【作者】 田庆华；

【导师】 尹岭；

【作者基本信息】 中国人民解放军军医进修学院 ， 神经病学， 2004， 硕士

【摘要】 目的 应用¹⁸FDG-PET和fMRI-BOLD脑功能成像方法，我们科观察到针刺后三里穴脑干的葡萄糖代谢和脑血流增加。本实验重点研究针刺大鼠后三里穴对脑干脑肠肽（BGP）表达的影响，探讨针刺后三里穴引起脑干功能变化的细胞分子生物学基础及可能的物质通路，通过对该区域的神经体液变化的研究将为进一步探索针刺的作用机制奠定基础。 方法 将78只健康雄性SD大鼠随机分成正常对照组，针刺后三里、非经非穴以及束缚后各2h、4h、6h、8h组。采用电针刺激后三里穴后不同时间点取脑干，应用RT-PCR、Southern Blot及免疫荧光组织化学方法观察不同时间点的大鼠脑干P物质（SP）、生长抑素（SOM）、胃泌素（GAS）的表达情况。应用Image-pro-plus图像分析系统进行半定量研究。实验结果采用SPSS10.0统计软件进行数据分析，统计学方法为单因素方差分析。 结果 1．针刺大鼠后三里穴对脑干P物质（SP）表达的影响 RT-PCR结果可见后三里穴组、非经非穴组在电针2h后脑干SPmRNA表达增加（P＜0.05），6h其表达两组间有差异（P＜0.05），8h后三里组的SPmRNA表达较另三组增高，有统计学意义（P＜0.05）。解放军总医院/军医进修学院硕士学位论文中文摘要Southem Blot的结果与之相符。免疫荧光组织化学染色实验中发现电针后三里穴脑干中SP阳性细胞较另三组明显增多，统计分析有意义 （P<0.05），阳性细胞染色加深，主要分布在中脑导水管周围灰质处腹内、外侧亚核、桥臂、以及中缝大核的广泛区域。 2.针刺大鼠后三里穴对脑干生长抑素（S OM）表达的影响 RT‘PCR结果可见针刺后三里穴组2h脑干SOMmRNA表达较另外三组降低（P<0 .05）。6h及8h电针后三里穴组 sOMmRNA表达较另三组增高（P<0.05）。Sonthem Blot的结果与之相符。应用免疫荧光组织化学方法发现电针后三里穴后脑干中尤其是中脑和脑桥处SOM阳性细胞比正常情况下明显增多（P<0.01），染色加深，主要分布在在中脑导水管周围、迷走神经背核、臂旁核、以及结合臂中缝等区域。 3.针刺大鼠后三里穴对脑干胃泌素（GAS）表达的影响 RT-PCR结果可见4h至8h电针后三里穴组脑干GAS mRNA表达增高（P<0.05）;免疫荧光组织化学方法发现电针后三里穴组GAS的阳性细胞数较另三组增加（P<0.05），主要分布在中脑导水管周围灰质、桥脑背外侧、中缝背核及中缝周围灰质等区域。结论 1.针刺大鼠后三里穴脑干SPmRNA表达增加、阳性细胞数明显增多，可见电针大鼠后三里穴激活了脑干SP能神经元，增加SP的基因和蛋白表达。 2.针刺大鼠后三里穴脑干SOMmRNA的表达先减少后增加，6h免疫阳性细胞数增加，可见电针大鼠后三里穴对脑干SOMmRNA表达有先抑制后促进的作用，并促进蛋白水平的表达。 3.电针大鼠后三里穴增加脑干GAS mRNA的表达，6h免疫阳性细胞数增加，可见电针大鼠后三里穴促进了脑干GAS的基因和蛋解放军总医院/军医进修学院硕士学位论文中文摘要白表达。 4.针刺后三里穴通过影响脑干脑肠肤（SP、SOM、GAS）的表达，可能参与电针后三里穴引起脑干的葡萄糖代谢和脑血流增加的过程。正是通过这些神经体液的改变引起脑干相应区域的变化进而影响机体各系统、器官的生理功能和代谢活动。更多还原

【Abstract】 ObjectiveBased on the traditional Chinese medicine, the meridian system make all the organs and tissues of the human body into an organic entity. A great amount of research has been done indicating that EA at ST36 can achieve positive and regulatory effects on gastrointestinal motility and eletroacupuncture analgesia.1. To investigate effects of electroacupuncture of Zusanli(ST36) on the expressions of SPmRNA, SOMmRNA, GASmRNA in rat brain stem. To study the relationship between neuropeptides and EA at ST36.2. To explore the effects of EA at ST36 on the protein levels of Substance P (SP), Somatostatin (SOM), Gastrin (GAS) expressed in rat brain stem.3. To explore the possible mechanism for gastrointestinal motility and eletroacupuncture analgesia and their signal transmission along the meridian.Methods1. Seventy-eight SD rats were evenly randomized into normal control, Zuanli-EA, non-acupoint and bondage groups (at 2h, 4h, 6h, 8h time piont after 30 minutes EA stimulation). RT-PCR and image processing were used to study the changes at gene level in brain stem.2. Southern-blot technique was used to observe the influence of acupuncture at ST36 on the expressions of SP, SOM, GAS in brain stem and the relationship between EA effect and time.3. Twenty-four SD rats were evenly randomized into normal control,Zuanli-EA, non-acupoint, and bondage groups(at 6h time piont after 30 minutes EA stimulation). Immunohistochemistry and image processing were used to study the changes of neuropeptides at protein level in brain stem. Results1. The increased SPmRNA expression was seen in zusanli group and non-acupoint group compared to the normal group at 2h time piont after 30 min EA stimulation(P<0.05), there was difference between the above two groups at 6h and 8h time piont (P<0.05), the resulte was coincident with the Southern-blot result.2. SOMmRNA expression was inhibited at 2h time piont after 30min EA stimulation in zusanli group (P<0.05) , but an increase was seen at 6h and 8h time piont (P<0.05), the resulte was the same as Southern-blot result.3. An significant increase in both SP-IR and SOM-IR was found in zusanli group in RM, PAG, DRG etc. at protein level.4. There was also change of GAS expression at mRNA at 4h,6h and 8h time piont (P<0.05),and protein level at 6h time piont (PO.05).ConclusionsEA stimulation elicited an accelerated rease of neuropeptide gene expression. Zusanli-EA stimulation was specific. The different changes of expression at mRNA and protein level may neflect the effect of EA at ST36. This may constitute one of the mechanisms for gastrointestinal motility and eletroacupuncture analgesia. Neuropeptides may act as the chemical components for signal transmission along the meridian.更多还原