Effect of all-trans-retinoic acid on expression of collagen Ⅲ, STAT3 and PIAS3 in TGF-β1-stimulated human lung fibroblasts
XIA Wu1, YANG Yu-ping1, CHEN Yong-feng2, CHENG Na1, LIU Ju-yuan1
1. Department of Pharmacology, School of Pharmacy, the Third Affiliated Hospital, Xinxiang Medical University, Xinxiang 453003, China;
2. Department of Respiratory Medicine, the Third Affiliated Hospital, Xinxiang Medical University, Xinxiang 453003, China
Abstract：AIM: To investigate the effects of all-trans-retinoic acid (ATRA) on the expression of collagen Ⅲ, signal transducer and activator of transcription 3(STAT3) and protein inhibitor of activated STAT3 (PIAS3) in TGF-β1-stimulated human lung fibroblasts (HFL-I). METHODS: Addition of TGF-β1 at dose of 5 μg/L were conducted to stimulate the cultured HFL-I cells. The mRNA expression of collagen Ⅲ, STAT3 and PIAS3 in the cells was determined by RT-PCR at the time of induction with TGF-β1 for 0 h, 6 h, 12 h, 24 h, 48 h and 72 h. Western blotting was also used to detect the protein levels of STAT3 and p-STAT3 at the time of induction with TGF-β1 for 1 d, 3 d and 5 d. After stimulated with TGF-β1, HFL-I cells were treated with ATRA at concentrations of 10 μmol/L, 1 μmol/L and 0.1 μmol/L. The mRNA expression of collagen Ⅲ, STAT3 and PIAS3 was detected by RT-PCR 24 h after ATRA treatment, and the protein levels of STAT3 and p-STAT3 were also assayed by Western blotting 3 d after ATRA treatment. RESULTS: The results of RT-PCR and Western blotting showed that TGF-β1 promoted the mRNA expression of collagen Ⅲ and STAT3, increased the protein levels of STAT3 and p-STAT3, and reduced the mRNA expression of PIAS3 in HFL-I cells (P<0.05). ATRA at different concentrations decreased the mRNA expression of collagen Ⅲ and STAT3, and protein levels of STAT3 and p-STAT3 in TGF-β1-stimulated HFL-I cells (P<0.05). Meanwhile, the mRNA expression of PIAS3 was obviously increased (P<0.05). CONCLUSION: ATRA prevents the progress of pulmonary fibrosis by down-regulating the expression of collagen Ⅲ and STAT3, and up-regulating the PIAS3 expression.
Gharaee-Kermani M, Gyetko MR, Hu B, et al. New insights into the pathogenesis and treatment of idiopathic pulmonary fibrosis:a potential role for stem cells in the lung parenchyma and implications for therapy[J]. Pharm Res, 2007, 24(5):819-841.
[2]
Vancheri C, Failla M, Crimi N, et al. Idiopathic pulmonary fibrosis:a disease with similarities and links to cancer biology[J]. Eur Respir J, 2010, 35(3):496-504.
[3]
Aluwihare P, Munger JS. What the lung has taught us about latent TGF-β activation[J]. Am J Respir Cell Mol Biol, 2008, 39(5):499-502.
[4]
Corvol H, Flamein F, Epaud R, et al. Lung alveolar epithelium and interstitial lung disease[J]. Int J Biochem Cell Biol, 2009, 41(8-9):1643-1651.
[5]
Vancheri C, Gili E, Failla M, et al. Bradykin in differentiates human lung fibroblasts to a myofibroblast phenotype via the B2 receptor[J]. J Allergy Clin Immunol, 2005, 116(6):1242-1248.
Guo W, Shan B, Klingsberg RC, et al. Abrogation of TGF-β1-induced fibroblast-myofibroblast differentiation by histone deacetylase inhibition[J]. Am J Physiol Lung Cell Mol Physiol, 2009, 297(5):L864-L870.
[8]
Bartram U, Speer CP. The role of transforming growth factor beta in lung development and disease[J]. Chest, 2004, 125(2):754-765.
[9]
Phan SH. The myofibroblast in pulmonary fibrosis[J]. Chest, 2002, 122(6): 286-289.
[10]
Lai CK,Wallace WD,Fishbein MC. Histopathology of pulmonary fibrotic disorders[J]. Semin Respir Crit Care Med, 2006, 27(6):613-622.
[11]
Ke S. Regulation of cytokine signaling pathways by PIAS proteins[J]. Cell Res, 2006, 16(2): 196-202.
[12]
Pang M, Ma L, Gong R, et al. A novel STAT3 inhibitor, S3I-201, attenuates renal interstitial fibroblast activation and interstitial fibrosis in obstructive nephropathy[J]. Kidney Int, 2010, 78(3):257-268.
[13]
Bromberg J, Wang TC.Inflammation and cancer:IL-6 and STAT3 complete the link[J]. Cancer Cell, 2009, 15(2):79-80.
