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目的:研究T型钙通道在逼尿肌过度活动(detrusor overactivity,DO)大鼠模型中的改变并探索其在发病机制中的作用。方法:部分结扎雌性大鼠近端尿道制作DO动物模型,假手术组作为对照;6周后使用尿流动力学技术膀胱测压筛选实验动物;应用钙荧光探针Fluo-4比较模型和对照大鼠培养逼尿肌静息钙浓度,观察比较加入T型钙通道阻断剂Mibefradil后细胞静息钙浓度的变化;利用穿孔膜片钳技术比较模型和对照组大鼠急性分离逼尿肌细胞中T型钙通道密度。结果:膀胱测压中出现非排尿收缩的为DO大鼠。结果表明近端尿道部分结扎可成功制作DO动物模型;钙探针激光共聚焦实验表明当T型钙通道被抑制后,DO、对照逼尿肌细胞的静息钙荧光强度均显著下降,其中自身前后荧光变化率DO组显著大于正常组;穿孔膜片钳结果表明DO大鼠逼尿肌中T型钙通道的密度显著大于对照。结论:T型钙通道活动在DO动物中显著上调,这可能是导致该疾病的重要分子机制。
Objective: To study the changes of T-type calcium channel in detrusor overactivity (DO) rat model and to explore its role in the pathogenesis. Methods: The DO rat model was established by partial ligation of the proximal urethra in rats, sham-operated group served as a control. Urine flow cytometry was used to screen the urinary bladder in 6 weeks. The resting calcium concentration of detrusor muscle was cultured and the changes of resting calcium concentration were observed and compared with the T-type calcium channel blocker Mibefradil. The perfusion patch clamp technique was used to compare the changes of T Calcium channel density. Results: Non-voiding contractions in bladder manometry were observed in DO rats. The results showed that part of the proximal urethra ligation can be successfully DO animal model; calcium probe laser confocal experiments showed that when the T-type calcium channel is inhibited, DO, control detrusor cells resting calcium fluorescence intensity decreased significantly, of which self The change of fluorescence before and after DO group was significantly greater than that of the normal group. The results of perforated patch-clamp showed that the density of T-type calcium channel in detrusor muscle of DO rats was significantly higher than that of the control group. CONCLUSION: T-type calcium channel activity is significantly up-regulated in DO animals, which may be the important molecular mechanism leading to this disease.