Aging Aggravated Liver Ischemia And Reperfusion Injury By Promoting Oxidized mtDNA Mediated-Macrophage Pyroptosis Through Acetylated MCU-Dependent Calcium Uptake

Aging Aggravated Liver Ischemia And Reperfusion Injury By Promoting Oxidized mtDNA Mediated-Macrophage Pyroptosis Through Acetylated MCU-Dependent Calcium Uptake | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Aging Aggravated Liver Ischemia And Reperfusion Injury By Promoting Oxidized mtDNA Mediated-Macrophage Pyroptosis Through Acetylated MCU-Dependent Calcium Uptake Jun-Hua Gong, Xin-Yi Wu, Rui Wang, Qi Zhang, Tao Liu, Jun-Yan Liu, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6717688/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 07 Oct, 2025 Read the published version in Cell Death Discovery → Version 1 posted 9 You are reading this latest preprint version Abstract The shortage of liver donors for liver transplantation is currently an urgent problem. Elderly donors have become an important source of donor livers, but they are more prone to ischemia reperfusion injury (IRI) in liver transplantation. Therefore, exploring the effects and mechanisms of aging on liver IRI will provide a new theoretical basis for improving the survival rate of liver transplant patients. We constructed a mouse model of liver ischemia for 90 minutes and reperfusion for 6 or 24 hours, and found that compared with young liver, the recovery of liver function in aged liver after IRI was slower. Detection of macrophage pyroptosis revealed that it was an important factor for aging deferring liver function restoration. Mechanistically, we demonstrated that aging triggered mitochondrial permeability transition pore (mPTP) channel opening to promote the release of Oxidized mtDNA (Ox-mtDNA), thereby inducing macrophage pyroptosis. Moreover, the activity of mPTP channel was mainly dependent on calcium uptake by acetylated mitochondrial calcium uniporter (MCU). These results illustrated that cytoplasmic Ox-mtDNA-induced macrophage pyroptosis was a key factor for aging exacerbating liver IRI. Calcium uptake via acetylated MCU triggered mPTP channels opening, which is an important mechanism for Ox-mtDNA release from mitochondria into the cytoplasm. Biological sciences/Immunology/Immune cell death Health sciences/Diseases/Immunological disorders/Inflammatory diseases/Crohn's disease Aging Liver ischemia reperfusion injury Macrophage Pyroptosis mtDNA Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Full Text Additional Declarations There is no conflict of interest Supplementary Files WBoriginaldata.pdf Dataset 1 Supplementaryfigures.pdf Supplementary figures and legends Cite Share Download PDF Status: Published Journal Publication published 07 Oct, 2025 Read the published version in Cell Death Discovery → Version 1 posted Editorial decision: revise 10 Jun, 2025 Review # 1 received at journal 08 Jun, 2025 Review # 2 received at journal 03 Jun, 2025 Reviewer # 2 agreed at journal 30 May, 2025 Reviewer # 1 agreed at journal 30 May, 2025 Reviewers invited by journal 30 May, 2025 Submission checks completed at journal 22 May, 2025 Editor assigned by journal 21 May, 2025 First submitted to journal 21 May, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6717688","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":464241667,"identity":"5c12e669-f001-4642-9da4-83abb8c6abf7","order_by":0,"name":"Jun-Hua Gong","email":"data:image/png;base64,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","orcid":"","institution":"The Second Affiliated Hospital of Chongqing Medical University,China","correspondingAuthor":true,"prefix":"","firstName":"Jun-Hua","middleName":"","lastName":"Gong","suffix":""},{"id":464241668,"identity":"dc03ed68-d818-42a2-8323-9ad7e42901f2","order_by":1,"name":"Xin-Yi Wu","email":"","orcid":"","institution":"The Second Affiliated Hospital of Chongqing Medical University,China","correspondingAuthor":false,"prefix":"","firstName":"Xin-Yi","middleName":"","lastName":"Wu","suffix":""},{"id":464241669,"identity":"0f06ee30-fb70-4c7f-ba1e-6a5810a3e526","order_by":2,"name":"Rui Wang","email":"","orcid":"","institution":"The Second Affiliated Hospital of Chongqing Medical University,China","correspondingAuthor":false,"prefix":"","firstName":"Rui","middleName":"","lastName":"Wang","suffix":""},{"id":464241670,"identity":"0cc96924-70fe-4979-ae62-319a62ec191c","order_by":3,"name":"Qi Zhang","email":"","orcid":"","institution":"The Second Affiliated Hospital of Chongqing Medical University,China","correspondingAuthor":false,"prefix":"","firstName":"Qi","middleName":"","lastName":"Zhang","suffix":""},{"id":464241671,"identity":"3ceda47b-6987-40aa-ac71-a439f1a5e206","order_by":4,"name":"Tao Liu","email":"","orcid":"","institution":"The Second Affiliated Hospital of Chongqing Medical University,China","correspondingAuthor":false,"prefix":"","firstName":"Tao","middleName":"","lastName":"Liu","suffix":""},{"id":464241672,"identity":"b9b2b1a0-e5a1-476c-a4c3-290a60aec36e","order_by":5,"name":"Jun-Yan Liu","email":"","orcid":"","institution":"The Second Affiliated Hospital of Chongqing Medical University,China","correspondingAuthor":false,"prefix":"","firstName":"Jun-Yan","middleName":"","lastName":"Liu","suffix":""},{"id":464241673,"identity":"eb538f70-4916-4ce0-84f1-fb2b4424929e","order_by":6,"name":"Xue-Song Xu","email":"","orcid":"","institution":"The Second Affiliated Hospital of Chongqing Medical University,China","correspondingAuthor":false,"prefix":"","firstName":"Xue-Song","middleName":"","lastName":"Xu","suffix":""}],"badges":[],"createdAt":"2025-05-21 14:55:56","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6717688/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6717688/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41420-025-02746-9","type":"published","date":"2025-10-07T04:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":83959601,"identity":"3ffa8748-54ec-437d-9d96-f04030d42c9c","added_by":"auto","created_at":"2025-06-05 04:38:57","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":4681187,"visible":true,"origin":"","legend":"\u003cp\u003eAging delayed the recovery of liver function after IRI. \u0026nbsp;A-B. The levels of P16 and P21 in young and aged mice were measured by WB. \u0026nbsp;Liver of young and aged mice were subjected to 90 min of warm ischemia followed by \u0026nbsp;6 or 24 h of reperfusion, respectively: C. Serum levels of ALT and AST were measured. \u0026nbsp;D-E. average Suzuki scores were based on H\u0026amp;E-stained liver sections from different \u0026nbsp;groups of mice (scale bar, 200 μm). All data are shown as the mean ± SD (n=6). ***p\u0026lt; 0.001, **p \u0026lt; 0.01 and *p \u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/9d42e52a043ec68582db363a.png"},{"id":83959604,"identity":"2a49d017-dc0c-4f88-9a65-c9739f81f997","added_by":"auto","created_at":"2025-06-05 04:38:57","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":3910875,"visible":true,"origin":"","legend":"\u003cp\u003eAging aggravated macrophage pyroptosis in liver IRI. \u0026nbsp;Liver in young and aged mice were subjected to ischemia for 90 min followed by \u0026nbsp;reperfusion for 24 h: A. Caspase1 was detected using IHC in liver tissue sections (scale \u0026nbsp;bar, 200 μm). B. TEM was used to observe the ultrastructural changes in macrophages \u0026nbsp;(magnification, 20000×). C-D. Liver macrophages in each group were isolated, and the \u0026nbsp;levels of Caspase1, Cleaved-Caspase1, GSDMD and GSDMD-N were measured by \u0026nbsp;WB. E. Liver macrophages in each group were isolated, and caspase1 activity was \u0026nbsp;determined with the caspase1 assay kit. F. Serum levels of LDH were measured. G. The \u0026nbsp;levels of serum inflammatory factors (IL-1β and IL-18) were tested by ELISA. All data \u0026nbsp;are shown as the mean ± SD (n=6). ***p\u0026lt; 0.001, **p \u0026lt; 0.01 and *p \u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/71c02847ba056328fcf1eae0.png"},{"id":83959895,"identity":"5b9aa3ff-78d1-4889-b4b7-0ffc9ca34f00","added_by":"auto","created_at":"2025-06-05 04:46:58","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":6296661,"visible":true,"origin":"","legend":"\u003cp\u003eIncreased Ox-mtDNA in the cytoplasm of aging-induced macrophages. \u0026nbsp;Young and aged mice were subjected to ischemia for 90 min followed by reperfusion \u0026nbsp;for 24 h. Liver macrophages in each group were isolated: A. Relative cytosolic mtDNA \u0026nbsp;amounts in each group. The relative ratios of D-loop mtDNA, Cox1 mtDNA, or Non-NUMT mtDNA are tested by qPCR. B. 8-OH-dG from the mitochondrial (left) or \u0026nbsp;cytosol (right) were quantified using 8-OH-dG ELISA Kit. RAW264.7 cells were co-cultured with the supernatant from normal or aged AML12 cells for 24 h, followed by \u0026nbsp;treatment with H/R: C. Relative cytosolic mtDNA amounts in each group. The relative \u0026nbsp;ratios of D-loop mtDNA, Cox1 mtDNA, or Non-NUMT mtDNA are tested by qPCR. D. 8-OH-dG from the mitochondrial (left) or cytosol (right) were quantified by ELISA. \u0026nbsp;E. The colocalization of Mito-mCherry (red) and dsDNA (green) was detected by confocal microscopy. All data are shown as the mean ± SD (n=6). ***p\u0026lt; 0.001, **p \u0026lt; \u0026nbsp;0.01 and *p \u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/a02ee3ee7e37e95ca92f0fce.png"},{"id":83959598,"identity":"57af37b2-6dca-4dbc-ae50-85d5024d7217","added_by":"auto","created_at":"2025-06-05 04:38:57","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":7756530,"visible":true,"origin":"","legend":"\u003cp\u003eInhibition of mPTP channel-mediated Ox-mtDNA release alleviated aging-induced macrophage pyroptosis during H/R. \u0026nbsp;RAW264.7 cells were co-cultured with the supernatant from normal or aged AML12 \u0026nbsp;cells in the absence or presence of CsA (1 μM) for 24 h followed by treatment with H/R: \u0026nbsp;A. Representative of immunofluorescence staining of Calcein (green) and \u0026nbsp;corresponding cell density (white light). B. Relative cytosolic mtDNA amounts in each \u0026nbsp;group. The relative ratios of D-loop mtDNA, Cox1 mtDNA, or Non-NUMT mtDNA \u0026nbsp;are tested by qPCR. C. 8-OH-dG in cytosol were quantified using ELISA. D. The \u0026nbsp;colocalization of Mito-mCherry (red) and dsDNA (green) was detected by confocal \u0026nbsp;microscopy. E-F. The levels of Caspase1, Cleaved-Caspase1, GSDMD and GSDMD-N were measured by WB. G. Caspase1 activity was determined with the caspase1 assay \u0026nbsp;kit. H. Supernatant levels of LDH were measured by LDH assay kit. I. Supernatant levels of IL-1β and IL-18 were tested by ELISA. J. TEM was used to observe the \u0026nbsp;ultrastructural changes in macrophages (magnification, 20000×). All data are shown as \u0026nbsp;the mean ± SD (n=6). ***p\u0026lt; 0.001, **p \u0026lt; 0.01 and *p \u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/0a01a2564e0fbb73067bcd2d.png"},{"id":83959606,"identity":"22742b65-badf-436e-abae-248faeb3b804","added_by":"auto","created_at":"2025-06-05 04:38:58","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":10629531,"visible":true,"origin":"","legend":"\u003cp\u003eAging enhanced MCU-mediated mitochondrial Ca2+ uptake to trigger \u0026nbsp;mPTP channel opening and Ox-mtDNA release in macrophages during H/R. \u0026nbsp;RAW264.7 cells were co-cultured with the supernatant from normal or aged AML12 \u0026nbsp;cells in the absence or presence of BAPTA-AM (10 μM), SKF (20 μM), 2APB (100 \u0026nbsp;μM), or RuR (10 μM) for 24 h followed by treatment with H/R: A. Representative of immunofluorescence staining of Calcein (green) and corresponding cell density (white \u0026nbsp;light). B. Relative cytosolic mtDNA amounts in each group. The relative ratios of D-loop mtDNA, Cox1 mtDNA, and Non-NUMT mtDNA are tested by qPCR. C. 8-OH-dG in cytosol were quantified using ELISA. D. The colocalization of Mito-EGFP (green) and Rhod2 (red) was detected by confocal microscopy. E. Representative of \u0026nbsp;immunofluorescence staining of Calcein (green) and corresponding cell density (white \u0026nbsp;light). F. Relative cytosolic mtDNA amounts in each group. The relative ratios of D-loop mtDNA, Cox1 mtDNA, or Non-NUMT mtDNA are tested by qPCR. G. 8-OH-dG \u0026nbsp;in cytosol were quantified using ELISA. All data are shown as the mean ± SD (n=6). \u0026nbsp;***p\u0026lt; 0.001, **p \u0026lt; 0.01 and *p \u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/b937b58a3f568fce037dfe36.png"},{"id":83959602,"identity":"dc481086-cd15-47d0-95cb-e8651d9d5dee","added_by":"auto","created_at":"2025-06-05 04:38:57","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1276681,"visible":true,"origin":"","legend":"\u003cp\u003eAging led to acetylation of MCU in macrophages during H/R. RAW264.7 cells were co-cultured with the supernatant from normal or aged AML12 \u0026nbsp;cells for 24 h, followed by treatment with H/R: A. The level of MCU was measured by \u0026nbsp;WB. B. Lysine-acetylation (K–Ac) expressions in whole-cell lysates was measured by \u0026nbsp;WB. C. Co-IP assay was performed to determine MCU acetylation. All data are shown \u0026nbsp;as the mean ± SD (n=6). ***p\u0026lt; 0.001, **p \u0026lt; 0.01 and *p \u0026lt; 0.05. D. PhosphSitePlus \u0026nbsp;predicted MCU acetylation at K331 site. E. GPS-PAIL predicted MCU acetylation at \u0026nbsp;K331 site.\u003c/p\u003e","description":"","filename":"figure6.png","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/091d6e5c7ac99c7757c3a09b.png"},{"id":83959605,"identity":"89c703b6-3109-4430-90ab-4a7df8f8517a","added_by":"auto","created_at":"2025-06-05 04:38:57","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1669054,"visible":true,"origin":"","legend":"\u003cp\u003eMCU deacetylation restrained mitochondrial Ca2+ uptake，mPTP channel \u0026nbsp;opening and Ox-mtDNA release in aging-induced macrophage pyroptosis during \u0026nbsp;H/R. RAW264.7 cells were infected with Lv-MCU-WT or Lv-MCU-K331R and co-cultured with the supernatant from normal or aged AML12 cells for 24 h, followed by \u0026nbsp;treatment with H/R: A. The colocalization of Mito-EGFP (green) and Rhod2 (red) was \u0026nbsp;detected by confocal microscopy. B. Representative of immunofluorescence staining of \u0026nbsp;Calcein (green) and corresponding cell density (white light). C. Relative cytosolic \u0026nbsp;mtDNA amounts in each group. The relative ratios of D-loop mtDNA, Cox1 mtDNA, \u0026nbsp;or Non-NUMT mtDNA are tested by qPCR. D. 8-OH-dG in cytosol were quantified \u0026nbsp;using ELISA. E. The colocalization of Mito-mCherry (red) and dsDNA (green) was \u0026nbsp;detected by confocal microscopy. All data are shown as the mean ± SD (n=6). ***p\u0026lt; \u0026nbsp;0.001, **p \u0026lt; 0.01 and *p \u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"figure7.png","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/a726374f61425c34e74b7d1b.png"},{"id":83959600,"identity":"80a99092-0638-4b3a-9bff-cb8f3c9848a5","added_by":"auto","created_at":"2025-06-05 04:38:57","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":1338183,"visible":true,"origin":"","legend":"\u003cp\u003eMCU deacetylation alleviated aging-induced macrophage pyroptosis during \u0026nbsp;H/R. RAW264.7 cells were infected with LV-MCU-WT or LV-MCU-K331R and co-cultured with the supernatant from normal or aged AML12 cells for 24 h, followed by \u0026nbsp;treatment with H/R: A-B. The levels of Caspase1, Cleaved-Caspase1, GSDMD and \u0026nbsp;GSDMD-N were detected by WB. C. Caspase1 activity was tested by the caspase1 \u0026nbsp;assay kit. D. The levels of IL-1β and IL-18 in the cell culture supernatant were measured \u0026nbsp;by ELISA. E. Supernatant LDH levels were measured. F. TEM was used to observe the \u0026nbsp;ultrastructural changes in macrophages (magnification, 12000×). All data are shown as \u0026nbsp;the mean ± SD (n=6). ***p\u0026lt; 0.001, **p \u0026lt; 0.01 and *p \u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"figure8.png","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/4f7c7e716e6512c94e49c734.png"},{"id":93009130,"identity":"0b83c9e0-f507-4526-857c-8d0aec556e12","added_by":"auto","created_at":"2025-10-08 07:07:14","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":7223233,"visible":true,"origin":"","legend":"Article File","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1_covered_35714f2f-5a30-458e-9ec9-bf69fdbdd634.pdf"},{"id":83959603,"identity":"443d77c9-bada-4744-9abb-fc53fa431c6a","added_by":"auto","created_at":"2025-06-05 04:38:57","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":283629,"visible":true,"origin":"","legend":"Dataset 1","description":"","filename":"WBoriginaldata.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/0b54166ab5df722a8cc28974.pdf"},{"id":83959599,"identity":"1c18d27d-248d-4fbb-9b40-8b99abdc6c6d","added_by":"auto","created_at":"2025-06-05 04:38:57","extension":"pdf","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":260820,"visible":true,"origin":"","legend":"Supplementary figures and legends","description":"","filename":"Supplementaryfigures.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6717688/v1/6708dac730182544fdf58c0c.pdf"}],"financialInterests":"There is no conflict of interest","formattedTitle":"Aging Aggravated Liver Ischemia And Reperfusion Injury By Promoting Oxidized mtDNA Mediated-Macrophage Pyroptosis Through Acetylated MCU-Dependent Calcium Uptake","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"cell-death-discovery","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"cddiscovery","sideBox":"Learn more about [Cell Death Discovery](http://www.nature.com/cddiscovery/)","snPcode":"41420","submissionUrl":"https://mts-cddiscovery.nature.com/","title":"Cell Death Discovery","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Aging, Liver ischemia reperfusion injury, Macrophage, Pyroptosis, mtDNA","lastPublishedDoi":"10.21203/rs.3.rs-6717688/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6717688/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The shortage of liver donors for liver transplantation is currently an urgent problem. Elderly donors have become an important source of donor livers, but they are more prone to ischemia reperfusion injury (IRI) in liver transplantation. Therefore, exploring the effects and mechanisms of aging on liver IRI will provide a new theoretical basis for improving the survival rate of liver transplant patients. We constructed a mouse model of liver ischemia for 90 minutes and reperfusion for 6 or 24 hours, and found that compared with young liver, the recovery of liver function in aged liver after IRI was slower. Detection of macrophage pyroptosis revealed that it was an important factor for aging deferring liver function restoration. Mechanistically, we demonstrated that aging triggered mitochondrial permeability transition pore (mPTP) channel opening to promote the release of Oxidized mtDNA (Ox-mtDNA), thereby inducing macrophage pyroptosis. Moreover, the activity of mPTP channel was mainly dependent on calcium uptake by acetylated mitochondrial calcium uniporter (MCU). These results illustrated that cytoplasmic Ox-mtDNA-induced macrophage pyroptosis was a key factor for aging exacerbating liver IRI. Calcium uptake via acetylated MCU triggered mPTP channels opening, which is an important mechanism for Ox-mtDNA release from mitochondria into the cytoplasm.","manuscriptTitle":"Aging Aggravated Liver Ischemia And Reperfusion Injury By Promoting Oxidized mtDNA Mediated-Macrophage Pyroptosis Through Acetylated MCU-Dependent Calcium Uptake","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-05 04:38:50","doi":"10.21203/rs.3.rs-6717688/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"revise","date":"2025-06-10T15:03:13+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"This content is not available.","date":"2025-06-08T13:47:14+00:00","index":1,"fulltext":"This content is not available."},{"type":"editorInvitedReview","content":"This content is not available.","date":"2025-06-03T14:22:31+00:00","index":2,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2025-05-30T18:51:04+00:00","index":2,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2025-05-30T15:14:35+00:00","index":1,"fulltext":"This content is not available."},{"type":"reviewersInvited","content":"","date":"2025-05-30T14:40:31+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-05-22T09:48:57+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-05-21T14:51:25+00:00","index":"","fulltext":""},{"type":"submitted","content":"Cell Death Discovery","date":"2025-05-21T14:51:24+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"cell-death-discovery","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"cddiscovery","sideBox":"Learn more about [Cell Death Discovery](http://www.nature.com/cddiscovery/)","snPcode":"41420","submissionUrl":"https://mts-cddiscovery.nature.com/","title":"Cell Death Discovery","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b439181b-05c2-401f-9b9f-5887b2d1e3d3","owner":[],"postedDate":"June 5th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":49466332,"name":"Biological sciences/Immunology/Immune cell death"},{"id":49466333,"name":"Health sciences/Diseases/Immunological disorders/Inflammatory diseases/Crohn's disease"}],"tags":[],"updatedAt":"2025-10-08T07:06:26+00:00","versionOfRecord":{"articleIdentity":"rs-6717688","link":"https://doi.org/10.1038/s41420-025-02746-9","journal":{"identity":"cell-death-discovery","isVorOnly":false,"title":"Cell Death Discovery"},"publishedOn":"2025-10-07 04:00:00","publishedOnDateReadable":"October 7th, 2025"},"versionCreatedAt":"2025-06-05 04:38:50","video":"","vorDoi":"10.1038/s41420-025-02746-9","vorDoiUrl":"https://doi.org/10.1038/s41420-025-02746-9","workflowStages":[]},"version":"v1","identity":"rs-6717688","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6717688","identity":"rs-6717688","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}