Objective To investigate the effects of caveolin-1 scaffolding domain peptide ( CSD-p)on expressions of extracellular matrix and Smads in human fetal lung fibroblasts. Methods Human fetal lung fibroblasts were cultured in vitro and divided into four groups. A control group: the cells were cultured in DMEMwithout TGF-β1 or CSD-p. A CSD-p treatment group: the cells were cultured in DMEMcontaining 5 μmol /L CSD-p. A TGF-β1 treatment group: the cells were cultured in DMEMcontaining 5 μg/L TGF-β1 .A TGF-β1 + CSD-p treatment group: the cells were cultured in DMEM containing 5 μg/L TGF-β1 and 5 μmol /L CSD-p. Caveolin -1 mRNA was detected by RT-PCR. Caveolin-1, collagen-Ⅰ, α-SMA, p-Smad2,p-Smad3 and Smad7 proteins were measured by Western blot.
Results Compared with the control group,the Caveolin -1 mRNA and protein expressions in the cells of TGF-β1 group significantly reduced ( mRNA:0. 404 ±0. 027 vs. 1. 540 ±0. 262; protein: 0. 278 ±0. 054 vs. 1. 279 ±0. 085; P lt; 0. 01) , and the expression levels of collagen-Ⅰ and α-SMA proteins significantly increased ( collagen-Ⅰ: 1. 127 ±0. 078 vs.0. 234 ±0. 048; α-SMA: 1. 028 ±0. 058 vs. 0. 295 ±0. 024) . Meanwhile, the expression levels of p-Smad2 ( 1. 162 ±0. 049 vs. 0. 277 ±0. 014) and p-Smad3 proteins ( 1. 135 ±0. 057 vs. 0. 261 ±0. 046) increased with statistical significance ( P lt; 0. 01) , but the expression level of Smad7 protein significantly reduced
( 0. 379 ±0. 004 vs. 1. 249 ±0. 046, P lt;0. 001) . In the CSD-p group, CSD-p had no significant effects on the expressions of above proteins compared with the control group. But in the TGF-β1 +CSD-p group, the overexpressions of collagen-Ⅰ, α-SMA, p-Smad2 and p-Smad3 induced by TGF-β1 were obviously inhibited by CSD-p ( collagen-Ⅰ: 0. 384 ±0. 040 vs. 1. 127 ±0. 078; α-SMA: 0. 471 ±0. 071 vs. 1. 127 ±0. 078;p-Smad2: 0. 618 ±0. 096 vs. 1. 162 ±0. 049; p-Smad3: 0. 461 ±0. 057 vs. 1. 135 ±0. 057; P lt; 0. 01) .Otherwise, the up-regulation of Smad7 ( 0.924 ±0. 065 vs. 0.379 ±0. 004) was found.
Conclusions CSD-p can reduce fibroblast collagen-I and α-SMA protein expressions stimulated by TGF-β1 , possibly through regulation of TGF-β1 /Smads signaling pathway. It is suggested that an increase in caveolin -1 function through the use of CSD-p may be an intervention role in pulmonary fibrosis.
Citation: CAO Ming,DING Hui,ZHOU Yihua,CAI Hourong.. Effects of Caveolin-1 Scaffolding Domain Peptide on Expressions of Extracellular Matrix and Smads in Human Fetal Lung Fibroblasts. Chinese Journal of Respiratory and Critical Care Medicine, 2011, 10(5): 442-445. doi: Copy