Department of Trauma and Acute Care Surgery, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Department of Trauma and Acute Care Surgery, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Cell Signal. 2024 Jan;113:110941. doi: 10.1016/j.cellsig.2023.110941. Epub 2023 Oct 27.
The mechanism by which neutrophil extracellular traps (NETs) may cause intestinal barrier dysfunction in response to trauma/hemorrhagic shock (T/HS) remains unclear. In this study, the roles and mechanisms of NETs in macrophage polarization were examined to determine whether this process plays a role in tissue damage associated with T/HS. Rat models of T/HS and macrophage polarization were developed and the levels of NETs formation in the intestinal tissue of T/HS rats were assessed. NET formation was inhibited in models of T/HS to examine the effect on intestinal inflammation and barrier injury. The proportions of pro-inflammatory and anti-inflammatory macrophages in the damaged intestinal tissues were measured. Finally, high-throughput sequencing was performed to investigate the underlying mechanisms involved in this process. The study revealed that the level of NETs formation was increased and that inhibition of NETs formation alleviated the intestinal inflammation and barrier injury. Moreover, the number of pro-inflammatory macrophages increased and the number of anti-inflammatory macrophages decreased. RNA sequencing analysis indicated that NETs formation decreased the expression of transforming growth factor-beta receptor 2 (TGFBR2), bioinformatic analyses revealed that TGFBR2 was significantly enriched in the transforming growth factor-beta (TGF-β) signaling pathway. Verification experiments showed that NETs impeded macrophage differentiation into the anti-inflammatory/M2 phenotype and inhibited TGFBR2 and TGF-β expression in macrophages. However, treatment with DNase I and overexpression of TGFBR2, and inhibition of TGF-β promoted and prevented this process, respectively. NETs may regulate the macrophage polarization process by promoting intestinal barrier dysfunction in T/HS rats through the TGFBR2-mediated TGF-β signaling pathway.
中性粒细胞胞外诱捕网（NETs）在创伤/失血性休克（T/HS）中引起肠道屏障功能障碍的机制尚不清楚。在这项研究中，研究了 NETs 在巨噬细胞极化中的作用和机制，以确定这一过程是否在与 T/HS 相关的组织损伤中发挥作用。建立了 T/HS 大鼠模型和巨噬细胞极化模型，并评估了 T/HS 大鼠肠道组织中 NETs 形成的水平。在 T/HS 模型中抑制 NETs 的形成，以观察其对肠道炎症和屏障损伤的影响。测量损伤肠道组织中促炎和抗炎巨噬细胞的比例。最后，进行高通量测序以研究该过程涉及的潜在机制。研究表明，NETs 形成的水平增加，抑制 NETs 形成可减轻肠道炎症和屏障损伤。此外，促炎巨噬细胞的数量增加，抗炎巨噬细胞的数量减少。RNA 测序分析表明，NETs 形成降低了转化生长因子-β受体 2（TGFBR2）的表达，生物信息学分析表明，TGFBR2 在转化生长因子-β（TGF-β）信号通路中显著富集。验证实验表明，NETs 阻碍巨噬细胞向抗炎/M2 表型分化，并抑制巨噬细胞中 TGFBR2 和 TGF-β 的表达。然而，DNase I 处理、TGFBR2 过表达和 TGF-β 抑制分别促进和阻止了这一过程。NETs 可能通过 TGFBR2 介导的 TGF-β 信号通路促进 T/HS 大鼠肠道屏障功能障碍，调节巨噬细胞极化过程。
