Dioleoylphosphatidylglycerol Accelerates Corneal Epithelial Wound Healing
Dioleoylphosphatidylglycerol Accelerates Corneal Epithelial Wound Healing
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DOI：
10.1167/iovs.61.3.29
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发表时间：
2020-03-01
影响因子：
4.4
通讯作者：
Watsky, Mitchell
中科院分区：
文献类型：
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作者：
Bollag, Wendy B.;Olala, Lawrence O.;Watsky, Mitchell
目的.在与外界环境接触时，角膜很容易受伤。尽管角膜伤口通常快速愈合，但与受损角膜相关的疼痛和感染风险增加以及在一些个体中观察到的愈合受损强调了对加速角膜愈合的新型治疗的需要。我们先前在表皮角质形成细胞中证明了甘油通道水通道蛋白3（AQP 3）与磷脂酶D2（PLD 2）相互作用以产生信号磷脂磷脂酰甘油（PG），其已被证明加速体内皮肤伤口愈合。我们假设，相同的信号通路可能是在角膜上皮细胞中运作。我们使用免疫共沉淀，免疫组织化学，划痕伤口愈合试验在体外，和角膜上皮伤口愈合试验在体内，以确定AQP 3/PLD 2/PG信号通路在角膜上皮细胞中的作用。AQP 3存在于人角膜原位，并且AQP 3和PLD 2从角膜上皮细胞裂解物中共免疫沉淀。这两种蛋白质也可以从同时感染AQP 3和PLD 2表达杆状病毒的昆虫细胞中共免疫沉淀，表明可能存在直接相互作用。一种特殊的PG，二油酰磷脂酰甘油（DOPG），在体外增强角膜上皮单层的划痕伤口愈合。DOPG还加速了野生型小鼠和角膜伤口愈合受损的小鼠模型（AQP 3基因敲除小鼠）体内角膜上皮伤口愈合。这些结果表明AQP 3/PLD 2/PG信号通路在角膜上皮细胞中的重要性，并建议开发DOPG作为药物治疗以促进患者角膜伤口愈合的可能性。
PURPOSE. In contact with the external environment, the cornea can easily be injured. Although corneal wounds generally heal rapidly, the pain and increased risk of infection associated with a damaged cornea, as well as the impaired healing observed in some individuals, emphasize the need for novel treatments to accelerate corneal healing. We previously demonstrated in epidermal keratinocytes that the glycerol channel aquaporin-3 (AQP3) interacts with phospholipase D2 (PLD2) to produce the signaling phospholipid phosphatidylglycerol (PG), which has been shown to accelerate skin wound healing in vivo. We hypothesized that the same signaling pathway might be operational in corneal epithelial cells.METHODS. We used co-immunoprecipitation, immunohistochemistry, scratch wound healing assays in vitro, and corneal epithelial wound healing assays in vivo to determine the role of the AQP3/PLD2/PG signaling pathway in corneal epithelium.RESULTS. AQP3 was present in human corneas in situ, and AQP3 and PLD2 were coimmunoprecipitated from corneal epithelial cell lysates. The two proteins could also be co-immunoprecipitated from insect cells simultaneously infected with AQP3- and PLD2-expressing baculoviruses, suggesting a likely direct interaction. A particular PG, dioleoylphosphatidylglycerol (DOPG), enhanced scratch wound healing of a corneal epithelial monolayer in vitro. DOPG also accelerated corneal epithelial wound healing in vivo, both in wild-type mice and in a mouse model exhibiting impaired corneal wound healing (AQP3 knockout mice).CONCLUSIONS. These results indicate the importance of the AQP3/PLD2/PG signaling pathway in corneal epithelial cells and suggest the possibility of developing DOPG as a pharmacologic therapy to enhance corneal wound healing in patients.