Background:: Recent studies in various animal models have suggested that anesthetics such as propofol, when administered early in life, can lead to neurotoxicity. These studies have raised significant safety concerns regarding the use of anesthetics in the pediatric population and highlight the need for a better model to study anesthetic-induced neurotoxicity in humans. Human embryonic stem cells are capable of differentiating into any cell type and represent a promising model to study mechanisms governing anesthetic-induced neurotoxicity.
Methods:: Cell death in human embryonic stem cell–derived neurons was assessed using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate in situ nick end labeling staining, and microRNA expression was assessed using quantitative reverse transcription polymerase chain reaction. miR-21 was overexpressed and knocked down using an miR-21 mimic and antagomir, respectively. Sprouty 2 was knocked down using a small interfering RNA, and the expression of the miR-21 targets of interest was assessed by Western blot.
Results:: Propofol dose and exposure time dependently induced significant cell death (n = 3) in the neurons and down-regulated several microRNAs, including miR-21. Overexpression of miR-21 and knockdown of Sprouty 2 attenuated the increase in terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate in situ nick end labeling–positive cells following propofol exposure. In addition, miR-21 knockdown increased the number of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate in situ nick end labeling–positive cells by 30% (n = 5). Finally, activated signal transducer and activator of transcription 3 and protein kinase B (Akt) were down-regulated, and Sprouty 2 was up-regulated following propofol exposure (n = 3).
Conclusions:: These data suggest that (1) human embryonic stem cell–derived neurons represent a promising in vitro human model for studying anesthetic-induced neurotoxicity, (2) propofol induces cell death in human embryonic stem cell–derived neurons, and (3) the propofol-induced cell death may occur via a signal transducer and activator of transcription 3/miR-21/Sprouty 2–dependent mechanism.
Propofol induced apoptosis of human embryonic stem cell–derived neurons and reduced expression of miR-21. Overexpression of miR-21 reduced this toxicity. Neurons derived from human embryonic stem cell represent a useful model for the study of anesthetic neurotoxicity in humans. miR-21 plays a role in propofol-induced toxicity, and manipulation of miR-21 may serve as a therapeutic approach for prevention of toxicity.
Anesthetic neurotoxicity has been repeatedly demonstrated in rodent and primate models. A model for evaluation of toxicity in human tissue is not currently available.
Neurons were derived from human embryonic stem cells and were grown in culture. The effect of propofol exposure on cell death was examined. In addition, the role of miR-21, a noncoding microRNA that inhibits target mRNA expression, was evaluated.
Propofol induced apoptosis of human embryonic stem cell–derived neurons and reduced expression of miR-21. Overexpression of miR-21 reduced this toxicity.
Neurons derived from human embryonic stem cell represent a useful model for the study of anesthetic neurotoxicity in humans. miR-21 plays a role in propofol-induced toxicity, and manipulation of miR-21 may serve as a therapeutic approach for prevention of toxicity.
Danielle M. Twaroski, Yasheng Yan, Jessica M. Olson, Zeljko J. Bosnjak, Xiaowen Bai; Down-regulation of MicroRNA-21 Is Involved in the Propofol-induced Neurotoxicity Observed in Human Stem Cell–derived Neurons. Anesthesiology 2014;121(4):786-800. doi: https://doi.org/10.1097/ALN.0000000000000345.
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