The effects of smoking on levels of endothelial progenitor cells and microparticles in the blood of healthy volunteers

Fariborz Mobarrez, Lukasz Antoniewicz, Jenny A Bosson, Jeanette Kuhl, David S Pisetsky, Magnus Lundbäck, Fariborz Mobarrez, Lukasz Antoniewicz, Jenny A Bosson, Jeanette Kuhl, David S Pisetsky, Magnus Lundbäck

Abstract

Background: Cigarette smoking, both active and passive, is one of the leading causes of morbidity and mortality in cardiovascular disease. To assess the impact of brief smoking on the vasculature, we determined levels of circulating endothelial progenitor cells (EPCs) and circulating microparticles (MPs) following the smoking of one cigarette by young, healthy intermittent smokers.

Materials and methods: 12 healthy volunteers were randomized to either smoking or not smoking in a crossover fashion. Blood sampling was performed at baseline, 1, 4 and 24 hours following smoking/not smoking. The numbers of EPCs and MPs were determined by flow cytometry. MPs were measured from platelets, leukocytes and endothelial cells. Moreover, MPs were also labelled with anti-HMGB1 and SYTO 13 to assess the content of nuclear molecules.

Results: Active smoking of one cigarette caused an immediate and significant increase in the numbers of circulating EPCs and MPs of platelet-, endothelial- and leukocyte origin. Levels of MPs containing nuclear molecules were increased, of which the majority were positive for CD41 and CD45 (platelet- and leukocyte origin). CD144 (VE-cadherin) or HMGB1 release did not significantly change during active smoking.

Conclusion: Brief active smoking of one cigarette generated an acute release of EPC and MPs, of which the latter contained nuclear matter. Together, these results demonstrate acute effects of cigarette smoke on endothelial, platelet and leukocyte function as well as injury to the vascular wall.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Study design.
Figure 1. Study design.
12 healthy volunteers were randomized to either smoking or not smoking (control) in a crossover fashion. A time period of at least 1 week separated the experiments. Blood sampling was performed at baseline, 1 h, 4 h and 24 hours.
Figure 2. Endothelial progenitor cells during smoking…
Figure 2. Endothelial progenitor cells during smoking and control.
Endothelial progenitor cells were measured as CD34+ KDR+ double positive cells during smoking and control visit. Data are presented as events of endothelial progenitor cells (mean±SE). Statistical differences (two-factor ANOVA) are indicated in the figure.
Figure 3. Microparticles during smoking and control.
Figure 3. Microparticles during smoking and control.
Microparticles were measured by flow cytometry and gated according to size and phosphatidylserine exposure (A) together with cell-specific antigens to determine cellular origin; endothelial-MPs (B), platelet-MPs (C) or leukocyte-MPs (D). Data is presented as mean±SE. Statistical differences (two-factor ANOVA) are indicated for each panel.
Figure 4. Nuclear molecule content of microparticles…
Figure 4. Nuclear molecule content of microparticles after smoking and control.
Microparticles were measured by flow cytometry and gated according to size and SYTO 13 binding alone (A) or together with HMGB1 exposure (B). Data is presented as mean±SE. Statistical differences (two-factor ANOVA) are indicated for each panel.

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