摘要
目的研究星型胶质细胞代谢抑制剂氟代柠檬酸(FC)对大鼠蛛网膜下腔出血(SAH)早期脑损伤的作用,探讨SAH后星型胶质细胞介导早期脑损伤的机制。方法雄性SD大鼠36只,随机分为假手术组、SAH 3d组和SAH3d+FC组,每组12只。采用颈内动脉穿刺法建立SAH模型,SAH 3d+FC组侧脑室注射FC,建模后3d,利用免疫组织学染色和电镜评估FC对皮层的神经保护作用和对星型胶质细胞、小胶质细胞活化的抑制作用,利用Western blot检测细胞核内NF-κB的含量及其磷酸化程度的变化,利用ELISA检测FC对炎性因子TNF-α、IL-1β、IL-6水平的影响,利用TUNEL检测细胞凋亡情况。结果与假手术组相比,SAH 3d组皮层神经元肿胀、核变形,微血管结构紊乱,内皮形态异常,SAH损伤标志物NSE、星型胶质细胞和小胶质细胞活化标志物GFAP、Iba-1的表达升高,凋亡细胞数、细胞核内NF-κB的含量及磷酸化程度、炎性因子TNF-α、IL-1β、IL-6的水平均增加(P<0.05)。而FC可减轻SAH 3d时的神经损伤,抑制NSE、GFAP、Iba-1的表达,减少细胞凋亡数、细胞核内NF-κB的含量及磷酸化程度,并降低炎性因子TNF-α、IL-1β、IL-6的水平(P<0.05)。结论星型胶质细胞在SAH后早期脑损伤中发挥重要作用,其可能通过激活NF-κB信号通路,释放TNF-α、IL-1β、IL-6等炎性因子,并激活小胶质细胞介导炎性损伤。抑制星型胶质细胞的过度活化对SAH后早期脑保护有重要意义。
Objective To study the effects of fluorocitrate（FC）,astrocytic metabolic inhibitor,on early brain injury after subarachnoid hemorrhage（SAH）in rats so as to explore the mechanism of early brain injury mediated by astrocyte after SAH.Methods Thirty six SD male rats were randomly divided into sham group,SAH3 d group and SAH 3 d＋FC group,with 12 rats in each group.The model of SAH was established by an endovascular perforation technique.FC was injected into the lateral ventricle.Three days after SAH,the expressions of GFAP,Iba-1 and TNF-αwere detected using immunohistochemistry in all rats.The expression and phosphorylation of NF-κB in the nucleus were detected by Western blot.The expressions of TNF-α,IL-1βand IL-6 were detected by ELISA.Cell apoptosis was detected by TUNEL.Results Neuronal swelling,nuclear anomalies,disorganization of micrangium,abnormality of endothelial cells appeared in SAH3 d group,but not in sham group.The expressions of NSE,GFAP and Iba-1 were significantly increased in SAH3 d group compared with those in sham group（all P〈0.05）.The number of apoptotic cells was increased.The expression and phosphorylation of NF-κB were significantly increased.The expressions of TNF-α,IL-1βand IL-6 were significantly increased.However,neuronal injuries were alleviated by injecting FC.The expressions of NSE,GFAP and Iba-1 in SAH3 d＋FC group were inhibited.The number of apoptotic cells in SAH3 d＋FC group was decreased compared with that in SAH3 d group.The expression and phosphorylation of NF-κB were decreased by FC.The expressions of TNF-α,IL-1βand IL-6 were significantly decreased by FC（P〈0.05）.Conclusion Astrocytes may play an important role in early brain injury after SAH,underlying the mechanism that they released TNF-α,IL-1βand IL-6 and activate microglia by activating NF-κB signal.The inhibition of excess activation of astrocytes may plays a protective role in early brain injury after SAH.
作者
行治国
赵君杰
宋锦宁
马旭东
黄廷钦
郭丹
XING Zhi-guo;ZHAO Jun-jie;SONG Jin-ning;MA Xu-dong;HUANG Ting-qin;GUO Dan(Department of Neurosurgery,the First Affiliated Hospital of Xi＇an Jiaotong University,Xi＇an 710061;Department of Neurosurgery,Weinan Central Hospital,Weinan 714000,China)
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2018年第4期530-536,共7页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
陕西省自然科学基础研究计划项目(No.S2015YFJM2049)~~
