Wang Jing, Zhang Zhiyu, Yu Mingxia, Xin LinYan
School of Biology and Food Engineering, Institute of Pharmaceutical Pharmacology, Research Center, Suzhou University, Suzhou, Anhui, China.
School of Biology and Food Engineering, Institute of Pharmaceutical Pharmacology, Research Center, Suzhou University, Suzhou, Anhui, China.
Mol Immunol. 2025 Jan;177:96-103. doi: 10.1016/j.molimm.2024.12.007. Epub 2025 Jan 3.
The aim of this study was to reveal the mechanism of cold stimulation (CS)-bronchial epithelial cells (BECs) derived exosomes (CS-BECs-exo) aggravated sepsis induced acute lung injury (SALI). CS-BECs-exo were separated by differential centrifugation and were characterized. Proteomics, immunoprecipitation, and RAGE knockout (RAGE) mice were used to investigate the mechanism of CS-BECs-exo aggravated SALI. The results of transmission electron microscope (TEM) showed that CS-BECs-exo showed a double-layer membrane structure like a saucer. Nanoparticle tracking analysis (NTA) particle size analysis showed that the average particle size of CS-BECs-exo was 123.6 nm. The results of proteomics showed that the expression level HMGB1 was significantly increased in CS-BECs-exo compared with BECs-exo. CS-BECs-exo significantly increased oxidative stress and inflammatory reaction of SALI. In addition, CS-BECs-exo significantly increased RAGE and decreased the levels of Nrf-2 and OH-1. RAGE knockout (RAGE KO) and silence of RAGE (RAGE siRNA) significantly canceled the effects of CS-BECs-exo on SALI. HMGB1 knockout (HMGB1) and silence of HMGB1 also significantly (HMGB1 siRNA) canceled the effects of CS-BECs-exo on SALI. In conclusion, CS-BECs-exo aggravated ALI in sepsis via HMGB1/RAGE/Nrf-2/OH-1 signal pathway.
本研究旨在揭示冷刺激(CS)-支气管上皮细胞(BECs)来源的外泌体(CS-BECs-exo)加重脓毒症诱导的急性肺损伤(SALI)的机制。通过差速离心分离CS-BECs-exo并对其进行表征。采用蛋白质组学、免疫沉淀和RAGE基因敲除(RAGE)小鼠来研究CS-BECs-exo加重SALI的机制。透射电子显微镜(TEM)结果显示,CS-BECs-exo呈现出碟状的双层膜结构。纳米颗粒跟踪分析(NTA)粒径分析表明,CS-BECs-exo的平均粒径为123.6 nm。蛋白质组学结果显示,与BECs-exo相比,CS-BECs-exo中HMGB1的表达水平显著升高。CS-BECs-exo显著增加了SALI的氧化应激和炎症反应。此外,CS-BECs-exo显著增加RAGE表达,并降低Nrf-2和OH-1的水平。RAGE基因敲除(RAGE KO)和RAGE沉默(RAGE siRNA)显著消除了CS-BECs-exo对SALI的影响。HMGB1基因敲除(HMGB1)和HMGB-1沉默(HMGB1 siRNA)也显著消除了CS-BECs-exo对SALI的影响。总之,CS-BECs-exo通过HMGB/RAGE/Nrf-2/OH-1信号通路加重脓毒症中的ALI。