电针对完全性脊髓损伤后神经源性膀胱大鼠脊髓组织中Caspase-9、细胞色素C及凋亡蛋白酶激活因子-1表达的影响

摘要

目的 观察电针对完全性脊髓损伤后神经源性膀胱大鼠膀胱功能及脊髓组织中Caspase-9、细胞色素C(Cyt-C)及凋亡蛋白酶激活因子-1(Apaf-1)表达的影响.方法 雌性Sprague-Dawley大鼠60只,随机抽取36只,采用改良T10脊髓横断法制作完全性脊髓损伤后神经源性膀胱模型,取24只成功模型分为模型组和电针组各12只,其余未造模大鼠24只随机分为假手术组和空白组各12只.于造模后第19天取次髎、中极、三阴交、大椎行电针,连续7 d后行尿流动力学检测,TUNEL法测定细胞凋亡率,Western blotting测定脊髓组织中Caspase-9、Cyt-C和Apaf-1蛋白的表达情况.结果 与空白组和假手术组比较,模型组和电针组膀胱最大容量及膀胱顺应性均明显降低(P<0.01),膀胱基础压力和漏尿点压力增加(P<0.05);脊髓组织TUNEL阳性率显著升高(P<0.001);脊髓组织中Caspase-9、Cyt-C和Apaf-1蛋白表达升高(P<0.05).与模型组比较,电针组的膀胱最大容量及膀胱顺应性明显增加(P<0.01),膀胱基础压力和漏尿点压力降低(P<0.05);脊髓组织TUNEL阳性率明显降低(P<0.01);脊髓组织中Caspase-9、Cyt-C及Apaf-1蛋白表达量降低(P<0.05).结论 电针次髎、中极、三阴交、大椎可改善完全性脊髓损伤后神经源性膀胱大鼠膀胱功能,其作用机制可能与脊髓组织中Caspase-9、Cyt-C和Apaf-1的表达下调,凋亡减少有关.%Objective To observe the effect of electroacupuncture on Caspase-9, cytochrome C (Cyt-C) and apoptotic protease activat-ing factor 1 (Apaf-1) protein in rats with neurogenic bladder after complete spinal cord injury. Methods A total of 60 female Sprague-Daw-ley rats were involved, in which 36 rats were randomly selected to establish the neurogenic bladder model with T10 spinal cord transected. 24 successful models selected were randomly divided into model group (n=12) and electroacupuncture group (n=12). The other 24 rats were al-so randomly divided into sham group (n=12) and blank group (n=12). The electroacupuncture group was electroacupunctured on Ciliao (BL32), Zhongji (RN3), Sanyinjiao (SP6) and Dazhui (GV14) for seven days. All of them were assessed with urodynamic test, TUNEL method was used to determine the apoptosis rate of the spinal cord, and Western blotting was used to detect the expression of Caspase-9, Cyt-C and Apaf-1 protein. Results Compared with the blank group and the sham group, the maximum bladder capacity and compliance de-creased (P<0.01), the urinary bladder pressure and leakage point pressure increased (P<0.05);the apoptosis rate of the spinal cord increased (P<0.001);the expression of Caspase-9, Cyt-C and Apaf-1 protein increased (P<0.05) in the model group and the electroacupuncture group. Compared with the model group, the maximum bladder capacity and compliance increased (P<0.01), the urinary bladder pressure and leak-age point pressure decreased (P<0.05); the apoptosis rate of the spinal cord decreased (P<0.05); the expression of Caspase-9, Cyt-C and Apaf-1 protein decreased (P<0.05) in the electroacupuncture group. Conclusion Electroacupuncture on Ciliao, Zhongji, Sanyinjiao and Da-zhui could improve the bladder function after spinal cord injury in rats. The mechanisms may be related with the down-regulation of Cas-pase-9, Cyt-C and Apaf-1 protein to reduce apoptosis.