Volume 20 Issue 1
Jan.  2022
Turn off MathJax
Article Contents
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.

     

    • FullText(HTML)
  • loading
    • References(15)
  • [1]
    HUANG L S, SUDHADEVI T, FU P, et al. Sphingosine Kinase 1/S1P signaling contributes to pulmonary fibrosis by activating Hippo/YAP pathway and mitochondrial reactive oxygen species in lung fibroblasts[J]. Int J Mol Sci, 2020, 21(6): 2064. doi: 10.3390/ijms21062064
    [2]
    GLASS D S, GROSSFELD D, RENNA H A, et al. Idiopathic pulmonary fibrosis: Molecular mechanisms and potential treatment approaches[J]. Respir Investig, 2020, 58(5): 320-335. doi: 10.1016/j.resinv.2020.04.002
    [3]
    MA J, SANCHEZ-DUFFHUES G, GOUMANS M J, et al. TGF-β-Induced endothelial to mesenchymal transition in disease and tissue engineering[J]. Front Cell Dev Biol, 2020, 8: 260. doi: 10.3389/fcell.2020.00260
    [4]
    SHIN J M, PARK J H, PARK I H, et al. Doxycycline inhibits TGF-β1-induced extracellular matrix production in nasal polyp-derived fibroblasts[J]. Int Forum Allergy Rhinol, 2016, 6(3): 256-263. doi: 10.1002/alr.21660
    [5]
    张凯, 马作旺, 刘瑞蒙, 等. 多西环素对慢性间歇缺氧所致的心房重构的改善作用[J]. 中华老年心脑血管病杂志, 2018, 20(10): 1074-1079 doi: 10.3969/j.issn.1009-0126.2018.10.017
    [6]
    WANG W, ZHANG K, LI X, et al. Doxycycline attenuates chronic intermittent hypoxia-induced atrial fibrosis in rats[J]. Cardiovasc Ther, 2018, 36(3): e12321. doi: 10.1111/1755-5922.12321
    [7]
    CHOI J S, KIM J M, KIM J W, et al. Prevention of tracheal inflammation and fibrosis using nitinol stent coated with doxycycline[J]. Laryngoscope, 2018, 128(7): 1558-1563. doi: 10.1002/lary.27028
    [8]
    HUA X F, LI X H, LI M M, et al. Doxycycline attenuates paraquat-induced pulmonary fibrosis by downregulating the TGF-β signaling pathway[J]. J Thorac Dis, 2017, 9(11): 4376-4386. doi: 10.21037/jtd.2017.10.42
    [9]
    周滟如, 庞昶, 陈思露, 等. 多西环素通过TGF-β1/Smad通路抑制百草枯诱导的肺纤维化研究[J]. 中华劳动卫生职业病杂志, 2019, 37(5): 337-341. doi: 10.3760/cma.j.issn.1001-9391.2019.05.004
    [10]
    ZHANG F, HU L, WU Y X, et al. Doxycycline alleviates paraquat-induced acute lung injury by inhibiting neutrophil-derived matrix metalloproteinase 9[J]. Int Immunopharmacol, 2019, 72: 243-251. doi: 10.1016/j.intimp.2019.04.015
    [11]
    MARSHALL D C, SALCICCIOLI J D, SHEA B S, et al. Trends in mortality from idiopathic pulmonary fibrosis in the European Union: An observational study of the WHO mortality database from 2001-2013[J]. Eur Respir J, 2018, 51(1): 1701603. doi: 10.1183/13993003.01603-2017
    [12]
    WONG A W, RYERSON C J, GULER S A. Progression of fibrosing interstitial lung disease[J]. Respir Res, 2020, 21(1): 32. doi: 10.1186/s12931-020-1296-3
    [13]
    ROUT-PITT N, FARROW N, PARSONS D, et al. Epithelial mesenchymal transition (EMT): A universal process in lung diseases with implications for cystic fibrosis pathophysiology[J]. Respir Res, 2018, 19(1): 136. doi: 10.1186/s12931-018-0834-8
    [14]
    FROESE A R, SHIMBORI C, BELLAYE P S, et al. Stretch-induced activation of transforming growth Factor-β1 in pulmonary fibrosis[J]. Am J Respir Crit Care Med, 2016, 194(1): 84-96. doi: 10.1164/rccm.201508-1638OC
    [15]
    ZEGLINSKI M R, HNATOWICH M, JASSAL D S, et al. SnoN as a novel negative regulator of TGF-β/Smad signaling: A target for tailoring organ fibrosis[J]. Am J Physiol Heart Circ Physiol, 2015, 308(2): H75-H82. doi: 10.1152/ajpheart.00453.2014
    • Relative Articles
    • Supplements(0)
    • Cited By
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Figures(4)

    Get Citation
    PDF
    XML

    Article Metrics

    Article views (1209) PDF downloads(29) Cited by()
    Proportional views
    Related

    Address：No. 287, Changhuai Road, Bengbu, Anhui Province, 233004, P.R.ChinaphoneNo：0552-3066635； 0552-3051890Email：zhqkyx@163.com

    Copyright © Chinese Journal of General Practice皖ICP备2020018345号-2

    Supported by: Beijing Renhe Information Technology Co., Ltd.    

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return