颅脑损伤后细胞骨架蛋白羰基化致轴索损伤研究  

Axonal injury induced by carbonylation of cytoskeletal brain proteins in rats after traumatic brain injury

作　　者：张秋生[1] 张猛[1] 黄贤键[1] 刘晓佳[1] 李维平[1] 

机构地区：[1]深圳市第二人民医院神经外科,518035

出　　处：《中华神经医学杂志》2015年第5期469-472,共4页Chinese Journal of Neuromedicine

基　　金：深圳市协同创新计划国际科技合作项目（GJHZ20120614154914623）;深圳市神经外科学重点实验室（ZDSYS20140509173142601）

摘　　要：目的 研究不同程度颅脑损伤（TBI）后脑组织氧化应激及细胞骨架蛋白羰基化水平变化及意义.方法 SD成年大鼠15只按随机数字表法分为轻度TBI组（n=5）、重度TBI组（n=5）和假手术组（n=5）,应用液压颅脑损伤仪根据相应参数制备轻度、重度及假手术组动物模型.应用酶联免疫吸附法和Western blotting检测TBI后24 h大鼠脑组织中还原型谷胱甘肽（GSH）、丙二醛（MDA）及细胞骨架蛋白[β-肌动蛋白、β-微管蛋白和胶质纤维酸性蛋白（GFAP）]羰基化水平,并应用Western blotting检测轴索损伤标志物磷酸化tau（p-tau）蛋白水平.结果 轻度、重度TBI组大鼠脑组织中MDA水平分别为（389.62±29.95） μmol/g、（642.50±37.56） μmol/g,较假手术组[（233.94±25.08） μmol/g]显著升高,差异均有统计学意义（P＜0.05）;GSH水平分别为（352.10±37.75） μmol/g、（153.27±43.49） μmol/g,较假手术组[（492.48±41.43） μmol/g]显著降低,差异均有统计学意义（P＜0.05）;β-肌动蛋白、β-微管蛋白和GFAP在轻度TBI组羰基化比例分别为0.099±0.104、0.194±0.114、0.643±0.037,重度TBI组分别为0.142±0.017、0.290±0.029、0.902±0.021,较假手术组（0.068±0.017、0.108±0.016、0.673±0.032）明显升高,差异均有统计学意义（P＜ 0.05）;p-tau比例在轻度、重度TBI组分别为0.289±0.014、0.373±0.012,较假手术组（0.185±0.009）差异均有统计学意义（P＜0.05）.结论 TBI后脑组织氧化应激及细胞骨架蛋白羰基化水平随着TBI程度的增加而增加,可造成神经轴索运输障碍而导致轴索损伤加重.Objective To investigate the oxidative stress and cytoskeleton protein carbonylation in rat brains after different severities of traumatic brain injury （TBI）.Methods Fluid percussion percussion device was used to establish the mild,severe and sham-operated Sprague-Dawley rat models （n=15）;24 h after that,enzyme linked immunosorbent assay was used to detect the levels of malondialdehyde （MDA） and glutathione （GSH）,and Western blotting was employed to detect the cytoskeletal proteins （β-actin,β-tublin and glial fibrillary acidic [GFAP]） carbonylation levels;phosphorylated tau （p-tau） protein expressions were examined by Western blotting.Results The expression levels of MDA in mild TBI group and severe TBI group were （389.62±29.95） μmol/g and （642.50±37.56） μmol/g,respectively,which was significantly increased as compared with the MDA level （[233.94±25.08] μmol/g） in sham-operated group （P〈0.05）.The expression level of GSH in mild TBI group and severe TBI group was （352.10±37.75） μmol/g and （153.27 ±43.49） μmol/g,respectively,which was significantly decreased as compared with the GSH level （[492.48 ±41.43] μmol/g） in sham-operated group （P〈0.05）.The β-actin,β-tublin and GFAP proteincarbonylation levels （0.099± 0.104,0.194±0.114 and 0.643±0.037;0.142±0.017,0.290±0.029 and 0.902±0.021） and p-tau level （0.289±0.014 and 0.373±0.012） in mild TBI group and severe TBI group were significantly higher than those in sham-operated group （0.068±0.017,0.108±0.016 and 0.673±0.032;0.185±0.009;P〈0.05）.Conclusions The oxidative stress and carbonylation of cytoskeleton proteins are significantly increased after TBI,and the expression levels are correlated with the severity of TBI.The carbonylation of cytoskeleton protein aggravates the axonal injury after TBI.

关 键 词：颅脑损伤 氧化应激 细胞骨架蛋白 羰基化 

分 类 号：R651.15[医药卫生—外科学]