电针上巨虚和足三里修复阳明腑实证大鼠小肠神经胶质细胞网络损伤的研究

The important treatment method of SIRS/MODS is recovery gastrointestinal motor dysfunction caused by Yangmingfushi.It is relatively clear that electro-acupuncturing Zusanli and Shangjuxu presents regulation of intestinal movement，But the mechanism of electroacupuncture Zusanli and Shangjuxu activates the intestinal motility remains unknown.Base our precious studies we investigated：Repair the network of enteric glial cells-endocrine through Electro-acupuncturing at the Zusanli and Shangjuxu.（1）investigate the frequency and the amplitude of slow wave in electromyography of intestine；（2）Immunofluorescence and transmission electron microscopy were used for detecting the distribution and overall expression of the morphological changes of EGCs；（3）The GDNF mRNA expression were detected using semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR), we analyzed the relationship neurotrophic and repair effect：（4）sulfur hydrogenated sodium as exogenous donor affect glial cells and neurons cells cultured in vitro;（5）To observe the effects of Ca2+ signaling pathways on EGC a metabolic pathway resulted by exogenous hydrogenated sulfide and in glial cells in vitro .Research the affection morphology of EGCs and the influence of endocrine network of he small intestine by acupuncture on zusanli and shangjuxu and then affect gastrointestinal motor function.To provides theoretical basis of treatment of gastrointestinal motor function obstacle by acupuncture and eridians-acu-point-internal organs research.

阳明腑实证所致的胃肠运动功能障碍是SIRS/MODS的始动因素。针刺同在阳明胃经上的足三里和上巨虚穴可有效治疗胃肠运动，但肠胶质细胞网络恢复胃肠运动功能尚不完全清楚。在已有的工作基础上，以阳明腑实证胃肠运动功能障碍大鼠为研究对象。通过（1）观察在体阳明腑实证大鼠小肠肌电慢波频率和振幅的变化；（2）免疫荧光和电子显微镜技术显示胶质神经纤维网络形态变化；（3）测定胶质细胞源性神经营养因子(GDNF)基因mRNA分析对神经元的营养支持和修复；（4）外源性神经递质H2S对小肠神经胶质细胞以及胶质细胞和神经元共同生长的影响（5）测定胶质神经细胞Ca2+离子通道的变化等方法。研究胃肠运动功能障碍小肠神经胶质细胞形态和神经传导网络的变化，以及针刺足三里和上巨虚穴对这种变化的影响，为针刺治疗胃肠运动功能障碍和经络-穴位-脏腑研究提供理论依据和实验资料。

提高胃肠动力是治疗阳明腑实证的根本。近年来研究表明针灸改善胃肠动力，促进胃肠道运动疗效显著。课题以大鼠小肠为研究对象，建立阳明腑实证大鼠模型，应用免疫荧光染色测定病理大鼠的小肠胶质神经细胞和神经元数量的变化，免疫组化和 GDNF 基因 mRNA 测定，以及不同浓度 H2S 对体外培养神经胶质细胞和神经元的影响和对 Ca2+离子通道的改变。实验结果：①阳明腑实证大鼠小肠运动功能明显异常，肌间神经丛EGCs网络结构改变，超微结构损伤。而电针上巨虚和足三里穴可提高小肠动力，修复肠神经胶质细胞网络损伤。②外源性硫化氢对肠神经胶质细胞增殖及凋亡呈双向调节作用，即低浓度硫化氢可抑制肠神经胶质细胞凋亡，促进其增殖；高浓度硫化氢可促进肠神经胶质细胞凋亡，抑制其增殖。综上研究表明针灸足三里和上巨虚改善阳明腑实证大鼠小肠动力，促进肠道运动。课题研究为进一步阐明针刺治疗胃肠运动功能障碍和经络-穴位-脏腑作用机制和推广临床应用提供了理论。