丹参酮ⅡA对骶上脊髓损伤后神经源性膀胱大鼠尿动力学的影响

Effect of tanshinone ⅡA on urodynamics in rats with neurogenic bladder after suprasacral cord injury

中文关键词:  脊髓损伤  丹参酮ⅡA  神经源性膀胱  尿动力学  大鼠

英文关键词:Spinal cord injury  Tanshinone ⅡA  Neurogenic bladder  Urodynamic  Rat

中文摘要:

  【摘要】 目的：观察丹参酮ⅡA对骶上脊髓损伤后神经源性膀胱大鼠尿动力学的影响，并探讨其可能的机制。方法：将40只SD雌性大鼠随机分为4组，每组10只，分别为对照组（Sham组）、模型组（SCI组）、丹参酮ⅡA组（TⅡA组）、甲泼尼龙组（MP组）。Sham组仅切除椎板，予尾静脉注射生理盐水1ml/d，连续7d；其他3组采用改良Allen法建立脊髓损伤后神经源性膀胱动物模型（用NYU脊髓打击器选择25mm高度打击T9节段脊髓背侧），SCI组造模成功后予尾静脉注射生理盐水1ml/d，连续7d；TⅡA组造模成功后予尾静脉注射丹参酮ⅡA磺酸钠注射液20mg/kg/d，连续给药7d；MP组造模成功后予尾静脉注射甲泼尼龙30mg/kg，仅给药1次。术后2周，每组大鼠取8只行尿动力学检查，记录膀胱内压力变化曲线并行相关参数测量；心脏灌注固定后取材T9节段脊髓组织并分离双侧L6-S1背根神经节，HE染色观察脊髓组织的结构并使用Image Pro-Plus（IPP）6.0病理分析软件测量脊髓背侧出血灶的面积占正常脊髓组织的比例，分离L6-S1背根神经节（dorsal root ganglion，DRG）后用TUNEL法（terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay）标记凋亡细胞，用IPP 6.0测得各组大鼠DRG中凋亡细胞的累积光密度（integrated optical density，IOD）并进行统计分析。结果：（1）术后2周，各损伤组与Sham组比较膀胱基础压、最大排尿压和排尿阈均显著升高（P<0.05）；TⅡA组、MP组与SCI组相比膀胱内基础压力明显下降（P<0.05），最大排尿压和排尿阈差异无统计学意义（P＞0.05）；TⅡA组与MP组比较差异无统计学意义（P＞0.05）。损伤各组与Sham组相比，残余尿量明显增加，排尿量明显减小，排尿效率显著降低（P<0.05）；TⅡA组、MP组与SCI组相比残余尿量明显减小，排尿量明显增加，排尿效率显著提高（P<0.05）；TⅡA组残余尿量较MP组明显下降，同时TⅡA组排尿效率较MP组显著升高（P<0.05）。损伤各组与Sham组相比收缩间期明显减小，无排尿性收缩明显增加（P<0.05）；TⅡA组、MP组与SCI组相比，收缩间期明显增加，无排尿性收缩明显减少（P<0.05）；TⅡA组、MP组相比收缩间期无显著性差异（P＞0.05），但TⅡA组较MP组无排尿性收缩明显减少（P<0.05）。（2）术后2周，损伤各组脊髓背侧出血灶面积占正常组织面积的比例分别为：SCI组0.048±0.001，TⅡA组0.006±0.003，MP组0.012±0.001，TⅡA组、MP组与SCI组比较损伤面积占比显著减小（P<0.05），TⅡA组和MP组比较差异无统计学意义（P＞0.05）。（3）术后2周，Sham组凋亡细胞IOD为0.67±0.37，SCI组为2.92±0.35，TⅡA组为1.81±0.27，MP组为2.16±0.18，损伤各组IOD与Sham组相比明显升高（P<0.05），TⅡA组和MP组IOD与SCI组相比显著减小（P<0.05），TⅡA组和MP组相比差异无统计学意义（P＞0.05）。结论：丹参酮ⅡA可以改善脊髓损伤后神经源性膀胱大鼠的尿动力学评价指标，改善膀胱功能并减少膀胱传入神经元细胞凋亡，其作用与甲泼尼龙相近，为脊髓损伤后神经源性膀胱的治疗提供了新的思路。

英文摘要:

  【Abstract】 Objectives: To observe the effect of tanshinone ⅡA(TⅡA) on the urodynamic of neurogenic bladder after suprasacral cord injury in rats, and to explore the possible mechanism. Methods: 40 female adult Sprague-Dawley rats were randomly divided into 4 groups: Sham group(laminectomy only, n=10), SCI group(T9 spinal cord contusive injury, n=10), TⅡA group[SCI with intravenous injection of TⅡA(20mg/kg) at 1 to 7 days post-injury at the same time, n=10], MP group[SCI with intravenous injection of methylprednisolone(30mg/kg) once at post-injury, n=10]. SCI was generated by using NYU injury device with a 10g weight dropped from a height of 25mm. Sham group and SCI group were treated with intravenous injection of 1ml normal saline at 1 to 7 days post-injury at the same time. Bladder pressure curve and associated parameters were measured by urodynamic examination at two weeks after injury. T9 segment of spinal cord was extracted and the injury area of dorsal hemorrhage was observed by light microscope after H&E staining. The L6-S1 dorsal root ganglions(DRGs) were isolated at the same time. And the levels of cell apoptosis in L6-S1 DRGs were examined by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay. The integrated optical density(IOD) of apoptotic cells in DRGs in each group was measured by Image Pro-Plus(IPP) 6.0 and statistical analyses were performed after that. Results: (1)At 2 weeks after surgery, urodynamic evaluation showed that the basal pressure(BP), micturition pressure (MP’) and threshold pressure(TP) in SCI group increased significantly compared with those in Sham group(P<0.05); BP, MP’ and TP in TⅡA group and MP group increased obviously compared with those in SCI group(P<0.05); there was no dramatically change of BP, MP’ or TP between TⅡA group and MP group. Secondly, SCI group, TⅡA group and MP group showed significantly changes in comparison of sham group, such as increase in residual volume(RV) and increase in voided volume(VV) and voiding efficiency(VE) (P<0.05); VV and VE in TⅡA group and MP group were obviously increased compared with those in SCI group(P<0.05); TⅡA group displayed marked decrease in RV and increase in VE in comparison with MP group(P<0.05). In addition, SCI group, TⅡA group and MP group showed more nonvoiding contractions(NVCs) and a reduction of intercontraction interval(ICI) compared with sham group(P<0.05); TⅡA group and MP group showed a marked change compared with SCI group, such as reduction in NVCs and increase in ICI(P<0.05); and there were less NVCs in TⅡA group than MP group(P<0.05). (2)At 2 weeks after surgery, the rate of dorsal hemorrhage area/normal tissue area was as following: 0.048±0.001 in SCI group, 0.006±0.003 in tanshinone ⅡA group, and 0.012±0.001 in MP group, the ratio in TⅡA group and MP group was decreased significantly compared with that in SCI group(P<0.05); and there was no obvious change between TⅡA group and MP group. (3)At 2 weeks after surgery, the IOD of apoptosis in L6-S1 DRGs was 0.67±0.37, 2.92±0.35, 1.81±0.27, 2.16±0.18 in Sham group, SCI group, TⅡA group and MP group, respectively. The IOD of apoptosis in SCI group, TⅡA group and MP group decreased dramatically compared with that in Sham group(P<0.05); but IOD of apoptosis in MP group, TⅡA group increased significantly compared with that in SCI group(P<0.05); there was no obvious difference between TⅡA and MP group. Conclusions: TⅡA may improve urodynamic parameters of neurogenic bladder after suprasacral cord injury, so that it improves bladder function. TⅡA has a similar effect of MP and it may provide a new idea for the treatment of neurogenic bladder after spinal cord injury.