Hepatocyte HSPA12A inhibits macrophage chemotaxis and activation to attenuate liver ischemia/reperfusion injury via suppressing glycolysis-mediated HMGB1 lactylation and secretion of hepatocytes.
Hepatocyte HSPA12A inhibits macrophage chemotaxis and activation to attenuate liver ischemia/reperfusion injury via suppressing glycolysis-mediated HMGB1 lactylation and secretion of hepatocytes.
复制标题
肝细胞HSPA12A抑制巨噬细胞趋化性和激活，通过抑制糖酵解介导的HMGB1 lactylation和肝细胞分泌来减轻肝缺血/再灌注损伤。
作者：
Du S;Zhang X;Jia Y;Peng P;Kong Q;Jiang S;Li Y;Li C;Ding Z;Liu L
基本原理：肝脏缺血再灌注（LI/R）损伤的特征在于两个相互关联的阶段：局部缺血引起肝细胞损伤以释放损伤相关分子模式（DAMP），DAMP募集免疫细胞以引发炎症级联反应以进一步损伤肝细胞。高迁移率族蛋白1（HMGB 1）是DAMP的代表。在巨噬细胞中的研究表明，HMGB 1在脓毒症期间的乳酸化后分泌。然而，乳酸化是否介导肝细胞在LI/R后分泌HMGB 1是已知的。热休克蛋白A12 A（Heat shock protein A12 A，HSPA 12 A）是HSP 70家族的一个非典型成员。方法：基因芯片分析和免疫印迹检测基因表达。用释放的ALT和AST活性测定分析肝损伤。肝巨噬细胞趋化性通过Transwell趋化性测定进行评价。通过免疫印迹法评价炎症介质。在外泌体或血清中检查HMGB 1分泌。采用免疫沉淀和免疫印迹法测定HMGB 1乳酸化。结果如下：在这里，我们报告说，LI/R降低HSPA 12 A在肝细胞中的表达，而肝细胞特异性HSPA 12 A过表达减弱LI/R诱导的肝功能障碍和小鼠死亡率。我们还注意到，肝细胞HSPA 12 A过表达抑制巨噬细胞趋化LI/R暴露的肝脏在体内和缺氧/复氧（H/R）暴露的肝细胞在体外。肝细胞HSPA 12 A过表达可抑制LI/R升高的小鼠血清HMGB 1水平和H/R升高的肝细胞HMGB 1的乳酸化和分泌水平。与此相反，HSPA 12 A基因敲除不仅促进H/R诱导的HMGB 1的乳酸化和肝细胞的分泌，而且还促进H/R-肝细胞对巨噬细胞趋化性和炎症激活的影响，而HSPA 12 A基因敲除的所有这些有害影响在肝细胞HMGB 1基因敲除后被逆转。进一步的分子分析表明，HSPA 12 A过表达减少了糖酵解产生的乳酸，从而减少了HMGB 1的乳酸化和肝细胞的分泌，从而不仅抑制了巨噬细胞的趋化性，而且抑制了随后的炎症级联反应，最终保护免受LI/R损伤。结论：总之，这些发现表明肝细胞HSPA 12 A是一种新的调节剂，通过抑制糖酵解介导的HMGB 1乳酸化和肝细胞分泌来抑制巨噬细胞趋化性和炎症活化，从而保护肝脏免受LI/R损伤。因此，靶向肝细胞HSPA 12 A可能在患者的LI/R损伤的管理中具有治疗潜力。
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.
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25.7
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通讯作者：
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