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Defective alterations in the collagen network to prostacyclin in COPD lung fibroblasts.

Larsson-Callerfelt AK, Hallgren O, Andersson-Sjöland A, Thiman L, Björklund J, Kron J, Nihlberg K, Bjermer L, Löfdahl CG, Westergren-Thorsson G - Respir. Res. (2013)

Bottom Line: The aim was to study the synthesis of prostacyclin and its effect on fibroblast activity and ECM production, and in particular collagen I and the collagen-associated proteoglycans biglycan and decorin.The prostacyclin analog iloprost was used to study the effect of prostacyclin on ECM protein synthesis, migration, proliferation and contractile capacity of fibroblasts.Iloprost reduced collagen I synthesis and fibroblast contractility but did not affect the collagen-associated proteoglycans or proliferation rate in fibroblasts from COPD patients.

View Article: PubMed Central - HTML - PubMed

Affiliation: Unit of Lung Biology, Department of Experimental Medical Sciences, BMC D12, Lund University, Lund 221 84, Sweden. Anna-Karin_L.Larsson@med.lu.se

ABSTRACT

Background: Prostacyclin analogs are potent vasodilators and possess anti-inflammatory properties. However, the effect of prostacyclin on extracellular matrix (ECM) in COPD is not well known. Collagen fibrils and proteoglycans are essential ECM components in the lung and fibroblasts are key players in regulating the homeostasis of ECM proteins. The aim was to study the synthesis of prostacyclin and its effect on fibroblast activity and ECM production, and in particular collagen I and the collagen-associated proteoglycans biglycan and decorin.

Methods: Parenchymal lung fibroblasts were isolated from lungs from COPD patients (GOLD stage IV) and from lungs and transbronchial biopsies from control subjects. The prostacyclin analog iloprost was used to study the effect of prostacyclin on ECM protein synthesis, migration, proliferation and contractile capacity of fibroblasts.

Results: TGF-β1 stimulation significantly increased prostacyclin synthesis in fibroblasts from COPD patients (p < 0.01), but showed no effect on fibroblasts from control subjects. Collagen I synthesis was decreased by iloprost in both control and COPD fibroblasts (p < 0.05). Conversely, iloprost significantly altered biglycan and decorin synthesis in control fibroblasts, but iloprost displayed no effect on these proteoglycans in COPD fibroblasts. Proliferation rate was reduced (p < 0.05) and contractile capacity was increased in COPD fibroblasts (p < 0.05) compared to control fibroblasts. Iloprost decreased proliferative rate in control fibroblasts (p < 0.05), whereas iloprost attenuated contraction capacity in both COPD (p < 0.01) and control fibroblasts (p < 0.05).

Conclusions: Iloprost reduced collagen I synthesis and fibroblast contractility but did not affect the collagen-associated proteoglycans or proliferation rate in fibroblasts from COPD patients. Enhanced prostacyclin production could lead to improper collagen network fibrillogenesis and a more emphysematous lung structure in severe COPD patients.

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Decorin production is altered by iloprost in fibroblasts from control subjects but not from COPD patients. Decorin production in distal lung fibroblasts from control subjects (n = 7; A) and patients with COPD (n = 7; B). Cells were stimulated with iloprost (10, 100 or 1000 nM) and in combination with TGF-β1 (10 ng/ml) in 0.4% fibroblast medium pretreated with indomethacin (3 μM). Changes in decorin production after TGF-β1 stimulation are expressed as fold change compared to non-stimulated fibroblasts in 0.4% medium from control subjects (C) and COPD patients (D). Data are presented as median with individual values and median values are represented as a line. *p < 0.05; **p < 0.01. Statistical analysis is performed with ANOVA on ranks followed by a post hoc test to compare differences between control and COPD fibroblasts and within the control and COPD fibroblasts after iloprost and TGF-β1 stimulation.
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Figure 4: Decorin production is altered by iloprost in fibroblasts from control subjects but not from COPD patients. Decorin production in distal lung fibroblasts from control subjects (n = 7; A) and patients with COPD (n = 7; B). Cells were stimulated with iloprost (10, 100 or 1000 nM) and in combination with TGF-β1 (10 ng/ml) in 0.4% fibroblast medium pretreated with indomethacin (3 μM). Changes in decorin production after TGF-β1 stimulation are expressed as fold change compared to non-stimulated fibroblasts in 0.4% medium from control subjects (C) and COPD patients (D). Data are presented as median with individual values and median values are represented as a line. *p < 0.05; **p < 0.01. Statistical analysis is performed with ANOVA on ranks followed by a post hoc test to compare differences between control and COPD fibroblasts and within the control and COPD fibroblasts after iloprost and TGF-β1 stimulation.

Mentions: Further studies were performed to evaluate if iloprost also affected the collagen-associated proteoglycans biglycan and decorin. There were no differences in either decorin or biglycan synthesis between fibroblasts from control subjects (n = 7) and patients with COPD (n = 7). Addition of TGF-β1 (10 ng/ml) did not change the decorin production (Figure 4C and 4D), whereas TGF-β1 enhanced the production of biglycan 6.6-fold in fibroblasts from control subjects (p < 0.01; Figure 5C) and 3.0-fold in fibroblasts from COPD patients (p < 0.05; Figure 5D) and the biglycan production after TGF-β1 stimulation was significantly higher in fibroblasts from control subjects compared to fibroblast from COPD patients (p < 0.05). Pretreatment with indomethacin did not affect proteoglycan production in lung fibroblasts from either control subjects or COPD patients (data not shown). Iloprost (100 nM and 1000 nM) significantly enhanced the synthesis of decorin (Figure 4A) and biglycan (Figure 5A) in fibroblasts from control subjects. Furthermore, after TGF-β1 treatment, iloprost significantly attenuated the synthesis of both decorin (Figure 4C) and biglycan (Figure 5C) in fibroblasts from control subjects. However, in fibroblasts from COPD patients there was no significant effect of iloprost treatment on either decorin (Figure 4B and D) or biglycan (Figure 5B and 5D) production before or after TGF-β1 stimulation.


Defective alterations in the collagen network to prostacyclin in COPD lung fibroblasts.

Larsson-Callerfelt AK, Hallgren O, Andersson-Sjöland A, Thiman L, Björklund J, Kron J, Nihlberg K, Bjermer L, Löfdahl CG, Westergren-Thorsson G - Respir. Res. (2013)

Decorin production is altered by iloprost in fibroblasts from control subjects but not from COPD patients. Decorin production in distal lung fibroblasts from control subjects (n = 7; A) and patients with COPD (n = 7; B). Cells were stimulated with iloprost (10, 100 or 1000 nM) and in combination with TGF-β1 (10 ng/ml) in 0.4% fibroblast medium pretreated with indomethacin (3 μM). Changes in decorin production after TGF-β1 stimulation are expressed as fold change compared to non-stimulated fibroblasts in 0.4% medium from control subjects (C) and COPD patients (D). Data are presented as median with individual values and median values are represented as a line. *p < 0.05; **p < 0.01. Statistical analysis is performed with ANOVA on ranks followed by a post hoc test to compare differences between control and COPD fibroblasts and within the control and COPD fibroblasts after iloprost and TGF-β1 stimulation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3585859&req=5

Figure 4: Decorin production is altered by iloprost in fibroblasts from control subjects but not from COPD patients. Decorin production in distal lung fibroblasts from control subjects (n = 7; A) and patients with COPD (n = 7; B). Cells were stimulated with iloprost (10, 100 or 1000 nM) and in combination with TGF-β1 (10 ng/ml) in 0.4% fibroblast medium pretreated with indomethacin (3 μM). Changes in decorin production after TGF-β1 stimulation are expressed as fold change compared to non-stimulated fibroblasts in 0.4% medium from control subjects (C) and COPD patients (D). Data are presented as median with individual values and median values are represented as a line. *p < 0.05; **p < 0.01. Statistical analysis is performed with ANOVA on ranks followed by a post hoc test to compare differences between control and COPD fibroblasts and within the control and COPD fibroblasts after iloprost and TGF-β1 stimulation.
Mentions: Further studies were performed to evaluate if iloprost also affected the collagen-associated proteoglycans biglycan and decorin. There were no differences in either decorin or biglycan synthesis between fibroblasts from control subjects (n = 7) and patients with COPD (n = 7). Addition of TGF-β1 (10 ng/ml) did not change the decorin production (Figure 4C and 4D), whereas TGF-β1 enhanced the production of biglycan 6.6-fold in fibroblasts from control subjects (p < 0.01; Figure 5C) and 3.0-fold in fibroblasts from COPD patients (p < 0.05; Figure 5D) and the biglycan production after TGF-β1 stimulation was significantly higher in fibroblasts from control subjects compared to fibroblast from COPD patients (p < 0.05). Pretreatment with indomethacin did not affect proteoglycan production in lung fibroblasts from either control subjects or COPD patients (data not shown). Iloprost (100 nM and 1000 nM) significantly enhanced the synthesis of decorin (Figure 4A) and biglycan (Figure 5A) in fibroblasts from control subjects. Furthermore, after TGF-β1 treatment, iloprost significantly attenuated the synthesis of both decorin (Figure 4C) and biglycan (Figure 5C) in fibroblasts from control subjects. However, in fibroblasts from COPD patients there was no significant effect of iloprost treatment on either decorin (Figure 4B and D) or biglycan (Figure 5B and 5D) production before or after TGF-β1 stimulation.

Bottom Line: The aim was to study the synthesis of prostacyclin and its effect on fibroblast activity and ECM production, and in particular collagen I and the collagen-associated proteoglycans biglycan and decorin.The prostacyclin analog iloprost was used to study the effect of prostacyclin on ECM protein synthesis, migration, proliferation and contractile capacity of fibroblasts.Iloprost reduced collagen I synthesis and fibroblast contractility but did not affect the collagen-associated proteoglycans or proliferation rate in fibroblasts from COPD patients.

View Article: PubMed Central - HTML - PubMed

Affiliation: Unit of Lung Biology, Department of Experimental Medical Sciences, BMC D12, Lund University, Lund 221 84, Sweden. Anna-Karin_L.Larsson@med.lu.se

ABSTRACT

Background: Prostacyclin analogs are potent vasodilators and possess anti-inflammatory properties. However, the effect of prostacyclin on extracellular matrix (ECM) in COPD is not well known. Collagen fibrils and proteoglycans are essential ECM components in the lung and fibroblasts are key players in regulating the homeostasis of ECM proteins. The aim was to study the synthesis of prostacyclin and its effect on fibroblast activity and ECM production, and in particular collagen I and the collagen-associated proteoglycans biglycan and decorin.

Methods: Parenchymal lung fibroblasts were isolated from lungs from COPD patients (GOLD stage IV) and from lungs and transbronchial biopsies from control subjects. The prostacyclin analog iloprost was used to study the effect of prostacyclin on ECM protein synthesis, migration, proliferation and contractile capacity of fibroblasts.

Results: TGF-β1 stimulation significantly increased prostacyclin synthesis in fibroblasts from COPD patients (p < 0.01), but showed no effect on fibroblasts from control subjects. Collagen I synthesis was decreased by iloprost in both control and COPD fibroblasts (p < 0.05). Conversely, iloprost significantly altered biglycan and decorin synthesis in control fibroblasts, but iloprost displayed no effect on these proteoglycans in COPD fibroblasts. Proliferation rate was reduced (p < 0.05) and contractile capacity was increased in COPD fibroblasts (p < 0.05) compared to control fibroblasts. Iloprost decreased proliferative rate in control fibroblasts (p < 0.05), whereas iloprost attenuated contraction capacity in both COPD (p < 0.01) and control fibroblasts (p < 0.05).

Conclusions: Iloprost reduced collagen I synthesis and fibroblast contractility but did not affect the collagen-associated proteoglycans or proliferation rate in fibroblasts from COPD patients. Enhanced prostacyclin production could lead to improper collagen network fibrillogenesis and a more emphysematous lung structure in severe COPD patients.

Show MeSH
Related in: MedlinePlus