Impact Of Cigarettes On Pulmonary Vascular Endothelial Cells’ Apoptosis And Expression Of VEGF/VEGFR2

Objective:The aim of this investigation was to reveal the impact of Cigarette Smoke Extract (CSE) on Human Pulmonary Micro vascular endothelial Cells’apoptosis and elucidate the differences of HPMEC apoptosis rate among CSE treatment of varied concentrations and time spans. Furthermore, mRNA and Protein expression levels of VEGF and VEGFR2in HPMEC were compared between CSE treatment group and control group which may imply a part of mechanism participated by cigarette smoke in HPMEC apoptosis and even pathogenesis of Chronic Obstructive Pulmonary Disease (COPD).Methods:HPMEC cultured in vitro were treated with CSE of different concentrations and periods of time before apoptosis rates were detected by double-staining flow cytometry. Levels of VEGF and VEGFR2mRNA and protein expression in HPMEC were compared to controls when incubated with the selected concentration and time span of CSE treatment from previous step using Real-time Polymerase Chain Reaction (RT-PCR) and western blot. All data were analyzed by SPSS19.0software and significance level was set at0.05.Results:Intervention of different CSE concentration(0%,0.5%,1%,2.5%,5%,10%) increased apoptosis rates of HPMEC compared to control group (P<0.01) and apoptosis rates were significantly different between each other group. Necrosis rates of HPMEC increased significantly in all groups treated with different CSE concentration compared to control group (P<0.01) except0.5%CSE group. Apoptosis rates of HPMEC increased significantly when treated with1%CSE for Ohour,3hours,6hours,12hours and24hours (P<0.01) and apoptosis rates were significantly different between each other group. Necrosis rates of HPMEC increased significantly in all groups treated with1%CSE for different periods compared to control group (P<0.01). CSE treatment did not have a negative influence on mRNA expression levels of VEGF and VEGFR2in HPMEC. CSE treatment significantly decreased protein expression of VEGF and VEGFR2in HPMEC.Conclusions:1. CSE induces concentration-dependent and span--dependent apoptosis of HPMEC.2. CSE does not have an impact on mRNA expression of VEGF and VEGFR2in HPMEC.3. CSE decreases protein expression of VEGF and VEGFR2in HPMEC...