Li Bo, Zani Augusto, Martin Zechariah, Lee Carol, Zani-Ruttenstock Elke, Eaton Simon, Pierro Agostino
Division of General and Thoracic Surgery, Physiology and Experimental Medicine Program, The Hospital for Sick Children, Toronto, ON, Canada.
UCL Institute of Child Health, London, United Kingdom.
J Pediatr Surg. 2016 May;51(5):775-8. doi: 10.1016/j.jpedsurg.2016.02.019. Epub 2016 Feb 12.
BACKGROUND/PURPOSE: Oxidative stress is implicated in the pathogenesis of necrotizing enterocolitis (NEC). Hydrogen sulfide (H2S) has been reported to have a protective function against oxidative stress in the gut. We hypothesize that administration of H2S can help decrease intestinal epithelial cell injury in vitro.
Intestinal epithelial cells (IEC-18) were treated with 200μM hydrogen peroxide (H2O2) for 21h. At 21h sodium hydrosulfide (NaHS), an H2S donor, was administered as a rescue treatment at two different concentrations: 0.1mM and 0.2mM. At 24h, cell viability was measured using a colorimetric assay (MTT). Oxidative stress was studied by glutathione peroxidase (GPx) activity and thiobarbituric acid reactive substances (TBARS). IL-6 and TNFα levels were tested to study inflammation. Data were presented as mean±SD and compared using one-way ANOVA with Bonferroni post-test.
Compared to control, H2O2-treated IEC-18 had reduced viability (p<0.01), lower GPx activity (p<0.01), higher TBARS levels (p<0.01), and increased IL6 and TNFα (p<0.001). Compared to H2O2-treated IEC-18, treatment with 0.2mM NaHS rescued viability (p<0.01), increased GPx activity (p<0.05), and reduced TBARS (p<0.01), IL6 and TNFα (p<0.001).
H2S successfully rescues epithelial cell damage induced by oxidative stress in vitro. This indicates that H2S could be a potential pharmacological intervention in conditions like NEC.
背景/目的：氧化应激与坏死性小肠结肠炎（NEC）的发病机制有关。据报道，硫化氢（H2S）对肠道氧化应激具有保护作用。我们假设给予H2S有助于在体外减少肠上皮细胞损伤。
用200μM过氧化氢（H2O2）处理肠上皮细胞（IEC-18）21小时。在21小时时，以两种不同浓度（0.1mM和0.2mM）给予硫化氢供体硫氢化钠（NaHS）作为挽救治疗。在24小时时，使用比色法（MTT）测量细胞活力。通过谷胱甘肽过氧化物酶（GPx）活性和硫代巴比妥酸反应性物质（TBARS）研究氧化应激。检测IL-6和TNFα水平以研究炎症。数据以平均值±标准差表示，并使用单因素方差分析和Bonferroni事后检验进行比较。
与对照组相比，H2O2处理的IEC-18活力降低（p<0.01），GPx活性降低（p<0.01），TBARS水平升高（p<0.01），IL-6和TNFα升高（p<0.001）。与H2O2处理的IEC-18相比，用0.2mM NaHS处理可挽救活力（p<0.01），增加GPx活性（p<0.05），并降低TBARS（p<0.01）、IL-6和TNFα（p<0.001）。
H2S成功挽救了体外氧化应激诱导的上皮细胞损伤。这表明H2S可能是NEC等病症的一种潜在药物干预措施。
