Figures
- Figure 1–
The effect of cigarette smoke extract (CSE) on cell viability. The cytotoxicity of CSE on a) the mitochondrial activity and b) lactate dehydrogenase (LDH) release of human airway smooth muscle (ASM) cells was measured by Thaizolyl blue tetrazolium bromide (MTT) and LDH assays at an absorbance of 570 nm/690 nm and 490 nm/690 nm, respectively. Human ASM cells were stimulated with serial dilution of CSE for 72 h (n=4). Data are presented as mean±sem. One-way ANOVA plus Bonferroni post-test was used to determine statistical significance. *: p<0.05; **: p<0.01; ***: p<0.001, compared with control.
- Figure 2–
Release of CXCL8 and CXCL1 from human airway smooth muscle (ASM) cells. The concentrations of a, b) CXCL8 and c, d) CXCL1 in supernatant from human ASM cells from subjects with (n=9) and without (n=9) chronic obstructive pulmonary disease (COPD) after 72 h stimulation with cigarette smoke extract (CSE) or transforming growth factor (TGF)-β1 were measured by ELISA. Data are presented as mean±sem. Two-way ANOVA plus Bonferroni post-test was used to determine statistical significance. **: p<0.01, ***: p<0.001, compared with control.
- Figure 3–
Production of matrix metalloproteinases (MMPs) from human airway smooth muscle (ASM) cells. The concentrations of a ,b) MMP-1, c, d) MMP-2, e, f) MMP-3, and g, h) MMP-10 in supernatant from human ASM cells from subjects with (n=8) and without (n=7) chronic obstructive pulmonary disease (COPD) after 72 h stimulation with cigarette smoke extract (CSE) or transforming growth factor (TGF-β1) were measured by ELISA. Data are presented as mean±sem. Two-way ANOVA plus Bonferroni post-test was used to determine statistical significance. *: p<0.05; **: p<0.01; ***: p<0.001, compared with control. #: p<0.05, comparison between two groups.
- Figure 4–
Production of active matrix metalloproteinase (MMP)-1 from human airway smooth muscle (ASM) cells. The concentrations of active MMP-1 in the supernatants from human ASM cells from subjects with (n=5) and without (n=5) chronic obstructive pulmonary disease (COPD) after 72 h stimulation with cigarette smoke extract (CSE) or transforming growth factor (TGF)-β1 were measured by ELISA. Data are presented as mean±sem. Two-way ANOVA plus Bonferroni post-test was used to determine statistical significance. *: p<0.05; **: p<0.01, compared with control.
- Figure 5–
The deposition of collagen VIII alpha 1 (COL8A1) from human airway smooth muscle (ASM) cells. The deposited COL8A1 in the extracellular matrix (ECM) from human ASM cells from subjects with (n=7) and without (n=7) chronic obstructive pulmonary disease (COPD) after 72 h stimulation with cigarette smoke extract (CSE) or transforming growth factor (TGF)-β1 was measured by ECM ELISA at an absorbance of 450 nm/570 nm. Data are presented as mean±sem. Two-way ANOVA plus Bonferroni post-test was used to determine statistical significance. *: p<0.05; ***: p<0.001, compared with control. #: p<0.05, comparison between two groups.
- Figure 6–
Collagen VIII alpha 1 (COL8A1) airway tissue staining. a) COL8A1 in airway bronchus from subjects with (n=10) and without (n=7) chronic obstructive pulmonary disease (COPD) was measured by immunohistochemistry. Specific staining was detected using a chemical chromophore, DAB (brown), and the cell nucleus was counterstained with haematoxylin (blue). Tissue structure was stained with haematoxylin and eosin. Scale bars=100 μm. b) Immunostaining of COL8A1 in non-COPD and COPD groups were quantified by positive staining area and corrected with isotype control. Data are presented as median. The Mann–Whitney test was used to determine statistical significance. *: p<0.05.
- Figure 7–
The effect of cigarette smoke extract (CSE) on 2-h cell attachment at an absorbance of 595 nm. Cell attachment on a CSE treated plate of human airway smooth muscle cells from subjects with (n=5) and without (n=5) chronic obstructive pulmonary disease (COPD) were measured by toluidine blue assay. Data are presented as mean±sem. Two-way ANOVA plus Bonferroni post-test was used to determine statistical significance. *: p<0.05; **: p<0.01; ***: p<0.001, compared with control.
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Supplementary material
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Figures S1
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Figures S3
Figures S4
Figures S5
Figures S6
Figures S7 - Supplementary methods -
Immunohistochemistry
Cell attachment assay
Would healing assay - Supplementary tables -
Table S1
- Supplementary figures -
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