Department of Neurology, The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen, China.
Department of Neurosurgery, First Affiliated Hospital of Guizhou Medical University, Guiyang, China.
J Cell Biochem. 2019 Jan;120(1):368-379. doi: 10.1002/jcb.27390. Epub 2018 Sep 23.
Traumatic brain injury (TBI), an acute degenerative pathology of the central nervous system, is a leading cause of death and disability. As the glial scar is a mechanical barrier to nerve regeneration, inhibitory molecules in the forming scar and methods to overcome them have suggested molecular modification strategies to allow neuronal growth and functional regeneration. Herein, we aim to investigate the effects of aquaporin-4 (AQP4) gene silencing on the glial scar formation after TBI by establishing rat models. After modeling, TBI rats were transfected with AQP4 small hairpin RNA [shRNA] (AQP4 gene silencing by lentiviral vector-delivered shRNA) and empty vectors, respectively. Neurological functions of the rats were evaluated after TBI. The hematoxylin and eosin staining was conducted to observe histomorphological changes in rat brain tissues. The messenger RNA (mRNA) and protein expressions of glial fibrillary acidic protein (GFAP), vimentin, fibronectin, laminin, and AQP4 were measured by reverse transcription-quantitative polymerase chain reaction and Western blot analysis. The ratio of positive expression area was calculated, and the glial scar was observed by immunohistochemistry. At the 7th, 14th, and 28th days after TBI, TBI rats treated with AQP4 shRNA showed improved neurological function and lessened histomorphological changes. AQP4 gene silencing mediated by lentivirus decreased the mRNA and protein expressions of GFAP, vimentin, fibronectin, and laminin, the number of positive cells, the ratio of positive expression area, and the glial scar. Our study demonstrates that lentivirus-mediated AQP4 gene silencing could inhibit the formation of glial scar after TBI, which is beneficial to the recovery of neurological function.
创伤性脑损伤（TBI）是一种中枢神经系统的急性退行性病变，是死亡和残疾的主要原因。由于神经胶质瘢痕是神经再生的机械屏障，因此形成瘢痕中的抑制性分子及其克服方法提示了允许神经元生长和功能再生的分子修饰策略。在此，我们旨在通过建立大鼠模型来研究水通道蛋白 4（AQP4）基因沉默对 TBI 后神经胶质瘢痕形成的影响。建模后，TBI 大鼠分别转染 AQP4 短发夹 RNA（shRNA）（通过慢病毒载体递送的 shRNA 沉默 AQP4 基因）和空载体。TBI 后评估大鼠的神经功能。进行苏木精和伊红染色以观察大鼠脑组织的组织形态学变化。通过逆转录-定量聚合酶链反应和 Western blot 分析测量神经胶质纤维酸性蛋白（GFAP）、波形蛋白、纤维连接蛋白、层粘连蛋白和 AQP4 的信使 RNA（mRNA）和蛋白表达。计算阳性表达面积的比值，并通过免疫组织化学观察神经胶质瘢痕。在 TBI 后第 7、14 和 28 天，用 AQP4 shRNA 处理的 TBI 大鼠表现出改善的神经功能和减轻的组织形态学变化。慢病毒介导的 AQP4 基因沉默降低了 GFAP、波形蛋白、纤维连接蛋白和层粘连蛋白的 mRNA 和蛋白表达、阳性细胞数、阳性表达面积的比值和神经胶质瘢痕。我们的研究表明，慢病毒介导的 AQP4 基因沉默可以抑制 TBI 后神经胶质瘢痕的形成，这有利于神经功能的恢复。
