缺氧诱导因子-1α对创伤性脑损伤大鼠炎性反应和血管生成因子表达的影响  

Effect of hypoxia-inducible factor-1α on inflammatory response and angiogenic factor expression in rats with traumatic brain injury

作　　者：靳春杰 方贵龙[2] 权伟[1] 江荣才[1] 张建宁[1] 

机构地区：[1]天津医科大学总医院神经外科,300052 [2]天津市北辰医院普通外科

出　　处：《中华创伤杂志》2016年第9期835-842,共8页Chinese Journal of Trauma

基　　金：中加国际合作项目（OCRIF,2011DFG33430）;天津市科技支撑项目（14ZCZDSY00179）;国家自然科学基金青年基金（81301629）;天津市卫生局科技基金（2012WSJ278）

摘　　要：目的探讨大鼠创伤性脑损伤（TBI）后缺氧诱导因子-1α（HIF-1α）表达对神经血管的保护作用。方法液压打击仪建立大鼠TBI模型，按随机数字表法将600只大鼠分为假手术组、TBI组、TBI＋HIF-1α沉默表达组及TBI＋对照病毒颗粒组，每组150只。TBI组利用液压打击造成大鼠脑组织脑挫裂伤模型；假手术组只开颅不进行打击；HIF—α沉默表达组打击前24h注射携带HIF-α沉默基因的病毒颗粒；对照病毒颗粒组打击前24h注射未携带HIF-α沉默基因的病毒颗粒。TBI后3，7，14d观察：（1）HE染色观察TBI脑组织损伤水肿面积和损伤区形态结构变化；（2）Westernblot检测大鼠血管内皮细胞标记物（vW下）和HIF-1α的表达变化；（3）酶联免疫吸附（ELISA）法检测外周血及脑组织血管内皮生长因子（VEGF）、基质金属蛋白酶-9（MMP-9）、核因子-KB（NF-KB）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）的表达变化；（4）使用改良大鼠神经功能缺损评分（mNSS）对大鼠的神经功能变化进行动态评估。结果（1）HE染色：HIF-1沉默组脑组织损伤水肿面积伤后3—14d均大于TBI组（P〈0．05）。（2）Westernblot检测：与假手术组、对照病毒颗粒组比较，TBI组HIF-1α表达逐渐升高，至伤后7，14d水平仍较高（P〈0．05）；与TBI组比较，HIF-1α沉默组3个时相点vWF表达均降低（P〈0．05），抑制血管形成。（3）ELISA法检测：与TBI组比较，HIF—1α沉默组VEGF显著降低（P〈0．05），而TNF-α、IL-6、NF—KB均表达升高（P〈0．05），MMP-9在7—14d表达亦升高（P〈0．05）。（4）mNSS评分：与TBI组比较，HIF-1α沉默组7-14d差异有统计学意义（P〈0．05）。结论TBI后大鼠HIF—1α高表达，利于血管形成和抑制炎性反应相关因子表达，从而减轻脑水肿和脑损伤。促HIF-1α表达可能为改善TBI后神经血管损伤提供新的手段。Objective To investigate the protective effect of hypoxia-inducible factor-lct （HIF- la） on the neurovascular unit in rats with traumatic brain injury （TBI）. Methods The fluid percussion model was applied to induce TBI in rats. A total of 600 rats were divided into sham operation group, TBI group, TBI ＋ HIF-1α silence group and TBI ＋ control virus group according to the random number table, with 150 rats in each. Virus-mediated HIF-lct silence gene and control virus were delivered 24 h before the fluid percussion injury. After 3, 7 and 14 d, brain injury area and morphological changes in injured region were detected by HE staining, expressions of vascular endothelial cell markers （vWF） and HIF-1α were detected by Western blot method, and expressions of vascular endothelial growth factor （VEGF）, matrix metalloproteinase-9 （ MMP-9 ） , tumor necrosis factor-α（ TNF-α ） , interleukin 6 （ IL-6 ） and nuclear factor-KB （NF -KB） in peripheral blood and brain tissue were detected by ELISA method. Rat neural function was dynamically assessed using the modified neurological severity score （mNSS）. Results （ 1 ） Brain injury area and edema area in TBI ＋ HIF-1α silence group were higher than those in TBI group at all time points （P 〈 0. 05 ）. （2） Compared with sham operation group and TBI ＋ control virus group, expression of HIF-1α in TBI group gradually increased and remained high at 7 and 14 d postinjury （P 〈0. 05）. Compared with TBI group, expression of vWF in TBI ＋ HIF-lccsilence group decreased at all time points （P 〈 0. 05 ） and inhibited angiogenesis. （3） TBI ＋ HIF-1α silence group versus TBI group showed remarkably decreased VEGF at all time points, increased expressions of TNF-α, IL-6 and NF -KB at all time point, and increased expression of MMP-9 at 7 and 14 d postinjury （ all P 〈 0. 05）. （4） TBI ＋ HIF-1α silence group versus TBI group showed significant difference in mNSS at 7 and 14 d postinjury （ all P 〈 0. 05 ）

关 键 词：脑损伤 缺氧诱导因子-1α亚基 炎症 血管生成 

分 类 号：R651.15[医药卫生—外科学]