{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T18:00:28Z","timestamp":1784916028399,"version":"3.55.0"},"posted":{"date-parts":[[2026,7,22]]},"group-title":"Neuroscience","reference-count":97,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T00:00:00Z","timestamp":1784678400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["No. 82171430"],"award-info":[{"award-number":["No. 82171430"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Natural Science Foundation key project of Yunnan Province","award":["No. 202301AS070075"],"award-info":[{"award-number":["No. 202301AS070075"]}]},{"name":"2025 STU-GTIIT Joint Research Grants","award":["No. 2025LKSFG07"],"award-info":[{"award-number":["No. 2025LKSFG07"]}]},{"name":"The Scientific Research Initiation Grant (SRIG) of Shantou University Medical College to Dr. Fan Li"},{"DOI":"10.13039\/100000065","name":"NINDS","doi-asserted-by":"crossref","award":["R21NS133533 to Xiangning Jiang"],"award-info":[{"award-number":["R21NS133533 to Xiangning Jiang"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,22]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Hypoxic-ischemic encephalopathy (HIE) is a major cause of neonatal mortality and neurodevelopmental impairments. Following brain hypoxia-ischemia (HI), microglia face substantial metabolic stress; and upon phagocytosis, they become overloaded with lipids derived from engulfed dead neurons and myelin debris. It is unclear how microglia respond to and process the lipid cargo, and whether lipid accumulation may affect microglia function following neonatal HI.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    The postnatal day 10 mice were subjected to HI using the Vannucci model. Lipid droplets (LD) were assessed by histology and immunofluorescent staining. Single-nucleus RNA sequencing (snRNA-seq) was performed using brain tissue from HI-injured and sham-operated mice at 72 hours after HI. LD-accumulating microglia (LDAM) were identified by a specific LD marker gene perilipin 2 (\n                    <jats:italic>Plin2<\/jats:italic>\n                    ). Differential gene expression was analyzed between\n                    <jats:italic>Plin2<\/jats:italic>\n                    -positive and\n                    <jats:italic>Plin2<\/jats:italic>\n                    -negative microglia after HI. Human HIE brain sections were also examined for LD accumulation. The dynamic changes of PLIN2-expressing microglia and infiltrating monocyte-derived macrophages (MDM) at 24 hours, 72 hours and 7 days after HI were compared using flow cytometry. In addition, mouse BV2 microglia were subjected to oxygen-glucose deprivation (OGD) to study phagocytosis and cytokine expression.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Lipid droplets accumulated primarily in microglia after HI in neonatal mice and in human HIE brain. LD were not found in astrocytes or neurons.\n                    <jats:italic>Plin2<\/jats:italic>\n                    -expressing LDAM emerged as new microglia clusters after HI. Compared with microglia without LD, LDAM showed a distinct transcriptional profile with upregulation of genes linked to microglial activation, enhanced cholesterol and lipid processing, and a shift towards phagocytic and pro-inflammatory state. Blocking LD biogenesis reduced elevated phagocytosis and IL-1\u03b2 expression in BV2 cells following OGD.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>Our study revealed that microglia accumulate lipid droplets as part of their metabolic responses to HI in the neonatal brain. Microglial lipid droplet formation is associated with a pro-inflammatory phenotype at early stage after HI, and increased phagocytosis in vitro. The lipid metabolic changes may regulate microglial function and influence HI outcomes.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.07.18.739219","type":"posted-content","created":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T22:50:11Z","timestamp":1784760611000},"source":"Crossref","is-referenced-by-count":0,"title":["Accumulation of Lipid Droplets in Microglia following Neonatal Brain Hypoxia-Ischemia"],"prefix":"10.64898","author":[{"given":"Fan","family":"Li","sequence":"first","affiliation":[{"name":"Department of Pediatrics, First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong Province 515041, China"},{"name":"Department of Pathophysiology, Shantou University Medical College, Shantou, Guangdong Province 515041, China"},{"name":"Key Laboratory of Immunopathology of Guangdong Provincial Universities, Guangdong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuqing","family":"Lei","sequence":"additional","affiliation":[{"name":"School of Medicine, Dentistry and Health Sciences, The University of Melbourne, Victoria 3010 Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sihao","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong Province 515041, China"},{"name":"Department of Pathophysiology, Shantou University Medical College, Shantou, Guangdong Province 515041, China"},{"name":"Key Laboratory of Immunopathology of Guangdong Provincial Universities, Guangdong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gui","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong Province 515041, China"},{"name":"Department of Pathophysiology, Shantou University Medical College, Shantou, Guangdong Province 515041, China"},{"name":"Department of Pathophysiology, Yunnan University Medical College, East Outer Ring South Road, Chenggong District, Kunming, Yunnan Province 650500, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinuo","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Pathophysiology, Yunnan University Medical College, East Outer Ring South Road, Chenggong District, Kunming, Yunnan Province 650500, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beiyan","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong Province 515041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donna M","family":"Ferriero","sequence":"additional","affiliation":[{"name":"Department of Neurology, University of California, San Francisco"},{"name":"Department of Pediatrics, University of California, San Francisco"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peipei","family":"Pan","sequence":"additional","affiliation":[{"name":"Department of Anesthesia and Perioperative Care, University of California, San Francisco"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhonghui","family":"Guan","sequence":"additional","affiliation":[{"name":"Department of Anesthesia and Perioperative Care, University of California, San Francisco"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangning","family":"Jiang","sequence":"additional","affiliation":[{"name":"Department of Anesthesia and Perioperative Care, University of California, San Francisco"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026072410001279000_2026.07.18.739219v1.1","doi-asserted-by":"crossref","first-page":"78","DOI":"10.3389\/fncel.2017.00078","article-title":"Neonatal Hypoxia Ischaemia: Mechanisms, Models, and Therapeutic Challenges","volume":"11","year":"2017","journal-title":"Front Cell Neurosci"},{"key":"2026072410001279000_2026.07.18.739219v1.2","doi-asserted-by":"crossref","first-page":"2444","DOI":"10.1038\/s41390-025-03986-2","article-title":"Neonatal encephalopathy and hypoxic-ischemic encephalopathy: the state of the art","volume":"98","year":"2025","journal-title":"Pediatr Res"},{"key":"2026072410001279000_2026.07.18.739219v1.3","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1001\/jamapediatrics.2014.3269","article-title":"Hypoxic-ischemic encephalopathy: a review for the clinician","volume":"169","year":"2015","journal-title":"JAMA Pediatr"},{"key":"2026072410001279000_2026.07.18.739219v1.4","doi-asserted-by":"crossref","unstructured":"Arnautovic T , Sinha S , Laptook AR : Neonatal Hypoxic-Ischemic Encephalopathy and Hypothermia Treatment. 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