Page 63 - F. Cell biology

Arachidonic acid metabolite, 17,18-epoxyeicosatetraenoic acid, attenuates profibrotic signalling
in idiopathic pulmonary fibrosis

1
Sujin Moon , Miae Kim , Gi Won Hwang , Hyun Ju Yoo , Jin Woo Song 1
1
2
1
1 Department of Pulmonary and Critical Care Medicine, and, Convergence Medicine, Asan Medical Institute of Convergence Science and
2
Technology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea
BACKGROUND AIM
17,18-Epoxyeicosatetraenoic acid (EpETE) is a lipid metabolites from This study explored the 17(18)-Epoxyeicosatetranoic acid (EpETE)
dietary omega-3 polyunsaturated fatty acids such as eicosapentaenoic has anti-fibrotic effects in pulmonary fibrosis.
acid (EPA). 17,18-EpETE is the major active metabolite produced by
cytochrome p450 and inhibits biological activity by soluble epoxide
hydrolase(sEH). Moreover, 17,18-EpETE is a identified class of
antiallergic and anti-inflammatory lipid metabolite of
eicosapentaenoic acid. However, the underlying mechanisms of
17,18-EpETE and the role of 17,18-EpETE in diseases such as idiopathic
pulmonary fibrosis(IPF) remain unexplored.

METHODS

EpETEs were analyzed with Agilent 7890/5975 GC/MSD system and HP-5 MS column in the human lung tissues (IPF and controls). The role of
17(18)-EpETE was evaluated using in vitro pulmonary fibrosis models. The levels of protein or mRNA in cell lysates was measured by western
blot assays or RT-qPCR .

RESULTS

Figure 1. Figure 2.
Quantification of 17,18-EpETE in IPF (n=29) and control lung tissues (n=15) was 17,18- EpETE were analyzed with LC-MS/MS in the human plasma. (A)
performed using Agilent 7890/5975 GC/MSD system and HP-5 MS column. (B) The level of 17,18-EpETE in humans plasma of IPF(n=76) and
Quantification of 17,18-EpETE in IPF-survival, IPF-death and control lung tissues control(n=39). (B) The level of 17,18-EpETE in human plasma of IPF
was performed using Agilent 7890/5975 GC/MSD system and HP-5 MS column. survivor, IPF death and control.

(A) *** (B) (A) (B)
***
8 8 6 *
6 6 4
fmol/mg 4 fmol/mg 4 Amount of 17(18)-EpETE ( fmol / ul )
2 2 2
0 0 0
normal IPF Normal IPF-survival IPF-death IPF Control

Figure 3. Figure 4.
17(18)-EpETE effects of reduced TGF-β1-induced protein expression of 17(18)-EpETE inhibits activation of effect of reduced TGF-β1-induced
epithelial mesenchymal transition(EMT) related gene (A) and inhibits activation mRNA on epithelial mesenchymal transition(EMT) related gene(SLUG).
of fibroblast to myofibroblasts (B).
(A) (B)
2.0 5
17(18)-EpETE 0 6 0 6
17(18)-EpETE 0 6 0 6 1.5 4
5ng/ml TGF - - + +
5ng/ml TGF - - + + E.cadherin/beta-actin 1.0 SLUG/beta-actin 3
Fibronectin 2
Fibronectin 0.5 1
Col-1
E-cadherin 0.0 6uM 0
a-SMA Control TGF+Control TGF+6uM Control 6uM TGF+Control TGF+6uM
Beta-actin
a-actinin
CONCLUSION

Our results suggest that 17(18)-EpETE may have anti-fibrotic effects on pulmonary fibrosis by suppressing activation of
fibroblasts and EMT of epithelial cells.