Volume 20 Issue 1
Jan.  2022
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Article Navigation >  Chinese Journal of General Practice  > 2022  >  20(1): 50-53
YANG Ming-feng, ZHANG Bin, YANG Yan-ying, WANG Yi-wen, GU Juan-fang, WANG Hong-zhi. Effect of doxycycline on the expression of alpha-SMA, fibronectin and collagen I in MRC-5 cell line induced by TGF-beta1[J]. Chinese Journal of General Practice, 2022, 20(1): 50-53. doi: 10.16766/j.cnki.issn.1674-4152.002274
Citation: YANG Ming-feng, ZHANG Bin, YANG Yan-ying, WANG Yi-wen, GU Juan-fang, WANG Hong-zhi. Effect of doxycycline on the expression of alpha-SMA, fibronectin and collagen I in MRC-5 cell line induced by TGF-beta1[J]. Chinese Journal of General Practice, 2022, 20(1): 50-53. doi: 10.16766/j.cnki.issn.1674-4152.002274
shu

Effect of doxycycline on the expression of alpha-SMA, fibronectin and collagen I in MRC-5 cell line induced by TGF-beta1

doi: 10.16766/j.cnki.issn.1674-4152.002274

  • Department of Rheumatology and Immunology, the First Hospital of Jiaxing (Affiliated Hospital of Jiaxing University), Jiaxing, Zhejiang 314000, China

Funds:

 LQ19H010001

 2019-ZC-03

  • Received Date: 2020-05-29
    Available Online: 2022-03-03
    Abstract
  •   Objective  To explore the effect of doxycycline (Doxy) on Transforming growth factor -β1 (TGF-β1) induced conversion in lung fibroblast, and to provide a theoretical basis for the treatment of pulmonary fibrosis.  Methods  Human embryonic lung fibroblast (MRC-5) were cultured in vitro, treated with different concentrations of doxycycline (0, 2.5, 5.0, 10.0, 20.0, 40.0, 80.0 μg/mL) for 72 hours, 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was used to detect the cell survival to determine the concentration of doxycycline. TGF-β1 was used to induce the conversion in MRC-5 cell, and treated with different concentrations of doxycycline (0, 2.5, 5.0, 10.0, 20.0 μg/mL) for 72 hours. Western blotting, RT-PCR, immunity fluorescence were used to detect the expression of α-smooth muscle actin (α-SMA), fibronectin (FN), collagen Ⅰ (Col Ⅰ).  Results  The MRC-5 cell viability decreased which the concentration of Doxy was 40.0 μg/mL and 80.0 μg/mL, relative to the control group were 0.93±0.03 and 0.83±0.08. qRT-PCR showed that the relative expressions of α-SMA, FN, and Col Ⅰ mRNA in the TGF-β1 10 ng/mL group were significantly higher than those in the control group (1.62±0.04 vs. 0.94±0.05, 4.33±0.28 vs. 0.98±0.06, 2.25±0.97 vs. 1.04±0.04, all P < 0.05), while the expressions of α-SMA, FN, and Col Ⅰ mRNA in the Doxy 10.0 μg/mL group and Doxy 20.0 μg/mL group were significantly lower than those in the TGF-β1 10 ng/mL group (0.70±0.15, 0.41±0.11 vs. 1.62±0.04; 1.19± 0.85, 0.79±0.06 vs. 4.33±0.28, 0.94±0.12, 0.52±0.07 vs. 2.25±0.97, all P < 0.05). Western blotting and immunofluorescence also found the same trend.  Conclusion  Doxy can exert anti-fibrosis effect by inhibiting the conversion of lung fibroblast induced by TGF-β1.

     

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