兔神经源性尿路障碍输尿管功能变化及BKca表达研究

发布时间：2018-02-09 03:28

  本文关键词： 神经源性尿路障碍 大电钙激活钾通道 影像尿动力学 输尿管功能　出处：《郑州大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的 神经源性尿路功能障碍(Neurogenic urinary tract dysfunction NUTD)是指任何与排尿有关中枢和神经受到损伤，出现的尿路功能障碍。有效持久保护上尿路和提高生活质量是NUTD基本治疗原则；传统研究表明，上尿路恶化很可能发生在排尿时尿道压力高或膀胱内的压力升高。NUTD的治疗的主要目标是实现低尿液储存压力，以保护上尿路功能。尽管口服抗胆碱能药物、逼尿肌注射A型肉毒毒素，并且结合清洁间歇导尿(CIC)，或进行外科干预（如自体膀胱扩大术或肠道膀胱扩大术），可以有效的减低膀胱内压，提高膀胱容量和顺应性，但部分患者仍出现反流性上尿路损害。提示除下尿路继发性尿动力学高危因素外，NUTD上尿路损害发生还可能与神经系统失去对输尿管的功能支配后输尿管发生自身功能障碍有关。本研究通过对骶髓损伤神经源性逼尿肌无收缩兔模型建立后两个月的输尿管功能及钙离子相关通道检测，初步探讨NUTD早期输尿管功能变化、病理学改变以及大电钙激活钾通道(large-conductance Ca2+-activated K+channels,BKca)的表达情况，以了解NUTD上尿路功能变化及其机制，为预防反流性上尿路损害发生和阻断其进展提供理论依据。 方法 1.动物模型的建立：将30只日本大耳白兔随机均分为NUTD组、实验对照组和空白对照组。NUTD组：以复方盐酸赛拉嗪注射液臀部肌肉注射（0.6ml/kg）麻醉后，俯卧固定，于背部沿L5-L7棘突行纵形切口，逐层分离皮暴露L6棘突，，在L6棘突两侧钝性分离腰大肌，单关节咬骨钳咬除L6棘突，暴露髓腔，眼科剪锐性横断脊髓，眼科镊进入髓腔钳夹捣毁横断面下骶髓，填充无菌明胶海绵，逐层关闭切口，2周后行脊柱MRI和影像尿动力学检查证实存在明确神经损害，膀胱表现为逼尿肌无收缩但无膀胱输尿管反流；实验对照组：仅咬除棘突，未破坏脊髓，2周后行脊柱MRI和尿动力学检查证实不存在明确神经损害，无明显膀胱尿道功能障碍；空白对照组：未进行任何手术处理。 2.建模后2月行影像尿动力学测定、输尿管功能评估、形态学观察及输尿管标本制作。 3.应用免疫组织化学法、荧光定量PCR和Western blot检测其输尿管平滑肌细胞（USMC）BKca mRNA和蛋白表达情况。 结果 1.发现动物模型建立后2月NUTD组表现为逼尿肌无收缩，无膀胱输尿管反流发生； 2.在1ml/kg/min恒定速度静脉滴注生理盐水条件下，NUTD组30min内产生的尿量为（18.5±3.7）ml、实验对照组为（19.5±2.4）ml、空白对照组为（17.8±3.2）ml，三组间的差异无统计学意义；但左侧输尿管蠕动次数NUTD组（39.0±3.0）显著低于实验对照组（44.0±5.0）和空白对照组（46.0±4.0），差异有统计学意义(P 0.05)； 3.光镜下NUTD组输尿管上皮细胞、固有层和肌层无明显变化，外膜有不同程度的充血和炎细胞浸润； 4. NUTD组USMC中BKcaα mRNA（2.89±0.67）和BKcaβ mRNA（4.59±1.22）表达相对于空白对照组mRNA（BKcaα：1.02±0.21；BKcaβ：1.03±0.28）显著上调，差异有统计学意义（P＜0.001）；NUTD组BKcaα（1.35±0.32）和BKcaβ（1.47±0.10）蛋白表达相对于空白对照组（BKcaα：0.89±010；BKcaβ：1.03±0.13）显著也上调，差异有统计学意义（P＜0.001）。 结论 1.神经源性尿路障碍(NUTD)存在原发性输尿管功能障碍； 2.原发性输尿管功能障碍在神经原性尿路功能障碍未发生膀胱输尿管反流和病理改变时已经发生； 3.输尿管平滑肌细胞中BKca表达水平升高，可能是其出现输尿管原发性功能障碍的原因之一。
[Abstract]:Purpose Neurogenic urinary tract dysfunction ( Neurogenic urinary tract dysfunction ) refers to any urinary tract dysfunction related to urinary tract dysfunction and urinary tract dysfunction . The main objective of this study is to reduce bladder internal pressure and improve bladder capacity and compliance . method 1 . Establishment of animal model : 30 Japanese white rabbits were randomly divided into two groups : Group D group , experimental control group and blank control group . 2 . Urodynamic measurement , ureter function evaluation , morphological observation and ureter specimen preparation were performed in February after modeling . 3 . Using immunohistochemical method , fluorescence quantitative PCR and Western blot to detect the mRNA and protein expression of the ureteral smooth muscle cells ( USMC ) . Results 1 . Two months after the establishment of the animal model , the group D showed no contraction and no vesicoureteral reflux . 2 . Under the condition of 1 ml / kg / min constant velocity intravenous drip , the urine volume was ( 18.5 卤 3.7 ) ml , the experimental control group was ( 19.5 卤 2.4 ) ml , the blank control group was ( 17.8 卤 3.2 ) ml , the difference between the three groups was not statistically significant , but the left ureter wriggling frequency was significantly lower than that in the experimental control group ( 44.0 卤 5.0 ) and the blank control group ( 46.0 卤 4.0 ) , the difference was statistically significant ( P 0.05 ) ; 3 . There was no obvious change of ureter epithelial cells , lamina propria and muscular layer in the group D group under light microscope , and the outer membrane had different degrees of congestion and inflammatory cell infiltration . 4 . The expression of BK ca 伪 mRNA ( 2.89 卤 0.67 ) and BK ca 尾 mRNA ( 4.59 卤 1.22 ) in group D group was significantly up - regulated with respect to blank control group mRNA ( BK ca . 伪 : 1.02 卤 0.21 ; BK ca . 尾 : 1.03 卤 0.28 ) . The difference was statistically significant ( P < 0.001 ) . NUTD缁

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