创伤性脑损伤后大鼠脑组织miRNA-9表达变化及意义

发布时间：2018-06-25 11:07

本文选题：创伤性脑损伤 + miRNA　；参考：《第三军医大学学报》2017年14期

【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(P0.01);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(P0.05);(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(P0.01),但miRNA-9过表达损伤模型组miRNA-9表达显著高于损伤模型组(P0.01);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(P0.01);(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(P0.01),Bcl-2/Bax值增加(P0.05),但Bax蛋白、cl-caspase-3蛋白表达降低(P0.05)。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]:Objective to observe the expression of miRNA-9 in rat cortical area after traumatic brain injury (traumatic brain injura) and to explore its protective effect on cerebral microvascular endothelial cells (MECs). Methods 60 adult male Sprague-Dawley rats were selected to make the model of (controlled cortical impaction injury. The expression of miRNA-9 and CD31 in the tissues around the injured focus was detected by fluorescence quantitative PCR and Western blot. Primary rat brain microvascular endothelial cells were cultured. Endothelial cells were divided into normal group, injury model group [etoposideside ETO injury model group] miRNA-9 overexpression model group, empty transfection injury model group, and CCK-8 were used to detect the cell viability of each group. The expression of B cell lymphoma-2Bcl 2 protein and B cell lymphoma / leukemia associated X protein (Bcl-2 associated XnBax) and activated cysteine aspartate specific protease 3 (cleaved cysteinyl aspartate specific proteinase 3) were detected by Western blot. Results (1) the expression of miRNA-9 was significantly increased in the perifocal area after traumatic brain injury. The expression level of CD31 protein in endothelial cells was continuously higher than that in normal group (P0.05); (2) on the 14th day after injury (P0.01). The results showed that the expression of miRNA-9 in the model group was significantly lower than that in the normal group (P0.01), but the expression of CD31 protein in the model group was significantly lower than that in the normal group (P0.01). The expression of miRNA-9 in the injury model group was significantly higher than that in the injury model group (P0.01). The results also showed that the cell viability of the injury model group was significantly higher than that of the injury model group (P0.01); (3) compared with the injury model group. The expression of Bcl-2 protein increased (P0.01) and Bcl-2 / Bax increased (P0.05), but the expression of Bax protein cl-caspase-3 decreased (P0.05). Conclusion the increased expression of miRNA-9 and overexpression of miRNA-9 can increase the viability of endothelial cells induced by etoposide after traumatic brain injury, suggesting that the increased expression of miRNA-9 is helpful to the development of cerebral vascular remodeling after traumatic brain injury.
【作者单位】：重庆医科大学附属第一医院神经外科;
【基金】：国家自然科学基金面上项目(81571159)~~
【分类号】：R651.15

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