【作者】 王海萍；

【导师】 朱兵；

【作者基本信息】 湖北中医药大学 ， 针灸推拿学， 2014， 博士

【摘要】 目的大量的临床资料和动物实验研究表明,针刺对胃肠运动具有双向调节效应,对治疗各种功能性胃肠疾病疗效显著。然而,针灸双向调节的确切机制并不十分明确。消化间期移行性复合运动(Migrating Motor Complex, MMC)是人类和哺乳动物胃肠固有的一种特征性的运动形式,也是反映胃肠运动状况的一种特异性的指标。本实验是在前期研究工作的基础上,采用肠道平滑肌浆膜下电极埋植以及平滑肌浆膜表面高灵敏度应变片传感器的缝合技术,在麻醉大鼠在体同步记录空肠平滑肌电活动与空肠的机械运动,在单元、集元经穴理论指导下,选取空肠的单元穴位天枢、大肠俞,及空肠集元穴位上巨虚、曲池,分别在MMC的Ⅰ、Ⅱ、Ⅲ时相针刺,在正常及游离肠管大鼠观察针刺对肠电(快、慢波)及肠运动频率与幅值的影响,探讨针刺不同穴位对肠运动的调节作用及机制。材料与方法实验选用成年雄性SD大鼠16只,体重250-300g。实验前大鼠禁食不禁水18h。10%乌拉坦(1-1.2g/kg)腹腔注射麻醉,手术取仰卧位,剑突下沿腹正中线作一约2～3cm的纵行切口,暴露空肠,Tretz韧带下10cm空肠浆膜表面缝置高灵敏度胃肠运动应变片传感器,另一端连接桥式放大器,肠壁张力信号输入Powerlab8/20数据采集系统。同时,空肠浆膜下放置自制的铂金丝针形双极电极,电极的另一端通过生物电放大器,信号输入到Powerlab8/20数据采集系统。手术后先记录基础蠕动的肠电和肠运动,待出现2个完整MMC后再行针刺操作。游离肠管大鼠模型的制作方法：在Tretz韧带上方,大约幽门下5cm的空肠肠管行游离术,肠系膜保持完整。游离肠管断面用生理盐水纱布覆盖。游离肠管后,先记录游离肠管的肠电与肠运动,待肠运动稳定后再行针刺操作。腧穴的定位和针刺操作：曲池(LI11)：桡骨近端,肘关节外侧前方之凹陷处,直刺4mm；上巨虚(ST37)：大鼠后肢足三里向下约5mm处,直刺5mm；天枢(ST25)：相当于脐中旁开5mm,直刺5mm；大肠俞(BL25)：在腰部,第4腰椎棘突下旁开5mm,直刺4mm。针刺操作手法：手动行针lmin,平补平泻,频率为120次/min。数据处理通过Chart7.0软件对肠电和肠运动信号进行分析。肠电分析的指标包括肠电快波、肠电慢波的频率(次/min)和幅值(mv),肠运动分析的指标包括收缩波的频率(次/min)、幅值(g)。针刺过程各项指标的均值与针刺前基础值比较,使用SPSS19.0统计分析软件包进行处理,采用配对t检验,计量资料以均数±标准差(x±SD)表示,P<0.05为差异有统计学意义。结果1针刺腧穴对正常大鼠空肠肠电及肠运动的影响1.1针刺腧穴对正常大鼠空肠MMCⅠ相肠电及肠运动的影响MMC Ⅰ相肠电几乎无快波,针刺曲池、上巨虚、天枢、大肠俞后,肠电快波的频率和幅值均无明显变化(P>0.05,P>0.05)。对于肠电慢波,针刺上巨虚和曲池均增加了其频率(P<0.01,P<0.05)和幅值(P<0.01,P<0.01)；针刺天枢穴降低了其频率和幅值(P<0.01,P<0.01)；针刺大肠俞对空肠肠电慢波的频率和幅值均无明显的影响(P<O.05,P<0.05)。对于肠运动,针刺上巨虚、曲池均增加了其频率(P<0.05,P<0.05)和幅值(P<0.01,P<0.01)；针刺天枢降低了其频率和幅值(P<0.01,P<0.01)；针刺大肠俞对其频率和幅值均无明显的影响(P<0.05,P>0.05)。可见,针刺集元经穴促进了MMCⅠ相空肠的肠电和肠运动,针刺单元经穴天枢起到了抑制性作用,大肠俞的效应有待进一步观察。1.2针刺腧穴对正常大鼠空肠MMCⅡ相肠电和肠运动的影响MMCⅡ相,对于肠电快波,针刺曲池、上巨虚增加了其频率(P<0.001,P<0.05)和幅值(P<0.01,P<0.01)；针刺天枢降低了其频率和幅值(P<0.01,P<0.01)；针刺大肠俞对空肠肠电快波的频率和幅值均无明显的影响(P>0.05,P>0.05)。对于肠电慢波,针刺上巨虚和曲池增加了其频率(P<0.01,P<0.05)和幅值(P<0.01,P<0.05)；针刺天枢穴降低了其频率和幅值(P<0.01,P<0.01)；针刺大肠俞对空肠肠电慢波的频率和幅值均无明显的影响(P>0.05,P>0.05)。对于肠运动,针刺上巨虚、曲池增加了其频率(P<0.01,P<0.05)和幅值(P<0.01,P<0.01)；针刺天枢降低了其频率和幅值(P<0.01,P<0.01)；针刺大肠俞对肠运动的频率和幅值无明显影响(P>0.05,P>0.05)。可见,针刺集元经穴促进了MMCⅡ相空肠的肠电和肠运动,针刺单元经穴天枢起到抑制作用,大肠俞的效应有待进一步观察。1.3针刺腧穴对正常大鼠空肠MMCⅢ相肠电和肠运动的影响MMCⅢ相,对于肠电快波,针刺曲池、上巨虚增加了其频率(P<0.05,P<0.01)和幅值(P<0.01,P<0.01)；针刺天枢降低了其频率和幅值(P<0.01,P<0.01)；针刺大肠俞对空肠肠电快波的频率和幅值均无明显的影响(P>0.05,P>0.05)。对于肠电慢波,针刺上巨虚和曲池增加了其频率(P<0.01,P<0.01)和幅值(P<0.01,P<0.01)；针刺天枢穴降低了其频率和幅值(P<0.01, P<0.01)；而针刺大肠俞对空肠肠电慢波的频率和幅值均无明显的影响(P>0.05,P>0.05)。对于肠运动,针刺上巨虚、曲池增加了其频率(P<0.05,P<0.05)和幅值(P<0.01,P<0.01)；针刺天枢降低了其频率和幅值(P<0.01,P<0.01)。针刺大肠俞对空肠肠运动的频率和幅值无明显影响(P>0.05,P>0.05)。可见,针刺集元经穴促进了MMCⅢ相空肠的肠电和肠运动,针刺单元经穴天枢起到了抑制作用,大肠俞的效应有待进一步观察。2针刺腧穴对游离肠管大鼠空肠肠电及肠运动的影响2.1针刺腧穴对游离肠管大鼠空肠MMC Ⅰ相肠电及肠运动的影响游离肠管后,HMCⅠ相肠电几乎无快波,针刺曲池、上巨虚、天枢、大肠俞后,肠电快波的频率和幅值均无明显变化(P>0.05,P>0.05)。对于肠电慢波,针刺上巨虚和曲池均增加了其频率(P<0.05,P<0.05)和幅值(P<0.01,P<0.01)；针刺天枢穴降低了其频率和幅值(P<0.01,P<0.05)；针刺大肠俞对空肠肠电慢波的频率和幅值均无明显的影响(D0.05,P>0.05)。对于肠运动,针刺上巨虚、曲池增加了其频率(P<0.05,P<0.05)和幅值(P<0.05,P<0.05)；针刺天枢降低了其频率和幅值(P<0.01,P<0.01)。针刺大肠俞对空肠肠运动的频率和幅值无明显影响(P>0.05,P>0.05)。可见,针刺集元经穴促进了空肠的肠电和肠运动,针刺单元经穴天枢起到抑制作用,大肠俞的效应有待进一步观察。2.2针刺腧穴对游离肠管大鼠空肠MMCⅡ-Ⅲ相肠电及肠运动的影响MMCⅡ-Ⅲ相,对于肠电快波,针刺曲池、上巨虚增加了其频率(P<0.05,P<0.01)和幅值(P<0.05,P<0.05)；针刺天枢降低了其频率和幅值(P<0.01,P<0.01)；针刺大肠俞对空肠肠电快波的频率和幅值均无明显的影响(P>0.05,P>0.05)。对于肠电慢波,针刺上巨虚和曲池增加了其频率(P<0.05,P<0.05)和幅值(P<0.01,P<0.01)；而针刺天枢穴降低了其频率和幅值(P<0.01,P<0.01)；针刺大肠俞对空肠肠电慢波的频率和幅值均无明显的影响(P>0.05,P>0.05)。对于肠运动,针刺上巨虚、曲池增加了其频率(P<0.05,P<0.05)和幅值(P<0.001,P<0.01)；针刺天枢降低了其频率和幅值(P<0.01,P<O.05)；针刺大肠俞对空肠肠运动的频率和幅值均无明显影响(P>0.05,P>0.05)。可见,针刺集元经穴促进了MMCⅡ-Ⅲ相空肠的肠电和肠运动,针刺单元经穴天枢起到抑制作用,大肠俞的效应有待进一步观察。结论本研究较系统地观察了针刺单元经穴和集元经穴对正常和游离肠管大鼠空肠MMC各个时相肠电、肠运动的影响,得出以下结论：(1)针刺集元经穴(与空肠异神经节段支配的腧穴)上巨虚、曲池对正常及游离肠管大鼠空肠MMC各个时相的肠电、肠运动均表现为兴奋性作用,这可能与针刺集元经穴激活副交感神经活动性有关。(2)针刺单元经穴天枢(与空肠同神经节段支配的腧穴)对正常及游离肠管大鼠空肠MMC各个时相的肠电、肠运动均表现为抑制性作用,这可能与针刺单元经穴天枢激活交感神经活动性有关。(3)针刺不同部位的穴位对肠运动有着不同的影响,且存在规律性调节。针刺对肠运动的影响与穴位所在的部位及肠的神经节段支配密切相关。(4)针刺对肠运动的双向调节效应与穴位特异生物学特性有关。单元经穴天枢对空肠运动产生抑制性调节,而集元经穴上巨虚、曲池对空肠运动产生兴奋性调节。二者对空肠运动的调节,并不受肠运动状态的影响,而与腧穴自身的生物学特性有关。更多还原

【Abstract】 ObjectMost of clinical and animal studies suggest that acupuncture has an effect of dual-directional regulation on gastrointestinal motility. There are preonunced effects of acupuncture on the treatment of various functional gastrointestinal disorders. However, the exact mechanism remains unclear for Dual effects of acupuncture on gastrointestinal motility.Interdigestive migrating motor complex (MMC), which is an inherent characteristic movement patterns in human and mammal’s gastrointestinal tract in the fasting state, is thought to be a specific index of reflecting gastrointestinal motility conditions.Based on the preliminary studies, Electromyography (EMG) and mechanical motilities were recorded by implanted electrode in the intestinal smooth muscle and sutured strain gauge sensors on the serosal surface of the normal and detached jejunum in anesthetized rats. Acupuncture was applied at ST37, LI11, ST25and BL25on different phases I, II and III of MMC respectively to explore the effect on EMG and mechanical motilities during different phases in normal and detached Jejunum rats respectly.Materials and MethodsSixteen healthy adult male Sprague-Dawley rats were used, weighing between250to300g. After an18h fast (with free access to water), rats were anesthetized with an intraperitoneal injection of10%urethane(1.0-1.2g/kg). A median abdominal incision (2-3cm) was made on surgical supine position. Then a strain gauge sensors was sutured on the smooth muscle serosal surface of the jejunum (10cm downstream from the Tretz Ligament) to record circular muscle contractions in anesthetized rats. The wires from the transducers were connected with the recording system. At the same time, one pair of bipolar platinum electrodes were implanted in the intestinal smooth muscle in Jejunum in anesthetized rats. The electrodes were connected to Powerlab8/20polygraph. The EMG and mechanical motilities were recorded by AD Instruments after Surgery. Acupuncture was carried on after two to three complete MMC Cycles were recorded about an hour later.Methods of detached Jejunum model:A detached surgery was operated on Jejunum (5cm proximal to the pylorus) with intact mesentery. A saline gauze was covered with detached Jejunum cross-sections. There was a recording of basic EMG and mechanical motilities after the detached surgery. Acupuncture was carried on after mechanical motilities recover to a stable condition.Acupoints positioning and Acupuncture operation: ST37(Shang ju xu),5mm lateral to anterior tubercle of the tibia and15mm below; LI11(Qu chi).locates in the midpoint between the lateral end of the transverse cubical crease and the lateral epicondyle of the humerus, inserting to a depth of4mm; ST25(Tian shu),2mm lateral to anterior median line, on the level of the navel, inserting to a depth of about5mm; BL25(Da chang shu), locates in the waist,5mm lateral to posterior midline under the fourth lumbar spines, inserting to a depth of about4mm. The needle was rotated clockwise and anti-clockwise for60s at2Hz at each acupoint.Statistical analysisStatistical analysis was performed with SPSS software for Windows,version19.0. All data were expressed as a mean±SD. Mean values were compared using the paired t test to compare the intestinal motility before and after acupuncture. The difference was considered significant at P<0.05.Signals of EMG and mechanical motilities were analyzed through Chart7.0software. The frequency (times/min) and amplitude (mv) of fast wave and slow wave were included in EMG analysis factors. The frequency (times/min) and amplitude (g) of contraction wave were included in mechanical motilities analysis factors. Statistical analysis was performed with SPSS software version19.0for Windows. All experiment data was expressed as a mean±SD. Mean values were compared using the paired T test to compare EMG and mechanical motilities before and after acupuncture. The difference was considered significant at P<0.05.Results1Effect of acupuncture at different acupoints on normal jejunum Electromyography (EMG) and Mechanical Motility during different MMC Phases in Anesthetized Rats1.1Effect of acupuncture at different acupoints on normal jejunum Electromyography(EMG) and Mechanical Motility during MMC Ⅰ Phases in Anesthetized RatsNo EMG fast wave during MMC Ⅰ Phases. There was no significant change (P>0.05,P>0.05) in the frequency and amplitude of jejunum EMG fast wave after acupuncture at ST37, LI11, ST25and BL25acupoints.For EMG slow wave, acupuncture at either ST37or LI11increased the frequency (P<0.01, P<0.05) and amplitude(P<0.01, P<0.01) of jejunum EMG slow wave significantly; whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of jejunum EMG slow wave significantly. Acupuncture at BL25had no significant effect (P>0.05,P>0.05) on the frequency and amplitude of jejunum EMG slow wave significantly.For mechanical motilities, acupuncture at either ST37or LI11increased the frequency (P<0.05, P<0.05) and amplitude (P<0.01, P<0.01) of mechanical motilities in jejunum significantly;whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of mechanical motilities in jejunum significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of mechanical motilities in jejunum significantly.1.2Effect of acupuncture at different acupoints on normal jejunum Electromyography (EMG) and mechanical motility during MMC Ⅱ Phases in Anesthetized RatsDuring MMC Ⅱ Phases,for EMG fast wave, acupuncture at either ST37or LI11increased the frequency (P<.001, P<0.05) and amplitude (P<0.01, P(0.01) of jejunum EMG fast wave significantly;whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of jejunum EMG fast wave significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of jejunum EMG fast wave significantly.For EMG slow wave, acupuncture at either ST37or LI11increased the frequency (P<0.01,P<0.05) and amplitude (P<0.01, P<0.05) of jejunum EMG slow wave significantly;whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of jejunum EMG slow wave significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of jejunum EMG slow wave significantly.For mechanical motilities, acupuncture at either ST37or LI11increased the frequency (P<0.01, P<0.05) and amplitude (P<0.01, P<0.01) of mechanical motilities in jejunum significantly; whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of mechanical motilities in jejunum significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of mechanical motilities in jejunum significantly.1.3Effect of acupuncture at different acupoints on normal jejunum Electromyography (EMG) and mechanical motility during MMC III Phases in Anesthetized RatsDuring MMC III Phases,for EMG fast wave,acupuncture at either ST37or LI11increased the frequency (P<0.05, P<0.01) and amplitude (P<0.01, P<0.01) of jejunum EMG fast wave significantly;whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of jejunum EMG fast wave significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of jejunum EMG fast wave significantly.For EMG slow wave, acupuncture at either ST37or LI11increased the frequency (P<0.01, P<0.01) and amplitude (P<0.01, P<0.01) of jejunum EMG slow wave significantly; whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of jejunum EMG slow wave significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of jejunum EMG slow wave significantly.For mechanical motilities,acupuncture at either ST37or LI11increased the frequency (P<0.05,P<0.05) and amplitude (P<0.01, P<0.01) of mechanical motilities in jejunum significantly; whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of mechanical motilities in jejunum significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of mechanical motilities in jejunum significantly.2Effect of Acupuncture at different acupoints on detached jejunum Electromyography (EMG) and mechanical motility during different MMC Phases in Anesthetized Rats2.1Effect of acupuncture at different acupoints on detached jejunum Electromyography (EMG)and mechanical motility during MMC Ⅰ Phases in Anesthetized RatsNo EMG fast wave still during MMC I Phases after the surgery of detached Jejunum. There was no significant change (P>0.05, P>0.05) in the frequency and amplitude of jejunum EMG fast wave after acupuncture at ST37, LI11, ST25and BL25acupoints.For EMG slow wave, acupuncture at either ST37or LI11increased the frequency (P<0.05, P<0.05) and amplitude(P<0.01, P<0.01) of jejunum EMG slow wave significantly; whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of jejunum EMG slow wave significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of jejunum EMG slow wave significantly.For mechanical motilities, acupuncture at either ST37or LI11increased the frequency (P<0.05, P<0.05) and amplitude (P<0.05, P<0.05) of mechanical motilities in jejunum significantly; whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of mechanical motilities in jejunum significantly. Acupuncture at BL25had no significant effect (P>0.05,P>0.05) on the frequency and amplitude of mechanical motilities in jejunum significantly.2.2Effect of acupuncture at different acupoints on detached jejunum Electromyography (EMG) and mechanical motility during MMC Ⅱ-Ⅲ Phases in Anesthetized RatsDuring MMC Ⅱ-Ⅲ Phases, for EMG fast wave, acupuncture at either ST37or LI11increased the frequency (P<0.05,P<0.01) and amplitude (P<0.05, P<0.05) of jejunum EMG fast wave significantly; whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of jejunum EMG fast wave significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of jejunum EMG fast wave significantly.For EMG slow wave,during MMC Ⅱ-Ⅲ Phases, acupuncture at either ST37or LI11increased the frequency (P<0.05,P<0.05) and amplitude (P<0.01,P<0.01) of jejunum EMG slow wave significantly, whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of jejunum EMG slow wave significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of jejunum EMG slow wave significantly.For mechanical motilities, during MMC Ⅱ-Ⅲ Phases, acupuncture at either ST37or LI11increased the frequency (P<0.05,P<0.05) and amplitude(P<0.001,P<0.01)of mechanical motilities in jejunum significantly, whereas acupuncture at ST25decreased the frequency (P<0.01) and amplitude (P<0.01) of mechanical motilities in jejunum significantly. Acupuncture at BL25had no significant effect (P>0.05, P>0.05) on the frequency and amplitude of mechanical motilities in jejunum significantly. ConclusionsThis study systematically investigated the effects of acupuncture at different acupoints on normal and detached jejunum Electromyography (EMG) and mechanical motility during MMC phases in anesthetized rats. We make the following conclusions according to the above results:(1)Acupuncture at heterotopic acupoints (ST37or LI11) has exciting effects on EMG and mechanical Motility during MMC phases in normal and detached Jejunum.(2)Acupuncture at homotopic acupoints ST25always produces inhibition effect on EMG and mechanical Motility during MMC phases in normal and detached Jejunum.(3)The effects of acupuncture at different acupoints on jejunum EMG and mechanical motility during MMC phases are different which are related to both stimulation acupoint and the segment innervation of gastrointestinal organ.(4)Dual-directional regulation effect of acupuncture depends on the acupoint specific biological characteristic. Heterotopic acupoints (ST37or LI11) mainly produce exciting effect on Jejunum mechanical motility, whereas homotopic acupoints (ST25) mainly produce inhibition effect on jejunum mechanical motility. The inhibitory effects of acupuncture at homotopic acupoint ST25and exciting effects of acupuncture at heterotopic acupoints ST37or LI11on jejnum mechanical motility are independent of intestinal movement, which are associated with acupoints characteristics.更多还原