Transplantation of sh-miR-199a-5p-Modified Olfactory Ensheathing Cells Promotes the Functional Recovery in Rats with Contusive Spinal Cord Injury.
Transplantation of sh-miR-199a-5p-Modified Olfactory Ensheathing Cells Promotes the Functional Recovery in Rats with Contusive Spinal Cord Injury.
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sh-miR-199a-5p修饰的嗅鞘细胞移植促进挫伤性脊髓损伤大鼠的功能恢复
DOI：
10.1177/0963689720916173
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发表时间：
2020-01
影响因子：
3.3
通讯作者：
Chang S
中科院分区：
文献类型：
--
作者：
Gao Z;Zhao Y;He X;Leng Z;Zhou X;Song H;Wang R;Gao Z;Wang Y;Liu J;Niu B;Li H;Ouyang P;Chang S
微小RNA（microRNAs，miRNAs）作为基因表达开关，参与脊髓损伤（spinal cord injury，SCI）的多种病理生理过程。嗅鞘细胞（OECs）可以减轻脊髓损伤后的病理损伤，促进功能恢复。然而，OECs恢复功能的机制还不清楚。本研究旨在确定沉默miR-199 a-5 p是否会增强OEC的有益作用。在这项研究中，我们测量了有和没有损伤的大鼠脊髓中的miR-199 a-5 p水平，有和没有OEC移植。然后，我们用sh-miR-199 a-5 p慢病毒载体转染OECs以降低miR-199 a-5 p表达，并通过Basso-Beattie-Bresnahan（BBB）运动评分、扩散张量成像（DTI）和组织学方法确定这些OECs在SCI大鼠中的作用。我们使用蛋白质印迹法测量Slit 1、Robo 2和srGAP 2的蛋白质水平。最后，我们使用双荧光素酶报告基因检测来评估miR-199- 5 p与Slit 1、Robo 2和srGAP 2表达之间的关系。我们发现SCI显著增加了miR-199 a-5 p的表达（P < 0.05），而OEC移植显著降低了miR-199 a-5 p的表达（P < 0.05）。OECs中miR-199 a-5 p的敲低对SCI大鼠具有更好的治疗效果，这通过更高的BBB评分和DTI的各向异性分数值以及组织学发现来指示。降低移植OECs中miR-199 a-5 p水平显著增加了Slit 1，Robo 2和srGAP 2的脊髓蛋白水平。我们的研究结果表明，移植sh-miR-199 a-5 p修饰的OEC促进了SCI大鼠的功能恢复，表明miR-199 a-5 p敲低更有利于OEC移植的治疗效果。这些发现为miRNAs介导的OECs治疗机制提供了新的见解，有助于我们开发基于miRNAs的治疗策略，优化SCI的细胞治疗。
MicroRNAs (miRNAs) function as gene expression switches, and participate in diverse pathophysiological processes of spinal cord injury (SCI). Olfactory ensheathing cells (OECs) can alleviate pathological injury and facilitate functional recovery after SCI. However, the mechanisms by which OECs restore function are not well understood. This study aims to determine whether silencing miR-199a-5p would enhance the beneficial effects of the OECs. In this study, we measured miR-199a-5p levels in rat spinal cords with and without injury, with and without OEC transplants. Then, we transfected OECs with the sh-miR-199a-5p lentiviral vector to reduce miR-199a-5p expression and determined the effects of these OECs in SCI rats by Basso–Beattie–Bresnahan (BBB) locomotor scores, diffusion tensor imaging (DTI), and histological methods. We used western blotting to measure protein levels of Slit1, Robo2, and srGAP2. Finally, we used the dual-luciferase reporter assay to assess the relationship between miR-199-5p and Slit1, Robo2, and srGAP2 expression. We found that SCI significantly increased miR-199a-5p levels (P < 0.05), and OEC transplants significantly reduced miR-199a-5p expression (P < 0.05). Knockdown of miR-199a-5p in OECs had a better therapeutic effect on SCI rats, indicated by higher BBB scores and fractional anisotropy values on DTI, as well as histological findings. Reducing miR-199a-5p levels in transplanted OECs markedly increased spinal cord protein levels of Slit1, Robo2, and srGAP2. Our results demonstrated that transplantation of sh-miR-199a-5p-modified OECs promoted functional recovery in SCI rats, suggesting that miR-199a-5p knockdown was more beneficial to the therapeutic effects of OEC transplants. These findings provided new insights into miRNAs-mediated therapeutic mechanisms of OECs, which helps us to develop therapeutic strategies based on miRNAs and optimize cell therapy for SCI.
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作者：
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通讯作者：
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影响因子：
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