MitoQ and CoQ10 supplementation mildly suppresses skeletal muscle mitochondrial hydrogen peroxide levels without impacting mitochondrial function in middle-aged men
Purpose Excess production of reactive oxygen species (ROS) from the mitochondria can promote mitochondrial dysfunction and has been implicated in the development of a range of chronic diseases. As such there is interest in whether mitochondrial-targeted antioxidant supplementation can attenuate mito...
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creator | Pham, Toan MacRae, Caitlin L. Broome, Sophie C. D’souza, Randall F. Narang, Ravi Wang, Hsiang W. Mori, Trevor A. Hickey, Anthony J. R. Mitchell, Cameron J. Merry, Troy L. |
description | Purpose
Excess production of reactive oxygen species (ROS) from the mitochondria can promote mitochondrial dysfunction and has been implicated in the development of a range of chronic diseases. As such there is interest in whether mitochondrial-targeted antioxidant supplementation can attenuate mitochondrial-associated oxidative stress. We investigated the effect of MitoQ and CoQ10 supplementation on oxidative stress and skeletal muscle mitochondrial ROS levels and function in healthy middle-aged men.
Methods
Skeletal muscle and blood samples were collected from twenty men (50 ± 1 y) before and following six weeks of daily supplementation with MitoQ (20 mg) or CoQ10 (200 mg). High-resolution respirometry was used to determine mitochondrial respiration and H
2
O
2
levels, markers of mitochondrial mass and antioxidant defences were measured in muscle samples and oxidative stress markers in urine and blood samples.
Results
Both MitoQ and CoQ10 supplementation suppressed mitochondrial net H
2
O
2
levels during leak respiration, while MitoQ also elevated muscle catalase expression. However, neither supplement altered urine F
2
-isoprostanes nor plasma TBARS levels. Neither MitoQ nor CoQ10 supplementation had a significant impact on mitochondrial respiration or mitochondrial density markers (citrate synthase, mtDNA/nDNA,
PPARGC1A
, OXPHOS expression).
Conclusion
Our results suggest that neither MitoQ and CoQ10 supplements impact mitochondrial function, but both can mildly suppress mitochondrial ROS levels in healthy middle-aged men, with some indication that MitoQ may be more effective than CoQ10. |
doi_str_mv | 10.1007/s00421-020-04396-4 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2407315055</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2407315055</sourcerecordid><originalsourceid>FETCH-LOGICAL-c428t-ae7e1654ba0154266cdcd5dace1b7be72574c5a137e70ea8ba5c28ddf42d578b3</originalsourceid><addsrcrecordid>eNp9kc1uFDEQhC0EIiHwAhyQJS5chvh3PDmiFQGkoCgSnC2P3bvr4LEH2xOy78OD4uyGIOXAya32V1UtFUKvKXlPCVGnhRDBaEcY6YjgZ30nnqBj2qau50w9fZjp2RF6Uco1IWRgdHiOjjgTcqCyP0a_v_qarrCJDq_SFSW4LPMcYIJYTfUp4skHF3b7dYZSoODyAwJUE_C0FBugETXZbYou-7bc7lxOG4h4hpxuvQMc4AZCwb983aalYj_NxlYfN4-E6yXafaK_C3UuQGc24HA75SV6tjahwKv79wR9P__4bfW5u7j89GX14aKzgg21M6CA9lKMhlApWN9bZ510xgId1QiKSSWsNJQrUATMMBpp2eDcWjAn1TDyE_Tu4Dvn9HOBUvXki4UQTIS0FM0EUZxKImVD3z5Cr9OSY7uuUZQzThnljWIHyuZUSoa1nrOfTN5pSvRdh_rQoW4d6n2HWjTRm3vrZZzAPUj-ltYAfgBK-4obyP-y_2P7B6dvq8I</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2413231213</pqid></control><display><type>article</type><title>MitoQ and CoQ10 supplementation mildly suppresses skeletal muscle mitochondrial hydrogen peroxide levels without impacting mitochondrial function in middle-aged men</title><source>MEDLINE</source><source>SpringerLink Journals - AutoHoldings</source><creator>Pham, Toan ; MacRae, Caitlin L. ; Broome, Sophie C. ; D’souza, Randall F. ; Narang, Ravi ; Wang, Hsiang W. ; Mori, Trevor A. ; Hickey, Anthony J. R. ; Mitchell, Cameron J. ; Merry, Troy L.</creator><creatorcontrib>Pham, Toan ; MacRae, Caitlin L. ; Broome, Sophie C. ; D’souza, Randall F. ; Narang, Ravi ; Wang, Hsiang W. ; Mori, Trevor A. ; Hickey, Anthony J. R. ; Mitchell, Cameron J. ; Merry, Troy L.</creatorcontrib><description>Purpose
Excess production of reactive oxygen species (ROS) from the mitochondria can promote mitochondrial dysfunction and has been implicated in the development of a range of chronic diseases. As such there is interest in whether mitochondrial-targeted antioxidant supplementation can attenuate mitochondrial-associated oxidative stress. We investigated the effect of MitoQ and CoQ10 supplementation on oxidative stress and skeletal muscle mitochondrial ROS levels and function in healthy middle-aged men.
Methods
Skeletal muscle and blood samples were collected from twenty men (50 ± 1 y) before and following six weeks of daily supplementation with MitoQ (20 mg) or CoQ10 (200 mg). High-resolution respirometry was used to determine mitochondrial respiration and H
2
O
2
levels, markers of mitochondrial mass and antioxidant defences were measured in muscle samples and oxidative stress markers in urine and blood samples.
Results
Both MitoQ and CoQ10 supplementation suppressed mitochondrial net H
2
O
2
levels during leak respiration, while MitoQ also elevated muscle catalase expression. However, neither supplement altered urine F
2
-isoprostanes nor plasma TBARS levels. Neither MitoQ nor CoQ10 supplementation had a significant impact on mitochondrial respiration or mitochondrial density markers (citrate synthase, mtDNA/nDNA,
PPARGC1A
, OXPHOS expression).
Conclusion
Our results suggest that neither MitoQ and CoQ10 supplements impact mitochondrial function, but both can mildly suppress mitochondrial ROS levels in healthy middle-aged men, with some indication that MitoQ may be more effective than CoQ10.</description><identifier>ISSN: 1439-6319</identifier><identifier>EISSN: 1439-6327</identifier><identifier>DOI: 10.1007/s00421-020-04396-4</identifier><identifier>PMID: 32458156</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Adult ; Antioxidants ; Antioxidants - pharmacology ; Biomedical and Life Sciences ; Biomedicine ; Catalase ; Chronic illnesses ; Citrate synthase ; Dietary Supplements ; Human Physiology ; Humans ; Hydrogen peroxide ; Hydrogen Peroxide - metabolism ; Isoprostanes ; Male ; Middle age ; Middle Aged ; Mitochondria ; Mitochondria - metabolism ; Mitochondrial DNA ; Muscle, Skeletal - metabolism ; Musculoskeletal system ; Occupational Medicine/Industrial Medicine ; Original Article ; Oxidative stress ; Oxidative Stress - drug effects ; Reactive oxygen species ; Reactive Oxygen Species - metabolism ; Respiration ; Skeletal muscle ; Sports Medicine ; Supplements ; Ubiquinone - analogs & derivatives ; Ubiquinone - metabolism</subject><ispartof>European journal of applied physiology, 2020-07, Vol.120 (7), p.1657-1669</ispartof><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2020</rights><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2020.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c428t-ae7e1654ba0154266cdcd5dace1b7be72574c5a137e70ea8ba5c28ddf42d578b3</citedby><cites>FETCH-LOGICAL-c428t-ae7e1654ba0154266cdcd5dace1b7be72574c5a137e70ea8ba5c28ddf42d578b3</cites><orcidid>0000-0001-8699-500X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00421-020-04396-4$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00421-020-04396-4$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32458156$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pham, Toan</creatorcontrib><creatorcontrib>MacRae, Caitlin L.</creatorcontrib><creatorcontrib>Broome, Sophie C.</creatorcontrib><creatorcontrib>D’souza, Randall F.</creatorcontrib><creatorcontrib>Narang, Ravi</creatorcontrib><creatorcontrib>Wang, Hsiang W.</creatorcontrib><creatorcontrib>Mori, Trevor A.</creatorcontrib><creatorcontrib>Hickey, Anthony J. R.</creatorcontrib><creatorcontrib>Mitchell, Cameron J.</creatorcontrib><creatorcontrib>Merry, Troy L.</creatorcontrib><title>MitoQ and CoQ10 supplementation mildly suppresses skeletal muscle mitochondrial hydrogen peroxide levels without impacting mitochondrial function in middle-aged men</title><title>European journal of applied physiology</title><addtitle>Eur J Appl Physiol</addtitle><addtitle>Eur J Appl Physiol</addtitle><description>Purpose
Excess production of reactive oxygen species (ROS) from the mitochondria can promote mitochondrial dysfunction and has been implicated in the development of a range of chronic diseases. As such there is interest in whether mitochondrial-targeted antioxidant supplementation can attenuate mitochondrial-associated oxidative stress. We investigated the effect of MitoQ and CoQ10 supplementation on oxidative stress and skeletal muscle mitochondrial ROS levels and function in healthy middle-aged men.
Methods
Skeletal muscle and blood samples were collected from twenty men (50 ± 1 y) before and following six weeks of daily supplementation with MitoQ (20 mg) or CoQ10 (200 mg). High-resolution respirometry was used to determine mitochondrial respiration and H
2
O
2
levels, markers of mitochondrial mass and antioxidant defences were measured in muscle samples and oxidative stress markers in urine and blood samples.
Results
Both MitoQ and CoQ10 supplementation suppressed mitochondrial net H
2
O
2
levels during leak respiration, while MitoQ also elevated muscle catalase expression. However, neither supplement altered urine F
2
-isoprostanes nor plasma TBARS levels. Neither MitoQ nor CoQ10 supplementation had a significant impact on mitochondrial respiration or mitochondrial density markers (citrate synthase, mtDNA/nDNA,
PPARGC1A
, OXPHOS expression).
Conclusion
Our results suggest that neither MitoQ and CoQ10 supplements impact mitochondrial function, but both can mildly suppress mitochondrial ROS levels in healthy middle-aged men, with some indication that MitoQ may be more effective than CoQ10.</description><subject>Adult</subject><subject>Antioxidants</subject><subject>Antioxidants - pharmacology</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Catalase</subject><subject>Chronic illnesses</subject><subject>Citrate synthase</subject><subject>Dietary Supplements</subject><subject>Human Physiology</subject><subject>Humans</subject><subject>Hydrogen peroxide</subject><subject>Hydrogen Peroxide - metabolism</subject><subject>Isoprostanes</subject><subject>Male</subject><subject>Middle age</subject><subject>Middle Aged</subject><subject>Mitochondria</subject><subject>Mitochondria - metabolism</subject><subject>Mitochondrial DNA</subject><subject>Muscle, Skeletal - metabolism</subject><subject>Musculoskeletal system</subject><subject>Occupational Medicine/Industrial Medicine</subject><subject>Original Article</subject><subject>Oxidative stress</subject><subject>Oxidative Stress - drug effects</subject><subject>Reactive oxygen species</subject><subject>Reactive Oxygen Species - metabolism</subject><subject>Respiration</subject><subject>Skeletal muscle</subject><subject>Sports Medicine</subject><subject>Supplements</subject><subject>Ubiquinone - analogs & derivatives</subject><subject>Ubiquinone - metabolism</subject><issn>1439-6319</issn><issn>1439-6327</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp9kc1uFDEQhC0EIiHwAhyQJS5chvh3PDmiFQGkoCgSnC2P3bvr4LEH2xOy78OD4uyGIOXAya32V1UtFUKvKXlPCVGnhRDBaEcY6YjgZ30nnqBj2qau50w9fZjp2RF6Uco1IWRgdHiOjjgTcqCyP0a_v_qarrCJDq_SFSW4LPMcYIJYTfUp4skHF3b7dYZSoODyAwJUE_C0FBugETXZbYou-7bc7lxOG4h4hpxuvQMc4AZCwb983aalYj_NxlYfN4-E6yXafaK_C3UuQGc24HA75SV6tjahwKv79wR9P__4bfW5u7j89GX14aKzgg21M6CA9lKMhlApWN9bZ510xgId1QiKSSWsNJQrUATMMBpp2eDcWjAn1TDyE_Tu4Dvn9HOBUvXki4UQTIS0FM0EUZxKImVD3z5Cr9OSY7uuUZQzThnljWIHyuZUSoa1nrOfTN5pSvRdh_rQoW4d6n2HWjTRm3vrZZzAPUj-ltYAfgBK-4obyP-y_2P7B6dvq8I</recordid><startdate>20200701</startdate><enddate>20200701</enddate><creator>Pham, Toan</creator><creator>MacRae, Caitlin L.</creator><creator>Broome, Sophie C.</creator><creator>D’souza, Randall F.</creator><creator>Narang, Ravi</creator><creator>Wang, Hsiang W.</creator><creator>Mori, Trevor A.</creator><creator>Hickey, Anthony J. R.</creator><creator>Mitchell, Cameron J.</creator><creator>Merry, Troy L.</creator><general>Springer Berlin Heidelberg</general><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7RV</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB0</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0001-8699-500X</orcidid></search><sort><creationdate>20200701</creationdate><title>MitoQ and CoQ10 supplementation mildly suppresses skeletal muscle mitochondrial hydrogen peroxide levels without impacting mitochondrial function in middle-aged men</title><author>Pham, Toan ; MacRae, Caitlin L. ; Broome, Sophie C. ; D’souza, Randall F. ; Narang, Ravi ; Wang, Hsiang W. ; Mori, Trevor A. ; Hickey, Anthony J. R. ; Mitchell, Cameron J. ; Merry, Troy L.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c428t-ae7e1654ba0154266cdcd5dace1b7be72574c5a137e70ea8ba5c28ddf42d578b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Adult</topic><topic>Antioxidants</topic><topic>Antioxidants - pharmacology</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Catalase</topic><topic>Chronic illnesses</topic><topic>Citrate synthase</topic><topic>Dietary Supplements</topic><topic>Human Physiology</topic><topic>Humans</topic><topic>Hydrogen peroxide</topic><topic>Hydrogen Peroxide - metabolism</topic><topic>Isoprostanes</topic><topic>Male</topic><topic>Middle age</topic><topic>Middle Aged</topic><topic>Mitochondria</topic><topic>Mitochondria - metabolism</topic><topic>Mitochondrial DNA</topic><topic>Muscle, Skeletal - metabolism</topic><topic>Musculoskeletal system</topic><topic>Occupational Medicine/Industrial Medicine</topic><topic>Original Article</topic><topic>Oxidative stress</topic><topic>Oxidative Stress - drug effects</topic><topic>Reactive oxygen species</topic><topic>Reactive Oxygen Species - metabolism</topic><topic>Respiration</topic><topic>Skeletal muscle</topic><topic>Sports Medicine</topic><topic>Supplements</topic><topic>Ubiquinone - analogs & derivatives</topic><topic>Ubiquinone - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pham, Toan</creatorcontrib><creatorcontrib>MacRae, Caitlin L.</creatorcontrib><creatorcontrib>Broome, Sophie C.</creatorcontrib><creatorcontrib>D’souza, Randall F.</creatorcontrib><creatorcontrib>Narang, Ravi</creatorcontrib><creatorcontrib>Wang, Hsiang W.</creatorcontrib><creatorcontrib>Mori, Trevor A.</creatorcontrib><creatorcontrib>Hickey, Anthony J. R.</creatorcontrib><creatorcontrib>Mitchell, Cameron J.</creatorcontrib><creatorcontrib>Merry, Troy L.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Nursing & Allied Health Database</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Nursing & Allied Health Premium</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><jtitle>European journal of applied physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pham, Toan</au><au>MacRae, Caitlin L.</au><au>Broome, Sophie C.</au><au>D’souza, Randall F.</au><au>Narang, Ravi</au><au>Wang, Hsiang W.</au><au>Mori, Trevor A.</au><au>Hickey, Anthony J. R.</au><au>Mitchell, Cameron J.</au><au>Merry, Troy L.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>MitoQ and CoQ10 supplementation mildly suppresses skeletal muscle mitochondrial hydrogen peroxide levels without impacting mitochondrial function in middle-aged men</atitle><jtitle>European journal of applied physiology</jtitle><stitle>Eur J Appl Physiol</stitle><addtitle>Eur J Appl Physiol</addtitle><date>2020-07-01</date><risdate>2020</risdate><volume>120</volume><issue>7</issue><spage>1657</spage><epage>1669</epage><pages>1657-1669</pages><issn>1439-6319</issn><eissn>1439-6327</eissn><abstract>Purpose
Excess production of reactive oxygen species (ROS) from the mitochondria can promote mitochondrial dysfunction and has been implicated in the development of a range of chronic diseases. As such there is interest in whether mitochondrial-targeted antioxidant supplementation can attenuate mitochondrial-associated oxidative stress. We investigated the effect of MitoQ and CoQ10 supplementation on oxidative stress and skeletal muscle mitochondrial ROS levels and function in healthy middle-aged men.
Methods
Skeletal muscle and blood samples were collected from twenty men (50 ± 1 y) before and following six weeks of daily supplementation with MitoQ (20 mg) or CoQ10 (200 mg). High-resolution respirometry was used to determine mitochondrial respiration and H
2
O
2
levels, markers of mitochondrial mass and antioxidant defences were measured in muscle samples and oxidative stress markers in urine and blood samples.
Results
Both MitoQ and CoQ10 supplementation suppressed mitochondrial net H
2
O
2
levels during leak respiration, while MitoQ also elevated muscle catalase expression. However, neither supplement altered urine F
2
-isoprostanes nor plasma TBARS levels. Neither MitoQ nor CoQ10 supplementation had a significant impact on mitochondrial respiration or mitochondrial density markers (citrate synthase, mtDNA/nDNA,
PPARGC1A
, OXPHOS expression).
Conclusion
Our results suggest that neither MitoQ and CoQ10 supplements impact mitochondrial function, but both can mildly suppress mitochondrial ROS levels in healthy middle-aged men, with some indication that MitoQ may be more effective than CoQ10.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>32458156</pmid><doi>10.1007/s00421-020-04396-4</doi><tpages>13</tpages><orcidid>https://orcid.org/0000-0001-8699-500X</orcidid></addata></record> |
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subjects | Adult Antioxidants Antioxidants - pharmacology Biomedical and Life Sciences Biomedicine Catalase Chronic illnesses Citrate synthase Dietary Supplements Human Physiology Humans Hydrogen peroxide Hydrogen Peroxide - metabolism Isoprostanes Male Middle age Middle Aged Mitochondria Mitochondria - metabolism Mitochondrial DNA Muscle, Skeletal - metabolism Musculoskeletal system Occupational Medicine/Industrial Medicine Original Article Oxidative stress Oxidative Stress - drug effects Reactive oxygen species Reactive Oxygen Species - metabolism Respiration Skeletal muscle Sports Medicine Supplements Ubiquinone - analogs & derivatives Ubiquinone - metabolism |
title | MitoQ and CoQ10 supplementation mildly suppresses skeletal muscle mitochondrial hydrogen peroxide levels without impacting mitochondrial function in middle-aged men |
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