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We hypothesized that mitophagy may play an important role in HOPE mitochondria protection. Experimental rat liver grafts were exposed to 30 min of in situ warm ischemia. Then, grafts were procured, followed by cold storage for 3 or 4 h to mimic the conventional preservation and transportation time in donation after circulatory death (DCD) in clinical contexts. Next, the grafts underwent hypothermic machine perfusion (HMP) or HOPE for 1 h through portal vein only perfusion. The HOPE-treated group showed a better preservation capacity compared with cold storage and HMP, preventing hepatocyte damage, nuclear injury, and cell death. HOPE can increase mitophagy marker expression, promote mitophagy flux via the PINK1\/Parkin pathway to maintain mitochondrial function, and reduce oxygen free radical generation, while the inhibition of autophagy by 3-methyladenine and chloroquine could reverse the protective effect. HOPE-treated DCD liver also demonstrated more changes in the expression of genes responsible for bile metabolism, mitochondrial dynamics, cell survival, and oxidative stress. Overall, HOPE attenuates hypoxia-ischemic injury in DCD liver by promoting mitophagy flux to maintain mitochondrial function and protect hepatocytes. Mitophagy could pave the way for a protective approach against hypoxia-ischemic injury in DCD liver.<\/jats:p>","DOI":"10.3390\/ijms24065403","type":"journal-article","created":{"date-parts":[[2023,3,13]],"date-time":"2023-03-13T05:36:21Z","timestamp":1678685781000},"page":"5403","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Hypothermic Oxygenated Machine Perfusion Promotes Mitophagy Flux against Hypoxia-Ischemic Injury in Rat DCD Liver"],"prefix":"10.3390","volume":"24","author":[{"given":"Jia","family":"Luo","sequence":"first","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"},{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiqing","family":"Hu","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"},{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinbiao","family":"Qiao","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"},{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoyu","family":"Li","sequence":"additional","affiliation":[{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiacheng","family":"Huang","sequence":"additional","affiliation":[{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kangdi","family":"Xu","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"},{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Jiang","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Wu","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"},{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyi","family":"Hu","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"},{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junjun","family":"Jia","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Zhou","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"},{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haiyang","family":"Xie","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"},{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianhui","family":"Li","sequence":"additional","affiliation":[{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"},{"name":"Department of Hepatobiliary and Pancreatic Surgery, Shulan (Hangzhou) Hospital, Zhejiang Shuren University School of Medicine, Hangzhou 310015, China"},{"name":"The Organ Repair and Regeneration Medicine Institute of Hangzhou, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1459-8261","authenticated-orcid":false,"given":"Shusen","family":"Zheng","sequence":"additional","affiliation":[{"name":"Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China"},{"name":"NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou 310003, China"},{"name":"Department of Hepatobiliary and Pancreatic Surgery, Shulan (Hangzhou) Hospital, Zhejiang Shuren University School of Medicine, Hangzhou 310015, China"},{"name":"Jinan Microecological Biomedicine Shandong Laboratory, Jinan 250117, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1111\/liv.13946","article-title":"Machine perfusion for liver transplantation in the era of marginal organs-New kids on the block","volume":"39","author":"Czigany","year":"2019","journal-title":"Liver Int."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1458","DOI":"10.1056\/NEJMe2102056","article-title":"Warming Up to Cold Perfusion","volume":"384","author":"Williams","year":"2021","journal-title":"N. 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