Department of Anesthesiology, The Affiliated Hospital of Beihua University, Jilin City, Jilin Province, 132000, China.
Department of Endocrinology, The Affiliated Hospital of Beihua University, Jilin City, Jilin Province, 132000, China.
Curr Neurovasc Res. 2020;17(2):140-146. doi: 10.2174/1567202617666200207124817.
Propofol is an intravenous drug commonly used in anesthesia procedures and intensive care in children. However, it also has neurotoxic effects on children. MicroRNA plays an important role in neurological diseases and neurotoxicity.
In this study, primary rat hippocampal neurons were used to investigate the role of miR- 582-5p in propofol-induced neurotoxicity. Cell viability was monitored by 3-(4,5-dimethylthiazolyl)- 2,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, while the expression of proteins was monitored by real-time quantitation polymerase chain reaction (RT-qPCR) and western blot. TargetScan and double luciferase report assay were used to predict the targeting relationship between miR-582-5p and Rho-associated serine-threonine protein kinase 1 (ROCK1).
In the present study, the viability of neurons and the expression of miR-582-5p were decreased in a time-dependent manner after propofol treatment. Besides, miR-582-5p overexpression significantly reduced the toxicity of propofol on neuron cells but had no significant effect on normal nerve cells. In addition, miR-582-5p overexpression significantly reversed the expression of apoptosis-related proteins (cleaved caspase 3 and cleaved caspase 9) induced by propofol but had no significant effect in normal nerve cells. TargetScan and Dual-luciferase report assay revealed that ROCK1 was a targeted regulatory gene for miR-582-5p, and propofol treatment up-regulated ROCK1 expression by inhibiting miR-582-5p expression. Notably, miR-582-5p overexpression significantly increased cell viability, while ROCK1 overexpression reversed the effect of miR-582- 5p.
Taken together, these findings suggest that miR-582-5p alleviated propofol-induced apoptosis of newborn rat neurons by inhibiting ROCK1.
丙泊酚是一种常用于儿童麻醉程序和重症监护的静脉内药物。然而，它也对儿童具有神经毒性作用。microRNA 在神经疾病和神经毒性中发挥重要作用。
在这项研究中，使用原代大鼠海马神经元来研究 miR-582-5p 在丙泊酚诱导的神经毒性中的作用。通过 3-(4,5-二甲基噻唑基)-2,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法监测细胞活力，而通过实时定量聚合酶链反应（RT-qPCR）和蛋白质印迹法监测蛋白质的表达。TargetScan 和双荧光素酶报告测定用于预测 miR-582-5p 和 Rho 相关丝氨酸-苏氨酸蛋白激酶 1（ROCK1）之间的靶向关系。
在本研究中，丙泊酚处理后神经元的活力和 miR-582-5p 的表达呈时间依赖性降低。此外，miR-582-5p 过表达显著降低了丙泊酚对神经元细胞的毒性，但对正常神经细胞没有显著影响。此外，miR-582-5p 过表达显著逆转了丙泊酚诱导的凋亡相关蛋白（cleaved caspase 3 和 cleaved caspase 9）的表达，但对正常神经细胞没有显著影响。TargetScan 和双荧光素酶报告测定显示，ROCK1 是 miR-582-5p 的靶向调节基因，丙泊酚通过抑制 miR-582-5p 的表达而上调 ROCK1 的表达。值得注意的是，miR-582-5p 过表达显著增加了细胞活力，而 ROCK1 过表达逆转了 miR-582-5p 的作用。
综上所述，这些发现表明，miR-582-5p 通过抑制 ROCK1 减轻了丙泊酚诱导的新生大鼠神经元凋亡。
