慢病毒介导的 CTGF 基因沉默抑制大鼠脊髓损伤后胶质瘢痕组织的形成。

Lentivirus-mediated silencing of the CTGF gene suppresses the formation of glial scar tissue in a rat model of spinal cord injury.

机构信息

Department of Orthopaedic, Shanghai Changzheng Hospital, No. 415, Fengyang Rd, Shanghai 200003, China.

出版信息

Spine J. 2018 Jan;18(1):164-172. doi: 10.1016/j.spinee.2017.01.004. Epub 2017 Jan 9.

DOI:10.1016/j.spinee.2017.01.004

PMID:28089819

Abstract

BACKGROUND CONTEXT

One of the many reactive changes following a spinal cord injury (SCI) is the formation of a glial scar, a reactive cellular process whereby glial cells accumulate and surround the central nervous system injury sites to seal in the wound. Thus, the inhibition of glial scar is of great importance for SCI recovery.

PURPOSE

This study aimed to explore the effect of lentivirus-mediated silencing of the CTGF gene on the formation of glial scar tissue in a rat model of SCI.

STUDY DESIGN

This is a prospective study.

STUDY SAMPLE

A total of 56 Wistar female rats aged 8 weeks were randomly selected for this study.

OUTCOME MEASURES

The motor function of the rats was assessed using the Basso, Beattie, and Bresnahan (BBB) functional scale, footprint analysis of gait, and the Basso Mouse Scale (BMS). Quantitative real-time polymerase chain reaction, Western blotting, and immunohistochemistry were performed to detect the mRNA and protein expressions of glial fibrillary acidic protein (GFAP), vimentin, fibronectin, and laminin in the spinal cord tissues.

METHODS

A rat model of SCI was successfully established. Fifty-six male Wistar rats were randomly selected and assigned into four groups (14 rats in each group): the sham operation group, the SCI model group, the negative control (NC) group (SCI rats transfected with empty vector plasmids), and the siRNA-CTGF group (SCI rats transfected with lentivirus CTGF siRNA).

RESULTS

The SCI rats showed decreased activity and were dragging their bodies while moving. Compared with the sham operation group, the BBB and BMS scores in the SCI model, NC, and siRNA-CTGF groups significantly decreased. However, the BBB and BMS scores in the siRNA-CTGF group were higher than those in the SCI model and NC groups. The mRNA and protein expressions of GFAP, vimentin, fibronectin, and laminin significantly increased in the SCI model, NC, and siRNA-CTGF groups in comparison with those in the sham operation group. Furthermore, the mRNA and protein expressions of GFAP, vimentin, fibronectin, and laminin in the siRNA-CTGF group were lower than those in the SCI model and NC groups 28 days after transfection.

CONCLUSIONS

These findings indicate that lentivirus-mediated silencing of the CTGF gene can suppress the formation of glial scar tissue after SCI.

摘要

背景

脊髓损伤（SCI）后会发生许多反应性变化，其中之一是胶质瘢痕的形成，这是一种反应性细胞过程，其中胶质细胞聚集并包围中枢神经系统损伤部位以封闭伤口。因此，抑制胶质瘢痕的形成对于 SCI 的恢复非常重要。

目的

本研究旨在探讨慢病毒介导的 CTGF 基因沉默对 SCI 大鼠模型中胶质瘢痕组织形成的影响。

研究设计

这是一项前瞻性研究。

研究样本

本研究共纳入 56 只 8 周龄 Wistar 雌性大鼠。

观察指标

采用 Basso、Beattie 和 Bresnahan（BBB）功能评分、步态足迹分析和 Basso 小鼠评分（BMS）评估大鼠的运动功能。通过定量实时聚合酶链反应、Western 印迹和免疫组织化学检测脊髓组织中神经胶质纤维酸性蛋白（GFAP）、波形蛋白、纤维连接蛋白和层粘连蛋白的 mRNA 和蛋白表达。

方法

成功建立了 SCI 大鼠模型。随机选择 56 只雄性 Wistar 大鼠，并将其分为四组（每组 14 只）：假手术组、SCI 模型组、阴性对照组（SCI 大鼠转染空载体质粒）和 siRNA-CTGF 组（SCI 大鼠转染慢病毒 CTGF siRNA）。

结果

SCI 大鼠活动减少，移动时拖着身体。与假手术组相比，SCI 模型组、NC 组和 siRNA-CTGF 组的 BBB 和 BMS 评分明显降低。然而，siRNA-CTGF 组的 BBB 和 BMS 评分高于 SCI 模型组和 NC 组。与假手术组相比，SCI 模型组、NC 组和 siRNA-CTGF 组中 GFAP、波形蛋白、纤维连接蛋白和层粘连蛋白的 mRNA 和蛋白表达显著增加。此外，转染后 28 天，siRNA-CTGF 组中 GFAP、波形蛋白、纤维连接蛋白和层粘连蛋白的 mRNA 和蛋白表达低于 SCI 模型组和 NC 组。