Overexpression of the steroidogenic acute regulatory protein increases the expression of ATP-binding cassette transporters in microvascular endothelial cells (bEnd.3)

Objective:To determine the effect of steroidogenic acute regulatory protein(StAR) overexpression on the levels of adenosine triphosphate(ATP)-binding cassette transporter A1(ABCA1) and ATP-binding cassette transporter G1(ABCG1) in an endothelial cell line(bEnd.3).Methods:The StAR gene was induced in bEnd.3 cells with adenovirus infection.The infection efficiency was detected by fluorescence activated cell sorter(FACS) and fluorescence microscopy.The expressions of StAR gene and protein levels were detected ...     

基于微阵列技术的缺氧/复氧诱导下血管内皮细胞转录组分析

目的:应用全转录组芯片研究缺氧/复氧诱导下人脐静脉内皮细胞(HUVEC)的转录组轮廓。创新点:血管内皮细胞(VEC)缺氧/复氧损伤被视定为许多生理和病理过程中导致器官功能障碍的重要驱动因素。然而,其详细病理生理机制和基因表达谱信息尚未阐明。本研究首次应用全转录组芯片技术研究VEC缺氧/复氧诱导下的转录组轮廓。方法:采用缺氧孵育3h后复氧1h的HUVEC为缺氧/复氧组,同时常氧孵育的HUVEC为常氧对照组。应用含58 339条探针的全转录组芯片检测每组三个样本。对差异表达基因进行生信分析和功能验证。结论:本研究发现372个有意义的差异表达基因探针。相关基因涵盖多种途径和功能,例如氧自由基的产生、钙超载、炎症、糖脂代谢、内皮细胞增殖、分化、细胞骨架及通透性调节、细胞裂解、凋亡和血管生成。另外,实验进一步表明,差异表达基因pleckstrin同源样域家族A成员1(PHLDA1)的m RNA和蛋白质表达结果与微阵列结果一致。STRING分析发现,PHLDA1可能与差异表达基因SLC38A3、SLC5A5、Lnc-SLC36A4-1和Lnc-PLEKHJ1-1具有物理性和/或功能性相互作用,这有望揭示VEC在缺氧/复氧环境下长链非编码RNA(lnc RNA)的相关机制。     

Adiponectin levels are associated with the number and activity of circulating endothelial progenitor cells in patients with coronary artery disease

Objective: To study the relationship between plasma adiponectin concentration and the functional activities of circulating endothelial progenitor cells (EPCs) in patients with coronary artery disease (CAD). Methods: Circulating EPCs were enumerated as AC133+/KDR+ cells via flow cytometry and identified by co-staining with Dii-acLDL and fluorescein isothiocy-anate (FITC)-conjugated lectin under a fluorescent microscope. The migratory capacity of EPCs was measured by modified Boyden chamber assay. Adhesion capacity was performed to count adherent cells after replating EPCs on six-well culture dishes coated with fibronectin. Results: The number of circulating EPCs (AC133+/KDR+ cells) decreased significantly in CAD patients, compared with control subjects [(74.2±12.3) vs (83.5±12.9) cells/ml blood, P<0.0\]. In addition, the number of EPCs also decreased in CAD patients after ex vivo cultivation [(54.4±8.6) vs (71.9±11.6) EPCs/field, P<0.01]. Both circulating EPCs and differentiated EPCs were positively correlated with plasma adiponectin concentration. The functional activities of EPCs from CAD patients, such as migratory and adherent capacities, were also impaired, compared with control subjects, and positively correlated with plasma adiponectin concentration. Conclusion: The study demonstrates that the impairment of the number and functional activities of EPCs in CAD patients is correlated with their lower plasma adiponectin concentrations.