基于胃黏膜细胞miRNA调控网络的加味七方胃痛颗粒治疗大鼠慢性萎缩性胃炎的机制研究

miRNA involved in biological value, differentiation, maturation, regulation of apoptosis and cancer and other pathological processes. The project team found that pre-flavored granules Qifangweitong atrophic gastritis with good clinical efficacy, can be on multiple targets and pathways to regulate gastric mucosal cells. But in the overall regulation of gene level studies have not been carried out, this project intends to establish a rat model of atrophic gastritis, gastric mucosal cells extracted, intervention with drug-containing serum, and then using TLDA miRNA microarray technology to observe flavored granules intervention Qifangweitong gastric mucosal cells miRNA expression profile changes after screening by differences in miRNA transfection and then validated intervention inhibitor, combined with bioinformatics software TargetScan, RNAhybrid predict their target genes, target proteins and regulatory network structure, and then again on the biological information in the real world Studies predict target genes, target proteins, signal transduction molecules HYDROBIOLOGICA verification. Establish flavored granules intervention Qifangweitong biological model of microRNA regulation of gastric mucosal cells of the network, from the system, the overall level of Chinese medicine to explain "more a result, multi-effect,multi-target" mechanism to lay the foundation.

miRNA参与生物体的增值、分化、成熟、凋亡以及肿瘤等病理过程的调控。本项目组前期发现加味七方胃痛颗粒对萎缩性胃炎具有良好临床疗效，可以在多个靶点和通路上对胃黏膜细胞进行调控。但是在总基因水平调控研究尚未开展，本课题拟建立萎缩性胃炎大鼠模型，提取胃黏膜细胞，用含药血清进行干预，然后采用TLDA miRNA芯片技术，观察加味七方胃痛颗粒干预胃黏膜细胞后miRNA表达谱的变化，筛选差异的miRNA然后通过转染和抑制剂干预进行验证，结合生物信息学软件TargetScan、RNAhybrid预测其靶基因、靶蛋白和调控网络结构，然后在真实世界再次对生物信息学预测的靶点基因、靶点蛋白、信号转导通路的分子生生物学验证。建立加味七方胃痛颗粒干预胃黏膜细胞后的microRNA 调控网络的生物模型，为从系统、整体水平解释中医药“多因、多效、多靶点”机制奠定基础。

背景：慢性萎缩性胃炎是一种长期以来威胁着人类健康的病症，由于其病理机制的复杂性，对于慢性萎缩性胃炎的机制及治疗的探讨始终研究的重点和难点。目前中医药治疗慢性萎缩性胃炎的疗效良好且无副作用，但其起效机制尚未明了，研究其机制可以为推拿镇痛提供科学依据。.主要研究内容：本项目通过培养CAG大鼠胃黏膜：①观察加味七方胃痛颗粒干预胃黏膜细胞后miRNA表达谱的变化。②选取部分miRNA进行qPCR生物学验证。.结果：①经加味七方胃痛颗粒干预后慢性萎缩性胃炎细胞miRNA出现了差异性的表达，14个miRNA出现下调，即：rno-miR-335、rno-miR-203a-3p、rno-miR-24-2-5p、novel_166、novel_226、novel_252、novel_120、novel_251、rno-miR-1949、novel_238、novel_240、novel_233、novel_230、novel_271；17个miRNA出现上调，即：rno-miR-125b-1-3p、rno-miR-130a-5p、rno-miR-148a-5p、rno-miR-96-5p、rno-miR-27b-5p、rno-let-7c-1-3p、rno-miR-92a-1-5p、rno-miR-582-3p、rno-miR-152-5p、rno-miR-19b-1-5p、rno-miR-582-5p、rno-let-7a-2-3p、rno-miR-24-1-5p、rno-miR-879-5p、rno-miR-6324、novel_114、novel_101；②RT-PCR结果显示：miRNA_146A_5P、miRNA_17_5P、miRNA_219_5P、miRNA_224_5P、miRNA_24_2_5P、miRNA_25B_1_5P、miRNA_96_5P空白组、模型组和给药组在各指标中均无显著差异。（P＞0.05）。.意义：经加味七方胃痛颗粒干预慢性萎缩性胃炎细胞后miRNA出现了差异性的表达，可能通过引起rno-miR-335等差异性表达，从而调控胃黏膜细胞。