Zhang Jie, Zhao Gui-Qiu, Qu Jing, Che Cheng-Ye, Lin Jing, Jiang Nan, Zhao Han, Wang Xue-Jun
Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.
Department of Administrative Management, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.
Int J Ophthalmol. 2016 Feb 18;9(2):191-7. doi: 10.18240/ijo.2016.02.02. eCollection 2016.
To explore the expression of S100B in corneal epithelial cells under Aspergillus stimulation both in vivo and in vitro.
Immortalized human corneal epithelial cells (HCECs) were exposed to inactive Aspergillus fumigatus (A. fumigatus) conidia at 0, 4, 8, 12, 16, and 24h respectively. The corneas of Wistar rats were exposed to active A. fumigatus at 0, 12, 24, 48h and the normal rat corneas were used for normal control. The mRNA level of S100B was evaluated by real time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). S100B protein expression in cornea epithelium was detected by immunohistochemical/immunocytochemical staining (IHC/ICC).
Histopathology revealed a significant inflammatory cell infiltration in fungal keratitis human and rat cornea. Corneal epithelial cells didn't express or rarely express S100B at baseline. A. fumigatus significantly induced S100B mRNA expression in cultured corneal epithelial cells in a time depended manner in vitro, the mRNA began to rise significantly at 8h in vitro (P<0.05) and continue to rise as time prolonged (P<0.01). In vivo, S100B mRNA level was low in the normal corneas. However, it was increased in keratitis corneas from 12h after infection (P<0.05) and reached to a peak at 24h (P<0.001). Immunochemistry revealed an obvious staining in fungal keratitis corneas as well as immortalized HCECs compared to the normal ones respectively, indicating an increased expression of S100B protein.
S100B exists in corneal epithelial cells and is over-expressed under A. fumigatus stimulation. S100B may play an important role in the innate immune response of the corneal epithelium during A. fumigatus infection.
探讨S100B在烟曲霉刺激下体内外角膜上皮细胞中的表达情况。
将永生化人角膜上皮细胞（HCECs）分别于0、4、8、12、16和24小时暴露于灭活的烟曲霉菌分生孢子。将Wistar大鼠角膜于0、12、24、48小时暴露于活性烟曲霉，正常大鼠角膜作为正常对照。通过实时定量逆转录-聚合酶链反应（qRT-PCR）评估S100B的mRNA水平。通过免疫组织化学/免疫细胞化学染色（IHC/ICC）检测角膜上皮中S100B蛋白的表达。
组织病理学显示真菌性角膜炎患者和大鼠角膜中有明显的炎性细胞浸润。角膜上皮细胞在基线时不表达或很少表达S100B。烟曲霉在体外以时间依赖性方式显著诱导培养的角膜上皮细胞中S100B mRNA表达，体外8小时时mRNA开始显著升高（P<0.05），并随时间延长持续升高（P<0.01）。在体内，正常角膜中S100B mRNA水平较低。然而，感染后12小时角膜炎角膜中其水平升高（P<0.05），并在24小时达到峰值（P<0.001）。免疫化学显示，与正常角膜和永生化HCECs相比，真菌性角膜炎角膜中有明显染色，表明S100B蛋白表达增加。
S100B存在于角膜上皮细胞中，在烟曲霉刺激下过度表达。S100B可能在烟曲霉感染期间角膜上皮的固有免疫反应中起重要作用。
