Brief Secondhand Smoke Exposure Depresses Endothelial Progenitor Cells Activity and Endothelial FunctionSustained Vascular Injury and Blunted Nitric Oxide Production
Christian Heiss, MD, Dr Med*,
Nicolas Amabile, MD*,
Andrew C. Lee, MD*,
Wendy May Real, BS*,
Suzaynn F. Schick, PhD ,
David Lao, MD*,
Maelene L. Wong, BS*,
Sarah Jahn, MB*,
Franca S. Angeli, MD*,
Petros Minasi, BA*,
Matthew L. Springer, PhD*,
S. Katharine Hammond, PhD ,
Stanton A. Glantz, PhD, FACC*,
William Grossman, MD, FACC*,
John R. Balmes, MD*, and
Yerem Yeghiazarians, MD, FACC*,*
* Division of Cardiology, Department of Medicine, University of California, San Francisco, California
Division of Occupational and Environmental Medicine, Department of Medicine, University of California, San Francisco, California
Division of Environmental Health Sciences, School of Public Health, University of California, Berkeley, California.
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Figure 1 Time Course of Plasma Cotinine
Cotinine levels significantly increased immediately after SHS exposure and remained significantly elevated until 24 h. No increase was seen after exposure to clean air in the chamber. Symbols are mean ± SEM. *p < 0.05 versus baseline of same day (exposure condition). #p < 0.05 comparing SHS and clean air at the same time. B = baseline; SHS = secondhand smoke.
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Figure 2 Mobilization of EPCs in Blood and EPC Survival After Culture
(A) Early less differentiated CD133+/KDR+ EPCs and (B) committed CD34+/KDR+ EPCs in blood were sequentially and significantly increased after SHS exposure but not after exposure to clean air only. (C) The survival of EPCs as determined by UEA-1+/acLDL+ after 7 days of culture was also significantly increased after SHS, mimicking the time course of CD34+/KDR+ EPC. Symbols are mean ± SEM. *p < 0.05 versus baseline of same day (exposure condition). #p < 0.05 comparing SHS and clean air at the same time. acLDL = acetylated low-density lipoprotein; EPC = endothelial progenitor cell; MNC = mononuclear cells; UEA-1 = Ulex europeus agglutinin-1; other abbreviations as in Figure 1.
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Figure 3 Impairment of Chemotaxis of EPCs and Increased Chemokinesis
The EPCs isolated from blood up to 24 h after SHS exposure showed abolished chemotactic response to a VEGF (50 ng/ml); no effect was seen after exposure to clean air in the chamber. Random cell movement was significantly increased at 2.5 h and 24 h. Symbols are mean ± SEM. *p < 0.05 versus baseline of same day (exposure condition). #p < 0.05 comparing SHS and clean air at the same time. VEGF = vascular endothelial growth factor; other abbreviations as in Figures 1 and 2.
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Figure 4 Vascular Injury: Increased Plasma VEGF, Acute Sustained MP Generation, and Acute Reversible Endothelial Dysfunction
(A) Plasma VEGF levels were increased up to 24 h after SHS exposure. (B to D) The microparticles (MPs) in plasma increased significantly immediately (0 h) after SHS exposure and remained elevated throughout the observation period at 24 h, but remained unchanged after exposure to clean air in the chamber. The effect was observed in the 3 subgroups of endothelial MPs studied [CD31+/CD41– MPs (B), CD144+ MPs (C), and CD62e+
(D)]. After 30 min of SHS exposure, flow-mediated dilation (E) was significantly decreased immediately (0 h) and at 1 h after, but was increased at 24 h; FMD remained unchanged after clean air control. Solid symbols indicate SHS exposure and open symbols depict clean air controls. Symbols are mean ± SEM. *p < 0.05 versus baseline of same day (exposure condition). #p < 0.05 comparing SHS and clean air at the same time. FMD = flow-mediated dilation; other abbreviations as in Figures 1 and 3.
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Figure 5 Plasma Isolated After SHS and Pure Cotinine Decreases VEGF-Stimulated NO Production in EPCs
(A) Representative fluorescence. (B) Quantitative results. *p < 0.05 versus control. NO = nitric oxide; other abbreviations as in Figures 1, 3, and 4.
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Figure 6 Effects of Post-Exposure Plasma and Cotinine on EPCs In Vitro
Plasma isolated after SHS exposure and pure cotinine decrease chemotaxis to VEGF (A), increase proliferation (B), and increase random cell movement (C) similar to NO synthase inhibition with L-NMMA. The EPCs were isolated from nonexposed subjects (n = 6) and incubated with L-NMMA, plasma isolated before (B, baseline), immediately after (0 h), and 24 h after SHS exposure. Right sides of graphs represent cotinine dose response. Dashed lines show plasma cotinine range after SHS exposure (approximately 0 to 1 ng/ ml). *p < 0.05 versus control. #p < 0.05 versus baseline plasma. L-NMMA = LG-monomethyl-arginine; other abbreviations as in Figures 1 and 3.
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