Zhou Xiang-tian, Zhang Li-hua, Lü Fan, Xie Ruo-zhong, Hu Dan-ning, Fu Xiao-ying, Qu Jia
School of Optometry and Ophthalmology, Wenzhou Medical College, Wenzhou 325003, China.
Zhonghua Yan Ke Za Zhi. 2005 Nov;41(11):995-9.
The aim of this study was to identify the presence of muscarinic acetylcholine receptors-1 (mAChRs-1) in human sclera in order to determine whether the sclera is a potential site of action for mAChR antagonists.
Cell lines of human scleral fibroblasts were cultured in Dulbecco Modified Eagle'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.
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.
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.
本研究旨在确定人巩膜中是否存在毒蕈碱型乙酰胆碱受体1（mAChRs - 1），以判定巩膜是否是mAChR拮抗剂的潜在作用部位。
人巩膜成纤维细胞系在杜尔贝科改良伊格尔培养基中培养。采用逆转录 - 聚合酶链反应（RT - PCR）检测成纤维细胞中M1受体的mRNA表达。免疫细胞化学用于检测细胞系中mAChRs的蛋白。免疫组织化学研究用于进一步检测冰冻巩膜切片中M1受体的存在情况。
在RT - PCR中，培养的成纤维细胞显示出M1受体的mRNA表达。通过免疫细胞化学和免疫组织化学染色检查，成纤维细胞中存在M1蛋白。
本研究在mRNA和蛋白水平均证实了人巩膜成纤维细胞中存在M1受体。这一发现表明，巩膜是目前用于预防人类近视的mAChR拮抗剂的潜在作用部位。
