[1]吴精川,李辉,何骏驰,等.创伤性脑损伤后大鼠脑组织miRNA-9表达变化及意义[J].第三军医大学学报,2017,39(14):1458-1463.
 WU Jingchuan,LI Hui,HE Junchi,et al.Expression of miRNA-9 in brain tissue and its role in promoting cerebral vascular remodeling in rats following traumatic brain injury[J].J Third Mil Med Univ,2017,39(14):1458-1463.
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创伤性脑损伤后大鼠脑组织miRNA-9表达变化及意义(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
39卷
期数:
2017年第14期
页码:
1458-1463
栏目:
基础医学
出版日期:
2017-07-30

文章信息/Info

Title:
Expression of miRNA-9 in brain tissue and its role in promoting cerebral vascular remodeling in rats following traumatic brain injury
作者:
吴精川李辉何骏驰钟建军张洪荣黄志坚孙晓川
重庆医科大学附属第一医院神经外科
Author(s):
WU Jingchuan LI Hui HE Junchi ZHONG Jianjun ZHANG Hongrong HUANG Zhijian SUN Xiaochuan 

Department of Neurosurgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
创伤性脑损伤miRNA内皮细胞血管重塑
Keywords:
traumatic brain injurymicroRNAendotheliumvascular remodeling
分类号:
R394.3;R446.8;R651.15
文献标志码:
A
摘要:

目的观察创伤性脑损伤(traumatic brain injury,TBI)后大鼠损伤皮质区miRNA9的表达变化规律,探讨其对脑微血管内皮细胞的保护作用。方法选取成年雄性SD大鼠60只制作控制性皮质撞击损伤模型(controlled cortical impact,CCI),分别应用荧光定量PCR和Western blot检测伤后6 h及1、3、7、14、28 d各时间点伤灶周围组织中miRNA9及CD31的表达情况。培养原代大鼠脑微血管内皮细胞,将内皮细胞分为正常组、损伤模型组[用依托泊苷(etoposide,ETO)损伤]、miRNA9过表达损伤模型组、空转染损伤模型组,应用CCK8检测各组细胞活力;应用Western blot检测各组B细胞淋巴瘤/白血病2蛋白(Bcell lymphoma2,Bcl2)、B细胞淋巴瘤/白血病2相关X蛋白(Bcl2 associated X,Bax)、活化半胱氨酸天冬氨酸特异性蛋白酶3蛋白(cleaved cysteinyl aspartate specific proteinase 3,clcaspase3)表达水平。结果(1)脑创伤后伤灶周围区域miRNA9表达明显增加,于伤后第14天达高峰(P<001);内皮细胞标志物CD31蛋白表达水平从伤后第3~28天持续高于正常组(P<005);(2)内皮细胞建模转染后,qPCR结果提示损伤模型组较正常组miRNA9表达显著降低(P<001),但miRNA9过表达损伤模型组miRNA9表达显著高于损伤模型组(P<001);CCK8结果同样显示miRNA9过表达损伤模型组细胞活力明显高于损伤模型组(P<001);(3)相比于损伤模型组,miRNA9 过表达损伤模型组内皮细胞Bcl2蛋白表达增加(P<001),Bcl2/Bax值增加(P<005),但Bax蛋白、clcaspase3蛋白表达降低(P<005)。结论创伤性脑损伤后伤灶周围区域miRNA9表达增多且过表达miRNA9可提高依托泊苷诱导损伤的内皮细胞活力,提示脑创伤后miRNA9表达增加有助于脑血管重塑的发生。

Abstract:
ObjectiveTo investigate the expression of miRNA9  in the brain tissue and its protective effect on cerebral microvascular endothelial cells in rats following traumatic brain injury(TBI). MethodsSixty adult male Sprague Dawley rats were  subjected to controlled cortical impact to induce TBI. Quantitative realtime PCR and Western blotting   were used to detect the expression of miRNA9 and CD31 protein in the brain tissues around the injured site at 6 h and 1, 3, 7, 14 and 28 d following the injury.Rat microvascular endothelial cells in primary culture were  transfected with a miRNA9 mimic or an empty plasmid before exposure to etoposide, and CCK8 assay was used to examine the viability of the cells; Western blotting was used to detect the expressions of Bcl2, Bax and clcaspase3 proteinsin the cells.  ResultsCompared with the normal rats, the rats with TBI showed significantly increased levels of miRNA9 in the brain tissue, which reached the peak level on day 14 following TBI (P<001);the expression of the endothelial cell marker CD31 protein   remained elevated from day 3 to the day 28 after the injury (P<005). In cultured primary rat microvascular endothelial cells, the expression of miRNA9 was decreased significantly following  the exposure to etoposide(P<001), and was  increased significantly after transfection with the miRNA9 mimic (P<001); the transfection obviously promoted the cell viability following etoposide exposure as compared to the nontransfected cells (P<001). Western blotting showed that compared with those in the nontransfected cells, the expression of Bcl2 and the Bcl2/Bax ratio were increased obviously and Bax and activated caspase 3 were decreased significantly in the cells transfected with the miRNA9 mimic following etoposide exposure(P<005). ConclusionThe expression of miRNA9 is increased after traumatic brain injury in rats, and miRNA9 overexpression can improve the viability of the microvascular endothelial cells with etoposideinduced injury.

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更新日期/Last Update: 2017-07-24