Department of Clinical Research Center, The Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou 310052, China.
Department of Intensive Care Medicine, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310016, China.
Mediators Inflamm. 2022 Jul 29;2022:7659282. doi: 10.1155/2022/7659282. eCollection 2022.
Human neutrophil peptides 1-3 (HNP1-3), also known as human -defensins, are the most abundant neutrophil granule proteins. The genes that encode HNP1-3, , exhibit extensive copy number variations, which correlate well with their protein levels. Human and mouse studies have shown that increased copy numbers of worsen sepsis outcomes. Additionally, high concentrations of HNP1-3 in body fluids have been reported in patients with sepsis. However, direct evidence for the pathogenic role of HNP1-3 proteins during sepsis progression is lacking. In current study, sepsis was induced by means of cecal puncture and ligation. Various doses of HNP-1 (low dose with 0.5 mg/kg body weight and high dose with 10 mg/kg body weight) or phosphate buffer saline were intraperitoneally administered to mice at six hours after sepsis onset. Survival rate was monitored, and vascular permeability, endothelial cell pyroptosis, and immunofluorescence of endothelial adherens junction protein vascular endothelial-cadherin were evaluated. The administration of a high dose of HNP-1 after sepsis onset led to increased mortality, more severe liver injury, and increased vascular permeability in the liver and mesentery. The injection of high dose of HNP-1 did not directly induce liver endothelial cell death but destroyed interendothelial junctions in the liver. Moreover, genetic deficiency of nucleotide-binding oligomerization domain-like receptor protein-3 or caspase-1 abrogated the high mortality and disrupted liver interendothelial junctions caused by high dose of HNP-1 during sepsis. This study directly demonstrates that neutrophil defensins play a key role in regulating endothelial stability during sepsis development.
人中性粒细胞肽 1-3（HNP1-3），也称为人防御素，是最丰富的中性粒细胞颗粒蛋白。编码 HNP1-3 的基因表现出广泛的拷贝数变异，与它们的蛋白水平密切相关。人类和小鼠研究表明，HNP1-3 基因拷贝数增加会导致脓毒症结局恶化。此外，脓毒症患者的体液中也报道了 HNP1-3 的高浓度。然而，缺乏 HNP1-3 蛋白在脓毒症进展过程中致病作用的直接证据。在本研究中，通过盲肠穿刺和结扎诱导脓毒症。在脓毒症发作后 6 小时，用不同剂量的 HNP-1（低剂量 0.5mg/kg 体重和高剂量 10mg/kg 体重）或磷酸盐缓冲盐水经腹腔注射到小鼠体内。监测存活率，并评估血管通透性、内皮细胞焦亡和内皮黏附连接蛋白血管内皮钙黏蛋白的免疫荧光。在脓毒症发作后给予高剂量 HNP-1 导致死亡率增加、更严重的肝损伤和肝及肠系膜血管通透性增加。高剂量 HNP-1 注射不会直接诱导肝内皮细胞死亡，但会破坏肝内的内皮细胞连接。此外，核苷酸结合寡聚化结构域样受体蛋白 3 或半胱氨酸天冬氨酸蛋白酶 1 的基因缺失消除了高剂量 HNP-1 在脓毒症期间引起的高死亡率和破坏肝内皮细胞连接。本研究直接证明了中性粒细胞防御素在调节脓毒症发展过程中内皮稳定性方面发挥着关键作用。
