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目的对比正常角膜与圆锥角膜中的Ⅰ型,Ⅵ型胶原及金属蛋白酶-2的定位与含量。方法应用酶免疫组化及间接免疫荧光技术检测正常与圆锥角膜中Ⅰ型、Ⅵ型胶原及金属蛋白酶-2(MMP毛)的定位。阳性结果应用图像分析系统进行定量分析。结果免疫组化结果显示所有角膜中Ⅰ型胶原存在于前弹力层和基质中;Ⅵ型胶原存在于上皮基底膜、前弹力层、基质、后弹力层;MMP-2存在于上皮、基质、内皮中。同正常角膜比,圆锥角膜中Ⅰ型、Ⅵ型胶原的含量无差别而 MMP-2的含量明显增多,差异有显著性。结论圆锥角膜的MMP-2含量增多,其所导致的胶原降解异常可能是圆锥角膜发病的关键。
Objective To compare the localization and content of type Ⅰ and type Ⅵ collagen and metalloproteinase-2 in normal cornea and keratoconus. Methods Enzyme immunohistochemistry (IHC) and indirect immunofluorescence staining were used to detect the type Ⅰ and type Ⅵ collagen and MMP - 2 in normal and keratoconus. Positive results using image analysis system for quantitative analysis. Results The results of immunohistochemistry showed that type Ⅰ collagen existed in the pre-elastic layer and matrix in all corneas. Type Ⅵ collagen existed in epithelial basement membrane, pre-elastic layer, stroma, and post-elastic layer. MMP-2 existed in epithelial, stromal and endothelial in. Compared with normal cornea, the content of type I and type VI collagen in keratoconus had no difference but the content of MMP-2 increased obviously, the difference was significant. Conclusions The content of MMP-2 in keratoconus is increased, and the abnormal collagen degradation caused by keratoconus may be the key to the development of keratoconus.