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目的探讨正常人眼巩膜成纤维细胞毒蕈碱受体Ⅰ型(M1)受体的表达与近视发生发展之间的关系。方法应用定点解剖及游走促进法分离培养人巩膜成纤维细胞,取3~5代生长良好的细胞,运用RT-PCR、间接免疫荧光试验检测培养的巩膜成纤维细胞中M1受体的表达;并制作巩膜组织的冷冻切片,以免疫组化法检测巩膜组织中巩膜成纤维细胞上M1受体亚型的表达。结果培养的巩膜成纤维细胞呈双极型或纺锤形,平均分裂时间为2~3d左右,细胞生长旺盛,近融合时细胞呈规则排列的纺锤形。RT-PCR和间接免疫荧光试验结果显示体外培养的人巩膜成纤维细胞表达M1受体。组织免疫组化法结果显示巩膜组织中的成纤维细胞表达M1受体。结论人巩膜成纤维细胞具有M1受体的分布。巩膜上M1受体的存在预示着M1受体拮抗剂延缓近视的发生发展的作用部位可能是位于巩膜上,而巩膜上的M1受体可能参与了近视的眼轴延长及近视屈光度的改变。(中华眼科杂志,2005,41:995-999)
Objective To investigate the relationship between the expression of muscarinic receptor type 1 (M1) receptors and the development of myopia in normal human scleral fibroblasts. METHODS: Human scleral fibroblasts were isolated and cultured by fixed-point anatomy and walking promotion method. The cells were cultured for 3-5 passages, and the expression of M1 receptor in scleral fibroblasts was detected by RT-PCR and indirect immunofluorescence assay. The frozen section of scleral tissue was made and the expression of M1 receptor subtype on scleral fibroblasts in scleral tissue was detected by immunohistochemistry. Results The cultured scleral fibroblasts were bipolar or spindle-shaped, the average time of division was about 2 ~ 3d, the cells grew vigorously and the cells were arranged in a regular spindle shape at the time of near fusion. RT-PCR and indirect immunofluorescence assay showed that human scleral fibroblasts cultured in vitro express M1 receptors. Tissue immunohistochemistry results showed that fibroblasts in the scleral tissue express M1 receptors. Conclusions Human scleral fibroblasts have M1 receptor distribution. The presence of M1 receptors on the sclera indicates that the M1 receptor antagonist may play a role in delaying the development of myopia. The scleral M1 receptor may be involved in the axial extension of myopia and the change of myopia diopter. (Chinese Journal of Ophthalmology, 2005,41: 995-999)