Hydrogen Sulfide Donor GYY4137 Acts Through Endothelial Nitric Oxide to Protect Intestine in Murine Models of Necrotizing Enterocolitis and Intestinal Ischemia
Hydrogen Sulfide Donor GYY4137 Acts Through Endothelial Nitric Oxide to Protect Intestine in Murine Models of Necrotizing Enterocolitis and Intestinal Ischemia
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DOI：
10.1016/j.jss.2018.08.048
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发表时间：
2019-02-01
影响因子：
2.2
通讯作者：
Markel, Troy A.
中科院分区：
文献类型：
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作者：
Drucker, Natalie A.;Jensen, Amanda R.;Markel, Troy A.
背景：早产儿坏死性小肠结肠炎（NEC）通常是一种预后不良的毁灭性外科疾病。GYY 4137是硫化氢的长效供体，硫化氢是一种气体递质，在实验性NEC中对肠损伤具有保护作用，可能是通过保护免受继发于缺血的损伤。我们假设GYY 4137的给药将改善肠系膜灌注，减少肠损伤，并减少实验性NEC和缺血再灌注损伤中的炎症反应，并且这些益处将通过内皮一氧化氮合酶依赖性途径介导。在C57 BL/6野生型（WT）和内皮型一氧化氮合酶（eNOS）敲除（eNOSKO）幼仔中通过母体分离、配方喂养、肠内脂多糖，以及间歇性缺氧和低温应激。幼仔每天腹腔注射50 mg/kg GYY 4137或磷酸盐缓冲盐水溶剂。在单独的组中，成年雄性WT和eNOSKO小鼠进行上级肠系膜动脉闭塞60分钟。在腹部闭合之前，向腹腔内给予50 mg/kg GYY 4137或磷酸盐缓冲盐水溶剂。激光多普勒成像用于评估幼仔在基线和出生后第9天的肠系膜灌注，以及成年小鼠在基线和缺血损伤后24小时的肠系膜灌注。人道处死后，将每只动物的回肠末端固定、石蜡包埋、切片并用苏木精和伊红染色。使用已发表的损伤评分对切片进行盲法分级。将肠组织均质化并通过ELISA测量细胞因子。使用Mann eWhitney U检验比较数据，P值
Background: Necrotizing enterocolitis (NEC) inpremature infants is often a devastating surgical condition with poor outcomes. GYY4137 is a long-acting donor of hydrogen sulfide, a gaso-transmitter that is protective against intestinal injury in experimental NEC, likely through protection against injury secondary to ischemia. We hypothesized that administration of GYY4137 would improve mesenteric perfusion, reduce intestinal injury, and reduce inflammatory responses in experimental NEC and ischemia-reperfusion injury, and that these benefits would be mediated through endothelial nitric oxide synthasee-dependent pathways.Methods: NEC was induced in C57BL/6 wild-type (WT) and endothelial nitric oxide synthase (eNOS) knockout (eNOSKO) pups via maternal separation, formula feeding, enteral lipopolysaccharide, and intermittent hypoxic and hypothermic stress. Pups received daily intraperitoneal injections of 50 mg/kg GYY4137 or phosphate buffered saline vehicle. In separate groups, adult male WT and eNOSKO mice underwent superior mesenteric artery occlusion for 60 min. Before abdominal closure, 50 mg/kg GYY4137 or phosphate buffered saline vehicle was administered into the peritoneal cavity. Laser doppler imaging was used to assess mesenteric perfusion of pups at baseline and on postnatal day 9, and the adult mice at baseline and 24 h after ischemic insult. After euthanasia, the terminal ileum of each animal was fixed, paraffin embedded, sectioned, and stained with hematoxylin and eosin. Sections were blindly graded using published injury scores. Intestinal tissue was homogenized and cytokines measured by ELISA. Data were compared using Mann eWhitney U test, and P-values