Faculty Collaboration Database

Down-regulation of microRNA-21 is involved in the propofol-induced neurotoxicity observed in human stem cell-derived neurons. Anesthesiology 2014 Oct;121(4):786-800

Date

06/21/2014

Pubmed ID

24950164

Pubmed Central ID

PMC4174986

DOI

10.1097/ALN.0000000000000345

Scopus ID

2-s2.0-84914165939 (requires institutional sign-in at Scopus site)   75 Citations

Abstract

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.

Author List

Twaroski DM, Yan Y, Olson JM, Bosnjak ZJ, Bai X

Authors

Xiaowen Bai PhD Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of Wisconsin
Jessica Olson PhD Director, Associate Professor in the Institute for Health and Humanity department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Anesthetics, Intravenous
Animals
Cell Differentiation
Coculture Techniques
Dose-Response Relationship, Drug
Down-Regulation
Humans
Mice
MicroRNAs
Neural Stem Cells
Neurons
Propofol