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The Fas Ligand/Fas Death Receptor Pathways Contribute to Propofol-Induced Apoptosis and Neuroinflammation in the Brain of Neonatal Rats

Abstract

A number of experimental studies have reported that exposure to common, clinically used anesthetics induce extensive neuroapoptosis and cognitive impairment when applied to young rodents, up to 2 weeks old, in phase of rapid synaptogenesis. Propofol is the most used general anesthetic in clinical practice whose mechanisms of neurotoxicity on the developing brain remains to be examined in depth. This study investigated effects of different exposures to propofol anesthesia on Fas receptor and Fas ligand expressions, which mediate proapoptotic and proinflammation signaling in the brain. Propofol (20 mg/kg) was administered to 7-day-old rats in multiple doses sufficient to maintain 2-, 4- and 6-h duration of anesthesia. Animals were sacrificed at 0, 4, 16 and 24 h after termination of anesthesia. It was found that propofol anesthesia induced Fas/FasL and downstream caspase-8 expression more prominently in the thalamus than in the cortex. Opposite, Bcl-2 and caspase-9, markers of intrinsic pathway activation, were shown to be more influenced by propofol treatment in the cortex. Further, we have established upregulation of caspase-1 and IL-1β cytokine transcription as well as subsequent activation of microglia that is potentially associated with brain inflammation. Behavioral analyses revealed that P35 and P60 animals, neonatally exposed to propofol, had significantly higher motor activity during three consecutive days of testing in the open field, though formation of the intersession habituation was not prevented. This data, together with our previous results, contributes to elucidation of complex mechanisms of propofol toxicity in developing brain.

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Acknowledgments

This work was supported by the Grant ON173056 from the Ministry of Education, Science and Technological Development of the Republic of Serbia.

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Correspondence to Desanka Milanovic.

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12640_2016_9629_MOESM1_ESM.tif

Supplementary Figure 1. Time-dependent changes of Bcl-xL, Bax and Bad mRNA expression in the cortex (A) and the thalamus (B) of P7 rats after propofol anesthesia were examined by using semi-quantitative RT-PCR. The results are presented for animals at different recovery time points (0, 4, 16, and 24 hr) after exposure to propofol anesthesia for 2, 4, and 6 hr. Histograms represent mRNA levels expressed as the fold change relative to the control value (100%) obtained from three separate experiments. The results are mean ±SEM. *P<0.05 vs. the control value, presented as a black line; #P<0.05 between treatments. (TIF 1,156 kb)

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Milanovic, D., Pesic, V., Loncarevic-Vasiljkovic, N. et al. The Fas Ligand/Fas Death Receptor Pathways Contribute to Propofol-Induced Apoptosis and Neuroinflammation in the Brain of Neonatal Rats. Neurotox Res 30, 434–452 (2016). https://doi.org/10.1007/s12640-016-9629-1

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Keywords

  • Propofol toxicity
  • FasL/Fas receptor
  • Bcl-2 gene family
  • Caspasa-1
  • IL-1β cytokine
  • Microglia activation