Snyder Katherine B, Calkins Chase L, Golubkova Alena, Leiva Tyler, Schlegel Camille, Hunter Catherine J
Division of Pediatric Surgery, Oklahoma City, OK, 73104, USA.
Department of Surgery, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA.
J Inflamm Res. 2024 Jan 17;17:331-341. doi: 10.2147/JIR.S436125. eCollection 2024.
Necrotizing enterocolitis (NEC) is the leading gastrointestinal cause of death of premature neonates. NEC is associated with prematurity, a hyperinflammatory response, and dysregulation of intestinal barrier function. We hypothesize that patients with NEC will have, and continue to have after recovery, an increased hyperinflammatory intestinal response compared to those patients without NEC.
Neonates with NEC, those that have recovered from NEC, and those without NEC undergoing intestinal resections had specimens collected and snap frozen or generated into enteroids. The enteroids were treated with 100ug/mL lipopolysaccharide (LPS) and subjected to 24 hr of hypoxia together, then compared with untreated controls. Expression of Tumor Necrosis Factor (TNF-α) and interleukin 8 (IL-8) were evaluated via RT-qPCR and ELISA to measure inflammatory response. ANOVA determined statistical significance (p<0.05).
There was no difference in inflammatory markers in recovered NEC tissue compared to non-NEC tissue on RTqPCR (p=0.701 TNF-α and 0.861 IL-8). However, recovered NEC enteroids demonstrate elevated levels of inflammatory markers after treatment compared to non-NEC enteroids after treatment on RTqPCR (p=0.0485 TNF-α, p=0.0057 IL-8) and ELISA (p=0.0354 TNF-α, p=0.0011 IL-8). Recovered NEC enteroids that underwent treatment demonstrated increased inflammatory markers compared to recovered NEC enteroids without treatment on RTqPCR (p=0.0045 TNF-α, p=0.0002 IL-8) and ELISA (p=0.034 TNF-α, p=0.0002 IL-8) suggesting a heightened inflammatory response to a second hit.
Intestinal tissue resected from neonates with NEC has an elevated hyperinflammatory response compared to neonates recovered from NEC and neonates without NEC. Enteroids generated from patients that have recovered from NEC have a heightened inflammatory response in response to NEC inducing stimuli compared to controls. This tendency towards an increased hyperinflammatory state may be correlated with an infant's proclivity to develop NEC and demonstrates the significance of a second hit on this tissue creating a heightened inflammatory response. This could be correlated with the impact and trajectory of an illness post recovery from NEC.
坏死性小肠结肠炎（NEC）是早产新生儿胃肠道死亡的主要原因。NEC与早产、过度炎症反应以及肠道屏障功能失调有关。我们假设，与无NEC的患者相比，NEC患者在患病期间以及恢复后，肠道炎症反应会增强。
收集接受肠道切除术的NEC新生儿、已从NEC恢复的新生儿以及无NEC的新生儿的标本，速冻或制成类器官。将类器官用100μg/mL脂多糖（LPS）处理并共同进行24小时缺氧处理，然后与未处理的对照进行比较。通过逆转录定量聚合酶链反应（RT-qPCR）和酶联免疫吸附测定（ELISA）评估肿瘤坏死因子（TNF-α）和白细胞介素8（IL-8）的表达，以测量炎症反应。方差分析确定统计学显著性（p<0.05）。
在RT-qPCR上，与非NEC组织相比，已恢复的NEC组织中的炎症标志物无差异（TNF-α p=0.701，IL-8 p=0.861）。然而，在RT-qPCR（TNF-α p=0.0485，IL-8 p=0.0057）和ELISA（TNF-α p=0.0354，IL-8 p=0.0011）上，与处理后的非NEC类器官相比，已恢复的NEC类器官在处理后炎症标志物水平升高。在RT-qPCR（TNF-α p=0.0045，IL-8 p=0.0002）和ELISA（TNF-α p=0.034，IL-8 p=0.0002）上，与未处理的已恢复的NEC类器官相比，接受处理的已恢复的NEC类器官炎症标志物增加，表明对二次打击的炎症反应增强。
与已从NEC恢复的新生儿和无NEC的新生儿相比，从患有NEC的新生儿切除的肠道组织炎症反应增强。与对照组相比，已从NEC恢复的患者产生的类器官对NEC诱导刺激的炎症反应增强。这种炎症反应增强状态的倾向可能与婴儿患NEC的倾向相关，并表明该组织受到二次打击会产生增强的炎症反应的重要性。这可能与NEC恢复后疾病的影响和发展轨迹相关。
