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. 2015 Jan;28(1):37-41.
doi: 10.1293/tox.2014-0053. Epub 2014 Dec 27.

Lipopolysaccharide induces expression of collagen VI in the rat lung

Sayuri Okawa  1 , Kana Unuma  1 , Atsushi Yamada  1 , Toshihiko Aki  1 , Koichi Uemura  1
Affiliations

Lipopolysaccharide induces expression of collagen VI in the rat lung

Sayuri Okawa et al. J Toxicol Pathol. 2015 Jan.

Erratum in

  • Errata (Printer's correction).
    [No authors listed] [No authors listed] J Toxicol Pathol. 2016 Jan;29(1):74. Epub 2016 Feb 17. J Toxicol Pathol. 2016. PMID: 26989306 Free PMC article.

Abstract

The involvement of the lung during the septic systemic inflammatory response elicited by administration of lipopolysaccharide (LPS) was investigated. Eight-week-old male Sprague-Dawley rats were injected i.p. with 15 mg/kg LPS. After 24 h, the lungs were excised to evaluate the cellular responses to LPS. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) analysis revealed that type VI collagen (ColVI) was extremely upregulated during sepsis in the rat lung within the first 24 h of LPS administration. Upregulation of ColVI protein and its mRNA was demonstrated by Western blot analysis, real time PCR, and immunohistochemistry. To the best of our knowledge, this is the first report demonstrating the activation of ColVI in the rat lung at the early stage of systemic inflammation. Activation of ColVI might be involved in sepsis-mediated lung fibrosis at an early stage.

Keywords: LPS; lung; rat; type VI collagen.

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Figures

Fig. 1.
Fig. 1.
Increase of ColVI protein in LPS-treated rat lungs. (a) Major changes in the protein composition of the rat lung treated with LPS (15 mg/kg for 24 h) as shown by conventional electrophoresis (SDS-PAGE) and protein staining with Coomassie Brilliant Blue. The arrow indicates the major LPS-responsible protein. (b) Western blot analysis of the rat lung treated with LPS (15 mg/kg for 24 h) to determine the levels of ColVI and ColI. GAPDH was used as a loading control. (c) Ratios of ColVI, ColI, and GAPDH determined using densitometry analysis. Each bar represents the mean ± SE (n = 4). **, p < 0.01 versus control by Student’s t-test.
Fig. 2.
Fig. 2.
Activation of ColVI genes. Relative mRNA levels of ColI alpha1 (a), ColVI alpha1 (b), ColVI alpha2 (c), and ColVI alpha3 (d) to GAPDH were analyzed by qPCR. Each graph represents the mean ± SE (n = 4). **, p < 0.05 versus control by Student’s t-test.
Fig. 3.
Fig. 3.
Histological analysis of the rat lung treated with or without LPS. (a, b) Representative photographs of HE staining of control (a) and LPS-treated. (b) rats. (c, d) Immunohistochemical analysis of ColVI expression in the lungs of rats treated with LPS (d) and without LPS (c) for 24 h. Scale bars, 50 µm. The inset in (b) indicates infiltrated neutrophils (scale bar: 5 µm), while that in (d) indicates ColVI-positive interstitial space (scale bar: 4 µm). (e, f) Immunohistochemical analysis of α-SMA expression in the lungs of rats treated with LPS (f) and without LPS (e) for 24 h. Scale bars, 30 µm.
See this image and copyright information in PMC

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