Wang Guoliang, Xu Lina, Shi Ruize, Ye Yingyue, Zeng Baihui, Yang Xiuqin, Liu Zeyu, Liu Zhen, Wang Shurong, Xue Yuhua, Li Cheng
School of Pharmaceutical Sciences, Eye Institute & Affiliated Xiamen Eye Center, Xiamen University, Xiamen, Fujian, 361102, China; Department of Ophthalmology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, 130033, China.
Department of Ophthalmology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, 130033, China; Department of Ophthalmology, The Second Hospital of Jilin University, Changchun, Jilin, 130041, China.
Ocul Surf. 2023 Oct;30:73-84. doi: 10.1016/j.jtos.2023.08.007. Epub 2023 Aug 22.
Meibomian glands (MGs) are crucial for maintaining tear film stability and ocular surface health. Here, we aim to establish a novel organotypic culture model of MGs and explore the risk factors of MG dysfunction (MGD).
We developed a novel organotypic culture model for MGs at the air-liquid interface. The viability and cell proliferation of MGs were assessed using CCK-8, immunofluorescence, and qPCR. Lipid accumulation was evaluated by Nile red staining and microscopic examination. Protein expression levels were evaluated by immunofluorescence and Western blot assay. EdU assay was employed to track the proliferation of acinar cells. The validity of the model was confirmed through culturing MGs from mice of different ages and incorporating certain drugs (Dex) into the culture system.
Utilizing the novel culture model, the MG tissue exhibited sustained viability, cellular division, and continuous production of lipids for a duration of 7 days. Lipid droplets formed were directly visualized using light field microscopy. Through the cultivation of aged mice's MGs, it was discovered that aging resulted in diminished proliferation and lipid synthesis, along with an aberrant increase in Krt10 expression. Further application of this model showed that Dex treatment diminished MG's proliferation and lipid synthesis. Finally, an in vivo study was conducted to provide additional confirmation of the phenomenon of Dex-induced abnormalities.
In this study, a stable organotypic culture model of the MGs was established. The organotypic culture model offers a valuable tool to investigate the pathophysiological mechanisms and facilitate drug screening for MG-related diseases.
睑板腺对维持泪膜稳定性和眼表健康至关重要。在此，我们旨在建立一种新型的睑板腺器官型培养模型，并探索睑板腺功能障碍（MGD）的危险因素。
我们在气液界面建立了一种新型的睑板腺器官型培养模型。使用CCK-8、免疫荧光和qPCR评估睑板腺的活力和细胞增殖。通过尼罗红染色和显微镜检查评估脂质积累。通过免疫荧光和蛋白质印迹分析评估蛋白质表达水平。采用EdU检测法追踪腺泡细胞的增殖。通过培养不同年龄小鼠的睑板腺并将某些药物（地塞米松）加入培养系统来证实该模型的有效性。
利用该新型培养模型，睑板腺组织在7天内表现出持续的活力、细胞分裂和脂质的持续产生。使用明场显微镜可直接观察到形成的脂滴。通过培养老年小鼠的睑板腺，发现衰老导致增殖和脂质合成减少以及角蛋白10（Krt10）表达异常增加。该模型的进一步应用表明，地塞米松处理会减少睑板腺的增殖和脂质合成。最后，进行了一项体内研究以进一步证实地塞米松诱导异常的现象。
在本研究中，建立了一种稳定的睑板腺器官型培养模型。该器官型培养模型为研究病理生理机制和促进与睑板腺相关疾病的药物筛选提供了有价值的工具。
