Kato Takuji, Chang Jin-Hong, Azar Dimitri T
Massachusetts Eye and Ear Infirmary and the Schepens Eye Research Institute, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
Invest Ophthalmol Vis Sci. 2003 Jan;44(1):78-85. doi: 10.1167/iovs.01-1257.
To determine the distribution of type XVIII collagen in mouse ocular tissues and to investigate the expression of type XVIII collagen during healing of corneal incisions and keratectomy wounds.
Immunohistochemical analysis of type XVIII collagen was performed in mouse ocular tissue, with polyclonal antibodies to the hinge domain. For wound-healing experiments, excimer laser keratectomy and single linear incisions were performed on mouse corneas. The corneas were harvested at various time points after wounding and processed for immunohistochemistry, in situ hybridization, competitive reverse transcription-polymerase chain reaction (RT-PCR), and Western blot analysis.
In the unwounded mouse cornea, type XVIII collagen was expressed by the corneal epithelial cells. Type XVIII collagen was immunolocalized to the mouse corneal epithelium, epithelial basement membrane, Descemet's membrane, ciliary epithelium, lens capsule, retinal inner limiting membrane, and Bruch's membrane. In the early stages of wound healing after excimer laser keratectomy (days 3 and 7), type XVIII collagen staining of the epithelial basement membrane was absent, whereas its localization to Descemet's membrane was unchanged. After linear corneal incisions, however, type XVIII collagen was clearly seen in the stroma and in the epithelial basement membrane. Type XVIII collagen immunolocalization to the subepithelial stromal wound region peaked at 1 week after wounding, and its mRNA showed a corresponding temporal increase in expression within the same region after linear corneal incisions.
The results suggest that type XVIII collagen is broadly expressed in ocular tissues and that it may have a role in wound healing, especially after incisional corneal wounds.
确定ⅩⅧ型胶原蛋白在小鼠眼组织中的分布，并研究角膜切开和角膜切除术伤口愈合过程中ⅩⅧ型胶原蛋白的表达情况。
使用针对铰链区的多克隆抗体对小鼠眼组织进行ⅩⅧ型胶原蛋白的免疫组织化学分析。对于伤口愈合实验，对小鼠角膜进行准分子激光角膜切除术和单一线性切口。在受伤后的不同时间点采集角膜，并进行免疫组织化学、原位杂交、竞争性逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹分析。
在未受伤的小鼠角膜中，ⅩⅧ型胶原蛋白由角膜上皮细胞表达。ⅩⅧ型胶原蛋白免疫定位在小鼠角膜上皮、上皮基底膜、Descemet膜、睫状体上皮、晶状体囊、视网膜内界膜和Bruch膜。在准分子激光角膜切除术后伤口愈合的早期阶段（第3天和第7天），上皮基底膜的ⅩⅧ型胶原蛋白染色缺失，而其在Descemet膜上的定位未改变。然而，在角膜线性切口后，在基质和上皮基底膜中可清楚看到ⅩⅧ型胶原蛋白。ⅩⅧ型胶原蛋白在伤口上皮下基质区域的免疫定位在受伤后1周达到峰值，并且其mRNA在角膜线性切口后在同一区域的表达呈现相应的时间性增加。
结果表明ⅩⅧ型胶原蛋白在眼组织中广泛表达，并且它可能在伤口愈合中起作用，尤其是在角膜切开伤口后。
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