Cellular mechanisms involved in the pathogenesis of airway remodeling in chronic lung disease

Figures & data

Table 1. Primers used for reverse transcription quantitative PCR.

Gene	Forward primer	Reverse primer
18s	5′-TGAAATATCCAGAACATCTTA-3′	5′-GCAAAATTTATTGTCCCATCAT-3′
CRP	5′-GTGTTTCCCAAAGAGTCGGATA-3′	5′-CCACGGGTCGAGGACAGTT-3′
SAA1	5′-ATCAGCGATGCCAGAGAGAAT-3′	5′-GTGATTGGGGTCTTTGCCA-3′
SAA2	5′-AGCCAATTACATCGGCTCAG-3′	5′-ATTTATTGGCAGCCTGATCG-3′
SAA4	5’-GTCCAACGAGAAAGCTGAGG-3’	5’-AGTGACCCTGTGTCCCTGTC-3’
MCP1	5´-CCCCAGTCACCTGCTGTTAT-3´	5´-TGGAATCCTGAACCCACTTC-3´
NF-κB1	5´-CCTGGATGACTCTTGGGAAA-3´	5´-TCAGCCAGCTGTTTCATGGTC-3´
MMP9	5´-TTGACAGCGACAAGAAGTGG-3´	5´-GCCATTCACGTCGTCCTTAT-3´
MMP12	5´-ACACATTTCGCCTCTCTGCT-3´	5´-CCTTCAGCCAGAAGAACCTG-3´
IL-8	5´-CAGGAATTGAATGGGTTTGC-3´	5´-AAACCAAGGCACAGTGGAAC-3´

18s: 18s ribosome; CRP: C-reactive protein; SAA: Serum Amyloid A; MCP1: Monocyte chemoattractant protein 1; NF-κB1: Nuclear factor kappa-B1; MMP: Matrix Metalloproteinase; IL-8: Interleukin-8.

Figure 1. Effect of IL-1β on inflammatory mediator expression in lung fibroblasts and airway epithelial cells. mRNA expression of IL-8 (panel A), mRNA expression of MCP-1 (panel B), protein levels of IL-8 (panel C) and protein levels of MCP-1 (panel D) produced by human lung fibroblasts without stimulation (control, white bars) and stimulated with IL-1β (10 ng/mL, black bars). mRNA expression of IL-8 (panel E), mRNA expression of MCP-1 (panel F), protein levels of IL-8 (panel G) and protein levels of MCP-1 (panel H) produced by human airway epithelial cells without stimulation (control, white bars) and stimulated with IL-1β (10 ng/mL, black bars). r.u. = Relative units versus control; *p < 0.05 versus control. Each bar in the figure is mean± SE n = 6.

Figure 2. Effect of IL-1β on SAA and CRP expression in lung fibroblasts and airway epithelial cells. mRNA expression of SAA1 (panel A), SAA2 (panel B), and SAA4 (panel C) produced by human lung fibroblasts without stimulation (white bars) and stimulated with IL-1β (10 ng/mL, black bars). mRNA expression of SAA (panel D), CRP (panel E) and SAA protein levels (panel D) produced by human airway epithelial cells without stimulation (white bars) and stimulated with IL-1β (10 ng/mL, black bars). r.u. = Relative units versus control; *p < 0.05 versus control. Each bar in the figure is mean± SE n = 6.

Figure 3. Upregulation of matrix metalloproteinase mRNA expression by IL-1β in lung fibroblasts and airway epithelial cells. Expression of MMP9 (panel A) and MMP12 (panel B) mRNA by human lung fibroblasts without stimulation (white bars) and stimulated with IL-1β (10 ng/mL, black bars). Expression of MMP9 (panel C) and MMP12 (panel D) mRNA by human lung airway epithelial cells without stimulation (white bars) and stimulated with IL-1β (10 ng/mL, black bars). r.u. = Relative units versus control; *p < 0.05 versus control. Each bar in the figure is mean± SE n = 6.

Figure 4. Effect of hypoxia on SAA expression in lung fibroblasts. mRNA expression of SAA1 and SAA2 produced by human lung fibroblasts, in normoxia (white bars) and stimulated with hypoxia (black bars). r.u. = Relative units versus control; *p < 0.05 versus normoxia. Each bar in the figure is mean± SE n = 6.

Figure 5. Effect of simvastatin on SAA expression in lung fibroblasts. mRNA expression of SAA1 and SAA2 produced by human lung fibroblasts, without stimulation (white bars) and stimulated with simvastatin (30 µM, black bars). r.u. = Relative units versus control; *p < 0.05 versus control. Each bar in the figure is mean± SE n = 6.

Figure 6. Responses of airway epithelial cells and lung fibroblasts after inflammatory stimuli associated with chronic lung disease. IL-1β, interleukin 1β; IL-8, interleukin 8; MCP1, monocyte chemoattractant protein-1; NF-κB1, factor-κB1; CRP, C-reactive protein; SAA, serum amyloid A; MM9 and MMP12, proteases matrix metalloproteinase 9 and 12.