【摘要】 目的探讨脱嘌呤/脱嘧啶核酸内切酶1(APE1)在脂多糖介导肺微血管内皮细胞(PMVECs)氧化反应中的调控作用。方法将体外培养人PMVECs分成4组:对照组(未加脂多糖的培养细胞)及各剂量脂多糖组(分别予以0.1、1、10μg/ml剂量脂多糖刺激培养细胞)。分别在脂多糖刺激6、12、24 h检测细胞内活性氧及一氧化氮的水平并进行比较。同时利用携带目的基因慢病毒载体转染PMVECs,将其分成对照组,脂多糖组,GFP组及APE1组。对照组为未加脂多糖的培养细胞,脂多糖组为1μg/ml剂量脂多糖刺激的培养细胞,GFP组为1μg/ml的脂多糖刺激培养慢病毒空载体转染的细胞,APE1组为1μg/ml的脂多糖刺激培养携带APE1基因慢病毒转染的细胞。在12 h检测细胞内活性氧、一氧化氮的水平以及细胞APE1的表达情况。结果与对照组比较,各剂量组脂多糖刺激6、12、24 h后一氧化氮及活性氧分泌明显上升(P均<0.05),且同一剂量组随着时间的推移,一氧化氮及活性氧分泌的比较,差异均具有统计学意义(P均<0.05)。在转染PMVECs12 h后,脂多糖组APE1的表达明显低于对照组,而APE1组细胞APE1的表达较脂多糖组显著提高(P均<0.05)。与对照组相比,脂多糖组能够诱导PMVECs的一氧化氮及活性氧生成显著增加,而APE1组能够一定程度抑制一氧化氮及活性氧的产生(P均<0.05)。结论 APE1可通过抑制一氧化氮及活性氧的分泌从而减轻脂多糖介导PMVECs的氧化应激反应。更多还原

【Abstract】 objective To investigate the modulation of apurinic / apyrimidinic endonuclease(APE1) on oxidative stress reaction of pulmonary microvascular endothelial cells(PMVECs)induced by lipopolysaccharides(LPS). Methods PMVECs were cultured in vitro and randomly divided into 4 groups: control group(Not polysaccharide of cultured cells), LPS group(polysaccharide of cultured cells with 0.1, 1, 10 μg / ml, respectively). The levels of reactive oxygen species(ROS) and NO were measured at 6, 12, 24 h after LPS stimulation. Meanwhile, the PMVECs were transfected with lentiviruses and randomly divided into 4 groups: control group(Not polysaccharide of cultured cells), LPS group(1 μg / ml polysaccharide of cultured cells), GFP group(lentivirus vector transfected empty cells with 1 μg / ml polysaccharide) and APE1 group(APE1 gene transfected lentivirus cells with 1 μg / ml polysaccharide). The levels of ROS, NO and APE1 expression cultured after 12 h were detected and compared. Results Compared with the control group, the levels of ROS and NO in the PMVECs induced by different dose of LPS for 6, 12, 24 h respectively were increased obviously(all P < 0.05), and at the same dose group,the levels of ROS and NO showed significant differences with the time lapse(all P < 0.05). After12 h of the transfection, the APE1 expression in the LPS group was lower than those in the control and APE1 groups(all P < 0.05). The levels of ROS, NO in the LPS group after stimulated with 1 μg / ml LPS for 12 h were higher than those in the control and APE1 groups(all P <0.05). Conclusion The APE1 can inhibit the levels of ROS and NO, and thereby relieve the oxidative stress reaction of PMVECs induced by LPS.更多还原