活性氧和细胞外信号调节激酶1%2f2信号通路对缺氧大鼠肺动脉平滑肌细胞增殖和凋亡影响的研究.pdfVIP

Effect of Reactive Oxygen Species and ERK1/2 Signaling Pathway on the Regulation of Cell Proliferation and Apoptosis in Hypoxic Pulmonary Arterial Smooth Muscle Cells of Rats Abstract Objective This work was designed to investigate the change of reactive oxygen species(ROS) production in hypoxic pulmonary arterial smooth muscle cells(PASMCs) of rats, the effect of ROS on the proliferation and apoptosis of rat PASMCs under hypoxic condition. In addition, this work to explore the role of extracellular signal-regulated kinase(ERK)1/2 signaling pathway in the imbalance of proliferation/apoptosis of PASMCs by ROS. Methods and Materials Primary cultures of PASMCs were established and cells between passages 2 to 4 were used for experiments.PASMCs were treated with ROS scavenger Tiron and ERK inhibitor PD98059 under normoxia or hypoxia condition.The ROS production was measured by DCFH-DA and NBT reduction.The expression of phosphorylated-ERK1/2 （p-ERK1/2 ） protein were detected by immunofluorescence.Cells proliferation were examined by MTT colorimetric assey and the expression of PCNA.Cells apoptosis were detected by TUNEL. The expression of Bcl-2 and Bax protein was detected by immunocytochemical. Results 1. Compared with control group （NBT:0.114±0.017 ，DCFH-DA:15.84±2.86 ）,the ROS levels of hypoxia group（NBT:0.214±0.024,DCFH-DA:39.33±3.27 ） were significantly increased(n=6 ，P0.01). 3 2. the proliferative capacity of hypoxia group (MTT:0.325±0.035,PCNA:53.83±4.48)was higher than control group(MTT: 0.228±0.019,PCNA:42.58±3.71) (n=6 ，P0.01) and the apoptosis index of hypoxia group (2.42±0.74)was lower than control group(4.50±0.45) (n=6,P0.01),