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（英文）

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。     

缺氧及复氧对hepG2细胞STC-1和STC-2的mRNA表达影响

目的:研究缺氧及复氧对hepG2细胞STC-1和STC-2的mRNA表达影响。方法:用化学缺氧剂氯化钴模拟缺氧环境,用RT-PCR法分别检测在hepG2细胞缺氧和复氧条件下STC-1、STC-2的mRNA表达情况。结果:在hepG2细胞中,STC-1mRNA无论在缺氧还是复氧情况下,几乎不表达;STC-2mRNA在缺氧条件下表达升高,并随着缺氧时间延长而升高,复氧后表达又降低,并随复氧时间延长而恢复至正常。结论:STC-2与肝癌的发生发展过程密切相关。     

转染瘦素人胎盘源间充质干细胞对经X射线辐照后人脐静脉内皮细胞的成血管潜能和周围炎症的影响

目的:放创复合伤是一种以血管损伤和促炎细胞因子缺乏为特征的难愈性创伤。瘦素(leptin)的直接应用在血管生成和炎症中起着重要作用。本研究构建了一种可持续稳定的leptin表达系统——leptin修饰的人胎盘来源间充质干细胞(HPMSCs/leptin),并探究其对经X射线辐照后的人脐静脉内皮细胞(HUVECs)的成血管潜能及周围炎症的影响和潜在机制。创新点:可持续稳定的leptin表达系统(HPMSCs/leptin)促进受X射线辐照后HUVECs的成血管潜能及外周炎症反应,有助于解决放创复合伤伤口愈合过程中血管损伤和促炎因子缺乏的问题。方法:利用慢病毒载体将leptin基因转染HPMSCs获得HPMSCs/leptin。采用X射线单次照射HUVECs,剂量为20 Gy。细胞迁移侵袭实验技术(Transwell)检测照射后HUVECs的迁移情况。在Transwell体系的基础上,建立HPMSCs与受辐照HUVECs共培养体系。CCK-8比色法测定细胞增殖。酶联免疫吸附法(ELISA)检测促炎细胞因子(粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-1α(IL-1α)、IL-6和IL-8)的分泌。实时荧光定量聚合酶链式反应(RT-qPCR)检测促血管生成因子(VEGF和bFGF)mRNA的表达。蛋白免疫印迹法(westernblot)检测核因子κB(NF-κB)和JAK/STAT信号通路的相关分子表达。结论:可持续稳定的leptin表达系统(HPMSCs/leptin)具有更好的细胞增殖、迁移和成血管潜能。HPMSCs/leptin单独培养和HPMSCs/leptin与受辐照HUVECs共培养体系中,促炎细胞因子的分泌增加与NF-κB和JAK/STAT信号通路的相互作用有关。HPMSCs/leptin可能促进X射线照射后HUVECs的成血管潜能和外周炎症反应。     

Piper sarmentosum as an antioxidant on oxidative stress in human umbilical vein endothelial cells induced by hydrogen peroxide

Endothelial cell death due to increased reactive oxygen species(ROS) may contribute to the initial endothelial injury,which promotes atherosclerotic lesion formation.Piper sarmentosum(PS),a natural product,has been shown to have an antioxidant property,which is hypothesized to inhibit production of ROS and prevent cell injury.Thus,the present study was designed to determine the effects of PS on the hydrogen peroxide(H2O2)-induced oxidative cell damage in cultured human umbilical vein endothelial cells(HUVECs).In this experiment,HUVECs were obtained by collagenase perfusion of the large vein in the umbilical cord and cultured in medium M200 supplemented with low serum growth supplementation(LSGS).HUVECs were treated with various concentrations of H2O2(0-1000 μmol/L) and it was observed that 180 μmol/L H2O2 reduced cell viability by 50% as denoted by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay.Using the above concentration as the positive control,the H2O2-induced HUVECs were concomitantly treated with various concentrations(100,150,250 and 300 μg/ml) of three different extracts(aqueous,methanol and hexane) of PS.Malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GPX) levels showed a significant increase(P0.05) in HUVECs compared to the negative control.However,PS extracts showed a protective effect on HUVECs from H2O2-induced cell apoptosis with a significant reduction in MDA,SOD,CAT and GPX levels(P0.05).Furthermore,PS had exhibited ferric reducing antioxidant power with its high phenolic content.Hence,it was concluded that PS plays a beneficial role in reducing oxidative stress in H2O2-induced HUVECs.