摘要
目的:蛛网膜下腔出血(subarachnoid hemorrhage,SAH)是一种严重的脑血管疾病。早期脑损伤(early brain injury,EBI)和脑血管痉挛是导致SAH患者预后不良的主要原因。组蛋白脱乙酰酶(histone deacetylase,HDAC)6特异性抑制剂tubastatin A(TubA)在多种急慢性中枢神经系统疾病的动物模型中被证实有明确的神经保护作用,但其对SAH的神经保护作用尚未明确。本研究旨在探讨HDAC6在SAH早期大脑皮质中的表达和细胞定位,并评估TubA对SAH后EBI和脑血管痉挛的保护作用及其机制。方法:选取成年健康雄性SD大鼠,采用改良的颈内动脉穿刺法建立大鼠SAH模型。第1部分实验,将大鼠随机分为假手术(sham)组、SAH-3 h组、SAH-6 h组、SAH-12 h组、SAH-24 h组、SAH-48 h组。分别在SAH建模后3、6、12、24 h,取各组大鼠损伤侧大脑皮质样本行蛋白质印迹法检测HDAC6的表达。另取SAH-24 h组大鼠,采用免疫荧光双染法测定HDAC6在损伤侧大脑皮质的分布。第2部分实验,将大鼠随机分为sham组、SAH组、SAH+TubA_(L)组(给予TubA 25 mg/kg)、SAH+TubA_(H)组(给予TubA 40 mg/kg)。建模后24 h,对各组大鼠行神经功能评分,检测各组大鼠脑组织含水量。第3部分实验,将大鼠随机分为SAH组、SAH+TubA组(给予TubA 40 mg/kg)。建模后24 h,取损伤侧大脑皮质组织行蛋白质印迹法检测HDAC6、内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)和诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的表达水平,末端脱氧核苷酸转移酶介导的d UTP缺口标记(terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling,TUNEL)染色检测凋亡,苏木精-伊红(hematoxylin and eosin,HE)染色检测大脑中动脉直径。结果:损伤侧大脑皮质中HDAC6的蛋白质表达水平在SAH后6 h开始升高(P<0.05),24 h时达到峰值(P<0.001),48 h时出现下降,但仍明显高于sham组(P<0.05)。HDAC6主要在神经元的细胞质中表达。与sham组大鼠�
Objective: Subarachnoid hemorrhage (SAH) is a serious cerebrovascular disease. Earlybrain injury (EBI) and cerebral vasospasm are the main reasons for poor prognosis of SAHpatients. The specific inhibitor of histone deacetylase 6 (HDAC6), tubastatin A (TubA), hasbeen proved to have a definite neuroprotective effect on a variety of animal models of acuteand chronic central nervous system diseases. However, the neuroprotective effect of TubAon SAH remains unclear. This study aims to investigate the expression and localization ofHDAC6 in the early stage of SAH, and to evaluate the protective effects of TubA on EBIand cerebral vasospasm after SAH and the underlying mechanisms.Methods: Adult male SD rats were treated with modified internal carotid artery puncture toestablish SAH model. In the first part of the experiment, rats were randomly divided into 6groups: a sham group, a SAH-3 h group, a SAH-6 h group, a SAH-12 h group, a SAH-24 hgroup, and a SAH-48 h group. At 3, 6, 12, and 24 h after SAH modeling, the injuredcerebral cortex of rats in each group was taken for Western blotting to detect the expressionof HDAC6. In addition, the distribution of HDAC6 in the cerebral cortex of the injured sidewas measured by immunofluorescence double staining in SAH-24 h group rats. In thesecond part, rats were randomly divided into 4 groups: a sham group, a SAH group, aSAH+TubAL group (giving 25 mg/kg TubA), and a SAH+TubAH group (giving 40 mg/kgTubA). At 24 h after modeling, the injured cerebral cortex tissue was taken for Westernblotting to detect the expression levels of HDAC6, endothelial nitric oxide synthase(eNOS), and inducible nitric oxide synthase (iNOS), terminal deoxynucleotidyl transferasemediateddUTP-biotin nick end labeling (TUNEL) staining to detect apoptosis, andhematoxylin and eosin (HE) staining to detect the diameter of middle cerebral artery.Results: The protein expression of HDAC6 began to increase at 6 h after SAH (P<0.05),peaked at 24 h (P<0.001), and decreased at 48 h, but there was still a difference com
作者
朱雨薇
郑海平
陈春丽
ZHU Yuwei;ZHENG Haiping;CHEN Chunli(Department of Neurology,Second Xiangya Hospital,Central South University,Changsha 410011,China)
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2023年第2期172-181,共10页
Journal of Central South University ：Medical Science
基金
湖南省自然科学基金(2021JJ40821)。
