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微小RNA-338与微小RNA-21共转染用于治疗大鼠坐骨神经损伤

MicroRNA-338 and microRNA-21 co-transfection for the treatment of rat sciatic nerve injury.

作者信息

Wang Jianyong, Muheremu Aikeremujiang, Zhang Ming, Gong Kai, Huang Chuyi, Ji Yuchen, Wei Yujun, Ao Qiang

机构信息

Institute of Clinical Medicine, Affiliated Hospital of Inner Mongolia University for Nationalities, No 92, Huolinhe Rd, Tongliao, 028000, Inner Mongolia, China.

Institute of Tissue Engineering, China Medical University, No. 77 Puhe Road, Shenyang North New Area, Shenyang, 110122, Liaoning, China.

出版信息

Neurol Sci. 2016 Jun;37(6):883-90. doi: 10.1007/s10072-016-2500-6. Epub 2016 Feb 24.

Abstract

The objective of this study is to find if co-transfecting microRNA-338 and microRNA-21 into the neurons in the spinal cord can promote functional recovery after peripheral nerve injury in rats. Animals were divided into three groups: 20 animals in the GFP control vector group (group A), 20 animals in the GFP experimental vector group (group B) and ten animals in the normal control group. Right sciatic nerves of animals in groups A and B were transected and were bridged with collagen nerve conduits with 10 mm distance between the stumps. 3 µl GFP control vector or 3 µl lentiviral vectors encoding the sequence of microRNA-338 and microRNA-21 were injected in the conduit. 8 weeks after the surgery, the treatment effect was evaluated by functional analysis, electrophysiological analysis, immunohistochemical analysis as well as transmitting electronic microscope observations in all the rats. Animals treated with microRNA-338 and microRNA-21 showed significantly better recovery than GFP control group animals by means of functional analysis (Sciatic nerve index -47.7 ± 2.5 vs -59.4 ± 3.7), electrophysiological analysis (Conduction velocity 20.5 ± 2.8 vs 10.5 ± 1.4 m/s), ratio of wet weight of the gastrocnemius muscles (0.83 ± 0.03 vs 0.55 ± 0.06), axon diameter (5.0 ± 1.8 µm vs 4.0 ± 2.2), myelin sheath thickness (1.4 ± 0.43 vs 0.80 ± 0.31 µm) and G-ratio (0.80 ± 0.06 vs 0.75 ± 0.04). Lentiviral vectors encoding microRNA 338 and 21 might be explored in the future as potential therapeutic intervention to promote nerve regeneration.

摘要

本研究的目的是探究将微小RNA - 338和微小RNA - 21共转染到大鼠脊髓神经元中是否能促进周围神经损伤后的功能恢复。将动物分为三组:绿色荧光蛋白(GFP)对照载体组20只动物(A组)、GFP实验载体组20只动物(B组)和正常对照组10只动物。A组和B组动物的右侧坐骨神经被切断,断端用胶原神经导管桥接,断端间距为10毫米。将3微升GFP对照载体或3微升编码微小RNA - 338和微小RNA - 21序列的慢病毒载体注入导管。术后8周,通过功能分析、电生理分析、免疫组织化学分析以及透射电子显微镜观察对所有大鼠的治疗效果进行评估。通过功能分析(坐骨神经指数 - 47.7 ± 2.5对 - 59.4 ± 3.7)、电生理分析(传导速度20.5 ± 2.8对10.5 ± 1.4米/秒)、腓肠肌湿重比(0.83 ± 0.03对0.55 ± 0.06)、轴突直径(5.0 ± 1.8微米对4.0 ± 2.2)、髓鞘厚度(1.4 ± 0.43对0.80 ± 0.31微米)和G比值(0.80 ± 0.06对0.75 ± 0.04),发现用微小RNA - 338和微小RNA - 21处理的动物比GFP对照组动物恢复得明显更好。编码微小RNA 338和21的慢病毒载体未来可能作为促进神经再生的潜在治疗干预手段进行探索。

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