腺苷受体在人巩膜成纤维细胞中的分布
Distribution of adenosine receptors in human sclera fibroblasts.
作者信息
Cui Dongmei, Trier Klaus, Chen Xiang, Zeng Junwen, Yang Xiao, Hu Jianmin, Ge Jian
机构信息
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
出版信息
Mol Vis. 2008 Mar 14;14:523-9.
PURPOSE
Systemic treatment with adenosine receptor antagonists has been reported to affect the biochemistry and ultrastructure of rabbit sclera. This study was conducted to determine whether adenosine receptors (ADORs) are present in human scleral fibroblasts (HSF).
METHODS
Primary HSF were cultured in vitro and identified with anti-vimentin, anti-keratin, anti-desmin, and anti-S-100 antibodies. Confocal fluorescence microscopy was used to study the distribution of ADORs in the HSF cell lines and in the frozen human scleral sections. ADOR protein expression in HSF and human sclera was confirmed by western blot analysis of cell lysates.
RESULTS
ADORs were expressed in both HSF and human sclera. This was confirmed by western blot. ADORA1 expression was concentrated in the nucleus. ADORA2A was concentrated mainly in one side of the cytoplasm, and ADORA2B was found both in the nucleus and the cytoplasm. ADORA3 was expressed weakly in the cytoplasm.
CONCLUSIONS
All four subtypes of ADOR were found in HSF and may play a role in scleral remodeling.
目的
据报道，用腺苷受体拮抗剂进行全身治疗会影响兔巩膜的生物化学和超微结构。本研究旨在确定人巩膜成纤维细胞（HSF）中是否存在腺苷受体（ADOR）。
方法
原代HSF在体外培养，并用抗波形蛋白、抗角蛋白、抗结蛋白和抗S-100抗体进行鉴定。共聚焦荧光显微镜用于研究ADOR在HSF细胞系和冷冻人巩膜切片中的分布。通过对细胞裂解物进行蛋白质印迹分析来确认HSF和人巩膜中ADOR蛋白的表达。
结果
ADOR在HSF和人巩膜中均有表达。蛋白质印迹证实了这一点。A1型腺苷受体（ADORA1）表达集中在细胞核中。A2A型腺苷受体（ADORA2A）主要集中在细胞质的一侧，而A2B型腺苷受体（ADORA2B）在细胞核和细胞质中均有发现。A3型腺苷受体（ADORA3）在细胞质中表达较弱。
结论
在HSF中发现了所有四种ADOR亚型，它们可能在巩膜重塑中发挥作用。
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