摘要
Objective To observe the effect of electroacupuncture on the morphological change of the bladder tissue and the protein expression levels of cytochrome C, Caspase-3 and Caspase-9 in the bladder tissue of rats with neurogenic bladder after suprasacral spinal cord injury, and to preliminarily explore its partial mechanism of action. Methods Forty-eight female SD rats were randomly divided into blank group（group B）, sham operation group（group S）, model group（group M） and electroacupuncture group（group EA） according to random number table method with 12 rats in each group. Neurogenic bladder models after suprasacral spinal cord injury were established by adopting modified spinal cord transection method. Electroacupuncture intervention was conducted at ＂Cìliáo（次髎 BL 32）＂＂Zhōngjí（中极 CV 3）＂ ＂Sānyīnjiāo（三阴交 SP 6）＂ and ＂Dàzhuī（大椎 GV 14）＂ on the 19 th day after modeling. After treatment for 7 consecutive days, the rats were sacrificed, and the bladder tissues were removed rapidly for microscopic observation of morphological change after HE stain and for determination of the protein expression levels of cytochrome C, Caspase-3 and Caspase-9 via Western blot method. Results① Under light microscope after HE stain, injury in varying degrees in muscle fiber arrangement and the shape and size of cells, etc. of bladder detrusor of rats in group M and group EA was found, and the degree of injury of detrusor in group M was severer than that in group EA;② comparison of the protein expression levels of cytochrome C, Pro-Caspase-9 and activated Caspase-9 between group M as well as group EA and group B as well as group S： the protein expression levels of cytochrome C, Pro-Caspase-9 and activated Caspase-9 in bladder tissue of rats in group M and group EA obviously increased（ P〈0.01, P〈0.05）; comparison between group EA and group M： the protein expression levels of cytochrome C, Pro-Caspase-9 and activated Caspase-9 in bladder tissue of rats in group EA obviously reduced（ P〈0.05）;③ comparison of the protein expression level of Caspase-3 between group M as well as group EA and group B as well as group S： the protein expression level of Pro-Caspase-3（35 KD） in group M and group EA reduced, and the protein expression level of activated Caspase-3（17 KD/19 KD） obviously increased（ P〈0.01, P〈0.05）; comparison between group EA andgroup M： the protein expression level of Pro-Caspase-3（35 KD） in group EA obviously increased, and the protein expression level of activated Caspase-3（17 KD/19 KD） obviously reduced（ P〈0.05）. Conclusion suprasacral spinal cord injury can increase the protein expression levels of cytochrome C, activated Caspase-3 and Caspase-9 in bladder tissues of rats, and electroacupuncture at BL 32, CV 3, SP 6 and GV 14 can down-regulate the protein expression levels of cytochrome C, activated Caspase-3 and Caspase-9 in bladder tissues of rats after suprasacral spinal cord injury and protect bladder tissue, which may be one of the mechanisms that electroacupuncture protects the involved bladder detrusor after suprasacral spinal cord injury.
目的:观察电针对骶上脊髓损伤后神经源性膀胱大鼠膀胱组织形态学的变化及膀胱组织中细胞色素C和caspase-3,caspase9蛋白表达量的影响,初步探讨其部分作用机制。方法:将48只雌性SD大鼠按照随机数字表方法随机分为空白组、假手术组、模型组、电针组,每组12只。采用改良的脊髓横断法制作骶上脊髓损伤后神经原性膀胱模型,于造模后第19天取"次髎""中极""三阴交""大椎"行电针干预,连续治疗7天后处死动物迅速剖取大鼠膀胱组织行HE染色后光镜观察膀胱组织形态学变化,Western blot法测定膀胱组织中细胞色素C和Caspase-3,caspase9蛋白表达情况。结果:(1)HE染色光镜下,模型组及电针组膀胱逼尿肌肌纤维排列、细胞形态大小等出现不同程度的损害,且电针组逼尿肌损害较模型组轻;(2)细胞色素C和procaspase-9及活化的caspase-9蛋白表达:与空白组及假手术组比较,模型组、电针组膀胱组织中细胞色素C和pro-caspase-9及活化的caspase-9蛋白表达明显升高(P<0.01,P<0.05);与模型组比较,电针组膀胱组织中的细胞色素C和pro-caspase-9及活化的caspase-9蛋白表达明显下降(P<0.05)。(3)Caspase-3蛋白的表达:与空白组及假手术组比较,模型组及电针组的pro-caspase-3(35 KD)蛋白表达量下降,活化的caspase-3(17 KD/19 KD)蛋白表达量明显增加(P<0.01,P<0.05);与模型组比较,电针组的pro-caspase-3(35 KD)蛋白表达明显增加,活化的caspase-3(17 KD/19 KD)蛋白表达量明显降低(P<0.05)。结论:骶上脊髓损伤可使大鼠膀胱组织中细胞色素C、活化的caspase-3,caspase9蛋白表达升高,电针"次髎""中极""三阴交""大椎"可下调骶上脊髓损伤后神经原性膀胱大鼠膀胱组织中细胞色素C、活化的caspase-3,9蛋白表达,保护膀胱组织,这可能是其保护骶上脊髓损伤后受累膀胱逼尿肌的作用机制之一。
基金
Supported by National Natural Science Foundation of China:81473753
