嘌呤受体P2X7在人眼小梁网细胞上的表达及分布  

Expression and distribution of purinergic receptor P2X7 in human trabecular meshwork cells

作　　者：祝芸芸[1,2] 左炜[1,2] 王恒[1,2] 张秀兰[3] 

机构地区：[1]深圳市第四人民医院 [2]广东医学院附属福田医院,广东省深圳市518033 [3]中山大学中山眼科中心,广东省广州市510060

出　　处：《眼科新进展》2015年第8期721-724,共4页Recent Advances in Ophthalmology

摘　　要：目的探讨嘌呤受体P2X7在体外培养正常人眼小梁网细胞上的表达及分布。方法体外培养、鉴定正常人眼小梁网细胞,利用RT-PCR、Western-blot和免疫荧光法分别检测正常人眼小梁网细胞上P2X7受体mRNA和蛋白的表达情况及受体蛋白的主要分布部位。结果光镜下观察原代培养的细胞具有人眼小梁网细胞的典型形态;免疫组织化学方法检测结果表明,培养的细胞纤维连接蛋白、层粘连蛋白、神经元特异性烯醇化酶及波形蛋白染色阳性,第Ⅷ因子染色阴性,鉴定为人眼小梁网细胞;RT-PCR扩增到了P2X7mRNA 152 bp大小的基因片段,Western-blot得到了相对分子质量约为74 000的P2X7的蛋白条带,免疫荧光法证实P2X7受体在小梁网细胞膜、细胞质及细胞核内均有广泛分布。结论人眼小梁网细胞上有P2X7受体表达,其在细胞内分布广泛。Objective To discuss the expression and distribution of the puriner- gic receptor P2X7 in human trabecular meshwork cells （HTMC） cultured in vitro. Methods HTMC （ obtained from donated bodies＇ eyeball）were cultured in vitro and identified. The expression and distribution of purinergic receptor P2X7 were detected by RT-PCR,Western-blot analysis and immunofluorescence. Results The typical mor- phological of HTMC were observed by optical microscope. Immunohistochemicaol assays confirmed that fibronectin, laminin, neuron-specific enolase, vimentin were positive,and factor Ⅷ were negative, which indicated that the primary cultures in vitro were indeed HTMC. RT-PCR amplified a 152 bp gene segment of P2X7 mRNA, and West- ern-blot analysis showed a protein band of P2X7 with relative molecular mass of 74 000. Immunofluorescence confirmed a wide range of distribution of P2X7 receptor in the cell membrane, cytoplasm and nucleus of HTMC. Conelusion The purinergic receptor P2X7 is expressed in HTMC ,and it is widely distributed in HTMC.

关 键 词：嘌呤受体 人眼小梁网细胞 P2X7 青光眼 

分 类 号：R446.6[医药卫生—诊断学]