下调15-LOX-2表达对缺氧状态下肺动脉平滑肌细胞的影响

Effect of 15-lipoxygenase-2 down-regulation on hypoxic pulmonary arterial smooth muscle cells

目的探讨下调15脂氧酶-2(15-LOX-2)对缺氧状态下肺动脉平滑肌细胞(PASMCs)的影响。方法将PASMCs随机分为四组:正常对照组、缺氧组、scramble组、15-LOX-2 si RNA组。实时定量荧光聚合酶链反应(Real time PCR)检测15-LOX-2表达;噻唑蓝比色法(MTT)检测PASMCs增殖情况;酶联免疫吸附测定(ELISA)法检测15-羟基-二十碳四烯酸(15-HETE);分析活性氧(ROS)和超氧化物歧化酶(SOD)的表达变化。结果缺氧增加15-LOX-2、15-HETE和ROS的产生,促进PASMCs增殖,减少SOD活性,与正常对照组比较,差异有统计学意义(P<0.05)。缺氧下转染15-LOX-2 si RNA可减少15-LOX-2、15-HETE和ROS的产生,抑制PASMCs增殖,增加SOD活性,与缺氧组比较,差异有统计学意义(P<0.05)。结论 15-LOX-2下调可通过减少15-HETE产生、调节氧化/抗氧化平衡、调控PASMCs的异常增殖,从而延缓缺氧性肺血管收缩的病理进程。

Objective To explore the effect of 15-1ipoxygenase-2 （15-LOX-2） down-regulation on hypoxic pulmonary arterial smooth muscle ceils （PASMCs）. Methods PASMCs were randomly divided into 4 groups, normal control group, hypoxic group, scramble group and 15-LOX-2 siRNA group. Real time PCR was used to detect the expression of 15- LOX-2. Proliferation of PASMCs was examined by MTF assay. EL1SA was employed to detect 15-hydroxyeicosatetraenoic acid （15-HETE）. The changes of reactive oxygen species （ROS） and superoxide dismutase （SOD） were analyzed. Results During hypoxia, the expression of 15-LOX-2, 15-HETE, ROS and PASMCs proliferation were in- creased, the activity of SOD reduced, there were statistically significant differences compared with normal control group （P 〈 0.05）. After transfection of 15-LOX-2 siRNA during hyoxia, 15-LOX-2, 15-HETE, ROS and PASMCs proliferation were down-regulated, while SOD activity increased, there were statistically significant differences compared with hypoxic group （P 〈 0.05）. Conclusion 15-LOX-2 down-regulation can inhibit 15-HETE formation and abnormal PASMCs proliferation, regulate oxidant / antioxidant balance, which delays pathological process of HPV.