基于microRNA靶向SCF/c-kit信号通路调控胃运动的半夏泻心汤拆方研究

SCF/c-kit signal pathway can cause the number and morphological changes of ICC, leading to motility disorder in digestive tract. Preliminary study shows that the effect of BanxiaXiexin Decoction and its decomposed Recipes can regulate ICC by SCF/c-kit signaling pathway. But the mechanism is still not clear. As the upstream regulators of gene expression, miRNA provides a new target on prevention and treatment of gastrointestinal motility disorders. We hereby propose the hypothesis that Banxiaxiexin Decoction may regulate gastric motion by means of miRNA targeting SCF/c-kit. By copying the rat model of electrogastric dysrhythmias, we screen the miRNAs targeting SCF/c-kit, and observe the effects of BanxiaXiexin Decoction on gastric motility disorder of miRNAs, using gene chip technology, combined with bioinformatics analysis, gene prediction. And then establish the cell models of miRNA over expression and suppression expression by transfecting miRNA Mimics and Inhibitor into ICC. Combined with the method of serum pharmacology, we explore the effects of Banxiaxiexin Decoction on regulating ICC proliferation, apoptosis and phenotypic transformation by miRNA targeting SCF/c-kit signaling pathway. Hope to further reveal its molecular mechanism of regulating gastric motion and prescription rule.

SCF/c-kit信号通路异常可引起胃肠道ICC数量和形态变化，导致动力障碍性疾病。前期研究表明，半夏泻心汤及其拆方具有调控ICC SCF/c-kit信号通路的作用，但其具体作用机制尚不清楚。作为基因表达的上游调控因子，miRNA为胃肠动力障碍性疾病的防治提供了新标靶。我们据此提出半夏泻心汤通过miRNA靶向SCF/c-kit途径调节胃运动的假说。通过复制大鼠胃电节律失常模型，采用基因芯片技术，结合生物信息学分析、靶基因预测，筛选靶向SCF/c-kit的miRNAs，观察半夏泻心汤对胃动力失常miRNAs的影响；并采用脂质体转染miRNA Mimics和Inhibitor至ICC中，建立miRNA过表达和表达受抑细胞模型，结合血清药理学方法，探索半夏泻心汤通过miRNA靶向SCF/c-kit信号通路调控ICC增殖、凋亡和表型转化的作用，以期进一步揭示其调节胃运动的细胞分子机制与组方规律。

SCF/c-kit信号通路异常可引起胃肠道ICC数量和形态变化，导致动力障碍性疾病。前期研究表明，半夏泻心汤及其拆方具有调控ICC SCF/c-kit信号通路的作用，但其具体作用机制尚不清楚。作为基因表达的上游调控因子，miRNA为胃肠动力障碍性疾病的防治提供了新标靶。我们据此提出半夏泻心汤通过miRNA靶向SCF/c-kit途径调节胃运动的假说。通过复制大鼠胃电节律失常模型，采用基因芯片技术，结合生物信息学分析、靶基因预测，筛选出靶向SCF/c-kit 的miR-451-5p，为胃肠动力紊乱疾病的治疗提供了新的干预靶点。以miR-451-5p为切入点，采用脂质体转染，建立miRNA过表达和表达受抑ICC细胞模型，结合血清药理学方法，探索半夏泻心汤对ICC增殖、凋亡和表型转化的影响。结果发现，半夏泻心汤汤能促进胃电节律失常模型大鼠和ICC细胞miR-451-5p的表达，改善miR-451-5p表达受抑模型的ICC细胞增殖、凋亡和表型维持，直接或间接增强SCF/c-kit信号通路表达，上调SCF/c-kit、Bcl-2 、IP3R、RyR mRNA和蛋白的表达，下调NF-κB、Caspase9 mRNA和蛋白的表达，同时增强c-Myc、JAK1、STAT1及其磷酸化蛋白的表达。为半夏泻心汤调节胃运动的细胞分子研究提供理论和研究思路。