毒蕈碱受体Ⅰ型受体在人眼巩膜成纤维细胞中的表达  

Expression of muscarinic acetylcholine receptor-1 in human scleral fibroblasts

作　　者：周翔天[1] 张立华[1] 吕帆[1] 谢若衷[1] 胡诞宁[2] 付小莹[1] 瞿佳[1] 

机构地区：[1]温州医学院眼视光学院,325003 [2]温州医学院近视研究所,纽约医学院纽约眼耳鼻喉科医院组织培养中心

出　　处：《中华眼科杂志》2005年第11期995-999,共5页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金资助项目(30371507);浙江省自然科学基金资助项目(302095;ZB0202);浙江省卫生厅重大课题基金资助项目(2002ZD009);浙江省科技厅科技攻关计划-重点科研项目-社会发展基金资助项目(2003C23005)

摘　　要：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2～3 d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 The aim of this study was to identify the presence of muscarinic acetylcboline receptors-1 （ mAChRs-1 ） in human sclera in order to determine whether the sclera is a potential site of action for mAChR antagonists. Methods Cell lines of human scleral fibroblasts were cultured in Dulbecco Modified Ealge＇s Medium. Reverse Transcription-Polymerase Chain Reaction （RT-PCR） was used to detect mRNA expression of M1 receptors in the fibroblasts. Immunocytochemistry was used to detect proteins of mAChRs in the cell lines. Immunohistochemical study was used to further detect the presence of M1 receptor in the frozen scleral sections. Results The cultured fibroblasts demonstrated mRNA expression of M1 receptor in RT-PCR. Protein of the M1 was present in the fibroblasts by examination of immunocytochemistry and immunohistochemistry staining. Conclusion This study demonstrated the presence of M1 receptor in human scleral fibroblasts at both mRNA and protein levels. This finding indicates that the sclera is a potential site of action for the currently used mAChR antagonists in prevention of human myopia.

关 键 词：近视 受体 毒蕈碱M1 巩膜 成纤维细胞 

分 类 号：R778.11[医药卫生—眼科]