睑板腺功能障碍(MGD)是一组由睑板腺功能异常相关的疾病。目前MGD发病机制的研究主要集中在睑板腺细胞,提供有关单细胞对实验操作的反应的信息,并且不维持完整睑板腺腺泡的结构和体内腺泡上皮细胞的分泌状态。在这项研究中,大鼠睑板腺外植体通过Transwell室辅助方法在空气-液体界面(气举)下体外培养96小时。组织学,生物标志物表达,用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物(MTT)和TUNEL测定法进行脂质积累,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),和蛋白质印迹(WB)。MTT,TUNEL,和H&E染色表明比先前研究中使用的浸没条件更好的组织活力和形态。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),连同氧化应激标志物,包括活性氧,丙二醛,和4-羟基-2-壬烯醛,随着培养时间的推移逐渐增加。在空运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先前研究报道的相似。提示腺泡细胞分化异常和腺上皮细胞角化过度可能与梗阻性MGD的发生有关。
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.