Cui Zhiming, Zhang Jinlong, Bao Guofeng, Xu Guanhua, Sun Yuyu, Wang Lingling, Chen Jiajia, Jin Huricha, Liu Jian, Yang Longfei, Feng Guijuan, Li Weidong
Department of Spine Surgery, The Second Affiliated Hospital of Nantong University, Nantong University, 226001, Nantong, Jiangsu, People's Republic of China,
J Mol Neurosci. 2014;54(2):252-63. doi: 10.1007/s12031-014-0282-y. Epub 2014 Mar 26.
Hypoxia and other adverse conditions are usually encountered by rapidly growing cells. The RNA-binding motif protein 3 (RBM3) is induced by low temperature and hypoxia. However, its expression and function in spinal cord injury are still unclear. To investigate the certain expression and biological function in the central nervous system, we performed an acute spinal cord contusion injury (SCI) model in adult rats. Western blot analysis indicated a striking expression upregulation of RBM3 after spinal cord injury (SCI). Double immunofluorescence staining prompted that RBM3 immunoreactivity was found in astrocytes and neurons. Interestingly, RBM3 expression was increased predominantly in astrocytes. Furthermore, colocalization of RBM3 with proliferating cell nuclear antigen (PCNA) was detected in astrocytes. To further understand whether RBM3 plays a role in astrocyte proliferation, we applied lipopolysaccharide (LPS) to induce astrocyte proliferation in vitro. Western blot analysis demonstrated that RBM3 expression was positively correlated with PCNA expression following LPS stimulation. Immunofluorescence analysis showed that the expression of RBM3 was also changed following the stimulation of astrocytes with LPS, which was parallel with the data in vivo. Additionally, knocking RBM3 down with small interfering RNA (siRNA) demonstrated that RBM3 might play a significant role in the proliferation of astrocytes treated by hypoxia in vitro. These results suggest that RBM3 may be involved in the proliferation of astrocytes after SCI. To summarize, we firstly uncover the temporal and spatial expression changes of RBM3 in spinal cord injury. Our data suggest that RBM3 might be implicated in central nervous system pathophysiology after SCI.
快速生长的细胞通常会遭遇缺氧及其他不利状况。RNA结合基序蛋白3(RBM3)可由低温和缺氧诱导产生。然而,其在脊髓损伤中的表达及功能仍不明确。为研究其在中枢神经系统中的具体表达及生物学功能,我们在成年大鼠中建立了急性脊髓挫伤损伤(SCI)模型。蛋白质免疫印迹分析表明,脊髓损伤(SCI)后RBM3的表达显著上调。双重免疫荧光染色显示,在星形胶质细胞和神经元中均发现了RBM3免疫反应性。有趣的是,RBM3的表达主要在星形胶质细胞中增加。此外,在星形胶质细胞中检测到RBM3与增殖细胞核抗原(PCNA)的共定位。为进一步了解RBM3是否在星形胶质细胞增殖中发挥作用,我们应用脂多糖(LPS)在体外诱导星形胶质细胞增殖。蛋白质免疫印迹分析表明,LPS刺激后RBM3的表达与PCNA的表达呈正相关。免疫荧光分析显示,用LPS刺激星形胶质细胞后,RBM3的表达也发生了变化,这与体内数据一致。此外,用小干扰RNA(siRNA)敲低RBM3表明,RBM3可能在体外缺氧处理的星形胶质细胞增殖中发挥重要作用。这些结果表明,RBM3可能参与了脊髓损伤后星形胶质细胞的增殖。总之,我们首次揭示了脊髓损伤中RBM3的时空表达变化。我们的数据表明,RBM3可能与脊髓损伤后中枢神经系统的病理生理过程有关。