Cigarette Smoking Cessation Temporarily Enhances the Release of Phosphorylated-HSP27 from Human Platelets

Figures

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  Figure 1.. Effect of smoking cessation on the collagen-induced release of phosphorylated-HSP27 from human platelets. The net increase in phosphorylated-HSP27 released from collagen-stimulated (3 μg/mL) human platelets obtained from 12 patients at the indicated point - before smoking cessation (0) and 4, 8 and 12 weeks after smoking cessation - are shown. Values above the upper limit (20,000 pg/mL) are plotted as 20,000 pg/mL. For the statistical analysis, in values exceeded the upper limit, 20,000 pg/mL was adopted instead of the real value. The Friedman test revealed no statistical significance. HSP27: heat shock protein 27

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  Figure 2.. Effect of smoking cessation on the collagen-induced release of phosphorylated-HSP27 from human platelets: change from the baseline. The collagen-induced net increase in the levels pre-smoking cessation is represented as 1, and the levels at 4, 8 and 12 weeks after smoking cessation are expressed as the fold increase compared to the pre-smoking cessation value. (A) The line graph shows the values before smoking cessation and at 4, 8 and 12 weeks after smoking cessation in each case (n=12). (B) Each boxplot shows the values before smoking cessation and the values at 4, 8 and 12 weeks after smoking cessation (n=12). The Friedman test revealed no statistical significance. (C) Each boxplot shows the pre-smoking cessation value, the average of 4 and 8 weeks after smoking cessation, and the values at 12 weeks after smoking cessation (n=12). The Friedman test revealed no statistical significance. N.S. indicates no significant difference between the indicated pairs. (D) Each boxplot shows the pre-smoking cessation value, the peak level at 4 or 8 weeks after smoking cessation, and the values at 12 weeks after smoking cessation (n=12). *p<0.05, significant difference between the indicated…

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  Figure 3.. Effect of smoking cessation on the collagen- induced release of phosphorylated-HSP27 from human platelets: A comparison between the pre-smoking cessation value and the peak level at 4 or 8 weeks after smoking cessation. The net increase in phosphorylated-HSP27 released from collagen- stimulated (3 μg/mL) human platelets obtained from 12 patients before smoking cessation and the peak value of phosphorylated-HSP27 at 4 or 8 weeks after smoking cessation are shown in a line graph (A) and box plots (B). Values above the upper limit (20,000 pg/mL) are plotted as 20,000 pg/mL. For the statistical analysis, in values exceeded the upper limit, 20,000 pg/mL was adopted instead of the real value. All 12 cases are included in the statistical analysis. *p<0.05, significant difference between the indicated pairs. HSP27: heat shock protein 27

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  Figure 4.. Effect of smoking cessation on the collagen-stimulated phosphorylation of HSP27 in human platelets. PRP obtained at indicated points was stimulated by 3 μg/mL of collagen or vehicle for 5 min, and the reaction was terminated by the addition of ice-cold EDTA solution in the series of the previous study. The lysate of platelets was harvested and subjected to SDS-PAGE with a Western blot analysis using antibodies against phospho-specific HSP27 (Ser-78) or HSP27. The representative results obtained from patient No. 6 are presented. HSP27: heat shock protein 27, PRP: platelet-rich plasma, EDTA: ethylenediamine tetra acetic acid, SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis