钙激活钾通道基因和蛋白水平在膀胱逼尿肌不稳定中的表达

摘要

目的 观察膀胱逼尿肌不稳定(DI)中钙激活钾通道(BKCa)基因和蛋白水平变化,探讨BKCa在DI发生机制中的作用.方法 通过膀胱出口部位梗阻法建立大鼠DI模型,取其膀胱平滑肌,提取总RNA和总蛋白,以β-actin为内参,分别采用定量PCR和WesternBlot化学发光法检测BKCa通道基因和蛋白水平.结果 DI组大鼠膀胱平滑肌BKCa通道基因和蛋白水平均明显下降,与对照组相比,DI组mRNA水平相对量(与内参照基因β-actin)由0.35±0.12(n=15)下降到0.21±0.09(n=12),两组比较有显著性差异(P＜0.05),BKCa通道蛋白水平相对量(与内参照蛋白β-actin)0.41±0.08(n=15)下降到0.23±0.06(n=12),两组相比有显著性差异(P＜0.01).结论 BKCa在膀胱逼尿肌不稳定时发生明显改变,可能参与了大鼠膀胱DI的发生,调控DI时BKCa通道活性可以作为治疗DI的靶点之一.%Objective To observe the change of gene and protein levels of large calcium activated potassium channels (BKCa) in rats with detrusor instability (DI) and discuss its role in DI. Methods Rats with DI were prepared by li-gating the proximal urethra in the perimeum. The bladder smooth muscle cell was acquired for exacting the total RNA and protein. Real-time PCR and Western blot were used for analyzing the mRNA and protein levels of BKCa channels, respectively. Results The mRNA and protein levels of BKCa were decreased in DI group, compared with the control. U-sing the β-actin as the housekeeping gene, the mRNA level of BKCa in DI group was decreased from 0. 35±0. 12 (Control, n=15) to 0. 21±0. 09 (DI, n=12) with P<0. 05. The protein level of BKCa, was reduced from 0. 41±0. 08 (control, n=15) to 0. 23±0. 06 (DI, n=12) with P<0. 01. Conclusion The mRNA and protein levels of BKc were decreased in rat with DI, which may be involved in the DI and BKCa may become one therapeutical target of DI.