灸法对逼尿肌反射亢进大鼠C纤维和膀胱胆碱受体作用机制的研究

逼尿肌反射亢进是脑血管病尿失禁患者最常见的尿动力学表现，艾灸疗法用于治疗逼尿肌反射亢进有较好疗效，但目前艾灸治疗逼尿肌反射亢进的起效途径尚不明确。M受体在逼尿肌中含量丰富，其兴奋是逼尿肌收缩的主要驱动因素。本研究使用逼尿肌反射亢进模型大鼠作为研究对象，以隔盐灸神阙穴为切入点，采用神经电生理技术观察大鼠排尿时C纤维神经放电情况，分析膀胱压力发生变化时神经电活动变化，观察艾灸神阙穴对C神经纤维的影响。通过免疫荧光双标、激光共聚焦显微镜观察的方法在蛋白水平测定艾灸对膀胱M2、M3 受体亚型的表达的影响，采用荧光定量PCR方法在基因水平对逼尿肌反射亢进大鼠膀胱黏膜的M2、M3受体mRNA标准化拷贝数进行比较，分析其与M受体亚型mRNA表达情况之间的相关性。围绕逼尿肌反射亢进大鼠膀胱M受体和膀胱C纤维的变化寻找艾灸治疗尿失禁的关键靶点和作用途径，为灸法治疗尿失禁提供科学实验依据。

The urodynamic detrusor hyperreflexia is most common in patients with cerebrovascular disease, urinary incontinence, moxibustion therapy has better curative effect for treatment of detrusor hyperreflexia, but the effect way of moxibustion treatment of detrusor hyperreflexia is not clear. M receptor rich content in detrusor, the excitement is the main driving factors of detrusor contraction. In this study, the use of detrusor hyperreflexia model rats as the research object, the salt separated moxibustion Shenque point, using electrophysiological techniques to observe the rat urinary C nerve fiber discharge conditions, analysis of bladder pressure changes of neural electrical activity changes, the influence of moxibustion God Shenque point on C nerve fibers. Methods by double immunofluorescence, confocal microscopy to observe the effect of moxibustion on Determination of expression of bladder M2, M3 receptor isoforms at the protein level, at the level of genes in bladder mucosa of rats with detrusor hyperreflexia M2, M3 receptor mRNA standard copy number are compared using fluorescence quantitative PCR method, analysis of the M receptor subtype mRNA expression correlation between. The key targets and pathways on the changes of bladder and bladder C fiber M receptor in rats with detrusor hyperreflexia for moxibustion treatment of urinary incontinence, and provide scientific experimental basis for moxibustion in the treatment of urinary incontinence.