Targeted Inhibition of Leucine-Rich Repeat and Immunoglobulin Domain-Containing Protein 1 in Transplanted Neural Stem Cells Promotes Neuronal Differentiation and Functional Recovery in Rats Subjected to Spinal Cord Injury*
Targeted Inhibition of Leucine-Rich Repeat and Immunoglobulin Domain-Containing Protein 1 in Transplanted Neural Stem Cells Promotes Neuronal Differentiation and Functional Recovery in Rats Subjected to Spinal Cord Injury*
复制标题
移植神经干细胞中富含亮氨酸重复序列和含有免疫球蛋白结构域的蛋白 1 的靶向抑制可促进脊髓损伤大鼠的神经元分化和功能恢复。
DOI：
10.1097/ccm.0000000000001351
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发表时间：
2016-03-01
影响因子：
8.8
通讯作者：
Wan, Yong
中科院分区：
文献类型：
--
作者：
Chen, Ningning;Cen, Jing-sheng;Wan, Yong
目的：富含亮氨酸重复序列和含免疫球蛋白结构域的蛋白（LINGO）-1在神经干细胞中表达，其中和作用导致神经元持续不成熟。因此，通过RNA干扰靶向抑制LINGO-1可以提高移植的神经干细胞存活和体内神经元分化。此外，LINGO-1 RNA干扰神经干细胞是一种潜在的治疗策略脊髓损伤设计：中山大学附属第一医院脊柱外科设置：转化医学中心研究实验室，中山大学附属第一医院，受试者：雌性Sprague-Dawley大鼠干预：动物分为三组，进行椎板切除术和脊髓全横断伴随移植对照RNA干扰治疗或LINGO-1-RNA干扰治疗的神经干细胞在体内损伤部位。体外培养的神经干细胞分为对照组、对照RNA干扰慢病毒组、LINGO-1 RNA干扰慢病毒组、LINGO-1互补DNA慢病毒组和广东省自然科学基金重点项目组（编号S2013020012818）。测量和主要结果：通过免疫荧光和Western印迹分析，在体外和体内检查每个处理组中的神经干细胞的细胞存活和神经元分化。轴突再生和组织修复进行了评估，通过逆行追踪使用荧光金，电子显微镜，苏木精-伊红染色和MRI。还根据脊髓诱发电位测量和Basso-Beattie-Bresnahan评分检查大鼠的功能恢复。LINGO-1-RNA干扰处理的神经干细胞移植促进大鼠损伤脊髓的组织修复和功能恢复。类似地，LINGO-1 RNA干扰增加了体外神经干细胞存活和神经元分化。结论：LINGO-1-RNA干扰治疗的神经干细胞移植可促进脊髓损伤后的功能恢复，为脊髓损伤的修复提供了新的思路。
Objective:Leucine-rich repeat and immunoglobulin domain-containing protein (LINGO)-1 is expressed in neural stem cells, and its neutralization results in sustained neuronal immaturity. Thus, targeted inhibition of LINGO-1 via RNA interference may enhance transplanted neural stem cell survival and neuronal differentiation in vivo. Furthermore, LINGO-1 RNA interference in neural stem cells represents a potential therapeutic strategy for spinal cord injury.Design:Department of Spine Surgery, First Affiliated Hospital of Sun Yat-sen University.Setting:Translational Medicine Center Research Laboratory, First Affiliated Hospital of Sun Yat-sen University.Subjects:Female Sprague-Dawley rats.Interventions:The animals were divided into three groups that underwent laminectomy and complete spinal cord transection accompanied by transplantation of control-RNA interference-treated or LINGO-1-RNA interference-treated neural stem cells at the injured site in vivo. In vitro, neural stem cells were divided into four groups for the following treatments: control, control RNA interference lentivirus, LINGO-1 RNA interference lentivirus and LINGO-1 complementary DNA lentivirusand the Key Projects of the Natural Science Foundation of Guangdong Province (No. S2013020012818).Measurements and Main Results:Neural stem cells in each treatment group were examined for cell survival and neuronal differentiation in vitro and in vivo via immunofluorescence and Western blot analysis. Axonal regeneration and tissue repair were assessed via retrograde tracing using Fluorogold, electron microscopy, hematoxylin-eosin staining and MRI. Rats were also examined for functional recovery based on the measurement of spinal cord-evoked potentials and the Basso-Beattie-Bresnahan score. LINGO-1-RNA interference-treated neural stem cell transplantation increased tissue repair and functional recovery of the injured spinal cord in rats. Similarly, LINGO-1 RNA interference increased neural stem cell survival and neuronal differentiation in vitro. The mechanism underlying the effect of LINGO-1 RNA interference on the injured rat spinal cord may be that the significant inhibition of LINGO-1 expression in neural stem cells inactivated the RhoA and Notch signaling pathways, which act downstream of LINGO-1.Conclusions:Our findings indicate that transplantation of LINGO-1-RNA interference-treated neural stem cells facilitates functional recovery after spinal cord injury and represents a promising potential strategy for the repair of spinal cord injury.