Li Yu, Jia Changxin, Zhang Dianlong, Ni Guangzhen, Miao Xia, Tu Ruirong
Department of Anesthesia, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
Department of Clinical Laboratory, People's Hospital of Weifang, Weifang, Shandong, China.
Cancer Biomark. 2017 Aug 23;20(2):175-181. doi: 10.3233/CBM-170167.
Studies in developing animals have demonstrated that when anesthetic agents, such as propofol, are early administered in life, it can lead to neuronal cell death and learning disabilities. However, the mechanisms causing these effects remains unknown. A recent report found that propofol could significantly upregulat miR-206 expression in the human ASCs. miR-206 could also induce apoptosis in human malignant cancers. Therefore, in this study, we hypothesized that propofol induces neurotoxicity in human embryonic stem cells (hESCs).
hESCs were exposed to propofol (50 μM) for 6 hr and cell death was assessed using TUNEL staining, and cleaved caspase-3 expression. miR-206 was knocked down using antagomir. PUMA was knocked down using a small interfering RNA. microRNA-206 (miR-206) expression was assessed using quantitative reverse transcription polymerase chain reaction (qRT-PCR). PUMA protein expression was detected using western blot assay.
hESCs exposed to propofol showed a significant increase in TUNEL positive cells and cleaved caspase-3 expression, followed by the upregulation of miR-206 and PUMA expression. Targeting PUMA inhibits propofol-induced cell apoptosis; miR-206 knockdown decreased propofol-induced cell apoptosis, cleaved caspase-3 and PUMA expression.
Propofol induce s cell death in hESC-derived neurons via activation of miR-206/PUMA signal pathway.
对发育中动物的研究表明，当在生命早期给予麻醉剂（如丙泊酚）时，可导致神经元细胞死亡和学习障碍。然而，导致这些影响的机制尚不清楚。最近的一份报告发现，丙泊酚可显著上调人脂肪干细胞中miR-206的表达。miR-206也可诱导人类恶性肿瘤细胞凋亡。因此，在本研究中，我们假设丙泊酚可诱导人胚胎干细胞（hESC）产生神经毒性。
将hESC暴露于丙泊酚（50μM）6小时，使用TUNEL染色和裂解的半胱天冬酶-3表达评估细胞死亡情况。使用抗miR抑制miR-206。使用小干扰RNA抑制PUMA。使用定量逆转录聚合酶链反应（qRT-PCR）评估微小RNA-206（miR-206）的表达。使用蛋白质免疫印迹法检测PUMA蛋白表达。
暴露于丙泊酚的hESC显示TUNEL阳性细胞和裂解的半胱天冬酶-3表达显著增加，随后miR-206和PUMA表达上调。靶向PUMA可抑制丙泊酚诱导的细胞凋亡；抑制miR-206可减少丙泊酚诱导的细胞凋亡、裂解的半胱天冬酶-3和PUMA表达。
丙泊酚通过激活miR-206/PUMA信号通路诱导hESC来源的神经元细胞死亡。
