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miR-129-5p 对急性脊髓损伤大鼠的保护作用。

Protective Effects of MiR-129-5p on Acute Spinal Cord Injury Rats.

机构信息

Department of Orthopedics, Lishui Municipal Central Hospital, Lishui, Zhejiang, China (mainland).

出版信息

Med Sci Monit. 2019 Nov 4;25:8281-8288. doi: 10.12659/MSM.916731.

DOI:10.12659/MSM.916731
PMID:31680116
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6854883/
Abstract

BACKGROUND Spinal cord injury (SCI) is a severe devastating condition associated with serious disability and neurologic deficits. Aberrant micro RNA (miRNA) expression has been related to a variety of central nervous system diseases including SCI. In the present study, we aimed to discover the role of miR-129-5p on SCI. MATERIAL AND METHODS An acute SCI rat model was induced, following the modified Allen method. A total of 36 rats were randomly assigned into 4 groups (n=9 in every group): Sham group; Model group (SCI+saline); SCI+NC group; and SCI+miR-129-5p group (100 nm solution, every 2 days). Basso-Beattie-Bresnahan (BBB) locomotor rating score was carried out to determine functional recovery. TUNEL (terminal dUTP nick-end labeling) staining was used to evaluate cell apoptosis. Hematoxylin and eosin staining was performed to assess the pathological state of spinal cord. Furthermore, western blot assay was conducted to measure the calpain1 and calpain2 expression. RESULTS Our data suggested that the expression level of miR-129-5p was markedly reduced in rats after SCI. Then miR-129-5p mimic was injected into the vertebral canal. We found that the SCI+miR-129-5p group had a high score in the BBB test compared with the SCI+NC group and the Model group. The overexpression of miR-129-5p obviously reduced tissue loss, damaged cells, and the number of TUNEL positive cells. Moreover, western blot assay exhibited that overexpression of miR-129-5p decreased calpain1, calpain2, and cleaved caspase-3 expression. CONCLUSIONS Our findings suggested that overexpression of miR-129-5p improved neurological function by promoting functional recovery, reducing tissue loss and cell apoptosis in rats in an SCI model, possibly through downregulation of calpain1 and calpain2.

摘要

背景

脊髓损伤（SCI）是一种与严重残疾和神经功能缺陷相关的严重破坏性疾病。异常的 microRNA（miRNA）表达与多种中枢神经系统疾病有关，包括 SCI。在本研究中，我们旨在发现 miR-129-5p 在 SCI 中的作用。

材料和方法

采用改良 Allen 法诱导急性 SCI 大鼠模型。共 36 只大鼠随机分为 4 组（每组 n=9）：假手术组；模型组（SCI+生理盐水）；SCI+NC 组；和 SCI+miR-129-5p 组（100nm 溶液，每 2 天一次）。采用 Basso-Beattie-Bresnahan（BBB）运动评分标准评估功能恢复情况。TUNEL（末端 dUTP 缺口末端标记）染色评估细胞凋亡。苏木精和伊红染色评估脊髓病理状态。此外，还进行了 Western blot 检测以测量钙蛋白酶 1 和钙蛋白酶 2 的表达。

结果

我们的数据表明，SCI 后大鼠 miR-129-5p 的表达水平明显降低。然后将 miR-129-5p 模拟物注入椎管。我们发现，与 SCI+NC 组和模型组相比，SCI+miR-129-5p 组在 BBB 测试中的得分较高。miR-129-5p 的过表达明显减少了组织丢失、受损细胞和 TUNEL 阳性细胞的数量。此外，Western blot 检测显示，miR-129-5p 的过表达降低了钙蛋白酶 1、钙蛋白酶 2 和 cleaved caspase-3 的表达。

结论

我们的研究结果表明，miR-129-5p 的过表达通过促进功能恢复、减少组织丢失和细胞凋亡来改善 SCI 模型大鼠的神经功能，可能通过下调钙蛋白酶 1 和钙蛋白酶 2 来实现。

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