LXA4 actions direct fibroblast function and wound closure.
LXA4 actions direct fibroblast function and wound closure.
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DOI：
10.1016/j.bbrc.2015.07.076
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发表时间：
2015-09-04
影响因子：
3.1
通讯作者：
Van Dyke TE
中科院分区：
文献类型：
--
作者：
Herrera BS;Kantarci A;Zarrough A;Hasturk H;Leung KP;Van Dyke TE
及时解决炎症对于正常伤口愈合至关重要。炎症的消退是由包括脂氧素和消退素的专门脂质介质调节的主动生物过程。消退活性的失败对慢性炎性疾病中的伤口愈合具有主要的负面影响，其表现为过度纤维化和瘢痕形成。脂氧素，包括脂氧素A4（LXA 4），具有已知的抗纤维化和抗瘢痕形成特性。本研究的目的是阐明LXA 4对成纤维细胞功能的影响。将小鼠成纤维细胞（3 T3 Mus musculus Swiss）在TGF-β1存在下培养72小时，以诱导成纤维细胞活化。通过流式细胞术测定外源性TGF-β1（1 ng/mL）对LXA 4受体表达（ALX/FPR 2）的影响。通过溴脱氧尿苷（BrdU）标记和在“划痕”试验伤口模型中的迁移来测量成纤维细胞增殖。Western blot检测α-平滑肌肌动蛋白（α-SMA）、I型和III型胶原的表达。我们观察到TGF-β1上调LXA 4受体表达，增强成纤维细胞增殖、迁移和划痕伤口闭合。α-SMA水平和I型和III型胶原沉积也增加。LXA 4在早期时间点（24小时）减缓成纤维细胞迁移和划痕伤口闭合，但在48和72小时伤口闭合等同于单独的TGF-β1。在两种浓度下，LXA 4倾向于减缓成纤维细胞增殖，但对TGF-β1刺激的成纤维细胞产生的α-SMA或胶原蛋白没有影响。在使用消退素D2（RvD 2）作为激动剂重复的实验中检查了消退分子作用的普遍性。RvD 2的活性在所有测定中通过尚未鉴定的受体模拟LXA 4的作用。结果表明，炎症消退的介质通过调节成纤维细胞功能部分地促进伤口愈合并限制纤维化。
Timely resolution of inflammation is crucial for normal wound healing. Resolution of inflammation is an active biological process regulated by specialized lipid mediators including the lipoxins and resolvins. Failure of resolution activity has a major negative impact on wound healing in chronic inflammatory diseases that is manifest as excess fibrosis and scarring. Lipoxins, including Lipoxin A4 (LXA4), have known anti-fibrotic and anti-scarring properties. The goal of this study was to elucidate the impact of LXA4 on fibroblast function. Mouse fibroblasts (3T3 Mus musculus Swiss) were cultured for 72 hours in the presence of TGF-β1, to induce fibroblast activation. The impact of exogenous TGF-β1 (1 ng/mL) on LXA4 receptor expression (ALX/FPR2) was determined by flow cytometry. Fibroblast proliferation was measured by bromodeoxyuridine (BrdU) labeling and migration in a “scratch” assay wound model. Expression of α-smooth muscle actin (α-SMA), and collagen types I and III were measured by Western blot. We observed that TGF-β1 up-regulates LXA4 receptor expression, enhances fibroblast proliferation, migration and scratch wound closure. α-SMA levels and Collagen type I and III deposition were also enhanced. LXA4 slowed fibroblast migration and scratch wound closure at early time points (24 hours), but wound closure was equal to TGF-β1 alone at 48 and 72 hours. LXA4 tended to slow fibroblast proliferation at both concentrations, but had no impact on α-SMA or collagen production by TGF-β1 stimulated fibroblasts. The generalizability of the actions of resolution molecules was examined in experiments repeated with resolvin D2 (RvD2) as the agonist. The activity of RvD2 mimicked the actions of LXA4 in all assays, through an as yet unidentified receptor. The results suggest that mediators of resolution of inflammation enhance wound healing and limit fibrosis in part by modulating fibroblast function.