Increased apoptosis and abnormal wound-healing responses in the heterozygous Pax6+/- mouse cornea
Increased apoptosis and abnormal wound-healing responses in the heterozygous Pax6+/- mouse cornea
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DOI：
10.1167/iovs.05-1028
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发表时间：
2006-05-01
影响因子：
4.4
通讯作者：
West, John D.
中科院分区：
文献类型：
--
作者：
Ramaesh, Thaya;Ramaesh, Kanna;West, John D.
目的.角膜伤口愈合涉及上皮和基质之间的级联相互作用。Pax 6上调，早期事件包括上皮细胞迁移和浅表角膜细胞凋亡。小鼠杂合Pax 6（Pax 6（+/-））角膜表型模拟人类无虹膜相关性角膜病变（方舟），并且伤口愈合的某些方面已被证明是异常的，包括基质金属蛋白酶（MMP）-9表达。本研究的目的是检测Pax 6（+/-）基因型是否影响角膜创伤愈合反应，包括基质细胞凋亡、上皮细胞迁移率和基质金属蛋白酶分泌。处死Pax 6（+/-）和野生型（Pax 6（+/+））小鼠，通过上皮清创术损伤它们的角膜。在器官培养中培养全眼，分别通过局部荧光素染色和TUNEL定量角膜上皮愈合率和角膜细胞凋亡。分离Pax 6（+/-）和野生型小鼠的角膜上皮细胞进行培养，并通过酶谱法测定分泌的MMP-9的活性。对于较大的伤口和Pax 6（+/-）角膜，前6小时的伤口愈合率明显更快。与野生型相比，受伤的Pax 6（+/-）眼睛表现出明显更多的基质细胞凋亡，培养的Pax 6（+/-）角膜上皮细胞产生较低的MMP-9活性。结论。以基质细胞凋亡增加和MMP-9分泌水平降低为特征的异常伤口愈合反应的累积效应可能导致Pax 6（+/-）小鼠的角膜变化。基质细胞凋亡和其他异常的伤口愈合反应升高方舟的可能贡献进行了讨论。
PURPOSE. Corneal wound healing involves a cascade of interactions between the epithelium and stroma. Pax6 is upregulated, and early events include epithelial cell migration and apoptosis of superficial keratocytes. The mouse heterozygous Pax6 (Pax6(+/-)) corneal phenotype mimics human aniridia-related keratopathy ( ARK), and some aspects of wound healing have been shown to be abnormal, including matrix metalloproteinase (MMP)-9 expression. The purpose of this study was to test whether the Pax6(+/-) genotype affects corneal wound-healing responses, including stromal cell apoptosis, epithelial cell migration rate, and MMP secretion in culture.METHOD. Pax6(+/-) and wild-type (Pax6(+/+)) mice were killed and their corneas wounded by epithelial debridement. Whole eyes were cultured in organ culture and corneal epithelial healing rates and keratocyte apoptosis were quantified by topical fluorescein staining and TUNEL, respectively. Dissociated corneal epithelial cells from Pax6(+/-) and wild-type mice were cultured, and the activities of secreted MMP-9 were determined by zymography.RESULTS. Wound-healing rates during the first 6 hours were significantly faster for larger wounds and for Pax6(+/-) corneas. Compared with wild-type, wounded Pax6(+/-) eyes showed significantly more stromal cell apoptosis, and cultured Pax6(+/-) corneal epithelial cells produced lower MMP-9 activity.CONCLUSIONS. The cumulative effect of abnormal wound-healing responses, characterized by increased stromal cell apoptosis and reduced levels of MMP-9 secretion may contribute to the corneal changes in the Pax6(+/-) mice. Possible contributions of elevated stromal cell apoptosis and other abnormal wound-healing responses to ARK are discussed.