Elsheikha Hany M, Alkurashi Mamdowh, Palfreman Suzy, Castellanos Marcos, Kong Kenny, Ning Evita, Elsaied Nashwa A, Geraki Kalotina, MacNaughtan William
Faculty of Medicine and Health Sciences, School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, Leicestershire LE12 5RD, UK.
Animal Production Department, College of Food and Agricultural Sciences, King Saud University, Riyadh 11451, Saudi Arabia.
Pathogens. 2020 Aug 28;9(9):710. doi: 10.3390/pathogens9090710.
In this work, the effects of the protozoan on the bioenergetics, chemical composition, and elemental content of human brain microvascular endothelial cells (hBMECs) were investigated. We showed that can impair cell mitochondrial (Mt) function and causes an arrest in host cell cycling at S and G2 phases. These adverse effects were also associated with altered expression of genes involved in Mt energy metabolism, suggesting Mt dysfunction caused by infection. Fourier Transform Infrared (FTIR) spectroscopy analysis of hBMECs revealed alterations in the FTIR bands as a function of infection, where infected cells showed alterations in the absorption bands of lipid (2924 cm), amide I protein (1649 cm), amide II protein (1537 cm), nucleic acids and carbohydrates (1092 cm, 1047 cm, and 939 cm). By using quantitative synchrotron radiation X-ray fluorescence (μSR-XRF) imaging and quantification of the trace elements Zn, Cu and Fe, we detected an increase in the levels of Zn and Cu from 3 to 24 h post infection (hpi) in infected cells compared to control cells, but there were no changes in the level of Fe. We also used Affymetrix array technology to investigate the global alteration in gene expression of hBMECs and rat brain microvascular endothelial cells (rBMVECs) in response to infection at 24 hpi. The result of transcriptome profiling identified differentially expressed genes involved mainly in immune response, lipid metabolism and apoptosis. These data further our understanding of the molecular events that shape the interaction between and blood-brain-barrier endothelial cells.
在这项研究中，我们调查了原生动物对人脑血管内皮细胞（hBMECs）的生物能量学、化学成分和元素含量的影响。我们发现，原生动物可损害细胞线粒体（Mt）功能，并导致宿主细胞周期在S期和G2期停滞。这些不良反应还与Mt能量代谢相关基因的表达改变有关，提示原生动物感染导致Mt功能障碍。傅里叶变换红外（FTIR）光谱分析显示，hBMECs的FTIR谱带随感染情况而改变，感染细胞的脂质（2924 cm）、酰胺I蛋白（1649 cm）、酰胺II蛋白（1537 cm）、核酸和碳水化合物（1092 cm、1047 cm和939 cm）吸收带均发生改变。通过使用定量同步辐射X射线荧光（μSR-XRF）成像和对微量元素锌、铜和铁进行定量分析，我们发现感染细胞在感染后3至24小时（hpi）内锌和铜的水平相对于对照细胞有所增加，但铁的水平没有变化。我们还使用Affymetrix芯片技术研究了hBMECs和大鼠脑血管内皮细胞（rBMVECs）在24 hpi时对原生动物感染的基因表达全局变化。转录组分析结果确定了主要参与免疫反应、脂质代谢和细胞凋亡的差异表达基因。这些数据进一步加深了我们对原生动物与血脑屏障内皮细胞相互作用所涉及分子事件的理解。
