Journal of inflammation research · 2025
Unveiling the Temporal Dynamics and Molecular Regulation Profiles of Neutrophil Extracellular Traps Following Spinal Cord Injury.
СтатьяНазвание и аннотация: Перевод пока не подготовлен
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Li Jinze, Chang Chao, Li Yanqiu, Cui Shengyu, Bai Jun, Zhang Can, Wang Xinyu, Li Kang, Jian Fengzeng
Аннотация
BACKGROUND: Spinal cord injury (SCI) initiates secondary inflammatory processes that exacerbate tissue damage, with neutrophil extracellular traps (NETs) playing a significant role in amplifying these cascades. This study aimed to explore the temporal dynamics and key regulatory genes of NET formation in SCI to identify therapeutic targets.
METHODS: We integrated two transcriptomic datasets from the GEO database to identify differentially expressed NETs-related genes (NRGs) in SCI. WGCNA identified SCI-related modules, while GSVA assessed NET formation dynamics. Publicly available single-cell RNA sequencing data from the GEO database determined cell-specific expression patterns of key NRGs. Findings were validated through immunofluorescence, Western blot, and qPCR in a mouse SCI model. Regulatory networks were constructed, and potential therapeutic compounds were predicted using DSigDB and molecular docking.
RESULTS: We identified seven key NRGs (Casp1, Ccl3, Fcgr2b, Itgam, Itgb2, Tlr2, Tlr4) in SCI. GSVA revealed peak NET score at day 1 post-injury, with attenuation at days 3 and 7. Single-cell transcriptome analysis demonstrated predominant expression of these key genes in neutrophils during the acute phase, most prominently at 1 day post-injury, which coincides with the most pronounced neutrophil infiltration. Immunofluorescence and Western blot analyses confirmed significantly elevated NET formation at 1 day post-SCI. qPCR verified the expression of all key NRGs. Regulatory network analysis identified CHD1 as an important transcription factor governing NET formation, while miRNA-mRNA network construction revealed sophisticated post-transcriptional regulation mechanisms. Drug prediction analysis identified atorvastatin as a promising therapeutic candidate with strong binding affinity to multiple key NET-related proteins.
CONCLUSION: Our study provides insights into the temporal dynamics and molecular mechanisms of NET formation after SCI, identifying potential therapeutic targets to mitigate neutrophil-mediated secondary injury and improve functional outcomes.
METHODS: We integrated two transcriptomic datasets from the GEO database to identify differentially expressed NETs-related genes (NRGs) in SCI. WGCNA identified SCI-related modules, while GSVA assessed NET formation dynamics. Publicly available single-cell RNA sequencing data from the GEO database determined cell-specific expression patterns of key NRGs. Findings were validated through immunofluorescence, Western blot, and qPCR in a mouse SCI model. Regulatory networks were constructed, and potential therapeutic compounds were predicted using DSigDB and molecular docking.
RESULTS: We identified seven key NRGs (Casp1, Ccl3, Fcgr2b, Itgam, Itgb2, Tlr2, Tlr4) in SCI. GSVA revealed peak NET score at day 1 post-injury, with attenuation at days 3 and 7. Single-cell transcriptome analysis demonstrated predominant expression of these key genes in neutrophils during the acute phase, most prominently at 1 day post-injury, which coincides with the most pronounced neutrophil infiltration. Immunofluorescence and Western blot analyses confirmed significantly elevated NET formation at 1 day post-SCI. qPCR verified the expression of all key NRGs. Regulatory network analysis identified CHD1 as an important transcription factor governing NET formation, while miRNA-mRNA network construction revealed sophisticated post-transcriptional regulation mechanisms. Drug prediction analysis identified atorvastatin as a promising therapeutic candidate with strong binding affinity to multiple key NET-related proteins.
CONCLUSION: Our study provides insights into the temporal dynamics and molecular mechanisms of NET formation after SCI, identifying potential therapeutic targets to mitigate neutrophil-mediated secondary injury and improve functional outcomes.