摘要
背景:近年来,越来越多的研究关注于睑板腺中腺泡细胞的增殖异常分化,认为这一过程与干眼症的发生和发展密切相关。睑板腺管腔阻塞和腺泡萎缩等结构和功能异常,可引起或加重干眼。因此,研究干眼中睑板腺变化对于深入理解干眼症的发病机制、寻找治疗和预防干眼症的新靶点具有重要的意义。目的:探究水液缺乏型干眼模型小鼠中睑板腺的变化。方法:选取6-8周雌性C57/B6小鼠32只,随机将其分成2组,每组16只。实验组小鼠采用同时切除眶外和眶内的泪腺的方法建造水液缺乏型干眼小鼠模型,对照组不做手术处理。正常饲养2周后于裂隙灯下观察两组小鼠角膜的改变,同时检测泪液分泌量。麻醉后脱颈处死两组小鼠后取睑板腺,观察睑板腺大体形态的变化,制成冰冻切片,苏木精-伊红染色观察睑板腺的结构,油红染色评价睑板腺的功能,免疫荧光染色和RT-qPCR观察两组小鼠睑板腺中祖细胞角蛋白14、细胞核增殖相关抗原(Ki67)以及异常分化的小脯氨酸丰富蛋白1B的表达情况。结果与结论:①造模2周后,实验组小鼠角膜可见片状缺损,角膜缘新生血管生成且侵入角膜中央;②实验组小鼠泪液分泌量较对照组明显减少(P<0.05);③显微镜照相显示,实验组睑板腺导管中断萎缩,排列紊乱;④苏木精-伊红结果显示,实验组小鼠睑板腺中的脂质空泡较对照组明显增多;⑤油红染色中可见实验组小鼠脂质沉积;⑥免疫荧光和RT-qPCR结果显示,实验组小鼠睑板腺中祖细胞角蛋白14、Ki67、小脯氨酸丰富蛋白1B的表达较对照组明显增多(P<0.05);⑦提示水液缺乏型干眼可导致睑板腺的代偿性肥大、增殖增加、脂质代谢异常,同时可发生睑板腺异常分化。
BACKGROUND:In recent years,increasing studies have focused on the abnormal proliferation and differentiation of acinous cells in the meibomian gland,suggesting that this process is closely related to the occurrence and development of dry eye.Structural and functional abnormalities such as blockage of the lumen of the meibomian gland and atrophy of the glands can cause or exacerbate dry eye.Therefore,the study of changes in the meibomian glands in dry eyes is important for understanding the pathogenesis of dry eyes in depth and finding new targets for the treatment and prevention of dry eyes.OBJECTIVE:To investigate the changes of the meibomian gland in a mouse model of aqueous deficient dry eyes.METHODS:Thirty-two female C57/B6 mice at 6-8 weeks were selected and randomly divided into experimental and control groups with 16 mice in each group.The mice in the experimental group were constructed by removing both the extra-orbital and intra-orbital lacrimal glands,while those in the control group were not treated.After 2 weeks of normal feeding,the corneal changes of both groups were observed under a slit lamp,and the tear secretion of both groups was measured.The meibomian glands of the two groups of mice were removed after decapitation.The changes in the gross morphology of the meibomian glands were observed and the meibomian glands were made into frozen sections.Hematoxylin-eosin staining was used to observe the structure of the meibomian glands,oil red staining was used to evaluate the function of the meibomian glands,and immunofluorescence staining and RT-qPCR were used to observe the expression of cytokeratin 14,Ki67 and abnormally differentiated small proline-rich protein 1B in the meibomian glands of mice.RESULTS AND CONCLUSION:Two weeks after modeling,lamellar defects were seen in the corneas of the experimental mice,and neovascularization of the limbal corneal was generated and invaded the central cornea.(2)Tear secretion volume was significantly reduced in the experimental group compared with the control group(P<0.05).Microscopic findings showed that the ducts of the meibomian glands in the experimental group were interrupted and atrophied,and their arrangement was disorganized.Hematoxylin-eosin staining results showed a significant increase in lipid vacuoles in the meibomian glands of the experimental mice compared with the control group.Lipid deposition was seen in oil red staining in the experimental group.Immunofluorescence and RT-qPCR results showed a significant increase in the expression of cytokeratin 14,Ki67 and small proline-rich protein 1B in the meibomian glands of mice in the experimental group compared with the control group(P<0.05).To conclude,aqueous deficient dry eye can lead to compensatory hypertrophy,increased proliferation,and abnormal lipid metabolism in the meibomian gland,as well as abnormal differentiation of the meibomian gland.
作者
周紫莹
欧尚坤
黄超
蒋浩
张丽颖
谷浩
Zhou Ziying;Ou Shangkun;Huang Chao;Jiang Hao;Zhang Liying;Gu Hao(Guizhou Medical University,Guiyang 550000,Guizhou Province,China;Department of Ophthalmology,Affiliated Hospital of Guizhou Medical University,Guiyang 550000,Guizhou Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2024年第11期1666-1671,共6页
Chinese Journal of Tissue Engineering Research
关键词
水液缺乏型干眼症
睑板腺功能障碍
睑板腺
干眼模型
动物模型
aqueous deficient dry eye
meibomian gland dysfunction
meibomian gland
dry eye model
animal model
