PMID- 37441587 OWN - NLM STAT- MEDLINE DCOM- 20230717 LR - 20240212 IS - 1838-7640 (Electronic) IS - 1838-7640 (Linking) VI - 13 IP - 11 DP - 2023 TI - Hepatocyte HSPA12A inhibits macrophage chemotaxis and activation to attenuate liver ischemia/reperfusion injury via suppressing glycolysis-mediated HMGB1 lactylation and secretion of hepatocytes. PG - 3856-3871 LID - 10.7150/thno.82607 [doi] AB - Rationale: Liver ischemia-reperfusion (LI/R) injury is characterized by two interconnected phases: local ischemia that causes hepatic cell damage to release damage-associated molecular pattern (DAMPs), and DAMPs that recruit immune cells to elicit inflammatory cascade for further injury of hepatocytes. High-mobility group box 1 (HMGB1) is a representative DAMP. Studies in macrophages demonstrated that HMGB1 is secreted after lactylation during sepsis. However, whether lactylation mediates HMGB1 secretion from hepatocytes after LI/R is known. Heat shock protein A12A (HSPA12A) is an atypical member of HSP70 family. Methods: Gene expression was examined by microarray analysis and immunoblotting. The hepatic injury was analyzed using released ALT and AST activities assays. Hepatic macrophage chemotaxis was evaluated by Transwell chemotaxis assays. Inflammatory mediators were evaluated by immunoblotting. HMGB1 secretion was examined in exosomes or serum. HMGB1 lactylation was determined using immunoprecipitation and immunoblotting. Results: Here, we report that LI/R decreased HSPA12A expression in hepatocytes, while hepatocyte-specific HSPA12A overexpression attenuated LI/R-induced hepatic dysfunction and mortality of mice. We also noticed that hepatocyte HSPA12A overexpression suppressed macrophage chemotaxis to LI/R-exposed livers in vivo and to hypoxia/reoxygenation (H/R)-exposed hepatocytes in vitro. The LI/R-increased serum HMGB1 levels of mice and the H/R-increased HMGB1 lactylation and secretion levels of hepatocytes were also inhibited by hepatocyte HSPA12A overexpression. By contrast, HSPA12A knockout in hepatocytes promoted not only H/R-induced HMGB1 lactylation and secretion of hepatocytes but also the effects of H/R-hepatocytes on macrophage chemotaxis and inflammatory activation, while all these deleterious effects of HSPA12A knockout were reversed following hepatocyte HMGB1 knockdown. Further molecular analyses showed that HSPA12A overexpression reduced glycolysis-generated lactate, thus decreasing HMGB1 lactylation and secretion from hepatocytes, thereby inhibiting not only macrophage chemotaxis but also the subsequent inflammatory cascade, which ultimately protecting against LI/R injury. Conclusion: Taken together, these findings suggest that hepatocyte HSPA12A is a novel regulator that protects livers from LI/R injury by suppressing glycolysis-mediated HMGB1 lactylation and secretion from hepatocytes to inhibit macrophage chemotaxis and inflammatory activation. Therefore, targeting hepatocyte HSPA12A may have therapeutic potential in the management of LI/R injury in patients. CI - (c) The author(s). FAU - Du, Shuya AU - Du S AD - Department of Geriatrics, Jiangsu Provincial Key Laboratory of Geriatrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. FAU - Zhang, Xiaojin AU - Zhang X AD - Department of Geriatrics, Jiangsu Provincial Key Laboratory of Geriatrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. FAU - Jia, Yunxiao AU - Jia Y AD - Department of Geriatrics, Jiangsu Provincial Key Laboratory of Geriatrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. FAU - Peng, Peipei AU - Peng P AD - Department of Anesthesiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. FAU - Kong, Qiuyue AU - Kong Q AD - Department of Anesthesiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. FAU - Jiang, Surong AU - Jiang S AD - Department of Geriatrics, Jiangsu Provincial Key Laboratory of Geriatrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. FAU - Li, Yuehua AU - Li Y AD - Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Nanjing 210029, China. FAU - Li, Chuanfu AU - Li C AD - Departments of Surgery, East Tennessee State University, Johnson City, TN 37614, USA. FAU - Ding, Zhengnian AU - Ding Z AD - Department of Anesthesiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. FAU - Liu, Li AU - Liu L AD - Department of Geriatrics, Jiangsu Provincial Key Laboratory of Geriatrics, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. AD - Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Nanjing 210029, China. LA - eng PT - Journal Article DEP - 20230703 PL - Australia TA - Theranostics JT - Theranostics JID - 101552395 RN - 0 (Heat-Shock Proteins) RN - 0 (HMGB1 Protein) SB - IM MH - Animals MH - Mice MH - Heat-Shock Proteins/metabolism MH - *HMGB1 Protein/metabolism MH - Chemotaxis MH - Liver/metabolism MH - Hepatocytes/metabolism MH - *Liver Diseases MH - Macrophages/metabolism MH - Glycolysis MH - *Reperfusion Injury/metabolism MH - Mice, Inbred C57BL PMC - PMC10334822 OTO - NOTNLM OT - Chemotaxis OT - Heat shock protein A12A (HSPA12A) OT - Hepatocyte OT - High-mobility group box 1(HMGB1) OT - Lactylation OT - Liver ischemia/reperfusion (LI/R) OT - Macrophage COIS- Competing Interests: The authors have declared that no competing interest exists. EDAT- 2023/07/13 19:15 MHDA- 2023/07/17 06:42 PMCR- 2023/01/01 CRDT- 2023/07/13 15:30 PHST- 2023/01/12 00:00 [received] PHST- 2023/05/26 00:00 [accepted] PHST- 2023/07/17 06:42 [medline] PHST- 2023/07/13 19:15 [pubmed] PHST- 2023/07/13 15:30 [entrez] PHST- 2023/01/01 00:00 [pmc-release] AID - thnov13p3856 [pii] AID - 10.7150/thno.82607 [doi] PST - epublish SO - Theranostics. 2023 Jul 3;13(11):3856-3871. doi: 10.7150/thno.82607. eCollection 2023.