Jeremias I C, Victorino V J, Machado J L, Barroso W A, Ariga S K, Lima T M, Soriano F G
Laboratório de Investigação Médica - LIM 51, Faculdade de Medicina, Universidade de São Paulo (FMUSP), São Paulo, Brazil.
Mol Neurobiol. 2016 Jul;53(5):3389-3399. doi: 10.1007/s12035-015-9290-3. Epub 2015 Jun 18.
Septic encephalopathy represents the most frequently observed form of encephalopathy in intensive care units. Interactions between the immune and nervous systems have been observed in experimental sepsis. Therefore, the aim of the current study was to characterize the effect of different severities of sepsis on encephalopathy and the inflammatory profile of the spleen. We hypothesized that different grades of sepsis severity would lead to variations in encephalopathy and activation of spleen cells. We induced sepsis of different severities in Balb/c mice by cecal ligature and puncture (CLP). Six and 12 h after CLP induction, behavioral impairment was assessed by the SmithKline/Harwell/Imperial College/Royal Hospital/Phenotype Assessment (SHIRPA) test. The animals were then killed, and the plasma, spleen, and hippocampus were removed. Levels of the encephalopathy marker S100β were measured in plasma. Spleens were weighed and then a characterization of splenic lymphocytes was performed by flow cytometry (cytotoxic T lymphocyte, T helper lymphocytes, B lymphocytes, T regulatory cells, and Th17 cells). Cytokine levels in the spleen and hippocampus were determined by enzyme-linked immunosorbent assay (ELISA), and cytokine levels in plasma were performed with MilliPlex® technology. Our results showed that behavioral impairment as measured by the SHIRPA test and elevation in plasma S100β levels were significant in moderate and severe CLP groups compared to those in the sham control group. Regarding immunological alterations, we were unable to observe changes in the weights of the spleen and the profile of lymphocytes 6 h after CLP. However, several cytokines, including IL-6, IL-10, and IL-1β, were increased in spleen and plasma. In conclusion, we observed variations in encephalopathy as measured by plasma S100β, which were mediated by the severity of sepsis; however, we did not observe a different activation of spleen cells 6 h post-CLP, despite evidence of inflammation. Taken together, our data indicate that the severity of sepsis impacts the brain in absence of a change in the spleen lymphocyte profile.
脓毒症性脑病是重症监护病房中最常见的脑病形式。在实验性脓毒症中已观察到免疫和神经系统之间的相互作用。因此,本研究的目的是描述不同严重程度的脓毒症对脑病和脾脏炎症特征的影响。我们假设不同等级的脓毒症严重程度会导致脑病和脾细胞激活的差异。我们通过盲肠结扎和穿刺(CLP)在Balb/c小鼠中诱导不同严重程度的脓毒症。CLP诱导后6小时和12小时,通过史克必成/哈韦尔/帝国理工学院/皇家医院/表型评估(SHIRPA)试验评估行为损伤。然后处死动物,取出血浆、脾脏和海马。测量血浆中脑病标志物S100β的水平。称取脾脏重量,然后通过流式细胞术(细胞毒性T淋巴细胞、辅助性T淋巴细胞、B淋巴细胞、调节性T细胞和Th17细胞)对脾淋巴细胞进行表征。通过酶联免疫吸附测定(ELISA)测定脾脏和海马中的细胞因子水平,血浆中的细胞因子水平采用MilliPlex®技术测定。我们的结果表明,与假手术对照组相比,中度和重度CLP组通过SHIRPA试验测得的行为损伤和血浆S100β水平升高具有显著性。关于免疫改变,CLP后6小时,我们未观察到脾脏重量和淋巴细胞特征的变化。然而,脾脏和血浆中的几种细胞因子,包括IL-6、IL-10和IL-1β有所增加。总之,我们观察到血浆S100β测定的脑病存在差异,这是由脓毒症的严重程度介导的;然而,尽管有炎症证据,但CLP后6小时我们未观察到脾细胞的不同激活。综上所述,我们的数据表明,脓毒症的严重程度在脾脏淋巴细胞特征未改变的情况下影响大脑。