从电针对FGIDs大鼠5-HT信号系统的整体调节探讨经穴-脏腑相关理论

In our previous research it was been shown that gastrointestinal motility and gastrointestinal bioelectric of functional gastrointestinal disorders FGIDs model of rats can be improved by electroacupuncture(EA), and the excessive expression of 5-HT in the brain, stomach and intestine was reduced. This study intends to copy the animal modl of FGIDs combined a variety of causes and based on the previous studies using cold - bondage, irregular diet and comprehensive methods. EA at Zusanli(ST 36), yinlingquan(SP 9), yingu(KI 10), non-acupoint after observing manifestations and symptoms and detecting gastrointestinal motility and visceral sensitivity. The content of 5-HT, MAO is to be detected by ELISA, the expression of 5-HT3R, 5-HT4R, 5-HT7R, SERT is to be detected by immunohistochemistry, and the mRNA quantification is to be detected by RT-PCR in each level parts of cerebro-gastroenteral axis such as the hypothalamus, hippocampus, spinal cord, stomach, and intestin It is to be studied that regulatory role of gastrointestinal motility and visceral sensitivity of FGIDs model rats by EA at different points. Exploring on the mechanism of influence gastrointestinal motility and visceral sensitivity through 5-HT signaling pathway on FGIDs being mediated by electroacupuncture. Comprehensively, exploration viscera function of FGIDs by electroacupuncture at different acupoint, and analysis the approach of acupoints and viscera of specific correlation in the concept of wholism,then to provide a scientific basis for the theory of correlation and specificity ways between acupoints and viscera.

我们之前的研究显示电针足三里穴能改善FGIDs模型大鼠胃肠运动，降低胃、肠和脑中5-HT的过度表达。本课题拟在前期研究基础上结合FGIDs多种病因采用寒冷-束缚、饥饱失常等综合法复制模型。电针足三里穴、阴陵泉穴、阴谷穴、非经非穴后观察症状体征并检测胃肠动力和内脏敏感性，并采用ELISA、免疫组化、RT-PCR等方法，检测下丘脑、海马、不同脊髓段、胃、肠等脑肠轴各水平部位的5-HT、MAO含量，5-HT3R、5-HT4R、5-HT7R、SERT的表达及mRNA，研究电针不同穴位对FGIDs模型大鼠胃肠动力异常、内脏高敏感性的调节作用，进一步探讨电针不同腧穴对5-HT信号通路介导FGIDs胃肠动力及内脏敏感性的影响，从整体上综合分析经穴-脏腑特异性相关联系的途径，为研究经穴-脏腑相关提供科学依据。

本项目旨在阐明电针不同穴位对功能性胃肠病（FGIDs）模型大鼠的胃肠功能的影响，从整体观念出发，以5-HT信号通路介导FGIDs的胃肠动力学、内脏敏感性为切入点，在正常、病理及治疗的不同状态下，观察脑-胃肠轴的各个不同部位、不同水平5-HT信号系统的影响，探讨电针不同的穴位对调节FGIDs的影响，并阐明其作用机制，从而在整体观念指导下，研究经穴、脏腑及两者之间的联系，是通过调节整个有机体来达到的，为其可能机制提供科学依据。.实验结果如下：.1. 针对FGIDs的病因、症状及病程上兼具本病的特点等，本研究采用寒冷-束缚、饥饱失常综合法、冰水灌胃等方法制作FGIDs大鼠模型，制定模型复制成功标准并进行评价。.2. FGIDs模型组大鼠胃肠电频率、振幅、胃排空率、小肠推进率较正常组有显著性差异（P<0.05）；电针足三里穴、阴陵泉穴能明显调节胃肠动力，较模型组、阴谷组、非经非穴组比较有显著性差异（P<0.05）。模型组AWR容量阈值明显升高，MC计数较正常组明显降低，电针足三里穴能明显改善腹腔高敏感性，较模型组、阴谷组、非经非穴组有显著性差异（P<0.05）。.3. 电针足三里穴、阴陵泉穴能明显降低FGIDs模型组脑肠轴各水平部位升高的5-HT、MAO含量，较阴谷组、非经非穴组有显著性差异（P<0.05）。模型组各组织中5- HT3R、5- HT4R、5- HT7R的表达与正常组比较都有显著性差异（P<0.05）；电针足三里穴、阴陵泉穴能明显降低各组织中5- HT3R、5- HT7R的表达，能明显升高各组织中5- HT4R的表达，较模型组、阴谷组、非经非穴组有显著性差异（P<0.05）。电针足三里穴、阴陵泉穴能明显降低模型组升高的下丘脑、脊髓、胃窦、结肠等组织的5- HT3R mRNA的表达，升高5- HT4R、SERT mRNA的表达，较阴谷组、非经非穴组的有显著性差异（P<0.05）；足三里组在胃窦、结肠中的表达变化较阴陵泉组有显著性差异（P<0.05）。.综上数据分析表明，足三里穴、阴陵泉穴具有一定的穴位特异性，提示足三里穴、阴陵泉穴与胃、脾、肠之间存在特异性关系，而且，足三里与胃、脾、肠之间的关系更为密切，这为穴位脏腑相关性理论提供科学依据。