Center for Pediatric Lung Research, Nemours/Alfred I. duPont Hospital for Children, Wilmington, DE.
Department of Pediatrics, Nemours/Alfred I. duPont Hospital for Children, Wilmington, DE.
J Neonatal Perinatal Med. 2020;13(4):469-476. doi: 10.3233/NPM-190364.
Hypothermia with xenon gas has been used to reduce brain injury and disability rate after perinatal hypoxia-ischemia. We evaluated xenon gas therapy effects in an in vitro model with or without hypothermia on cultured human airway epithelial cells (Calu-3).
Calu-3 monolayers were grown at an air-liquid interface and exposed to one of the following conditions: 1) 21% FiO2 at 37°C (control); 2) 45% FiO2 and 50% xenon at 37°C; 3) 21% FiO2 and 50% xenon at 32°C; 4) 45% FiO2 and 50% xenon at 32°C for 24 hours. Transepithelial resistance (TER) measurements were performed and apical surface fluids were collected and assayed for total protein, IL-6, and IL-8. Three monolayers were used for immunofluorescence localization of zonula occludens-1 (ZO-1). The data were analyzed by one-way ANOVA.
TER decreased at 24 hours in all treatment groups. Xenon with hyperoxia and hypothermia resulted in greatest decrease in TER compared with other groups. Immunofluorescence localization of ZO-1 (XY) showed reduced density of ZO-1 rings and incomplete ring-like staining in the 45% FiO2- 50% xenon group at 32°C compared with other groups. Secretion of total protein was not different among groups. Secretion of IL-6 in 21% FiO2 with xenon group at 32°C was less than that of the control group. The secretion of IL-8 in 45% FiO2 with xenon at 32°C was greater than that of other groups.
Hyperoxia and hypothermia result in detrimental epithelial cell function and inflammation over 24-hour exposure. Xenon gas did not affect cell function or reduce inflammation.
氙气低温疗法已被用于减少围产期缺氧缺血后脑损伤和残疾率。我们评估了体外模型中氙气治疗对培养的人气道上皮细胞（Calu-3）的影响，包括低温治疗和不低温治疗两种情况。
Calu-3 单层细胞在气-液界面生长，并暴露于以下条件之一：1）37°C 时 21% FiO2（对照）；2）37°C 时 45% FiO2 和 50%氙气；3）32°C 时 21% FiO2 和 50%氙气；4）32°C 时 45% FiO2 和 50%氙气 24 小时。进行跨上皮电阻（TER）测量，并收集顶端表面液体，用于总蛋白、IL-6 和 IL-8 的测定。对紧密连接蛋白-1（ZO-1）进行免疫荧光定位分析，使用三个单层细胞。数据采用单因素方差分析进行分析。
所有治疗组在 24 小时时 TER 均降低。与其他组相比，高氧和低温条件下的氙气导致 TER 降低最大。在 32°C 时，与其他组相比，45% FiO2-50%氙气组的 ZO-1 环的密度降低，且不完全呈环状染色。各组之间总蛋白的分泌没有差异。在 32°C 时，21% FiO2 中含氙气组的 IL-6 分泌少于对照组。在 32°C 时，45% FiO2 中含氙气组的 IL-8 分泌大于其他组。
在 24 小时暴露中，高氧和低温导致上皮细胞功能和炎症的损害。氙气气体对细胞功能没有影响，也没有减轻炎症。
