大鼠胸10节段脊髓损伤致膀胱顺应性改变及形态学研究

发布时间：2018-04-17 06:25

本文选题：胸10节段脊髓损伤 + 膀胱　；参考：《天津医科大学》2014年硕士论文

【摘要】：目的：通过建立大鼠胸10节段脊髓横断模型,诱导出神经源性膀胱,观察胸10节段脊髓横断致大鼠膀胱顺应性改变后膀胱重量、逼尿肌漏尿点压、膀胱顺应性、膀胱壁组织结构及ICC细胞分布、数量的变化。探讨不同顺应性膀胱的病理生理改变,明确ICC细胞在神经源性低顺应性膀胱发病中的作用,为进一步丰富和完善神经源性低顺应性膀胱发病机理及治疗提供必要的理论和实验基础,同时,为后续进一步实验提供前期支持。方法：正常成年雌性Wistar大鼠40只,体重在210-220g,随机分为两组：A组8只、B组32只。A组为假手术对照组,B组为胸10节段脊髓横断组。主要步骤为：称重、麻醉后,鼠板俯卧位固定,定位：根据浮肋连接的第13胸椎作为骨性标志定位至胸10节段脊髓(对应脊柱第8胸椎),确定损伤部位后背部备皮、消毒,取背部正中切口,依次切开皮肽、皮下筋膜、向两侧钝性分离竖脊肌,破坏椎体,直至暴露脊髓,用眼科手术刀将脊髓快速切断(可见大鼠后肢痉挛性抽搐数次后软瘫,用显微剪剪除一段约1mm长脊髓,保证脊髓完全横断)。对照组只暴露脊髓,不切断,逐层缝合切口。各组均于手术后6周行尿动力学检测,检测完成后处死大鼠并留取膀胱标本,行膀胱称重。常规HE染色观察膀胱壁平滑肌、纤维结缔组织变化；VG染色及Vaehoeff染色,光镜下观察膀胱中肌层厚度、胶原纤维及弹力纤维量的变化；应用免疫荧光技术观察ICC细胞在膀胱壁的分布及数量,每个标本随机选取5个视野,得到的数码照片用Image pro图像处理软件分析,计算每个视野ICC细胞数量,进行统计分析。结果： 1.成功制作了大鼠胸10节段脊髓横断动物模型并顺利地进行了大鼠膀胱尿流动力学测定。 2.膀胱顺应性：至行尿流动力学检测时,32只实验组大鼠,死亡10只,尿流动力学结果显示,存活的22只大鼠中,神经源性高顺应性膀胱大鼠12只,神经源性低顺应性膀胱大鼠6只,正常顺应性膀胱大鼠4只。神经源性高顺应组膀胱顺应性与假手术对照组相比明显升高,有统计学意义(P0.05)；神经源性低顺应组膀胱顺应性与假手术对照组相比明显降低,有显著差异(P0.01)。 3.形态学研究显示：胸10节段脊髓横断6周后膀胱结构和功能发生了明显改变,膀胱重量明显增加,实验组膀胱重量明显比对照组重,有显著差异(P0.01)；膀胱/体重比实验组明显高于对照组,有显著差异(P0.01)。光镜观察到胸10节段脊髓横断6周后逼尿肌出现不同程度的肌萎缩,肌束间胶原纤维浸润显著增多；低顺应组纤维结缔组织的百分含量明显大于对照组及高顺应组,有显著差异(P0.01)；高顺应组纤维结缔组织百分含量大于对照组,有统计学意义(P0.05)。荧光显微镜观察显示不同组间ICC细胞有所变化,高顺应组平均每视野下ICC数量较正常组明显减少(P0.01),差异显著；低顺应组平均每个视野下ICC数量较正常组明显增加(P0.01),差异显著。结论： 1胸10节段脊髓损伤对膀胱功能的影响在膀胱顺应性上可表现为高顺应性及低顺应性两类,具体类型应根据尿流动力学结果划分。 2神经源性膀胱尿流动力学表现为：低顺应组膀胱最大容量降低、逼尿肌漏尿点压力增高、膀胱顺应性降低；高顺应组膀胱最大容量增大、逼尿肌漏尿点压力降低、膀胱顺应性升高。 3胸10节段脊髓损伤后,膀胱壁组织发生重塑,逼尿肌出现不同程度的肌萎缩,肌束间胶原纤维浸润,纤维结缔组织含量发生变化,影响膀胱顺应性。 4胸10节段脊髓损伤后,不同顺应性组别间ICC数量发生变化,可能与膀胱顺应性发生变化有关。
[Abstract]:Purpose :

Objective To investigate the changes of bladder weight , pressure of urinary bladder , bladder compliance , bladder wall structure and the distribution and quantity of ICC cells after spinal cord transection in thoracic 10 segments of rats . The changes of bladder weight , urinary bladder pressure , bladder compliance , bladder wall structure and ICC cell distribution and quantity were observed .

Method :

Forty - four normal adult female Wistar rats weighing 210 - 220g were randomly divided into two groups : Group A : Group A : Group A : Group A : Group A : Group A : Group A : Group A : The spinal cord of the thoracic 10 segments ( corresponding to the eighth thoracic vertebra ) . In the control group , only the spinal cord was exposed , the incision was not cut and the incision was sutured layer by layer . After 6 weeks after surgery , the rats were sacrificed and the bladder specimens were removed and the bladder was weighed . Routine HE staining was used to observe the changes of bladder wall smooth muscle and fibrous connective tissue ;
VG staining and Vaehoeff staining were used to observe the changes of muscle layer thickness , collagen fiber and elastic fiber quantity in the bladder under light microscope .
The distribution and quantity of ICC cells in the bladder wall were observed by immunofluorescence technique . Five visual fields were randomly selected for each specimen . The digital photographs obtained were analyzed by image pro image processing software , and the number of ICC cells in each field of view was calculated and analyzed statistically .

Results :

1 . The rat model of spinal cord transection was successfully established and the urodynamic measurement of bladder urodynamic in rats was performed successfully .

2 . Bladder compliance : At the time of urodynamic examination , 32 experimental rats died 10 rats . The results of urodynamic showed that in 22 rats survived , 12 were neurogenic high - compliance bladder rats , 6 in neurogenic low - compliance bladder , 4 in normal - compliant bladder .
Compared with the sham - operated control group , the bladder compliance in the low - compliance group was significantly lower than that in the sham operation group ( P0.01 ) .

3 . The morphology study shows :

The bladder structure and function changed significantly after 6 weeks of spinal cord transection in thoracic 10 segments . The bladder weight was significantly increased , and the bladder weight in the experimental group was significantly higher than that of the control group ( P0.01 ) .
Compared with the control group , the bladder / body weight ratio was significantly higher than that of the control group ( P0.01 ) .

After 6 weeks of transection of the spinal cord in the thoracolumbar spinal cord , different degrees of muscle atrophy were observed , and there was a significant increase in the infiltration of collagen fibers between the muscle bundles ;
The percentage of fibrous connective tissue in low - compliance group was significantly higher than that in control group and high - compliance group ( P0.01 ) .
The percentage of ICC in the high - compliance group was higher than that of the control group ( P0.05 ) .
The average number of ICC in the low - compliance group was significantly higher than that in the normal group ( P0.01 ) , and the difference was significant .

Conclusion :

The effect of spinal cord injury on bladder function in thoracic 10 segments can be seen as high compliance and low compliance in bladder compliance , and the specific types should be divided according to the results of urodynamic results .

The urodynamic characteristics of neurogenic bladder were as follows : the lower urinary bladder capacity of the low - compliance group , the increase of the pressure of the urethral sphincter and the decrease of the bladder compliance ;
The bladder maximum capacity of the high - compliance group was increased , the pressure of the urethral sphincter decreased , and the bladder compliance increased .

After spinal cord injury in 10 segments of the thoracic cavity , the bladder wall tissue was remodeled , the muscle atrophy , the infiltration of collagen fibers between the muscle bundles , the changes of connective tissue content in the muscle bundles and the bladder compliance were affected .

After spinal cord injury in thoracic 10 segments , the number of ICC changes in different compliance groups may be related to changes in bladder compliance .

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R651.2

【参考文献】