摘要
目的探讨丝裂原激活蛋白激酶/细胞外信号调节激酶(MEK/ERK)1/2信号通路在大鼠实验性蛛网膜下腔出血(SAH)后早期脑损伤中的作用。方法取成年雄性SD大鼠60只,随机分为对照组,SAH造模后1、6、12、24、48、72 h组,SAH+MEK抑制剂U0126干预24、48、72 h组,共10组,每组6只。除对照组外,另9组大鼠于枕大池注血制备SAH模型,于眶下静脉丛取血。采用酶联免疫吸附法测定各组血清白细胞介素6(IL-6)、IL-1β、肿瘤坏死因子α(TNF-α)含量,脑组织伊文思蓝含量测定评定血-脑屏障损伤,Western-blot法测定基底动脉组织中磷酸化细胞外信号调节激酶(p-ERK1/2)、基质金属蛋白酶9(MMP-9)蛋白的水平并加以比较。结果 SAH组大鼠造模后6、12、24、48、72 h,血清IL-6、IL-1β水平与对照组同一时间点比较,差异均有统计学意义(均P<0.05);造模后24、48、72 h,SAH组大鼠血清TNF-α水平均高于对照组,差异均有统计学意义(均P<0.05)。造模后12、24、48、72 h,SAH组大鼠基底动脉组织p-ERK1/2蛋白表达水平分别为0.73±0.09、0.85±0.12、0.94±0.09、0.96±0.09,均明显高于对照组,差异均有统计学意义(均P<0.05)。SAH组造模后48、72 h,MMP-9蛋白水平明显高于对照组(1.27±0.15比0.68±0.08、2.41±0.11比0.71±0.14)。造模后72 h,SAH组脑组织伊文思蓝含量明显高于对照组[(15.3±2.2)μg/g比(2.7±0.4)μg/g]。给予MEK抑制剂U0126干预后,造模后24、48、72 h血清IL-6、IL-1β、TNF-α水平,造模后48、72 h p-ERK1/2、MMP-9蛋白表达水平(p-ERK1/2:0.76±0.07、0.81±0.06;MMP-9:0.92±0.14、1.79±0.16),以及造模后72 h脑组织伊文思蓝含量[(8.9±1.7)μg/g]均明显低于SAH组,差异均有统计学意义(均P<0.05)。结论 MEK/ERK1/2信号通路与大鼠实验性SAH后炎性反应及血-脑屏障损伤密切相关,提示干预MEK/ERK1/2信号通路可能为预防SAH后早期脑损伤的潜在靶点。
Objective To investigate the effect of mitogen activated protein kinase / extracellular signal-regulated kinase （MEK / ERK）1 / 2 signaling pathway on early brain injury （EBI）following experimental subarachnoid hemorrhage （SAH）in rats. Methods Sixty male SD rats were randomly divided into a control group and a 1,6,12,24,48,or 72 h group after SAH modeling. SAH ＋ MEK inhibitor U0126 was used to intervene the 24,48,and 72 h groups （a total of 10 groups;n = 6 in each group）. In 〈br〉 addition to the control group,blood was injected into the cisterna magna of the rats to induce a SAH model in another 9groups. The blood samples were taken from infraorbital venous plexus. Enzyme-linked immune sorbent assay （ELISA）was used to detect the levels of interleukin-6 （IL-6）,IL-1β,and tumor necrosis factor α（TNF-α）in each group. Evans blue content in brain tissue was used to evaluate the blood-brain barrier damage. Western blot was used to detect the levels of phosphorylated extracellular signal-regulated kinase （p-ERK1/ 2）and matrix metalloproteinase-9 （MMP-9）proteins in basilar artery tissue,and compared them. Results Compared with the control group at the same time points,there were significant differences in the levels of IL-6 and IL-1β at 6,12,24,48,and 72h after modeling in the SAH group （all P ＆lt;0. 05）. At 12,24, 48,and 72 h after modeling,the expression levels of p-ERK1/ 2 protein of the basilar artery tissue of the SAH group were 0. 73 ± 0. 09,0. 85 ± 0. 12,0. 94 ± 0. 09,and 0. 96 ± 0. 09,respectively,they were significantly higher than those of the control group （all P ＆lt; 0. 05）. At 48 and 72 h after modeling in the SAH group,the level of MMP-9 protein was significantly higher than that in the control group （1. 27 ± 0. 15 vs. 0. 68 ± 0. 08,2. 41 ± 0. 11 vs. 0. 71 ± 0. 14）. At 72 h after modeling,the Evans blue content in brain tissue of the SAH group was significantly higher than that of the control group （15. 3 ± 2. 2 μg/ g vs. 2. 7 ± 0. 4 �
出处
《中国脑血管病杂志》
CAS
CSCD
北大核心
2015年第4期192-198,共7页
Chinese Journal of Cerebrovascular Diseases
基金
中国人民解放军神经创伤防治重点实验室2013年开放课题资助项目(NTP2013005)
国家科技惠民计划项目(2013GS500101-15)
