Evaluation of intestinal injury, inflammatory response and oxidative stress following intracerebral hemorrhage in mice
Evaluation of intestinal injury, inflammatory response and oxidative stress following intracerebral hemorrhage in mice
复制标题
DOI：
10.3892/ijmm.2018.3755
复制
发表时间：
2018-10-01
影响因子：
5.4
通讯作者：
Zhao, Weiguo
中科院分区：
文献类型：
--
作者：
Cheng, Yijun;Zan, Jieyu;Zhao, Weiguo
肠损伤是脑出血后常见的并发症，可导致营养不良、免疫功能低下和预后不良。先前的研究已经揭示了创伤性脑损伤后肠损伤的发病机制，使用缺血性中风模型。然而，ICH对肠损伤的影响仍然未知。本研究旨在探讨小鼠脑出血后肠道损伤的病理学改变、分子机制及时间过程。将雄性C57 BL/6小鼠随机分为以下7组（n=6只小鼠/组）：对照组，进行假手术，6个ICH组（2、6、12和24 h，以及第3和7天）。脑出血模型通过两个阶段立体定向注射自体血到脑内来诱导。随后，用苏木精和伊红对肠组织进行染色以进行组织病理学检查。采用炭餐试验检测小肠运动功能，通过检测血浆内毒素水平评价肠屏障功能障碍。进行定量聚合酶链反应（qPCR）、荧光化学和ELISA分析，以评价肠组织和血清中炎性细胞因子[白细胞介素（IL）-1 β、IL-6、肿瘤坏死因子-α、细胞间粘附分子1、单核细胞趋化蛋白1和趋化因子（C-C基序）配体-5]的mRNA和蛋白表达水平。此外，通过测量髓过氧化物酶活性检测肠白细胞浸润。通过qPCR和蛋白质印迹分析测量活性氧相关标志物（丙二醛含量和超氧化物歧化酶活性测定）和抗氧化基因[核因子（红细胞衍生2）样2、锰超氧化物歧化酶和血红素加氧酶1]的mRNA和蛋白质表达水平，间接检测氧化应激。结果表明，肠粘膜的显着破坏，延迟小肠运动，肠屏障功能障碍，并增加炎症反应和氧化应激迅速发生响应ICH。这些症状早在ICH后2 h出现，并持续7天。这些结果表明，ICH可能会引起肠道结构和屏障功能的即时和持续损害，这可能与炎症和氧化应激标志物的上调有关。
Intestinal injury is a common complication following intracerebral hemorrhage (ICH), which leads to malnutrition, impaired immunity and unsatisfactory prognosis. Previous studies have revealed the pathogenesis of intestinal injury following traumatic brain injury using ischemic stroke models. However, the effects of ICH on intestinal injury remain unknown. The present study aimed to investigate the pathological alterations and molecular mechanism, as well as the time course of intestinal injury following ICH in mice. Male C57BL/6 mice were randomly divided into the following seven groups (n=6 mice/group): Control group, which underwent a sham operation, and six ICH groups (2, 6, 12 and 24 h, and days 3 and 7). The ICH model was induced by stereotactically injecting autologous blood in two stages into the brain. Subsequently, intestinal tissue was stained with hematoxylin and eosin for histopathological examination. Small intestinal motility was measured by charcoal meal test, and gut barrier dysfunction was evaluated by detecting the plasma levels of endotoxin. Quantitative polymerase chain reaction (qPCR), immunohisto-chemistry and ELISA analysis were performed to evaluate the mRNA and protein expression levels of inflammatory cytokines [interleukin (IL)-1 beta, IL-6, tumor necrosis factor-alpha, intercellular adhesion molecule 1, monocyte chemotactic protein 1 and chemokine (C-C motif) ligand-5] in intestinal tissue and serum. Furthermore, intestinal leukocyte infiltration was detected by measuring myeloperoxidase activity. Oxidative stress was indirectly detected by measuring reactive oxygen species-associated markers (malondialdehyde content and superoxide dismutase activity assays) and the mRNA and protein expression levels of antioxidant genes [nuclear factor (erythroid-derived 2)-like 2, manganese superoxide dismutase and heme oxygenase 1] by qPCR and western blot analysis. The results demonstrated that significant destruction of the gut mucosa, delayed small intestinal motility, intestinal barrier dysfunction, and increased inflammatory responses and oxidative stress occurred rapidly in response to ICH. These symptoms occurred as early as 2 h after ICH and persisted for 7 days. These findings suggested that ICH may induce immediate and persistent damage to gut structure and barrier function, which may be associated with upregulalion of inflammation and oxidative stress markers.