Abstract：
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.
摘要：
中性粒细胞胞外陷阱（NETs）可能导致创伤/出血性休克（T/HS）肠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,以确定该过程是否在与T/HS相关的组织损伤中起作用.建立了T/HS和巨噬细胞极化的大鼠模型，并评估了T/HS大鼠肠组织中NETs的形成水平。在T/HS模型中，NET的形成受到抑制，以检查其对肠道炎症和屏障损伤的影响。测量受损肠组织中促炎和抗炎巨噬细胞的比例。最后,我们进行了高通量测序,以研究参与这一过程的潜在机制.研究表明，NETs的形成水平增加，抑制NETs的形成减轻了肠道炎症和屏障损伤。此外，促炎巨噬细胞数量增加,抗炎巨噬细胞数量减少.RNA测序分析表明，NETs的形成降低了转化生长因子-β受体2（TGFBR2）的表达，生物信息学分析显示,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阻碍巨噬细胞分化为抗炎/M2表型，并抑制巨噬细胞中TGFBR2和TGF-β的表达。然而,用DNaseI和TGFBR2的过表达以及TGF-β的抑制促进和阻止了这一过程，分别。NETs可能通过TGFBR2介导的TGF-β信号通路促进T/HS大鼠肠屏障功能障碍,从而调节巨噬细胞极化过程。
