人眼Tenon囊成纤维细胞缝隙连接细胞间通讯的检测

目的:观察体外培养的人眼Tenon囊成纤维细胞(human Tenon’s capsule fibroblasts,HTFs)缝隙连接细胞间通讯(gap junctional intercellular communication,GJIC)功能及其构成蛋白connexin43(Cx43)的表达及分布。方法:锐刀划痕罗氏黄(lucifer yellow)荧光染料传输法观察GJIC功能;RT-PCR及Western blot分别检测Cx43 mRNA及蛋白表达;免疫细胞化学方法检测Cx43的分布。结果:荧光染料自划痕线向邻近未损伤细胞传输达6级细胞以上,同时检测到Cx43 mRNA与蛋白的表达;免疫细胞化学法观察到Cx43分布于细胞膜。结论:培养的HTFs具有较强的GJIC功能,通过Cx43介导GJIC赋予细胞能够相互之间传递信息的能力,提示在创伤愈合过程中GJIC对成纤维细胞功能同步化方面发挥重要作用。

Detection of gap junctional intercellular communication in cultured human Tenon's capsule fibroblasts

AIM:To explore gap junctional intercellular communication(GJIC) in cultured human Tenon's capsule fibroblasts(HTFs) and detect the expression and distribution of connexin43(Cx43).METHODS:GJIC were demonstrated by scrape-loading and Lucifer yellow dye transfer methods.RT-PCR and Western blot were carried out to detect mRNA and protein expression of Cx43.Immunocytochemistry was performed to observe the distribution of Cx43.RESULTS:Cultured HTFs demonstrated functional GJIC in vitro.The transporting degree of ...