LPS-induced macrophage HMGB1-loaded extracellular vesicles trigger hepatocyte pyroptosis by activating the NLRP3 inflammasome.
LPS-induced macrophage HMGB1-loaded extracellular vesicles trigger hepatocyte pyroptosis by activating the NLRP3 inflammasome.
复制标题
LPS诱导巨噬细胞负载HMGB1的细胞外囊泡通过激活NLRP3炎性体触发肝细胞焦亡
DOI：
10.1038/s41420-021-00729-0
复制
发表时间：
2021-11-06
影响因子：
7
通讯作者：
Li X
中科院分区：
文献类型：
--
作者：
Wang G;Jin S;Huang W;Li Y;Wang J;Ling X;Huang Y;Hu Y;Li C;Meng Y;Li X
细胞外小泡(EVS)是细胞间对话的重要载体。高迁移率族蛋白1(HMGB1)是一种典型的损伤相关分子模式(DAMP)分子，具有细胞毒性，可导致细胞死亡和组织损伤。EVS是否参与了脂多糖(LPS)诱导的急性肝损伤中HMGB1的释放，有待进一步研究。通过透射电子显微镜、纳米颗粒跟踪分析(NTA)和Western blotting鉴定EVS。共聚焦显微镜检测HMGB1、RAGE、EEA1、Rab5、Rab7、LAMP1和转铁蛋白的共定位。免疫共沉淀法研究HMGB1与RAGE的相互作用。用PKH67标记EVS并用于摄取实验。用FLICA 660-YVAD-FMK检测细胞死亡。免疫印迹和免疫组织化学方法检测NLRP3(含3个结节样受体家族结构域)炎性小体的表达。用商品化试剂盒检测血清HMGB1、ALT(丙氨酸氨基转移酶)、AST(天冬氨酸氨基转移酶)、LDH(乳酸脱氢酶)和MPO(髓过氧化物酶)。脓毒症患者血清中检测到胞外囊泡HMGB1。巨噬细胞通过EVS促进HMGB1的释放。HMGB1-RAGE相互作用参与了HMGB1进入EVS的过程。这些EV通过转铁蛋白介导的内吞作用将HMGB1运送到靶细胞，通过激活NLRP3炎性小体导致肝细胞下垂。此外，脓毒症患者血清EVS-HMGB1水平与临床肝损害呈正相关。这一发现为开发急性肝损伤的新诊断和治疗策略提供了见解。
Extracellular vesicles (EVs) have emerged as important vectors of intercellular dialogue. High mobility group box protein 1 (HMGB1) is a typical damage-associated molecular pattern (DAMP) molecule, which is cytotoxic and leads to cell death and tissue injury. Whether EVs are involved in the release of HMGB1 in lipopolysaccharide (LPS)-induced acute liver injuries need more investigation. EVs were identified by transmission electron microscopy, nanoparticle tracking analysis (NTA), and western blotting. The co-localization of HMGB1, RAGE (receptor for advanced glycation end-products), EEA1, Rab5, Rab7, Lamp1 and transferrin were detected by confocal microscopy. The interaction of HMGB1 and RAGE were investigated by co-immunoprecipitation. EVs were labeled with the PKH67 and used for uptake experiments. The pyroptotic cell death was determined by FLICA 660-YVAD-FMK. The expression of NLRP3 (NOD-like receptor family pyrin domain containing 3) inflammasomes were analyzed by western-blot or immunohistochemistry. Serum HMGB1, ALT (alanine aminotransferase), AST (aspartate aminotransferase), LDH (lactate dehydrogenase) and MPO (myeloperoxidase) were measured using a commercial kit. The extracellular vesicle HMGB1 was detected in the serums of sepsis patients. Macrophages were found to contribute to HMGB1 release through the EVs. HMGB1-RAGE interactions participated in the loading of HMGB1 into the EVs. These EVs shuttled HMGB1 to target cells by transferrin-mediated endocytosis leading to hepatocyte pyroptosis by the activation of NLRP3 inflammasomes. Moreover, a positive correlation was verified between the sepsis serum EVs-HMGB1 level and clinical liver damage. This finding provides insights for the development of novel diagnostic and therapeutic strategies for acute liver injuries.
登录
查看更多内容
影响因子：
5.6
作者：
Tsutsui H;Nishiguchi S
通讯作者：
Nishiguchi S
影响因子：
13.5
作者：
Wree, Alexander;Eguchi, Akiko;McGeough, Matthew D.;Pena, Carla A.;Johnson, Casey D.;Canbay, Ali;Hoffman, Hal M.;Feldstein, Ariel E.
通讯作者：
Feldstein, Ariel E.
影响因子：
10.6
作者：
Kang, Rui;Chen, Ruochan;Zhang, Qiuhong;Hou, Wen;Wu, Sha;Cao, Lizhi;Huang, Jin;Yu, Yan;Fan, Xue-gong;Yan, Zhengwen;Sun, Xiaofang;Wang, Haichao;Wang, Qingde;Tsung, Allan;Billiar, Timothy R.;Zeh, Herbert J., III;Lotze, Michael T.;Tang, Daolin
通讯作者：
Tang, Daolin
影响因子：
64.8
作者：
Shi, Jianjin;Zhao, Yue;Shao, Feng
通讯作者：
Shao, Feng
影响因子：
38.9
作者：
Karlsson, Sari;Pettila, Ville;Ruokonen, Esko
通讯作者：
Ruokonen, Esko