Bolognese Alexandra C, Yang Weng-Lang, Hansen Laura W, Denning Naomi-Liza, Nicastro Jeffrey M, Coppa Gene F, Wang Ping
Elmezzi Graduate School of Molecular Medicine, Manhasset, NY; Department of Surgery, Zucker School of Medicine at Hofstra/Northwell, Hempstead, NY.
Elmezzi Graduate School of Molecular Medicine, Manhasset, NY; Department of Surgery, Zucker School of Medicine at Hofstra/Northwell, Hempstead, NY; Center for Immunology and Inflammation, The Feinstein Institute for Medical Research, Manhasset, NY.
Surgery. 2018 Apr 27. doi: 10.1016/j.surg.2018.02.017.
Neonatal sepsis represents a unique therapeutic challenge owing to an immature immune system. Necroptosis is a form of programmed cell death that has been identified as an important mechanism of inflammation-induced cell death. Receptor-interacting protein kinase 1 plays a key role in mediating this process. We hypothesized that pharmacologic blockade of receptor-interacting protein kinase 1 activity would be protective in neonatal sepsis.
Sepsis was induced in C57BL/6 mouse pups (5-7 days old) by intraperitoneal injection of adult cecal slurry. At 1 hour after cecal slurry injection, the receptor-interacting protein kinase 1 inhibitor necrostatin-1 (10 µg/g body weight) or vehicle (5% dimethyl sulfoxide in phosphate buffered saline) was administered via retro-orbital injection. At 20 hours after cecal slurry injection, blood and lung tissues were collected for various analyses.
At 20 hours after sepsis induction, vehicle-treated pups showed a marked increase in serum levels of interleukin 6, interleukin 1-beta, and interleukin 18 compared to sham. With necrostatin-1 treatment, serum levels of interleukin 6, interleukin 1-beta, and interleukin 18 were decreased by 77%, 81%, and 63%, respectively, compared to vehicle. In the lungs, sepsis induction resulted in a 232-, 10-, and 2.8-fold increase in interleukin 6, interleukin 1-beta, and interleukin 18 mRNA levels compared to sham, while necrostatin-1 treatment decreased these levels to 40-, 4-, and 0.8-fold, respectively. Expressions of the neutrophil chemokines keratinocyte chemoattractant and macrophage-inflammatory-protein-2 were also increased in the lungs in sepsis, while necrostatin-1 treatment decreased these levels by 81% and 61%, respectively, compared to vehicle. In addition, necrostatin-1 treatment significantly improved the lung histologic injury score and decreased lung apoptosis in septic pups. Finally, treatment with necrostatin-1 increased the 7-day survival rate from 0% in the vehicle-treated septic pups to 29% (P = .11).
Inhibition of receptor-interacting protein kinase 1 by necrostatin-1 decreases systemic and pulmonary inflammation, decreases lung injury, and increases survival in neonatal mice with sepsis. Targeting the necroptosis pathway might represent a new therapeutic strategy for neonatal sepsis.
由于免疫系统不成熟，新生儿败血症是一个独特的治疗挑战。坏死性凋亡是一种程序性细胞死亡形式，已被确定为炎症诱导细胞死亡的重要机制。受体相互作用蛋白激酶1在介导这一过程中起关键作用。我们假设，药物性阻断受体相互作用蛋白激酶1的活性对新生儿败血症具有保护作用。
通过腹腔注射成年小鼠盲肠匀浆诱导C57BL/6幼鼠（5 - 7日龄）发生败血症。在注射盲肠匀浆1小时后，通过眶后注射给予受体相互作用蛋白激酶1抑制剂坏死素-1（10μg/g体重）或溶剂（磷酸盐缓冲盐水中5%的二甲基亚砜）。在注射盲肠匀浆20小时后，采集血液和肺组织进行各项分析。
败血症诱导20小时后，与假手术组相比，溶剂处理的幼鼠血清白细胞介素6、白细胞介素1-β和白细胞介素18水平显著升高。与溶剂处理组相比，坏死素-1处理使血清白细胞介素6、白细胞介素1-β和白细胞介素18水平分别降低了77%、81%和63%。在肺组织中，与假手术组相比，败血症诱导使白细胞介素6、白细胞介素1-β和白细胞介素18的mRNA水平分别升高了232倍、10倍和2.8倍，而坏死素-1处理使这些水平分别降至40倍、4倍和0.8倍。中性粒细胞趋化因子角质形成细胞趋化因子和巨噬细胞炎性蛋白-2在败血症小鼠肺组织中的表达也增加，而与溶剂处理组相比，坏死素-1处理使这些水平分别降低了81%和61%。此外，坏死素-1处理显著改善了败血症幼鼠的肺组织学损伤评分并减少了肺细胞凋亡。最后，坏死素-1处理使7天生存率从溶剂处理的败血症幼鼠的0%提高到29%（P = .11）。
坏死素-1抑制受体相互作用蛋白激酶1可降低全身和肺部炎症，减轻肺损伤，并提高新生败血症小鼠的生存率。靶向坏死性凋亡途径可能是新生儿败血症的一种新治疗策略。
