Table 1.
Characteristics of volunteers.
Figure 1.
TGF-β1 increased the deposited and cellular FBLN-1 in human ASM cells.
After 72 hours stimulation with 10 ng/ml TGF-β1, the deposition of FBLN-1 by human ASM cells obtained from COPD (black bar, n = 10) and non-COPD (grey bar, n = 7) was measured by ELISA and data were expressed as absorbance at 450 nm–570 nm (panel A). Cellular FBLN-1 and GAPDH from COPD and non-COPD ASM cells were detected by western blot (panel B). Data from COPD (black bar, n = 8) and non-COPD (grey bar, n = 7) group were normalized to GAPDH and expressed as fold change relative to control (panel C). Data were expressed as mean ± SEM, two-way ANOVA with Bonferroni post tests, **P<0.01, ***P<0.001, compared with control.
Figure 2.
TGF-β1 decreased soluble FBLN-1 from human ASM cells.
Soluble FBLN-1 released from human ASM cells was detected by western blot, following 72 hours stimulation with 10 ng/ml TGF-β1 (panel A). Data from COPD (black bar, n = 9) group and non-COPD (grey bar, n = 9) group were expressed as fold change relative to control (panel B). Data were expressed as mean ± SEM and analysed by two-way ANOVA with Bonferroni post tests, ***P<0.001, compared with control.
Figure 3.
TGF-β1 decreased FBLN-1 gene expression in human ASM cells.
COPD (black bar, n = 7) and non-COPD (grey bar, n = 5) human ASM cells were stimulated with 10 ng/ml TGF-β1, and FBLN-1 mRNA expression was detected during the time course by real time PCR. Results were normalized to the endogenous control (18S rRNA), and expressed as fold change in FBLN-1 mRNA compared with time matched control. Data were expressed as mean ± SEM and analysed by two-way ANOVA with Bonferroni post tests, *P<0.01, ***P<0.001, compared with time matched control.
Figure 4.
Cycloheximide inhibited soluble FBLN-1 synthesis and down-regulated the deposition of FBLN-1 induced by TGF-β1.
After stimulation with 10 ng/ml TGF-β1 in the presence or absence of 0.5 µg/ml cycloheximide, the soluble FBLN-1 released from human ASM cells was detected by western blot (panel A). Data were normalized to cell number and results were expressed as the fold change compared with vehicle (panel B, n = 5). The deposition of FBLN-1 from human ASM cells in different treatments was measured by ECM ELISA and data were expressed as absorbance at 450 nm–570 nm (panel C, n = 5). Data were expressed as mean ± SEM and analysed by one-way ANOVA with Bonferroni’s multiple comparison test, **P<0.01, ***P<0.001. V: vehicle; CHX: cycloheximide; T: TGF-β1.
Figure 5.
FBLN-1 from a soluble source is incorporated into the ECM.
Cycloheximide treated and untreated human ASM cells were stimulated with or without 10 ng/ml TGF-β1 in quiescing medium or soluble FBLN-1 containing medium for 72 hours (n = 7). The incorporation of FBLN-1 into the matrix was detected by ECM ELISA and data were expressed as absorbance at 450 nm–570 nm. Data were expressed as mean ± SEM and analysed by one-way ANOVA with Bonferroni’s multiple comparison test, *P<0.05, ***P<0.001. V: vehicle; CHX: cycloheximide; S: soluble FBLN-1 containing medium; T: TGF-β1.