In Vivo Effects of Neostigmine and Physostigmine on Neutrophil Functions and Evaluation of Acetylcholinesterase and Butyrylcholinesterase as Inflammatory Markers during Experimental Sepsis in Rats

Mediators Inflamm. 2019 Jan 20:2019:8274903. doi: 10.1155/2019/8274903. eCollection 2019.

Abstract

Introduction: Recent studies have shown that acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) may serve as important diagnostic and therapeutic targets in sepsis. Since polymorphonuclear neutrophils (PMNs) play a pivotal role in the early phase of sepsis, we evaluated the potential therapeutic effects of cholinesterase inhibitors on PMN functions during cecal ligation and puncture- (CLP-) induced sepsis and investigated the roles of AChE and BChE as inflammatory markers under standardized experimental conditions.

Methods: Sham surgery or CLP was performed in male Wistar rats (n = 60). Animals were randomized into four groups: physostigmine, 100 μg/kg; neostigmine, 75 μg/kg; 0.9% saline (control group); and sham group, each applied four times over 24 h. The levels of reactive oxygen species (ROS) production and CD11b/CD62l expression were quantified by flow cytometry at t = 0, 6, 15, 20, and 24 h. Blood gas analysis as well as AChE and BChE activity levels was measured by validated point-of-care measurements. Clinical scores and survival times were determined.

Results: CLP induced a significant increase in ROS production and CD11b upregulation by rat PMNs. Treatment with physostigmine or neostigmine significantly reduced ROS production and CD11b upregulation by PMNs 20 h after CLP induction. In physostigmine-treated animals, survival times were significantly improved compared to the control animals, but not in neostigmine-treated animals. While AChE activity significantly decreased in the control animals at t > 6 h, AChE activity did not change in the sham group. BChE activity decreased at t > 20 h in the control animals.

Conclusion: While AChE activity may serve as an acute inflammatory marker, BChE activity shows a delayed decrease. Administration of centrally acting physostigmine in CLP-induced sepsis in rats has protective effects on PMN functions and improves survival times, which may be of interest in clinical practice.

MeSH terms

  • Acetylcholinesterase / metabolism*
  • Animals
  • Biomarkers / metabolism*
  • Blood Gas Analysis
  • Butyrylcholinesterase / metabolism*
  • Male
  • Neostigmine / therapeutic use*
  • Neutrophils / drug effects*
  • Physostigmine / therapeutic use*
  • Rats
  • Rats, Wistar
  • Reactive Oxygen Species / metabolism
  • Sepsis / drug therapy*
  • Sepsis / metabolism*

Substances

  • Biomarkers
  • Reactive Oxygen Species
  • Neostigmine
  • Physostigmine
  • Acetylcholinesterase
  • Butyrylcholinesterase