不稳定逼尿肌中Cx43的表达及其对细胞间通讯的影响

The expression of Cx43 and its effects on the gap junction intercellular communication in unstable bladder detrusor cells

目的研究Cx43在不稳定逼尿肌细胞中的表达及其对不稳定逼尿肌细胞间缝隙连接通讯的影响,探讨Cx43与逼尿肌不稳定(DI)的关系。方法采用Western blot法检测正常及不稳定逼尿肌细胞中Cx43蛋白表达。构建Cx43反义真核表达载体,脂质体介导转染体外培养不稳定逼尿肌细胞(转染组),空载体做对照转染(空载组)。利用划痕标记荧光染料示踪(SLDT)技术观察Cx43表达降低后不稳定逼尿肌细胞间缝隙连接通讯改变。结果不稳定逼尿肌细胞中Cx43表达显著增高(P<0.01),转染反义Cx43组不稳定逼尿肌细胞间缝隙连接通讯较空载组明显减弱(P<0.05)。结论抑制Cx 43表达可明显降低不稳定逼尿肌细胞间缝隙连接通讯功能,Cx43表达增高可能是不稳定逼尿肌发生的重要原因。

Objective To investigate the expression of Cx43 and its effects on the gap junction intercelluar communication （GJIC） in cultured unstable detrusor cells. Methods Western blot was used to detect the expression of Cx43 protein in cultured detrusor cells. A Cx43 antisense vector, ASCx43/pCI-neo, was constructed, and then cultured unstable detrusor cells were transient transfected with ASCx43/pCI-neo and blank pCI-neo vectors mediated by Lipofectamine 2000, and Scrape loading dye transfer （SLDT） was used to monitor the changes of GJIC activity in detrusor cells. Results The expression of Cx43 increased markedly in unstable detrusor cells, and transfection of antisense Cx43 cDNA resulted in dramatic downregulation of Cx43 protein （P〈0.01）, which may cause remarkable reduction of GJIC activity in cultured unstable detrusor cells （P〈 0.05）. Conclusion These results show a significant increase of Cx43 expression in unstable detrusor cells. When the expression is inhibited, the GJIC activity in unstable detrusor cells decreases significantly, which implies that Cx43 may play an important role in the development of detrusor instability （DI）.