电针对T10脊髓横断后神经源性膀胱大鼠尿流动力学及Wnt/β-catenin信号通路的影响

Effects of electro-acupuncture on urodynamics and Wnt/β-catenin signaling pathway in rats with neurogenic bladder after T10 spinal cord transection

目的:观察电针对T10脊髓横断(SCT)所致神经源性膀胱大鼠的干预作用,并阐明其作用机制。方法:48只雌性SD大鼠随机分为假手术组(12只)和模型组(36只)。通过手术建立T10 SCT模型,采用脊髓损伤行为学(BBB)评分和尿流动力学指标对术后大鼠进行模型评价。术后第15天,将模型复制成功大鼠再次随机分为模型对照组(12只)、电针组(12只)和电针对照组(12只)。电针组以“大椎”和“次髎”穴(“次髎”穴隔日左右更替)电针干预;电针对照组于相同时间以“大椎”和“次髎”穴远脊柱侧(“次髎”穴用法同上)1 cm处为针刺点,进行电针干预。电针采用疏密波(疏波10 Hz/9 s,密波50 Hz/5 s),电针20 min,强度以针刺处出现规律性收缩抽动为宜,每日1次,连续1周。末次电针干预后各组大鼠行尿流动力学检测,采用RT-PCR和免疫荧光法检测各组大鼠脊髓组织中Wnt-1和β-catenin mRNA及蛋白表达水平。结果:与假手术组比较,模型对照组、电针对照组和电针组大鼠膀胱基础压力和最大压力升高(P<0.05或P<0.01),膀胱最大容量和顺应性明显降低(P<0.01);模型对照组和电针对照组大鼠膀胱漏尿点压力升高(P<0.01)。与模型对照组比较,电针组大鼠膀胱基础压力、最大压力和漏尿点压力明显降低(P<0.01),膀胱最大容量和顺应性升高(P<0.01)。与电针对照组比较,电针组大鼠膀胱基础压力、最大压力和漏尿点压力降低(P<0.05或P<0.01),膀胱最大容量和顺应性升高(P<0.05或P<0.01)。与假手术组比较,模型对照组、电针对照组和电针组大鼠脊髓组织中Wnt-1和β-catenin mRNA及蛋白表达水平明显升高(P<0.01);与模型对照组比较,电针组大鼠脊髓组织中Wnt-1和β-catenin mRNA及蛋白表达水平明显升高(P<0.01);与电针对照组比较,电针组大鼠脊髓组织中Wnt-1和β-catenin mRNA及蛋白表达水平明显升高(P<0.01)。结论:电针“大椎”和“次髎”穴对T10 SCT后神经源性膀胱大鼠尿流动力学具有明显改善作用,其作用机制与调控Wnt/β-catenin信号通路活化有关联。

Objective:To observe the intervention effect of electro-acupuncture in the rats with neurogenic bladder caused by T10 spinal cord transection(SCT),and to clarify its mechanism.Methods:Forty-eight female SD rats were randomly divided into sham operation group(12 rats)and model group(36 rats).The T10 SCT models were induced by surgery,and the models were evaluated by Basso Beattie Bresnahan(BBB)score and urodynamics indexes.On the 15th day after surgery,the successful replication model rats were randomly divided into model control group(12 rats),electro-acupuncture group(12 rats)and electro-acupuncture control group(12 rats).The rats in electro-acupuncture group were treated with electro-acupuncture at“Dazhui”(DU14)and“Ciliao”(BL32);the rats in electro-acupuncture control group were treated with electro-acupuncture at the 1 cm acupuncture point next to the“Dazhui”(DU14)and“Ciliao”(BL32).The electro-acupuncture used dilatational wave(10 Hz/9 s,50 Hz/5 s)for 20 min,and the intensity was suitable for regular contraction and twitching at the acupuncture point,once a day for a week in a row.After the last electro-acupuncture intervention,the urodynamic test was performed in the rats in each group,and the expression levels of Wnt-1 andβ-catenin mRNA and protein in spinal cord tissue of the rats in various groups were detected by RT-PCR and immunofluorescence methods.Results:Compared with sham operation group,the bladder base pressures and maximum pressures of the rats in model control group,electro-acupuncture control group and electro-acupuncture group were increased(P<0.05 or P<0.01),and the bladder maximum capacities and compliance were decreased(P<0.01);the pressure of bladder leak point of the rats in model control group and electro-acupuncture control group were increased(P<0.01).Compared with model control group,the bladder base pressure,maximum pressure and leak point pressure of the rats in electro-acupuncture group were increased(P<0.01),and the bladder maximum capacity and compliance were increased(P<0.01).Compared with electro-acupuncture control group,the bladder basic pressure,maximum pressure and leak point pressure of the rats in electro-acupuncture group were decreased(P<0.05 or P<0.01),and the bladder maximum capacity and compliance were increased(P<0.05 or P<0.01).Compared with sham operation group,the expression levels of Wnt-1 andβ-catenin mRNA and proteins in spinal cord tissue of the rats in model control group,electro-acupuncture control group and electro-acupuncture group were significantly increased(P<0.01).Compared with model control group,the expression levels of wnt-1 andβ-catenin mRNA and proteins in spinal cord tissue of the rats in electro-acupuncture group were significantly increased(P<0.01).Compared with electro-acupuncture control group,the expression levels of Wnt-1 andβ-catenin mRNA and proteins in the spinal cord tissues of the rats in electro-acupuncture group were significantly increased(P<0.01).Conclusion:electro-acupuncture on“Dazhui”(DU14)and“Ciliao”(BL32)has a significant improvement effect on the urodynamics of the rats with neurogenic bladder after T10 SCT,and its mechanism is related to regulating the activation of Wnt/β-catenin signal pathway.