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12-Hydroxyheptadecatrienoic acid promotes epidermal wound healing by accelerating keratinocyte migration via the BLT2 receptor.

Liu M, Saeki K, Matsunobu T, Okuno T, Koga T, Sugimoto Y, Yokoyama C, Nakamizo S, Kabashima K, Narumiya S, Shimizu T, Yokomizo T - J. Exp. Med. (2014)

Bottom Line: Leukotriene B4 (LTB4) receptor type 2 (BLT2) is a G protein-coupled receptor (GPCR) for 12(S)-hydroxyheptadeca-5Z,8E,10E-trienoic acid (12-HHT) and LTB4.Despite the well-defined proinflammatory roles of BLT1, the in vivo functions of BLT2 remain elusive.These results identify a novel mechanism underlying the action of the 12-HHT/BLT2 axis in epidermal keratinocytes and accordingly suggest the use of BLT2 agonists as therapeutic agents to accelerate wound healing, particularly for intractable wounds, such as diabetic ulcers.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry, Juntendo University School of Medicine, Tokyo 113-8421, Japan Department of Medical Biochemistry, Kyushu University, Fukuoka 812-8582, Japan.

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12-HHT and a synthetic BLT2 agonist enhance primary keratinocyte migration. (A) BrdU incorporation was assessed in primary epidermal keratinocytes obtained from WT and BLT2 KO mice (n = 5 mice per group). (B–D) Keratinocytes were cultured to confluency, mechanically wounded by scratching, and then incubated in medium containing the indicated reagents. (B) Representative fields show the wound gap filled by WT and BLT2 KO primary keratinocytes cultured in the presence of 1 nM 12-HHT at 0 and 60 h after scratching. Lines indicate remaining gap. (C and D) Quantification of mouse primary keratinocyte (C, n = 9 experimental replicates) and NHEK (D, n = 12 experimental replicates) migration. Data represent the mean ± SEM. **, P < 0.01; N.S., not significant (A, unpaired Student’s t test; C and D, two-way ANOVA). All the results are representative of at least two independent experiments.
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fig5: 12-HHT and a synthetic BLT2 agonist enhance primary keratinocyte migration. (A) BrdU incorporation was assessed in primary epidermal keratinocytes obtained from WT and BLT2 KO mice (n = 5 mice per group). (B–D) Keratinocytes were cultured to confluency, mechanically wounded by scratching, and then incubated in medium containing the indicated reagents. (B) Representative fields show the wound gap filled by WT and BLT2 KO primary keratinocytes cultured in the presence of 1 nM 12-HHT at 0 and 60 h after scratching. Lines indicate remaining gap. (C and D) Quantification of mouse primary keratinocyte (C, n = 9 experimental replicates) and NHEK (D, n = 12 experimental replicates) migration. Data represent the mean ± SEM. **, P < 0.01; N.S., not significant (A, unpaired Student’s t test; C and D, two-way ANOVA). All the results are representative of at least two independent experiments.

Mentions: To assess the role of the 12-HHT/BLT2 axis in keratinocyte migration during wound repair, an in vitro scratch assay was performed by using mouse and human primary keratinocytes. BLT2 deficiency did not affect primary keratinocyte proliferation (Fig. 5 A). Furthermore, in the absence of 12-HHT (Fig. 5 C, left), the wound closure rate was similar between BLT2 WT and BLT2 KO keratinocytes. However, BLT2 WT keratinocytes exhibited enhanced migration in the presence of 1 nM 12-HHT, whereas BLT2 KO keratinocytes did not (Fig. 5, B and C, right; Videos 1 and 2). This finding was confirmed in normal human epidermal keratinocytes (NHEKs). Both 12-HHT and a synthetic BLT2 agonist, CAY10583 (Iizuka et al., 2005), significantly enhanced NHEK migration relative to control, untreated cells (Fig. 5 D).


12-Hydroxyheptadecatrienoic acid promotes epidermal wound healing by accelerating keratinocyte migration via the BLT2 receptor.

Liu M, Saeki K, Matsunobu T, Okuno T, Koga T, Sugimoto Y, Yokoyama C, Nakamizo S, Kabashima K, Narumiya S, Shimizu T, Yokomizo T - J. Exp. Med. (2014)

12-HHT and a synthetic BLT2 agonist enhance primary keratinocyte migration. (A) BrdU incorporation was assessed in primary epidermal keratinocytes obtained from WT and BLT2 KO mice (n = 5 mice per group). (B–D) Keratinocytes were cultured to confluency, mechanically wounded by scratching, and then incubated in medium containing the indicated reagents. (B) Representative fields show the wound gap filled by WT and BLT2 KO primary keratinocytes cultured in the presence of 1 nM 12-HHT at 0 and 60 h after scratching. Lines indicate remaining gap. (C and D) Quantification of mouse primary keratinocyte (C, n = 9 experimental replicates) and NHEK (D, n = 12 experimental replicates) migration. Data represent the mean ± SEM. **, P < 0.01; N.S., not significant (A, unpaired Student’s t test; C and D, two-way ANOVA). All the results are representative of at least two independent experiments.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4042643&req=5

fig5: 12-HHT and a synthetic BLT2 agonist enhance primary keratinocyte migration. (A) BrdU incorporation was assessed in primary epidermal keratinocytes obtained from WT and BLT2 KO mice (n = 5 mice per group). (B–D) Keratinocytes were cultured to confluency, mechanically wounded by scratching, and then incubated in medium containing the indicated reagents. (B) Representative fields show the wound gap filled by WT and BLT2 KO primary keratinocytes cultured in the presence of 1 nM 12-HHT at 0 and 60 h after scratching. Lines indicate remaining gap. (C and D) Quantification of mouse primary keratinocyte (C, n = 9 experimental replicates) and NHEK (D, n = 12 experimental replicates) migration. Data represent the mean ± SEM. **, P < 0.01; N.S., not significant (A, unpaired Student’s t test; C and D, two-way ANOVA). All the results are representative of at least two independent experiments.
Mentions: To assess the role of the 12-HHT/BLT2 axis in keratinocyte migration during wound repair, an in vitro scratch assay was performed by using mouse and human primary keratinocytes. BLT2 deficiency did not affect primary keratinocyte proliferation (Fig. 5 A). Furthermore, in the absence of 12-HHT (Fig. 5 C, left), the wound closure rate was similar between BLT2 WT and BLT2 KO keratinocytes. However, BLT2 WT keratinocytes exhibited enhanced migration in the presence of 1 nM 12-HHT, whereas BLT2 KO keratinocytes did not (Fig. 5, B and C, right; Videos 1 and 2). This finding was confirmed in normal human epidermal keratinocytes (NHEKs). Both 12-HHT and a synthetic BLT2 agonist, CAY10583 (Iizuka et al., 2005), significantly enhanced NHEK migration relative to control, untreated cells (Fig. 5 D).

Bottom Line: Leukotriene B4 (LTB4) receptor type 2 (BLT2) is a G protein-coupled receptor (GPCR) for 12(S)-hydroxyheptadeca-5Z,8E,10E-trienoic acid (12-HHT) and LTB4.Despite the well-defined proinflammatory roles of BLT1, the in vivo functions of BLT2 remain elusive.These results identify a novel mechanism underlying the action of the 12-HHT/BLT2 axis in epidermal keratinocytes and accordingly suggest the use of BLT2 agonists as therapeutic agents to accelerate wound healing, particularly for intractable wounds, such as diabetic ulcers.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry, Juntendo University School of Medicine, Tokyo 113-8421, Japan Department of Medical Biochemistry, Kyushu University, Fukuoka 812-8582, Japan.

Show MeSH
Related in: MedlinePlus