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目的观察早产儿氧暴露后,外周血单个核细胞(PBMC)内去乙酰化酶沉默接合型信息调节因子2同源蛋白1(SIRT1)与活性氧(ROS)产生的关系。方法将诊断为呼吸窘迫综合征且需要吸氧的胎龄<32周的早产儿,根据吸入氧体积分数(Fi O2)分为低剂量吸氧组(Fi O2<300 m L/L),中剂量吸氧组(Fi O2300 m L/L~400 m L/L),高剂量吸氧组(Fi O2>400 m L/L)。在吸氧48 h后,采集外周血分离PBMC及血清,Mito SOXTMRed标记结合激光共聚焦显微镜检测细胞内活性氧(ROS)的生成量,全光谱分光光度仪检测血清中丙二醛(MDA)含量,免疫荧光技术检测SIRT1定位,Western blot法检测PBMC中SIRT1蛋白水平。结果随着吸入氧体积分数的增加,PBMC内的ROS、MDA含量逐渐增加,SIRT1转位率逐渐增加,SIRT1蛋白水平明显降低。结论氧暴露诱导早产儿PBMC中产生大量的ROS,并促使SIRT1核质穿梭,抑制SIRT1的抗氧化应激能力。
Objective To investigate the relationship between sIRT1 and reactive oxygen species (ROS) production in peripheral blood mononuclear cells (PBMCs) after oxygen exposure in preterm infants. Methods Preterm infants with gestational age <32 weeks, who were diagnosed as respiratory distress syndrome and required oxygen inhalation, were divided into low dose oxygen inhalation group (Fi O2 <300 m L / L) according to Fi O2 fraction, FiO22300 m L / L to 400 m L / L, and Fi O2> 400 m L / L. PBMC and serum were collected from peripheral blood 48 h after oxygen inhalation. Mito SOX TMRed labeling and laser scanning confocal microscopy were used to detect the production of intracellular reactive oxygen species (ROS). The content of malondialdehyde (MDA) in serum was detected by spectrophotometer. , SIRT1 localization was detected by immunofluorescence and SIRT1 protein level in PBMC was detected by Western blot. Results With the increase of inhaled oxygen concentration, the content of ROS and MDA in PBMC gradually increased, the translocation rate of SIRT1 increased gradually and the level of SIRT1 protein decreased significantly. Conclusion Oxygen exposure can induce a large amount of ROS in PBMCs of premature infants and promote the shuttling of SIRT1 nucleosomes and the anti-oxidative stress of SIRT1.