Justet Cristian, Evans Frances, Torriglia Alicia, Chifflet Silvia
Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Gral. Flores 2125, 11800, Montevideo, Uruguay.
Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Gral. Flores 2125, 11800, Montevideo, Uruguay.
Cell Tissue Res. 2015 Dec;362(3):557-68. doi: 10.1007/s00441-015-2223-7. Epub 2015 Jun 18.
Tissue injury triggers a complex network of cellular and molecular responses. Although cell migration and proliferation are the most conspicuous, several other responses, such as apoptosis and increased protease activity, are necessary for a proper restitution of the tissue. In this work, we study the leukocyte elastase inhibitor (LEI) expression during wound healing of bovine corneal endothelial monolayers in culture. LEI is a multifunctional protein with anti-protease and anti-apoptotic activity. When properly cleaved, it is transformed into L-DNase II, a pro-apoptotic enzyme and translocated to the nucleus. We found that early after injury LEI increases its protein and mRNA expressions, without nuclear translocation and returns to basal levels immediately after wound closure. This increase is blocked by N-acetylcysteine, suggesting that production of reactive oxygen species immediately after wounding is involved in the LEI increase. Another finding of this work is that there is an acidification of the cells at the wound border which, in contrast to other cell types, does not determine nuclear translocation of the protein. Taken together, the results of this work suggest that the function of LEI during wound healing is related to its activity as a protease inhibitor and/or to its anti-apoptotic activity.
组织损伤引发细胞和分子反应的复杂网络。尽管细胞迁移和增殖最为显著，但其他几种反应，如细胞凋亡和蛋白酶活性增加，对于组织的正常修复也是必要的。在这项研究中，我们研究了培养的牛角膜内皮单层伤口愈合过程中白细胞弹性蛋白酶抑制剂（LEI）的表达。LEI是一种具有抗蛋白酶和抗凋亡活性的多功能蛋白。当被适当切割时，它会转化为L-DNase II，一种促凋亡酶并转移至细胞核。我们发现，损伤后早期LEI的蛋白质和mRNA表达增加，无核转位现象，伤口闭合后立即恢复至基础水平。这种增加被N-乙酰半胱氨酸阻断，表明受伤后立即产生的活性氧参与了LEI的增加。这项研究的另一个发现是伤口边缘的细胞出现酸化，与其他细胞类型不同，这并未导致该蛋白的核转位。综上所述，这项研究结果表明，LEI在伤口愈合过程中的功能与其作为蛋白酶抑制剂的活性和/或抗凋亡活性有关。
