【摘要】 目的探讨肺动脉内皮细胞线粒体定位的eNOS减少与新生儿持续性肺动脉高压(persistent pulmonaryhypertension of the new born,PPHN)的相关性。方法 PPHN的胎羊模型并分离肺动脉内皮细胞,运用免疫荧光、Western Blot和特异性靶向eNOS(endothelial nitric oxide synthase)等方法,比较正常胎羊和持续性肺动脉高压的胎羊来源肺动脉内皮细胞的eNOS线粒体定位与胞内活性氧ROS(reactive oxygen species)和一氧化氮NO(nitric oxide)的关系。结果在持续性肺动脉高压来源的肺动脉内皮细胞中,Western Blot结果显示线粒体定位的eNOS减少;用Mito-HE荧光探针标记肺动脉内皮细胞中ROS,显示在PPHN的肺动脉内皮细胞中ROS增加;在293HEK细胞过表达不同靶向的eNOS,在线粒体过表达eNOS可减少胞内ROS含量。结论线粒体定位的eNOS可能通过ROS调控胞内NO的量,从而影响血管的舒张。我们的研究揭示了PPHN发病新的机制并为PPHN治疗提供了潜在的靶向位点。更多还原

【Abstract】 Objective To investigate the relationship between the decrease of endothelial nitric oxide synthase(eNOS) inpulmonary artery endothelial cells and persistent pulmonary hypertension of the new born(PPHN). Methods Afetal sheepmodel of PPHN was constructed and the pulmonary artery endothelial cells were isolated, and by usingthe immunofluorescence,Western blot and specific targeting eNOS, the relationship was compared between the mitochondrial localization eNOS andintracellular reactive oxygen species(ROS)/nitric oxide(NO) in pulmonary artery endothelial cells derived from normal fetalsheep and from PPHN fetal sheep. Results Western blot indicated that the mitochondrial targeting eNOS was decreased inthe pulmonary artery endothelial cells derived from PPHN sheep. Mito-HE fluorescent probe was used to label the intracellularROS and results showed that the intracellular ROS increased in the pulmonary artery endothelial cells derived from PPHNsheep. Different targeting eNOS were constructed and over expressing of mitochondrial eNOS in HEK293 T cells significantlydecreased the intracellular level of ROS.Conclusion PPHN may be associated with a reduction of mitochondrial localizationeNOS, which provides a potential target for the treatment of PPHN.更多还原