Ouyang Mao, Liu Hengdao, Yang Kan, Jiang Weihong, Ding Qi, Yu Xunzhang, Chen Wu
Department of Cardiology, The Third Xiangya Hospital of Central South University Changsha, Hunan 410013, China.
Int J Clin Exp Med. 2014 Apr 15;7(4):879-85. eCollection 2014.
This study aimed to investigate the molecular mechanism underlying the myeloperoxidase (MPO) induced apoptosis of human umbilical vein endothelial cells (HUVECs).
HUVEC-12 cells were treated with myeloperoxidase at different concentrations (0.1 μ/ml, 0.2 μ/ml, 0.4 μ/ml and 0.6 μ/ml) and apoptotic cells were detected by flow cytometry. Then, cells were harvested for the detection of mRNA and protein expression of caspase-3 and Bax by reverse transcription PCR and Western blot assay, respectively.
When compared with negative control group, the apoptosis index in 0.2 μ/ml MPO group, 0.4 μ/ml MPO group and 0.6 μ/ml MPO group increased markedly (P<0.05). When compared with negative control group, the mRNA expression of caspase-3 in 0.6 μ/ml MPO group and positive control group increased dramatically (P<0.05). When compared with negative control group, the protein expression of pre-caspase-3 and activated caspase-3 elevated significantly in 0.4 μ/ml MPO group, 0.6 μ/ml MPO group and positive control group (P<0.05). When compared with negative control group, the mRNA and protein expression of Bax elevated dramatically in 0.4 μ/ml MPO group, 0.6 μ/ml MPO group and positive control group (P<0.05).
MPO at certain extents may induce the apoptosis of HUVEC-12. The MPO induced apoptosis of HUVEC-12 may be dependent on capase-3 signaling pathway, and Bax is also involved in the MPO induced apoptosis of HUVEC-12.
本研究旨在探讨髓过氧化物酶（MPO）诱导人脐静脉内皮细胞（HUVECs）凋亡的分子机制。
用不同浓度（0.1μ/ml、0.2μ/ml、0.4μ/ml和0.6μ/ml）的髓过氧化物酶处理HUVEC-12细胞，通过流式细胞术检测凋亡细胞。然后，分别通过逆转录PCR和蛋白质免疫印迹法检测细胞中半胱天冬酶-3（caspase-3）和Bax的mRNA及蛋白表达。
与阴性对照组相比，0.2μ/ml MPO组、0.4μ/ml MPO组和0.6μ/ml MPO组的凋亡指数显著升高（P<0.05）。与阴性对照组相比，0.6μ/ml MPO组和阳性对照组中caspase-3的mRNA表达显著增加（P<0.05）。与阴性对照组相比，0.4μ/ml MPO组、0.6μ/ml MPO组和阳性对照组中前半胱天冬酶-3和活化的半胱天冬酶-3的蛋白表达显著升高（P<0.05）。与阴性对照组相比，0.4μ/ml MPO组、0.6μ/ml MPO组和阳性对照组中Bax的mRNA和蛋白表达显著升高（P<0.05）。
一定程度的MPO可能诱导HUVEC-12细胞凋亡。MPO诱导HUVEC-12细胞凋亡可能依赖于半胱天冬酶-3信号通路，且Bax也参与了MPO诱导的HUVEC-12细胞凋亡。
