基于miR-203a-3p调控靶基因ATG14介导ICC自噬探讨芪石升降归元饮对胃食管反流病的作用机制

胃食管反流病（GERD）与胃肠动力障碍密切相关，我们前期研究发现胃肠动力起搏细胞ICC自噬是GERD的关键，证明了“任督二脉经气失衡致脾胃枢机失衡是本病病机本质”及“以枢调枢”法可以改善ICC自噬治疗GERD。现本团队基于“以枢调枢”理论研创了芪石升降归元饮，具有调节任督经气以调脾胃枢机的作用。目前基于自噬的基因调控在中医药治疗GERD的分子机制层面相关研究仍欠缺。我们通过预测分析自噬基因及相关miRNA后提出：芪石升降归元饮可能基于miR-203a调控ATG14抑制ICC自噬从而治疗GERD。因此，本项目通过体内外实验，采用芪石升降归元饮干预，电镜观察ICC自噬体，WB、qPCR检测miR-203a、ATG14、自噬因子表达；双荧光素酶基因报告验证ATG14为miR-203a下游靶基因及二者负反馈调控关系，探究miR-203a调控ATG14介导ICC自噬的效应，揭示该方作用的分子机制。

Gastroesophageal reflux disease (GERD) is closely related to gastrointestinal motility disorder. Our previous study found that ICC autophagy of gastrointestinal motility pacemaker cells is the key to GERD, which proved that " the intersectional imbalance of meridian qi of Ren and Du caused the imbalance of the motions of the spleen and stomach qi, this is the essence of the pathogenesis of GERD" and "adjusting cardinal with cardinal" method can improve ICC autophagy treatment of GERD. Based on the theory of "adjusting pivot by pivot", our team has developed Qi-Shi-Sheng-Jiang-Gui-Yuan-Yin, which has the function of regulating Ren and Du's meridian qi to regulate the cardinal of spleen and stomach. At present, the research of gene regulation based on autophagy in the molecular mechanism of GERD is still lacking. Through the prediction and analysis of autophagy gene and related miRNA, we suggest that Qi-Shi-Sheng-Jiang-Gui-Yuan-Yin may be based on miR-203a regulation of ATG14 mediated ICC autophagy to treat GERD. Therefore, in this project, after in vivo and in vitro experiments, Qi-Shi-Sheng-Jiang-Gui-Yuan-Yin was used to observe the ICC autophagy, WB or qPCR were used to detect the expression of mir-203a, ATG14 and autophagy factors; double luciferase gene report verified that ATG14 was the downstream target gene of miR-203a and the negative feedback regulation relationship between them, to explore the effect of miR-203a on ATG14 mediated ICC autophagy, and to reveal the molecular mechanism of the effect of miR-203a.