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目的:研究缝隙连接蛋白-43在体外培养不稳定膀胱组织Cajal间质细胞中的表达,初步探讨其与逼尿肌不稳定的关系。方法:建立膀胱出口梗阻豚鼠模型,用胶原酶消化法进行逼尿肌不稳定豚鼠膀胱组织中Cajal间质细胞及逼尿肌细胞的原代培养,利用免疫荧光染色法进行两种细胞特征性蛋白c-kit及SMA染色观察;RT-PCR法进行两种细胞的缝隙连接蛋白-43的检测。结果:豚鼠膀胱组织中Cajal间质细胞c-kit染色阳性,SMA染色阴性;逼尿肌细胞SMA染色阳性,c-kit染色阴性。逼尿肌不稳定豚鼠膀胱组织中Cajal间质细胞缝隙连接蛋白-43表达较逼尿肌细胞强(P<0.05)。结论:体外培养不稳定膀胱组织中Cajal间质细胞缝隙连接蛋白-43表达增强,提示Cajal间质细胞间兴奋传递增强与逼尿肌不稳定的发生关系密切。
AIM: To investigate the expression of connexin-43 in interstitial cells of Cajal of unstable bladder tissue cultured in vitro and to explore the relationship between the expression of connexin-43 and detrusor instability. Methods: Guinea pigs with bladder outlet obstruction were established. The primary culture of Cajal interstitial cells and detrusor myocytes in the bladder tissue of detrusor instability guinea pigs were performed by collagenase digestion. Two kinds of cell-specific proteins were identified by immunofluorescence staining c-kit and SMA staining were observed; RT-PCR method for the detection of two kinds of cells, connexin-43. Results: The c-kit staining of Cajal interstitial cells in guinea pig bladder tissue was negative and SMA staining was negative. The SMA staining of detrusor muscle cells was negative and the c-kit staining was negative. The expression of Connexin-43 in Cajal interstitial cells in detrusor instability guinea pig bladder tissue was higher than that in detrusor muscle cells (P <0.05). CONCLUSIONS: The expression of connexin-43 in interstitial cells of Cajal in unstable bladder tissue in vitro is increased, which suggests that the enhanced transmission of Ca 2+ in interstitial cells is closely related to the detrusor instability.