Zheng Ma, Cui-Yun Li, Li-Juan Wang, Yan Xia, Cheng-An Feng, Yu-Fang Peng, Yan-Bing Han, Yan Fan, Ying-Chun Ba
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引用次数: 0
Abstract
Introduction: Dysregulation of spinal cord development can lead to serious neuronal damage and dysfunction, causing significant health problems in newborns. MiRNA-138 appears to be crucial for proliferation, differentiation, and apoptosis of cells. However, the regulation of miRNA-138 and downstream molecules in embryonic spinal cord development remain elusive. The aim of this experiment is to determine whether overexpression of miRNA-138 or RNA interference (RNAi) can regulate the development of spinal cord in fetal rats.
Methods: Two plasmid vectors including pLenti-III-mico-GFP (miRNA-138 open reading frame [ORF]) and pLenti-III-miR-Off (miRNA-138 short hairpin) were constructed and injected into the tail vein of rats on the 14th day of pregnancy. Hematoxylin-eosin (HE) staining was used to observe the cell morphology. QRT-PCR, Western blot, and immunostaining confirmed the regulatory relationship between miRNA-138 and downstream molecules sonic hedgehog (Shh).
Results: Overexpression of miRNA-138 increased neuron regeneration significantly and decreased neuronal apoptosis when compared with the control. Silencing of miRNA-138 increased neuronal apoptosis and spinal cord atrophy significantly. Furthermore, miRNA-138 ORF treatment effectively increased the expression level of miRNA-138 and also upregulated the level of Shh. Comparatively, knockdown of miRNA-138 downregulated Shh levels in myelodysplastic regions.
Conclusion: These findings indicated that miRNA-138 overexpression could protect the spinal cord development of fetal rats, and the underlying mechanisms were associated with Shh expression. The present study provides a novel strategy to promote the molecular mechanism of embryonic spinal cord development.
脊髓发育失调可导致严重的神经元损伤和功能障碍，对新生儿造成严重的健康问题。MiRNA-138似乎对细胞的增殖、分化和凋亡至关重要。然而，miRNA-138及其下游分子在胚胎脊髓发育中的调控作用尚不明确。本实验旨在确定miRNA-138过表达或RNA干扰(RNAi)是否可以调节胎鼠脊髓的发育。方法:构建mgm - iii - micro - gfp (miRNA-138开放阅读框[ORF])和mgm - iii - mir - off (miRNA-138短发夹)两个质粒载体，于妊娠第14天注入大鼠尾静脉。苏木精-伊红(HE)染色观察细胞形态。QRT-PCR、Western blot和免疫染色证实了miRNA-138与下游分子sonic hedgehog (Shh)之间的调控关系。结果:与对照组相比，过表达miRNA-138可显著促进神经元再生，减少神经元凋亡。miRNA-138的沉默显著增加了神经元凋亡和脊髓萎缩。此外，miRNA-138 ORF处理有效地增加了miRNA-138的表达水平，也上调了Shh的水平。相比之下，miRNA-138的敲低下调了骨髓增生异常区域的Shh水平。结论:miRNA-138过表达对胎鼠脊髓发育具有保护作用，其机制可能与Shh表达有关。本研究为促进胚胎脊髓发育的分子机制提供了一种新的策略。
期刊介绍：
Articles in ''Pediatric Neurosurgery'' strives to publish new information and observations in pediatric neurosurgery and the allied fields of neurology, neuroradiology and neuropathology as they relate to the etiology of neurologic diseases and the operative care of affected patients. In addition to experimental and clinical studies, the journal presents critical reviews which provide the reader with an update on selected topics as well as case histories and reports on advances in methodology and technique. This thought-provoking focus encourages dissemination of information from neurosurgeons and neuroscientists around the world that will be of interest to clinicians and researchers concerned with pediatric, congenital, and developmental diseases of the nervous system.
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