Ye H Q, Maeda M, Yu F S, Azar D T
Schepens Eye Research Institute and the. Massachusetts Eye and Ear Infirmary Department of Ophthalmology, Harvard Medical School, Boston 02114, USA.
Invest Ophthalmol Vis Sci. 2000 Sep;41(10):2894-9.
Several members of the matrix metalloproteinase (MMP) group have been identified in the rat cornea during corneal wound healing. The aim of the present study was to identify additional members of the MMP gene family in the rat cornea and localize the expression of membrane type-1 matrix metalloproteinase (MT1-MMP; MMP-14) and collagenase III (MMP-13) in normal and wounded corneas.
Adult rats underwent laser keratectomy on the right eye. Unwounded left eyes were normal controls. Corneas were collected and processed at different times post-wounding. Reverse transcription-polymerase chain reaction (RT-PCR) and DNA sequencing were used to discover the MMP genes expressed in the corneas. In situ hybridization was performed to localize the mRNA expression of MMP-14 and MMP-13.
MMP-13 mRNA was detected in epithelial cells of wounded corneas, but not in normal controls; MMP-14 was found in both normal and wounded corneas. MMP-14 mRNA was expressed predominantly in the stromal keratocytes and rarely in the basal epithelial cells in normal and wounded corneas. MMP-13 mRNA was localized exclusively to basal cells of the epithelium at the wounded area from 6 hours to 3 days after wounding.
MMP-14 and MMP-13 expression in rat corneas parallels that of gelatinases A and B, respectively. MMP-13 may play an important role in the gelatinase B-associated proteolytic cascade that allows rapid turnover of the extracellular matrix (ECM) components during corneal wound healing. MMP-14 may contribute to removing abnormal ECM components through activation of gelatinase A in rat corneas.
在角膜伤口愈合过程中,已在大鼠角膜中鉴定出基质金属蛋白酶(MMP)家族的几个成员。本研究的目的是鉴定大鼠角膜中MMP基因家族的其他成员,并定位膜型1基质金属蛋白酶(MT1-MMP;MMP-14)和胶原酶III(MMP-13)在正常和受伤角膜中的表达。
成年大鼠右眼接受激光角膜切除术。未受伤的左眼作为正常对照。在受伤后的不同时间收集角膜并进行处理。采用逆转录-聚合酶链反应(RT-PCR)和DNA测序来发现角膜中表达的MMP基因。进行原位杂交以定位MMP-14和MMP-13的mRNA表达。
在受伤角膜的上皮细胞中检测到MMP-13 mRNA,但在正常对照中未检测到;在正常和受伤角膜中均发现了MMP-14。在正常和受伤角膜中,MMP-14 mRNA主要在上皮基质细胞中表达,在基底上皮细胞中很少表达。在受伤后6小时至3天,MMP-13 mRNA仅定位在受伤区域上皮的基底细胞中。
大鼠角膜中MMP-14和MMP-13的表达分别与明胶酶A和B的表达平行。MMP-13可能在与明胶酶B相关的蛋白水解级联反应中起重要作用,该级联反应允许在角膜伤口愈合过程中细胞外基质(ECM)成分快速更新。MMP-14可能通过激活大鼠角膜中的明胶酶A来促进异常ECM成分的清除。