microRNA转录后调控 VEGF 信号通路及MMP9活性和痔病之间关系以及中药干预的机制

MiRNAs are short chain non-coding RNA which post-transcriptional regulate gene expression in Epigenetics.Lots study found that abnormal miRNAs regulation was closely associated with vascular injury,repair and angiogenesis. Hemorrhoids angiogenesis incidence theory currently is a hot point in hemorrhoidal diease research. But the relationship between post-transcriptional regulate and the development of hemorrhoids was unclear. Based on previous studies we found that VEGF, MMP9 highly expressed in the hemorrhoid tissue, and ZHIXUENING Mixture could well interpose this change.We propose "TCM intervention on hemorrhoidal tissue with post-transcriptional regulation of VEGF Expression and MMP9 activity by MicroRNA plays a key role in the process. " To confirm this hypothesis, this study uses RNA chips and bioinformatics methods, firstly by drawing expression profiling miRNAs, through analysis and comparison among the groups established target miRNA, predict the effect of potential target genes and signaling pathways of the target protein, immunohistochemical analysis and verification. To predict and confirm the relationship between post-transcriptional Regulation of VEGF Expression and MMP9 activity by MicroRNA and its TCM molecular mchanism on hemorrhoidal disease.

miRNAs是表观遗传学中重要的起转录后基因表达调控作用的非编码短链RNA。研究发现， miRNAs调控异常与血管的损伤修复、新生密切相关。《素问·生气通天论》记载“筋脉横解，肠澼为痔”，目前痔病的血管新生发病理论正成为研究的热点。但miRNAs转录后调控和痔病发生发展的关系还不明确。立足于本课题组前期研究发现VEGF、MMP9 在痔组织中高表达，痔血宁合剂能够很好的干预这种变化机制，我们提出“中药影响miRNAs靶向干预VEGF 信号通路及MMP9活性在痔组织中血管增生进程中起关键作用”。为证实该假说，本研究拟采用RNA芯片和生物信息学技术等方法，首先通过绘制miRNAs表达图谱，通过各组间分析比较确立目标miRNA,预测其作用的潜在靶基因及目标蛋白的信号通路，并进行免疫组化分析验证。以期预测和证实湿热下注型痔病与miRNAs转录后调控VEGF、MMP9表达之间的关系以及中药干预机制。

miRNAs是表观遗传学中重要的起转录后基因表达调控作用的非编码短链RNA。研究发现，miRNAs调控异常与血管的损伤修复、新生密切相关。《素问·生气通天论》记载“筋脉横解，肠澼为痔”，目前痔病的血管新生发病理论正成为研究的热点。但miRNAs转录后调控和痔病发生发展的关系还不明确。立足于本课题组前期研究发现VEGF、MMP9在痔组织中高表达，痔血安合剂（原痔血宁合剂）能够很好的干预这种变化机制，我们提出“中药影响miRNAs靶向干预VEGF信号通路及MMP9活性在痔组织中血管增生进程中起关键作用”。为证实该假说，本研究采用了RNA芯片和生物信息学技术等方法，首先通过绘制miRNAs表达图谱，通过各组间分析比较确立目标miRNA,预 测其作用的潜在靶基因及目标蛋白的信号通路，并进行免疫组化分析验证。最终证明mircroRNA可以作为湿热下注型混合痔的表观遗传学标志物，其中miRNA412-5p通过靶向结合XPO1 mRNA 3’UTR来降解XPO1 mRNA，激活p53-p66 Shc -p16通路，从而削弱血管内皮细胞的功能。过表达miR-4729，通过沉默靶基因METTL14的表达，诱导整体mRNA甲基化和TIE1 mRNA 3'UTR特定位点甲基化修饰下降，并降低TIE1 mRNA的稳定性，下调TIE1/VEGFA信号分子环路的表达，削弱血管增生能力。痔血安合剂可以通过上述途径削弱湿热下注型混合痔的血管增生能力。