reductive stress and its correlation with disease severity in leigh syndrome a pilot study using patient fibroblasts and a mouse model |
| topic | LC-MS/MS NADH reductive stress Leigh syndrome mitochondrial diseases <i>Ndufs4</i>-KO mice |
| url | https://www.mdpi.com/2218-273X/15/1/38 |
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