翼状胬肉是常见眼表疾病，有上皮细胞间质化（EMT）的特征表现。近来miRNA在其发病中的作用逐渐受到重视，但缺乏对特定miRNA的深入研究。前期我们采用芯片筛选出了显著高表达的miR-199家族，并进行了临床样本的验证，初步确定了miR-199家族的靶基因DUSP5和MAP4K3，均在与EMT密切相关的MAPK通路上。因此设想miR-199家族靶向MAPK通路致翼状胬肉EMT的发生。拟：1.收集翼状胬肉组织和正常对照的结膜大样本，确认miR-199家族在翼状胬肉组织中的作用；2. 体外实验确认miR-199和靶基因的相互作用；3. 将miR-199转染至人结膜上皮细胞中，检测其DUSP5、MAP4K3水平及p38-MAPK通路、MAP4K3蛋白的磷酸化水平，比较EMT和上皮细胞特征分子的表达；4.建立兔眼翼状胬肉模型，进行腺病毒包装的miRNA感染实验。以期为翼状胬肉寻找新的治疗靶点。

Pterygium is the commonest ocular surface disease with epithelioid tumor biological characteristics, presents epithelial-mesenchymal transition (EMT). Recently, the role of miRNA in pterygium gets more and more attention, but there is few researches on the detail function of certain miRNA in pterygium formation. We adopted miRNA chip to screen the significant high expression of miR-199 family in pterygium, which was verified in part of the clinical samples. Then we identified the target gene DUSP5 of miR-199a/b-3p, which is the key element in p38-MAPK pathway. And the target gene of miR-199a-5p was reported to be MAP4K3, which was expressed in pterygium. MAPK pathway was reported to play an important role in EMT. So we assume that: miR-199a/b -3p and miR-199a-5p were upregulated in pterygium and targeted DUSP5 and MAP4K3 genes to mediate MAPK pathway, eventually leading to EMT. So, we proposed the following experiments: 1. To expand the pterygium and normal conjunctiva contrast samples, further verify the higher expression of miR-199a/b-3p and miR-199a-5p and the lower expression of target genes in big samples; 2. In vitro experiments to confirm the interaction between miRNA and target genes; 3. After the transfection of human conjunctival epithelial cells with miR-199, to detect the DUSP5 and MAP4K3 levels, and p38, MAPK, MAP4K3 phosphorylation protein levels, and to compare the molecules of EMT and epithelial characteristics; 4. To establish the pterygium model in rabbit eyes, and carry on infection experiments of miRNA. Our purpose is to find a new target for pterygium therapy.