基于钙离子振荡介导下的内质网应激-自噬反应探讨“以俞调枢”法对Cajal间质细胞的调控效应机制

Cajal细胞(ICC)是胃肠平滑肌的起搏细胞，对胃肠运动起到绝对调控作用。前期研究表明，以俞调枢法能够改善任督二脉经气的运行，并通过调控ICC细胞数量和超微结构的表达起到改善胃肠动力的作用，但其对ICC的调控机制不明。新近研究发现，细胞内Ca2+稳态失衡会导致内质网应激-自噬反应并对细胞数量和形态结构的表达起到双重调控作用。据此我们推测以俞调枢法可能是通过改善任督二脉经气的运行影响了Ca2+振荡介导下的内质网应激-自噬反应对ICC的调控效应，进而改善了胃肠动力。为此，项目组拟在前期研究成果基础上进一步利用血清药理学、共聚焦显微镜、透射电镜等现代技术，观察以俞调枢法对IP3R/RyR介导的Ca2+振荡的影响、内质网应激-自噬反应/内质网自噬体表达的影响以及ICC细胞数量/活性和形态结构表达的影响，并深入探讨各指标之间的相关性，以揭示该疗法的“经络-分子”作用机制，丰富中医外治法的理论内涵。

Cajal mesenchymal cells are the pacing cells of gastrointestinal motility, which play an absolute regulatory role in the function of gastrointestinal motility.Previous studies show that,the“Regulating the pivot through the shu points” method can improve Ren and Du Meridian by operation of the Qi, and by regulating the number of ICC cells and the expression of ultramicrostructure to improve the gastrointestinal motility function,but the regulation mechanism of ICC is unknown.Recent studies have found that the Ca2+ homeostasis in cells are disorder leads to endoplasmic reticulum stress autophagy and plays a dual regulatory role in the expression of cell number and morphology.On this basis, we speculate that“Regulating the pivot through the shu points” method is likely to improve the operation of the Qi on Ren and Du Meridian and influence endoplasmic reticulum stress - mediated autophagy Ca2+ oscillation reaction regulation effect on ICC, and then improve the gastrointestinal motility.To this end,the project team intend to further utilize modern technologies such as serum pharmacology,confocal microscopy and transmission electron microscopy to observe the infuence of IP3R/RyR mediated Ca2+ oscillation, endoplasmic reticulum stress - autophagy reactions/endoplasmic reticulum autophagy body expression and the ICC cell number/activity and morphological structure expression which after using the“Regulating the pivot through the shu points”method .And explore the correlation between the indexes in order to reveal the mechanism of "meridian-molecular" regulation effect of the treatment, enrich the connotation of the theory of Chinese medicine external treatment.