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HCN1通道基因敲除下调小鼠膀胱Cajal间质细胞中BK通道的表达及功能

, PP. 1281-1287

Keywords: ICCs,超极化激活环核苷酸门控阳离子通道,大电导钙激活钾通道,小鼠,基因敲除,膀胱

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Abstract:

目的观察小鼠HCN1通道基因敲除对其膀胱Cajal间质细胞(interstitialcellsofCajal,ICCs)中BK通道的表达和功能的影响,探讨这种影响对膀胱兴奋性调控的意义。方法健康清洁成年的野生型C57BL/6J小鼠和HCN1通道基因敲除的C57BL/6J小鼠各48只(雌雄各半)分别记为正常组和敲基因组,反转录PCR(RT-PCR)、荧光定量PCR(Q-PCR)、Westernblot和免疫荧光双标检测其膀胱ICCs中BK通道各亚基的表达变化,离体逼尿肌肌条实验检测加入BK通道阻滞剂IBTX前后肌条收缩的变化,激光共聚焦显微镜下检测分别加入BK通道激动剂NS1619、阻滞剂IBTX前后小鼠膀胱ICCs内钙荧光的变化。结果Q-PCR显示敲基因组小鼠膀胱中BK通道α、β1、β2、β3、β4各亚基表达均降低(P<0.01);Westernblot显示敲基因组小鼠膀胱中BK通道α、β1、β2、β3、β4亚基表达均降低(其中α、β3、β4:P<0.01;β1、β2:P<0.05);免疫荧光双标显示敲基因组小鼠膀胱ICCs中BK通道α亚基表达降低(P<0.01);离体逼尿肌肌条实验显示加入IBTX后敲基因组和正常组肌条收缩幅度均变大(P<0.01,P<0.05),且敲基因组肌条收缩幅度的变化值小于正常组(P<0.05);激光共聚焦显微镜下可见加入NS1619后两组ICCs内钙荧光均降低(P<0.05)、加入IBTX后两组ICCs内钙荧光均增强(P<0.01),且不论加入激动剂或阻滞剂,敲基因组加药前后ICCs内钙荧光的变化值均小于正常组(P<0.01)。结论小鼠HCN1通道基因敲除下调了其膀胱ICCs中BK通道的表达及功能,这种下调可能是对HCN1通道基因敲除后膀胱收缩减弱的一种代偿。

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