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 neu...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.展开更多
基金Supported by National Natural Science Foundation of China:81473753
文摘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.
