Down-regulation of microRNA-21 is involved in the propofol-induced neurotoxicity observed in human stem cell-derived neurons.
Down-regulation of microRNA-21 is involved in the propofol-induced neurotoxicity observed in human stem cell-derived neurons.
复制标题
microRNA-21的下调参与人类干细胞衍生的神经元中观察到的丙泊酚诱导的神经毒性。
DOI：
10.1097/aln.0000000000000345
复制
发表时间：
2014-10
期刊：
影响因子：
8.8
通讯作者：
Bai X
中科院分区：
文献类型：
--
作者：
Twaroski DM;Yan Y;Olson JM;Bosnjak ZJ;Bai X
最近在各种动物模型中的研究表明，麻醉剂如丙泊酚在生命早期给药时可能导致神经毒性。这些研究提出了关于在儿科人群中使用麻醉剂的重大安全性问题，并强调需要一个更好的模型来研究人体麻醉诱导的神经毒性。人胚胎干细胞（hESC）能够分化成任何细胞类型，是研究麻醉诱导神经毒性机制的一个有前途的模型。使用TUNEL染色评估hESC衍生的神经元中的细胞死亡，并使用定量逆转录聚合酶链反应（qRTPCR）评估microRNA（miR）表达。分别使用miR-21模拟物和Escheromir过表达和敲低miR-21。使用小干扰RNA敲低Sprouty 2，并通过Western印迹评估目标miR-21靶标的表达。丙泊酚剂量和暴露时间依赖性地诱导神经元中的显著细胞死亡（n = 3），并下调几种microRNA，包括miR-21。miR-21的过表达和Sprouty 2的敲低减弱了丙泊酚暴露后TUNEL阳性细胞的增加。此外，miR-21敲低使TUNEL阳性细胞的数量增加了30%（n = 5）。最后，激活的信号转导和转录激活因子3（STAT 3）和蛋白激酶B（Akt）下调，Sprouty 2上调丙泊酚暴露后（n = 3）。这些数据表明：（1）hESC衍生的神经元代表了用于研究麻醉剂诱导的神经毒性的有前景的体外人模型，（2）丙泊酚诱导hESC衍生的神经元中的细胞死亡，以及（3）丙泊酚诱导的细胞死亡可能通过STAT 3/miR-21/Sprouty 2依赖性机制发生。
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 by which to study anesthetic-induced neurotoxicity in humans. Human embryonic stem cells (hESCs) are capable of differentiating into any cell type and represent a promising model to study mechanisms governing anesthetic-induced neurotoxicity. Cell death in hESC-derived neurons was assessed using TUNEL staining and microRNA (miR) expression was assessed using quantitative reverse transcription polymerase chain reaction (qRTPCR). miR-21 was overexpressed and knocked down using a 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. Propofol dose and exposure time-dependently induced significant cell death (n = 3) in the neurons and downregulated several microRNAs, including miR-21. Overexpression of miR-21 and knockdown of Sprouty 2 attenuated the increase in TUNEL-positive cells following propofol exposure. In addition, miR-21 knockdown increased the number of TUNEL-positive cells by 30% (n = 5). Finally, activated Signal Transducer and Activator of Transcription 3 (STAT3) and protein kinase B (Akt) were downregulated and Sprouty 2 was upregulated following propofol exposure (n = 3). These data suggest that: (1) hESC-derived neurons represent a promising in vitro human model for studying anesthetic-induced neurotoxicity, (2) propofol induces cell death in hESC-derived neurons and (3) the propofol-induced cell death may occur via a STAT3/miR-21/Sprouty2-dependent mechanism.