Capcha Jose Manuel Cóndor, Moreira Roberto S, Rodrigues Camila E, Silveira Marcelo A D, Andrade Lucia, Gomes Samirah A
Laboratory of Basic Research, University of São Paulo School of Medicine, São Paulo, Brazil.
Laboratory of Genetics, Cellular Biology, and Molecular Biology, University of São Paulo School of Medicine, São Paulo, Brazil.
Bio Protoc. 2021 Apr 5;11(7):e3979. doi: 10.21769/BioProtoc.3979.
Sepsis is a dysregulated hyperinflammatory disease caused by infection. Sepsis leads to multiple organ dysfunction syndrome (MODS), which is associated with high rates of mortality. The cecal ligation and puncture (CLP) model has been widely used in animals and has become the gold-standard method of replicating features of sepsis in humans. Despite several studies and modified CLP protocols, there are still open questions regarding the multifactorial determinants of its reproducibility and medical significance. In our protocol, which is also aimed at mimicking the sepsis observed in clinical practice, male Wistar rats are submitted to CLP with adequate fluid resuscitation (0.15 M NaCl, 25 ml/kg BW i.p.) immediately after surgery. At 6 h after CLP, additional fluid therapy (0.15 M NaCl, 25 ml/kg BW s.c.) and antibiotic therapy with imipenem-cilastatin (single dose of 14 mg/kg BW s.c.) are administered. The timing of the fluid and antibiotic therapy correspond to the initial care given when patients are admitted to the intensive care unit. This model of sepsis provides a useful platform for simulating human sepsis and could lay the groundwork for the development of new treatments.
脓毒症是一种由感染引起的炎症调节失调的疾病。脓毒症会导致多器官功能障碍综合征(MODS),这与高死亡率相关。盲肠结扎穿孔(CLP)模型已在动物中广泛应用,并成为复制人类脓毒症特征的金标准方法。尽管有多项研究以及改良的CLP方案,但关于其可重复性和医学意义的多因素决定因素仍存在未解决的问题。在我们的方案中,同样旨在模拟临床实践中观察到的脓毒症,雄性Wistar大鼠在手术后立即接受CLP并进行充分的液体复苏(0.15 M NaCl,25 ml/kg体重,腹腔注射)。CLP后6小时,给予额外的液体治疗(0.15 M NaCl,25 ml/kg体重,皮下注射)以及亚胺培南-西司他丁抗生素治疗(单剂量14 mg/kg体重,皮下注射)。液体和抗生素治疗的时间与患者入住重症监护病房时给予的初始治疗相对应。这种脓毒症模型为模拟人类脓毒症提供了一个有用的平台,并可为新治疗方法的开发奠定基础。