Xie Guang-Bin, Wang Chun-Xi, Zhou Chen-Hui, Li Hua, Zhang Xiang-Sheng, Zhou Xiao-Ming, Zhang Li, Hang Chun-Hua, Zhou Meng-Liang, Shi Ji-Xin
Department of Neurosurgery, Jinling Hospital, School of Medicine, Nanjing University, 305 East Zhongshan Road, Nanjing, 210002, Jiangsu Province, China.
Cell Mol Neurobiol. 2015 Jul;35(5):723-31. doi: 10.1007/s10571-015-0168-8. Epub 2015 Mar 6.
Convincing evidence indicates that apoptosis contributes to the unfavorable prognosis of subarachnoid hemorrhage (SAH), a significant cause of morbidity and case fatality throughout the world. Gelsolin (GSN) is a Ca(2+)-dependent actin filament severing, capping, and nucleating protein, as well as multifunctional regulator of cell structure and metabolism, including apoptosis. In the present study, we intended to investigate the expression pattern and cell distribution of GSN in rat brain after experimental SAH. GSN expression was examined in sham group and at 3, 6, 12 h, day 1 (1 day), 2, 3, 5, and 7 days after SAH by Western blot analysis as well as real-time polymerase chain reaction. Immunohistochemistry and immunofluorescence were performed to detect the localization of GSN. The level of GSN protein expression was significantly decreased in SAH group and reached a bottoming point on 1 day after SAH. GSN mRNA level was significantly decreased in SAH groups in comparison with the sham group, and reached a minimum value at 12 h after SAH. Immunohistochemistry showed that GSN was constitutively and obviously expressed in the cortex of the normal rat brain and significantly decreased in the rat cortex after SAH. In addition, immunofluorescence results revealed that GSN expression could be found in both neurons and microglias, as well as in glialfibrillary acidic protein-positive astrocytes. The decreased expression of GSN could mainly be found in neurons and astrocytes as well, and GSN-positive microglias showed different cell morphological characteristics. Interestingly, the protein and gene levels of GSN seemed to be constant in the rat hippocampus of sham and SAH groups. These findings suggested a potential role of GSN in the pathophysiology of the brain at the early stage of SAH.
有说服力的证据表明,细胞凋亡导致蛛网膜下腔出血(SAH)预后不良,SAH是全球发病和病死率的一个重要原因。凝溶胶蛋白(GSN)是一种依赖钙离子的肌动蛋白丝切断、封端和成核蛋白,也是细胞结构和代谢(包括细胞凋亡)的多功能调节因子。在本研究中,我们旨在探讨实验性SAH后大鼠脑内GSN的表达模式和细胞分布。通过蛋白质免疫印迹分析以及实时聚合酶链反应,检测假手术组和SAH后3、6、12小时、1天、2、3、5和7天的GSN表达。采用免疫组织化学和免疫荧光法检测GSN的定位。SAH组GSN蛋白表达水平显著降低,在SAH后1天降至最低点。与假手术组相比,SAH组GSN mRNA水平显著降低,在SAH后12小时达到最小值。免疫组织化学显示,GSN在正常大鼠脑皮质中持续且明显表达,SAH后大鼠皮质中显著降低。此外,免疫荧光结果显示,GSN表达可见于神经元和小胶质细胞,以及胶质纤维酸性蛋白阳性星形胶质细胞。GSN表达降低也主要见于神经元和星形胶质细胞,GSN阳性小胶质细胞表现出不同的细胞形态特征。有趣的是,假手术组和SAH组大鼠海马中GSN的蛋白和基因水平似乎保持不变。这些发现提示GSN在SAH早期脑病理生理学中可能发挥作用。