Laboratory of Ocular Atopic Diseases, Department of Ophthalmology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-Ku, Tokyo, 113-8431, Japan.
Sci Rep. 2020 Sep 14;10(1):15036. doi: 10.1038/s41598-020-72036-0.
To clarify the early alterations of gene expression using a mouse model of glaucoma filtration surgery, we carried out microarray expression analysis. Using BALB/c mice, a filtration surgery model was made by incision of the limbal conjunctiva, followed by the insertion of a 33G needle tip into the anterior chamber, and 11-0 nylon sutures. Subgroups of mice were treated intraoperatively with 0.4 mg/ml mitomycin-C (MMC). At day 3 after surgery the bleb was maintained. The bleb region tissue was sampled 3 days after the filtration surgery, and gene expression analysis was carried out using a mouse Agilent 8 × 60 K array. We found 755 hyperexpressed transcripts in the bleb region compared to control conjunctiva. The hyperexpressed transcripts included epithelial cell metaplasia-related (Il1b, Krt16, Sprr1b), inflammation-related (Ccl2, Il6) and wound healing-related (Lox, Timp1) genes. We also found downregulation of a goblet cell marker gene (Gp2) in the bleb conjunctiva. MMC treatment suppressed elastin (Eln) gene expression and enhanced keratinization-related gene expression (Krt1, Lor) in the bleb region. Our results suggest the importance of epithelial wound healing after filtration surgery, and this filtration surgery model will be a useful tool for further pathophysiological analysis.
为了阐明青光眼滤过性手术小鼠模型中基因表达的早期改变,我们进行了微阵列表达分析。我们使用 BALB/c 小鼠,通过角膜缘结膜切开,然后将 33G 针尖插入前房,并使用 11-0 尼龙缝线进行了滤过性手术模型的制作。部分小鼠在手术中接受了 0.4mg/ml 丝裂霉素 C(MMC)治疗。手术后第 3 天维持了囊泡。在滤过性手术 3 天后,采集囊泡区域组织,并使用小鼠 Agilent 8×60K 阵列进行基因表达分析。与对照结膜相比,我们在囊泡区域发现了 755 个高表达的转录本。高表达的转录本包括上皮细胞化生相关(Il1b、Krt16、Sprr1b)、炎症相关(Ccl2、Il6)和伤口愈合相关(Lox、Timp1)基因。我们还发现囊泡结膜中杯状细胞标记基因(Gp2)的下调。MMC 处理抑制了囊泡区域中的弹性蛋白(Eln)基因表达,并增强了角蛋白相关基因表达(Krt1、Lor)。我们的结果表明滤过性手术后上皮细胞伤口愈合的重要性,并且这种滤过性手术模型将成为进一步病理生理分析的有用工具。