Department of Surgery, University of Texas Health Science Center at Houston, Houston, Texas 77030, USA.
Shock. 2012 Jul;38(1):57-62. doi: 10.1097/SHK.0b013e31825a188a.
Syndecan 1 is the predominant heparan sulfate proteoglycan found on the surface of epithelial cells and, like glutamine, is essential in maintaining the intestinal epithelial barrier. We therefore hypothesized that loss of epithelial syndecan 1 would abrogate the gut-protective effects of enteral glutamine. Both an in vitro and in vivo model of gut ischemia-reperfusion (IR) was utilized. In vitro, intestinal epithelial cells underwent hypoxia-reoxygenation to mimic gut IR with 2 mM (physiologic) or 10 mM glutamine supplementation. Permeability, caspase activity, cell growth, and cell surface and shed syndecan 1 were assessed. In vivo, wild-type and syndecan 1 knockout (KO) mice received ± enteral glutamine followed by gut IR. Intestinal injury was assessed by fluorescent dye clearance and histopathology, permeability as mucosal-to-serosal clearance ex vivo in everted sacs, and inflammation by myeloperoxidase (MPO) activity. In an in vitro model of gut IR, glutamine supplementation reduced epithelial cell permeability and apoptosis and enhanced cell growth. Shed syndecan 1 was reduced by glutamine without an increase in syndecan 1 mRNA. In vivo, intestinal permeability, inflammation, and injury were increased after gut IR in wild-type mice and further increased in syndecan 1 KO mice. Glutamine's attenuation of IR-induced intestinal hyperpermeability, inflammation, and injury was abolished in syndecan 1 KO mice. These results suggest that syndecan 1 plays a novel role in the protective effects of enteral glutamine in the postischemic gut.
硫酸乙酰肝素蛋白聚糖 1 是上皮细胞表面主要的肝素硫酸蛋白聚糖,与谷氨酰胺一样,对于维持肠道上皮屏障至关重要。因此,我们假设上皮细胞硫酸乙酰肝素蛋白聚糖 1 的缺失会消除肠内谷氨酰胺的肠道保护作用。我们利用了肠缺血再灌注(IR)的体外和体内模型。在体外,肠上皮细胞经历缺氧复氧以模拟肠道 IR,同时补充 2 mM(生理)或 10 mM 谷氨酰胺。评估通透性、半胱天冬酶活性、细胞生长以及细胞表面和脱落的硫酸乙酰肝素蛋白聚糖 1。在体内,野生型和硫酸乙酰肝素蛋白聚糖 1 敲除(KO)小鼠接受或不接受肠内谷氨酰胺,然后进行肠道 IR。通过荧光染料清除和组织病理学评估肠道损伤,通过外翻肠囊的黏膜-浆膜清除评估通透性,通过髓过氧化物酶(MPO)活性评估炎症。在肠道 IR 的体外模型中,谷氨酰胺补充可降低上皮细胞通透性和凋亡,并增强细胞生长。谷氨酰胺可减少脱落的硫酸乙酰肝素蛋白聚糖 1,而不会增加硫酸乙酰肝素蛋白聚糖 1 mRNA。在体内,野生型小鼠的肠道 IR 后肠道通透性、炎症和损伤增加,而硫酸乙酰肝素蛋白聚糖 1 KO 小鼠的这些指标进一步增加。在硫酸乙酰肝素蛋白聚糖 1 KO 小鼠中,谷氨酰胺对 IR 诱导的肠道高通透性、炎症和损伤的缓解作用被消除。这些结果表明,硫酸乙酰肝素蛋白聚糖 1 在肠内谷氨酰胺对缺血后肠道的保护作用中发挥了新的作用。