| Part1: Changes of Cav-1, collagen-Ⅰandα-SMA proteins expression in patients with idiopathic pulmonary fibrosis【Abstract】Objective By investigating the different expression of Caveolin-1 (Cav-1) mRNA and protein, myofibroblast markersα-smooth muscle actin (α- smooth muscle actin,α-SMA), a product of the extracellular matrix: collagen -Ⅰprotein between idiopathic pulmonary fibrosis (Idiopathic pulmonary fibrosis, IPF) patients and normal control group, we preliminary study the relationship between Cav-1 and IPF.Methods IPF was diagnosed based on findings from history, physical examination, pulmonary function studies, chest high-resolution computed tomography and corroborated by open lung biopsy at the DrumTower Hospital., which were fulfilled the criteria of the American Thoracic Society and European Respiratory Society. The pathological diagnosis of IPF was usual interstitial pnermonia (UIP), which was characterized by diffuse interstitial fibrosis with only mild inflammation, honeycomb cysts and fibroblast foci. Six normal lung tissues were also obtained from patients with lung cancer by surgical resecetions. Expression of Cav-1 was detected by immunohistochemistry. In order to observe the relationship between Cav-1 and cell phenotypes, we determined the myofibroblasts makerα-SMA expression. Cav-1 mRNA was detected from total RNA extracted in lung tissues by RT-PCR. The expression of Cav-1α-SMA and Collagen-Ⅰproteins was measured by Western bolt.Results Cav-1 mRNA (0.05±0.018 vs 0.65±0.19) and Cav-1 protein expression in lungs of patients with IPF were markedly decreased (0.16±0.05 vs 0.81±0.10) (P <0.05). However, compared with the control groups, exprsession of collagen -Ⅰ(0.85±0.109vs0.16±0.045) andα-SMA (0.78±0.084 vs 0.14±0.053) proteins in lung tissue of IPF patients were significantly increased (P <0.05). Conclusion Cav-1 mRNA and protein expression in lung tissue of IPF patients was significantly decreased, while collagen -Ⅰandα-SMA expression increased, which suggested the decreased Cav-1 may be related to the development and progress of pulmonary fibrosis.Part2: Expression of caveolin-1 and extracellular matrix induced by transforming growth factor beta 1 in human fetal lung fibroblasts【Abstract】Objective By investigating the expression of caveolin-1, collagen-Ⅰa ndα-SMA in human fetal lung fibroblasts induced by transforming growth factor beta1(transforming growth factor beta1, TGF-β1), we in order to find the probable target for the treatment of pulmonary fibrosis.Methods With no TGF-β1-stimulated cells for the control group, human fetal lung fibroblasts (hman fetal lung fibroblast, HFLF) were cultured in vitro. Cultured cells were exposed to TGF-β1 with different final concentration (2, 5, 10μg/L). Caveolin-1 mRNA and protein, collagen-Ⅰa ndα-SMA proteins in cell (at least 5*10~8) lyses, measured by integral optical density, were detected at different points after the treatment of TGF-β1 by RT-PCR, Western blot and Immunohistochemistry. Each experiment was repeated three times.Results TGF-β1 reduced caveolin-1 mRNA and protein expressions in a dose- and time-dependent manner, measured by integral optical density. Compared with the control group (1.22±0.12 and 1.45±0.06), caveolin-1 mRNA and protein expressions (0.59±0.06 and 0.53±0.04) were significantly reduced, with statistical significance (F=29.279 and F=95.786, P<0.01), in cells exposed to TGF-β1 (5μg/L) for 12h. The collagen-Ⅰa ndα-SMA proteins expression (1.35±0.09 and 0.75±0.06) levels measured by integral optical density increased after stimulation, peaked at 24h in 5μg/L TGF-β1 group. At that point, the collagen-Ⅰandα-SMA proteins expressions were 4.2 and 4.8 times of the control group (0.28±0.04 and 0.18±0.04), (F=81.221 and F=65.477, P<0.01). Caveolin-1 expression had a negative correlation with collagen-Ⅰ(r=-0.9225, P<0.05) andα-SMA protein expression (r=-0.7929, P<0.05).Conclusion The reduced caveolin-1, which was negatively correlation with expressions of collagen-Ⅰa ndα-SMA proteins in HFLF cells exposed to TGF-β1, may be related to the development and progress of pulmonary fibrosis. The evidence between Cav-1 and IPF was offered by the experiments. |
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