基于红外热成像技术及SCF/c-kit/ICC信号通路探讨“以俞调枢”法改善胃肠动力的作用机制

The function of gastrointestinal smooth muscle cell is controled by the interstitial cells of Cajal (ICC)，and the SCF/c-kit signal pathway is the main way to regulate the expression of ICC.So seeking for a treatment scheme which can regulate the SCF/c-kit/ICC signaling pathway is the key to solve the disorders of gastrointestinal motility.The pioneer study found that the acupoint temperature of Ren and Du Meridian is low in the people with disorders of gastrointestinal motility diseases such as gastroesophageal refluxdisease、lowtransitconstipation. We also found that "Regulating the pivot through the shu points" has the remarkable curative effect to disorders of gastrointestinal motility diseases, and it also can improve the skin temperature of the human body where the Ren and Du Meridian acupoints located. The change of the acupoint temperature of meridian reflects the movement of meridian-QI. So it is speculated that "Regulating the pivot through the shu points" can improve the circulation of the movement of Ren and Du meridian-QI,then deliver the corresponding extracellular signal effect generated by "Regulating the pivot" to the SCF/c-kit signaling pathway, which will regulate and control the expression of ICC and finally to improve the gastrointestinal motility. We'll treat ICC-loss-model rats by the way of "Regulating the pivot through the shu points", to discuss its influence to the skin temperature where the Ren and Du Meridian points located, to the gastrointestinal dynamics related monitoring indicators(including gastric emptying, the intestinal propulsive rate, gastrointestinal hormone, etc.), to the SCF/c-kit signaling pathway of the rats and analysis their correlation, reveal the mechanism of "Regulating the pivot through the shu points", promote the development of Chinese medicine external treatment.

胃肠平滑肌细胞的运动功能离不开Cajal细胞(ICC)的调控，而SCF/c-kit信号通路是调控ICC表达的主要途径，故寻找能够调控SCF/c-kit/ICC信号通路的治疗方案是改善胃肠动力的关键。研究表明，胃食管反流病、慢传输型便秘等胃肠动力障碍性疾病患者的任督二脉穴位皮温偏低，而“以俞调枢”法对该类疾病疗效显著，并能够改善其任督二脉的穴位皮温。经络穴位皮温的变化反映了经气运行状态的改变，据此我们推测“以俞调枢”法能通过改善任督二脉经气的运行，传导指针治疗产生的相应胞外信号至SCF/c-kit信号通路，影响该通路对ICC的调控而改善胃肠动力。为此，我们将通过“以俞调枢”法干预ICC缺失模型大鼠，探讨其对大鼠任督二脉穴位皮温、胃肠动力学监测指标（胃排空、小肠推进比、胃肠激素水平等）、SCF/c-kit/ICC信号通路的影响及其相关性，揭示该疗法改善胃肠动力的作用机制，促进中医外治法的发展。

在本研究中，项目组采用抗c-kit单克隆抗体制备ICC缺失模型大鼠，观察了ICC缺失模型大鼠造模前后任督二脉穴位皮温的变化，ICC缺失模型大鼠与正常大鼠任督二脉穴位皮温的差异性，及“以俞调枢”法干预前后大鼠任督二脉穴位皮温的变化。同时也观察了“以俞调枢”法干预前后模型大鼠胃肠动力学主要监测指标（胃电图、胃内残留率、小肠推进比、胃动素、胃泌素、血管活性肠肽、生长抑素等）的变化；及其对SCF/c-kit/ICC信号通路（包括SCF蛋白、c-kit蛋白、SCF mRNA、c-kit mRNA、ICC细胞数量、ICC细胞超微结构等）的影响，并深入地探讨任督二脉经络皮温的变化同胃肠动力学主要监测指标、SCF/c-kit/ICC信号通路三者之间的相关性。研究结果表明：①ICC缺失模型大鼠其任脉和督脉的主要穴位皮温、任督二脉的经络均温均较正常大鼠偏低，而通过“以俞调枢”法干预治疗后，ICC缺失模型大鼠任脉和督脉的主要穴位皮温、任督二脉的经络均温均得到了显著地提高；②“以俞调枢”法能够显著改善ICC缺失模型大鼠的胃电活动主频和平均振幅，提高其胃排空能力和小肠推进能力；③“以俞调枢”法对模型大鼠的胃动素、胃泌素、血管活性肠肽、生长抑素等胃肠激素具有良好的调控作用；④“以俞调枢”法可显著地提高模型大鼠消化道各节段（食道下段、胃运动起搏区、小肠、结肠等）平滑肌组织中的SCF蛋白、c-kit蛋白、SCF mRNA、c-kit mRNA的表达；⑤“以俞调枢”法可以显著提高模型大鼠消化道各节段平滑肌组织中ICC细胞的数量，电镜下进一步观察ICC细胞的超微结构显示其细胞的胞浆未见溶解，可见有完整的基膜，细胞器较丰富、有较多的线粒体，细胞核染色质分布较均匀，内质网正常或轻度扩张。综上，“以俞调枢”法干预前后大鼠任督二脉经络穴位皮温的变化同胃肠动力学主要监测指标的变化、SCF/c-kit/ICC 信号通路的表达之间显示出了良好的正相关性，故我们推测“以俞调枢”法很可能是通过指针疗法作用于背俞穴所产生的物理效应，经得气后转化成相应的生物电信号或生化信号，激发并改善了任督二脉经气的运行，进而影响了SCF/c-kit信号通路对ICC的调控，最终起到改善胃肠动力的“经络-分子”作用机制。本项目研究突破了既往胃肠动力障碍性疾病脏腑气机升降理论的局限性，为“以俞调枢”法在临床中的应用奠定了理论基础。