摘要
目的探讨钩藤碱对骶上脊髓损伤(SSCI)致神经源性膀胱大鼠逼尿肌反射亢进的治疗作用及其机制。方法将10周龄健康雌性SD大鼠30只(SPF级)按照随机数字表法分为对照组、模型组、治疗组,每组10只。对照组不做任何手术处理,模型组和治疗组采用脊髓横断法制备神经源性膀胱模型。对照组和模型组腹腔注射0.9%氯化钠溶液,治疗组注射钩藤碱(5 mg/kg)。对照组10只大鼠全部存活,模型组8只造模成功,治疗组8只造模成功。4周后,大鼠膀胱行尿动力学检测,离体逼尿肌条电生理学指标检测,膀胱病理切片HE染色观察,蛋白质印迹法检测c-kit蛋白的表达,激光共聚焦显微镜下观察Cajal间质细胞(ICC)数量变化,实时荧光PCR检测ICC中T型钙通道蛋白亚型α1G基因表达水平。结果与模型组相比,治疗组膀胱最大容量增大(q=8.251,P<0.05),膀胱漏尿点压(q=3.764,P<0.05)和膀胱充盈压均减小(q=5.687,P<0.05),逼尿肌收缩频率降低(q=9.661,P<0.05),最小张力增大(q=5.217,P<0.05),c-kit蛋白表达降低(q=13.688,P<0.05),ICC数量减少(q=7.060,P<0.05),α1G基因表达量降低(q=8.762,P<0.05),差异均有统计学意义。病理切片下观察到治疗组逼尿肌无断裂,肌间隙增宽减少。结论钩藤碱通过减少神经源性膀胱逼尿肌ICC数量,抑制ICC T型钙通道蛋白亚型α1G基因的表达来抑制ICC起搏兴奋活性,从而减轻逼尿肌的过度亢进,使逼尿肌兴奋性趋于正常。
Objective To investigate the effects of phynchophylline on detrusor overactivity induced by sacral spinal cord injury （ SSCI） in rats and the underlying mechanism .Methods Using random number table method , we divided 30 10-week female healthy SD rats into control group , model group and treatment group , with 10 rats in each group.No surgical intervention was made in control group , neurogenic bladder models were made in model group and treatment group by spinal cord transaction.Control group and model group were administrated with 0.9%sodium chloride solution by intraperitoneal infection . Treatment group was administrated with phynchophylline （5 mg/kg）.10 rats in control group were all alive, successful model building was made in 8 rats in model group and 8 rats in treatment group . Four weeks later , urine dynamics testing was undertaken with the bladder of rats , in vitro test of the electrophysiological indicators of detrusor was conducted , the pathological sections of bladder were observed by HE staining method , c-kit protein expression was tested by western blotting method , the change in the number of ICC was observed under laser scanning confocal microscope , and the expression level of α1G gene of subunit of T-type calcium channels in ICC was detected by real -time PCR.Results Compared with the model group , the treatment group had larger maximum capacity of bladder （q=8.251, P〈0.05）, lower bladder leak point pressure （q=3.764, P〈0.05）, lower bladder filling pressure （q =5.687, P 〈0.05）, lower detrusor contraction frequency （q =9.661, P 〈0.05）, higher minimum tension （q=5.217, P〈0.05）, lower expression level of c -kit protein （q=13.688, P〈0.05）, lower number of ICC （q =7.060, P 〈0.05） and lower α1G gene expression （q =8.762, P 〈0.05）.Conclusion Rhynchophylline can reduce the number of ICC cells in neurogenic detrusor and inhibit the pacemaker activity of ICC cells by inhibiting the expression of α1G gene of subunit of T -type calcium channels
出处
《中国全科医学》
CAS
CSCD
北大核心
2015年第23期2817-2821,共5页
Chinese General Practice
基金
辽宁省科技厅计划项目(2013225305)
