Department of Ophthalmology, Kunming Medical University, Kunming, China.
Department of Ophthalmology, The Affiliated Hospital of Yunnan University, Kunming, Yunnan, China.
J Histotechnol. 2023 Sep;46(3):101-113. doi: 10.1080/01478885.2023.2199370. Epub 2023 May 22.
Meibomian gland dysfunction (MGD) is a group of disorders linked by functional abnormalities of the meibomian glands. Current studies on MGD pathogenesis focus on meibomian gland cells, providing information on a single cell's response to experimental manipulation, and do not maintain the architecture of an intact meibomian gland acinus and the acinar epithelial cells' secretion state in vivo. In this study, rat meibomian gland explants were cultured by a Transwell chamber-assisted method under an air-liquid interface (airlift) in vitro for 96 h. Analyses for tissue viability, histology, biomarker expression, and lipid accumulation were performed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and TUNEL assays, hematoxylin and eosin (H&E) staining, immunofluorescence, Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), transmission electron microscopy (TEM), and western blotting (WB). MTT, TUNEL, and H&E staining indicated better tissue viability and morphology than the submerged conditions used in previous studies. Levels of MGD biomarkers, including keratin 1 (KRT1) and 14 (KRT14) and peroxisome proliferator-activated receptor-gamma (PPAR-γ), along with oxidative stress markers, including reactive oxygen species, malondialdehyde, and 4-hydroxy-2-nonenal, gradually increased over culture time. The MGD pathophysiological changes and biomarker expression of meibomian gland explants cultured under airlift conditions were similar to those reported by previous studies, indicating that abnormal acinar cell differentiation and glandular epithelial cell hyperkeratosis may contribute to obstructive MGD occurrence.
睑板腺功能障碍 (MGD) 是一组与睑板腺功能异常相关的疾病。目前关于 MGD 发病机制的研究主要集中在睑板腺细胞上，提供了关于单个细胞对实验操作的反应的信息，而不能维持完整的睑板腺小叶和腺上皮细胞在体内的分泌状态。在这项研究中，通过 Transwell 室辅助方法在体外气液界面（气举）下培养大鼠睑板腺外植体 96 小时。使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐 (MTT) 和 TUNEL 检测、苏木精和伊红 (H&E) 染色、免疫荧光、定量实时逆转录聚合酶链反应 (qRT-PCR) 、透射电子显微镜 (TEM) 和 Western blot (WB) 进行组织活力、组织学、生物标志物表达和脂质积累分析。MTT、TUNEL 和 H&E 染色表明组织活力和形态比以前研究中使用的浸没条件更好。MGD 生物标志物的水平，包括角蛋白 1 (KRT1) 和 14 (KRT14) 和过氧化物酶体增殖物激活受体-γ (PPAR-γ)，以及氧化应激标志物，包括活性氧、丙二醛和 4-羟基-2-壬烯醛，随着培养时间的延长逐渐增加。在气举条件下培养的睑板腺外植体的 MGD 病理生理变化和生物标志物表达与以前的研究报告相似，表明异常的腺泡细胞分化和腺上皮细胞过度角化可能导致阻塞性 MGD 的发生。
