Abstract. The aim of the present study was to investigate the role of homocysteine (Hcy) in the pathogenesis of pulmonary embolism (PE) and the associated molecular mechanisms in human umbilical vein endothelial cells (HUVECs). Hcy contents were detected with high-performance liquid chromatography. Apoptosis was detected by flow cytometry using Annexin-V staining. Cytochrome c oxidase (COX) activity was assessed with an enzyme activity assay, and the expression levels of COX 17 were determined by western blot analysis. Intracellular reactive oxygen species levels were measured using a microplate reader with a fluorescence probe. The results demonstrated that, compared with the control group, the serum Hcy levels were significantly elevated in the PE group, suggesting that Hcy may be an indicator for PE. Following treatment with Hcy, the apoptosis rate was markedly elevated in HUVECs. Moreover, Hcy decreased COX activity and downregulated the expression of COX 17 in HUVECs. Furthermore, Hcy increased the ROS levels in these endothelial cells. However, all the above-mentioned physiopathological changes induced by Hcy in HUVECs could be restored by folic acid. In conclusion, the results of the present study demonstrated that Hcy inhibited COX activity, downregulated COX 17 expression, increased intracellular ROS levels and enhanced apoptosis in endothelial cells.
IntroductionPulmonary embolism (PE) is a frequent clinical syndrome with abnormal pulmonary circulation, which is predominantly caused by the blockage of pulmonary and/or bronchial arteries by thrombus shedding (1). Due to the sudden disease onset, PE is difficult to treat and usually results in the patient succumbing to the disease (2,3). Homocysteine (Hcy) is an intermediate product of methionine metabolism, and is an independent risk factor for cardiovascular diseases (4,5). Previous studies reported that serum Hcy levels are markedly elevated in patients with PE, which is implicated in the disease pathogenesis (6-8).Vessel wall injury is one of the important factors for PE, as the pathological changes in endothelial cells may result in the formation of thrombosis (9,10). It has been demonstrated that high concentrations of Hcy induce apoptosis in endothelial cells (11,12), in which mitochondria are involved. Cytochrome c oxidase (COX) is a key enzyme in mitochondrial function (13)(14)(15). Enhanced COX activity may increase intracellular reactive oxygen species (ROS) levels, resulting in cellular apoptosis (16)(17)(18)(19)(20)(21). Based on these reports, it would be of interest to define the association between Hcy, COX, and ROS in apoptosis, specifically in the context of PE pathogenesis.In the present study, the effects of Hcy on endothelial cells and the associated molecular mechanisms were investigated.
Materials and methods
Patients.A total of 10 patients with PE, 4 males and 6 females, aged 50-67 years (with an average age of 60.4 years), were included in this study. These PE cases were diagnosed based on the clinical manifestations and the resu...