Li Jing, Li Cheng, Wang Guoliang, Liu Zhen, Chen Pei, Yang Qichen, Dong Nuo, Wu Huping, Liu Zuguo, Li Wei
Eye Institute of Xiamen University Xiamen, Fujian, China 2Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China.
Xiamen University Affiliated Xiamen Eye Center, Xiamen, Fujian, China.
Invest Ophthalmol Vis Sci. 2016 Feb;57(2):444-52. doi: 10.1167/iovs.15-17519.
Squamous metaplasia is a common pathologic condition in ocular surface diseases for which there is no therapeutic medication in clinic. In this study, we investigated the effect of a small molecule, APR-246/PRIMA-1(Met), on squamous metaplasia in human conjunctival epithelium.
Human conjunctival explants were cultured for up to 12 days under airlifting conditions. Epithelial cell differentiation and proliferation were assessed by Cytokeratin 10 (K10), K14, K19, Pax6, MUC5AC, and p63 immunostaining patterns. β-catenin and TCF-4 immunofluorescent staining and real-time PCR characterized Wnt signaling pathway involvement. Pterygium clinical samples were cultured under airlifting conditions with or without APR-246 for 4 days. p63, K10, β-catenin, and TCF-4 expression in pterygial epithelium was determined by immunofluorescent staining and real-time PCR.
Airlift conjunctival explants resulted in increased stratification and intrastromal epithelial invagination. Such pathology was accompanied by increases in K10, K14, and p63 expression, whereas K19 and Pax6 levels declined when compared to those in freshly isolated tissue. On the other hand, APR-246 reversed all of these declines in K10, K14, and p63 expression. Furthermore, K19 and Pax6 increased along with rises in goblet cell density. These effects of APR-246 were accompanied by near restoration of normal conjunctival epithelial histology. APR-246 also reversed squamous metaplasia in pterygial epithelium that had developed after 4 days in ex vivo culture.
Reductions in squamous metaplasia induced by APR-246 suggest it may provide a novel therapeutic approach in different squamous metaplasia-associated ocular surface diseases.
鳞状化生是眼表疾病中的一种常见病理状况，临床上尚无治疗药物。在本研究中，我们调查了小分子APR - 246/PRIMA - 1(Met)对人结膜上皮鳞状化生的影响。
将人结膜外植体在气提条件下培养长达12天。通过细胞角蛋白10（K10）、K14、K19、Pax6、MUC5AC和p63免疫染色模式评估上皮细胞分化和增殖。β-连环蛋白和TCF-4免疫荧光染色以及实时PCR确定Wnt信号通路的参与情况。翼状胬肉临床样本在有或无APR - 246的气提条件下培养4天。通过免疫荧光染色和实时PCR测定翼状胬肉上皮中p63、K10、β-连环蛋白和TCF-4的表达。
气提结膜外植体导致分层增加和基质内上皮内陷。这种病理变化伴随着K10、K14和p63表达的增加，而与新鲜分离组织相比，K19和Pax6水平下降。另一方面，APR - 246逆转了K10、K14和p63表达的所有这些下降。此外，K19和Pax6随着杯状细胞密度的增加而增加。APR - 246的这些作用伴随着结膜上皮组织学接近恢复正常。APR - 246还逆转了离体培养4天后翼状胬肉上皮中出现的鳞状化生。
APR - 246诱导的鳞状化生减少表明它可能为不同的鳞状化生相关眼表疾病提供一种新的治疗方法。
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